Oxidative Medicine and Cellular Longevity

Oxidative Medicine and Cellular Longevity / 2019 / Article
Special Issue

Genetic and Chemical Effects on Somatic and Germline Aging

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Review Article | Open Access

Volume 2019 |Article ID 1607304 | 28 pages | https://doi.org/10.1155/2019/1607304

Triclosan: An Update on Biochemical and Molecular Mechanisms

Academic Editor: Demetrios Kouretas
Received30 Nov 2018
Revised28 Feb 2019
Accepted01 Apr 2019
Published02 May 2019

Abstract

Triclosan (TCS) is a synthetic, chlorinated phenolic antimicrobial agent commonly used in commercial and healthcare products. Items made with TCS include soaps, deodorants, shampoos, cosmetics, textiles, plastics, surgical sutures, and prosthetics. A wealth of information obtained from in vitro and in vivo studies has demonstrated the therapeutic effects of TCS, particularly against inflammatory skin conditions. Nevertheless, extensive investigations on the molecular aspects of TCS action have identified numerous adversaries associated with the disinfectant including oxidative injury and influence of physiological lifespan and longevity. This review presents a summary of the biochemical alterations pertaining to TCS exposure, with special emphasis on the diverse molecular pathways responsive to TCS that have been elucidated during the present decade.

1. Introduction

Triclosan (TCS), or 5-chloro-2-(2,4-dichlorophenoxy)phenol, is a synthetic broad-spectrum antimicrobial developed in the 1960s. As a polychlorinated bisphenolic compound, TCS has a perceptible aromatic odor and is weakly soluble in water. It dissolves well in organic solvents including ethanol, dimethylsulfoxide (DMSO), and methanol [1], and the type of solvent and detergent availability seem to influence TCS activity [24]. For example, TCS dissolved in oils (e.g., olive oil) and alkali (e.g., sodium carbonate) exhibits markedly reduced efficacy when compared to other solvents such as glycerol and polyethylene glycol (PEG) [3, 5]. In fact, using propylene glycol (PG) as a solvent renders TCS more effective than using PEG, which is probably due to micellar solubilization of TCS in the larger PEG molecules [3]. Recently, we have shown that the presence of nonionic detergents (e.g., Tween 20) inhibits TCS activity in vivo, most likely due to micelle formation [6]. In contrast, sodium dodecyl sulfate (SDS) has been reported to potentiate the antibacterial effect of TCS in vitro [7].

TCS has gained enormous popularity in commerce and in healthcare owing to its antibacterial, antiviral, and antifungal properties [810]. This efficacy has led to the widespread use of TCS as a preservative in a variety of consumer products, including cosmetics, soaps, mouthwashes, antiperspirants, kitchen utensils, clothing textiles, bedclothes, electronics, plastics, and toys (Triclosan White Paper prepared by the Alliance for the Prudent Use of Antibiotics (APUA)). In clinical practice, TCS is used as a disinfectant and an antiseptic in surgical sutures, scrubs, implants, and medical devices [11, 12]. Annual global production of TCS was estimated at 1500 tons [13], and a total of 132 million liters of TCS-containing products was consumed in a single year in the United States (Safety and Effectiveness of Consumer Antiseptics; Topical Antimicrobial Drug Products for Over-the-Counter Human Use; Proposed Amendment of the Tentative Final Monograph. 2013 https://www.fda.gov/downloads/AboutFDA/ReportsManualsForms/Reports/EconomicAnalyses/UCM379555.pdf).

The high demand for TCS has consequently led to substantial buildup in drinking and wastewater sources and, more alarmingly, accumulation in body fluids [1420], establishing the antimicrobial as an environmental pollutant. Pharmacokinetic studies in man show that TCS reaches the systemic circulation by rapid absorption through the skin and mucous membranes of the oral cavity and gastrointestinal tract, and variations in the bioavailability of TCS unsurprisingly affect the rate of urinary excretion [21, 22]. TCS content in commercial products may reach as high as 17 mM and comprise up to 1% of ingredients [12, 19, 23]. Moreover, absorption of up to 25% of applied TCS has been recorded [24], and metabolic studies in rats and mice revealed sulfation, glucuronidation, and hydroxylation products in tissues and excreta [25, 26].

Since the advent of TCS, early studies on the antiseptic have shown evidence of symptomatic relief from acne [27, 28] and contact dermatitis [29, 30] with fewer, or at least comparable, side effects to other therapeutic alternatives [31]. Later, TCS was found to be effective against crural ulcer [32] and chemically induced dermatitis and desquamation [33, 34], which could be attributed to its anti-inflammatory [35], hypoallergenic [36], and analgesic [37] properties. Moreover, a battery of studies collectively indicate that TCS is not a skin or oral mucosal irritant, has a very low sensitization potential (0.1-0.3% of 14,000 subjects), and is unlikely to be phototoxic to human skin (http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_166.pdf). This is in contrast to the reversible skin and eye irritation caused by up to 10% TCS reported in animals (http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_166.pdf). Also, in initial studies by Lyman and Furia, it was suggested that TCS is carcinogenic when orally administered to rats [38, 39]. Subsequent investigations in rats and mice disclosed that TCS perturbs microsomal detoxification [40], causes nephrotoxicity and hepatotoxicity [41], reduces prenatal and postnatal survival [42], and leads to central nervous system suppression [43] and hypothermia [44]. In humans, the earliest description of an adverse TCS reaction probably comes from a case report of two patients who developed contact dermatitis following application of deodorants containing 0.12% and 0.2% TCS [45]. Since then, several case reports of the same ailment have thus far been in congruence [4649]. It is important to mention that, as is the case with healthy subjects, in patients diagnosed with, or suspected to have, contact dermatitis, TCS was similarly found to have a very low sensitization potential (0.6-0.8% of 11,887 patients) (http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_166.pdf).

In light of the dichotomous debate surrounding TCS, the US Food and Drug Administration (FDA), following extensive examination of available data, has effectively banned antiseptic products containing TCS since September 2016 [19]. In Europe, TCS was approved for use in cosmetics by the European Community Cosmetic Directive in 1986 (http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp_o_166.pdf). However, the European Commission disapproved the use of TCS for hygienic purposes in 2017, but maintained its legality as a preservative in select cosmetics and mouthwashes in concentrations up to 0.3% and 0.2%, respectively (http://ec.europa.eu/health/scientific_committees/consumer_safety/docs/sccs_o_054.pdf; http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=OJ:L:2014:107:FULL&from=EN). Furthermore, the Scientific Committee on Consumer Safety (SCCS) expressed its concern over the continued use of TCS in cosmetics, but not in antiseptics, mainly due to the cumulative pattern of exposure (http://ec.europa.eu/health/scientific_committees/consumer_safety/docs/sccs_o_054.pdf). Importantly, the European Chemicals Agency (ECHA) classifies TCS, under the classification, labeling, and packaging (CLP) regulation, as an eye irritant 2 (causes serious eye irritation), skin irritant 2 (causes skin irritation), aquatic acute 1 (very toxic to aquatic life), and aquatic chronic 1 (very toxic to aquatic life with long-lasting effects) (https://echa.europa.eu/documents/10162/21680461/bpc_opinion_triclosan_pt1_en.pdf/efc985e4-8802-4ebb-8245-29708747a358). Because of the previously mentioned ecotoxic properties, TCS is currently a candidate for substitution under the Biocides European Union regulation (Reg 528/2012/EC) (https://echa.europa.eu/potential-candidates-for-substitution-previous-consultations/-/substance-rev/12/term?_viewsubstances_WAR_echarevsubstanceportlet_SEARCH_CRITERIA_EC_NUMBER=222-182-2&_viewsubstances_WAR_echarevsubstanceportlet_DISS=true).

Our aim in this review is to provide an update on current knowledge regarding TCS therapeutic and toxic potential. Emphasis is placed on the biochemical and molecular alterations, either brought about by, or in response to, TCS exposure. Data from both in vitro and in vivo studies, obtained from humans and other organisms, are incorporated into the analysis, with special attention being given to reports published during the present decade.

2. Membrane and Cytoskeletal Damage

Perhaps the earliest report describing the antimicrobial activity of TCS was by Vischer and Regös [50] which was shown through topical application. In a follow-up study, TCS was found to be more effective with the broadest spectrum against bacteria and fungi when compared to other antimicrobials such as gentamicin and clotrimazole [10]. Subsequent efforts, which continue to this day, have focused on dissecting the diverse action mechanisms and cellular targets of TCS. Initially, it was thought that TCS interacts with the prokaryotic cell membrane nonspecifically [9]. This was corroborated by the resistance of Gram-negative bacteria to TCS, which was ascribed to their outer membrane [51, 52]. Investigating the genetic response of Mycobacterium tuberculosis to TCS, Betts et al. [53] identified perturbations in a wide assortment of genes involved in cell wall, transport, detoxification, and DNA replication and transcription. Also, Klebsiella pneumoniae with inactive efflux pump KpnGH exhibit pronounced susceptibility to multiple antibiotics including TCS [54]. Several genes in the membrane stress response pathway were also studied in Escherichia coli and Rhodospirillum rubrum S1H [5557]. During the electro-Fenton transformation of TCS, significant changes in expression patterns of genes involved in cell wall and membrane structure, cell envelope, flagella, and multidrug efflux were observed (Table 1). These findings complement an earlier report describing enhanced resistance to TCS due to overexpressed acrAB multidrug efflux pump [58]. It was recently suggested that TCS binds to the transcriptional repressor AcrR, causing conformational changes and preventing its binding to the efflux pump AcrA promoter in Agrobacterium tumefaciens [59].


ModelTargetResponse
Gene/proteinMolecular identity

K. pneumoniaeKpnGHEfflux pumpSensitive to TCS

E. coliAcrABEfflux pumpsUpregulated by TCS
acrEUpregulated by TCS
mdtEUpregulated by TCS
acrFUpregulated by TCS
mdtBUpregulated by TCS
mdtCUpregulated by TCS
yddAUpregulated by TCS
emrAUpregulated by TCS
emrEUpregulated by TCS
sanACell wall/membrane structureUpregulated by TCS
dacBUpregulated by TCS
amiCCell envelopeUpregulated by TCS
clsAUpregulated by TCS
ompXMembrane porinDownregulated by TCS
motAFlagellarUpregulated by TCS
flgMUpregulated by TCS

R. rubrum S1HsugESmall multidrug resistance proteinUpregulated by TCS
mexFRND efflux system, inner membrane transporterUpregulated by TCS
mexBUpregulated by TCS
mexERND efflux system, membrane fusion proteinsSensitive to TCS
mexAUpregulated by TCS
mexMUpregulated by TCS
oprMRND efflux system, outer membrane transporterUpregulated by TCS
glmMCell envelope; phosphoglucosamine mutaseUpregulated by TCS
exoDCell envelope; exopolysaccharide synthesis protein DUpregulated by TCS
wbpMCell envelope; polysaccharide biosynthesis protein MUpregulated by TCS

A. tumefaciens C58AcrARND efflux system, periplasmic adaptor proteinUpregulated by TCS

Human erythrocytesNa+,K+,Mg2+-ATPaseMembrane ion transporterSensitive to TCS

C. elegansPmp-3Membrane ABC transporterDownregulated by TCS

D. rerioActin, cytoplasmic 2CytoskeletonDownregulated by TCS
Actin α1, skeletal muscleDownregulated by TCS
Light polypeptide 3Downregulated by TCS
DesminCytoskeleton; muscular filament structureUpregulated by TCS
Fast skeletal muscle myosinSensitive to TCS
Keratin, type I cytoskeletal 18Upregulated by TCS
Tropomyosin α-1 chainDownregulated by TCS
Type II cytokeratinUpregulated by TCS
Lamin B1Cytoskeleton; nuclear laminaDownregulated by TCS

D. polymorphaTubulin β-2/α-4 chainCytoskeletonUpregulated by TCS
Tubulin β-4 chainUpregulated by TCS
Myosin light chainCytoskeleton; muscular filament structureUpregulated by TCS

Abbreviation: RND: resistance-nodulation-division; ABC: ATP-binding cassette.

The interaction of TCS with the cell membrane was also studied in human red blood cells (RBCs; erythrocytes). TCS exposure led to K+ leakage and overt hemolysis, indicating membrane damage, while antagonizing hypotonic lysis, which may be due to membrane expansion [60]. TCS also inhibited membrane-bound Na+,K+,Mg2+-ATPase enzymatic activity [61]. These observations suggest that TCS causes membrane destabilization, perturbs monovalent ion transport, and modulates the overall osmoregulation of erythrocytes. Evidence for membrane damage is further confirmed in numerous studies by means of compromised stability and permeability [62]. To directly observe how TCS interacts with the cell membrane, Guillén and coworkers utilized nuclear magnetic resonance (NMR) spectroscopy to demonstrate that TCS intercalates within hydrophobic pockets in the lipid bilayer, perpendicularly to phospholipid molecules [63]. Furthermore, using neutral red to evaluate membrane integrity, diminished uptake of the dye in hemocytes of the clam Ruditapes philippinarum and mussel Mytilus galloprovincialis was related to TCS-induced suppression of pinocytosis and disturbed phagocytosis [64, 65].

Along those lines, our recent findings indicate that TCS blunts the expression of the pmp3 membrane transporter in Caenorhabditis elegans nematodes and that pmp3(ok1087) mutants exhibit increased sensitivity to the disinfectant [66]. Finally, a proteomic analysis of zebrafish (Danio rerio) larvae and gills of freshwater mussel Dreissena polymorpha revealed alterations in cytoskeletal protein levels following TCS exposure (summarized in Table 1) [67, 68].

There is a consensus in the literature regarding the membranotropic nature of TCS in different membrane models across various species. The cell membrane is a primary target for TCS and among the first cellular obstacles that must be overcome by the antiseptic to exert its effects. Although evidence implicating membrane-associated efflux pumps as part of the cellular response to TCS is strong, there is paucity in reports describing TCS modulation of structural or functional membrane components in human-based systems. Similarly lacking is an understanding of the role of membrane receptors not only in pumping out TCS molecules but also in transducing both inter- and intracellular signals as a consequence to TCS presence.

3. Cellular Longevity

The interest in TCS and ultimate cell fate has originally stemmed from its use in oral hygiene products, which is reflected in two seminal studies on human gingival cells [69, 70]. TCS was shown to be cytotoxic to gingival fibroblasts and epithelial cells, identifying it as a novel stimulator of apoptosis in the latter.

Investigations have thus far followed a more comprehensive approach, relating cell death induced by TCS to other cellular adversaries, utilizing both human and non-human model systems. When TCS was treated to human choriocarcinoma placental cells (JEG-3), multiple dose- and time-dependent responses were observed [71]. While there was a proportional increase in estradiol and progesterone secretion, β-human chorionic gonadotropin (β-hCG) release was nevertheless inhibited with increasing TCS concentrations [71]. In addition to blunted proliferation, significant cell death was recognized as apoptotic in nature evidenced by activated caspase-3 and Hoechst 33342-stained fragmented DNA [71]. Similarly, using anoikis-resistant H460 human lung cancer cells, Winitthana et al. demonstrated that 24-hour exposure to 10 μM TCS causes cell death and apoptosis. Nontoxic levels (≤7.5 μM), however, enhanced cell growth (increased colony number and reduced size) without altering proliferation. TCS also promoted epithelial-to-mesenchymal transition (EMT), along with the migratory and invasive abilities of the cells [72].

A research group performed a series of in vivo and in vitro studies on the effect of TCS on growth and proliferation of human BG-1 ovarian cancer cells. Results from these studies indicate that TCS increases cellular proliferation and both gene expression and protein levels of cyclin D1 and decreases p21 and Bax gene expression and protein levels [73]. These effects were significantly antagonized by the estrogen receptor (ER) antagonist ICI 182,780, implicating ER in TCS-induced cell cycle progression and in its antiapoptotic role. Investigators from the same group also reported a similar response to TCS by MCF-7 breast cancer cells and LNCaP prostate cancer cells. In MCF-7 cells, 1 μM TCS enhanced growth and proliferation during a six-day period, which was associated with increased cyclin D1 and reduced p21 expression levels. When mice were treated with TCS for 8 weeks, brdU-positive breast tumor cells were significantly increased compared to the control group treated with corn oil [74]. Similar to BG-1 cells, TCS-promoted proliferation of MCF-7 cells was mediated through ERα signaling, demonstrated as antagonism by kaempferol and 3,3-diindolylmethane (DIM), two phytoestrogens [75]. In addition to cyclin D1 and p21, TCS caused an increase in cyclin E and a decrease in Bax and induced metastasis through elevated cathepsin D protein expression. These observations were paralleled in vivo using xenografted mouse models. Researchers from this report expanded their findings to VM7Luc4E2 cells, a variant of the MCF-7 model, to show that TCS (0.1-10 μM) is pro-proliferative and antiapoptotic by inhibiting oxidative stress, with both effects being antagonized by kaempferol [76]. In LNCaP cells exposed to concentrations of TCS ranging from 0.01 to 10 μM for up to 5 days showed enhanced proliferation and migration and reduced p21 protein expression [77]. In primary human syncytiotrophoblasts, TCS at 0.001 to 10 μM induced apoptosis as seen by condensed nuclei and fragmented DNA [78]. TCS also reduced 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) via a caspase-dependent mechanism. Other targets included both Bax and Bcl-2 proteins.

Similar to human cells, both pro- and antiapoptotic properties were observed in rodent cells treated with TCS. Beside its cytotoxicity, TCS caused caspase-dependent apoptosis in rat neural stem cells along with elevated Bax and reduced Bcl-2 [79]. In a series of studies, [8082] mouse neurons were used to show that TCS is apoptotic through the Fas receptor (FasR), aryl hydrocarbon receptor (AhR), and caspase activation involving N-methyl-D-aspartate receptors (NMDARs). In agreement with the cytotoxicity data, TCS-treated mouse lung epithelial cells were deformed with reduced viability [83]. Conversely, TCS stimulated the proliferation of mouse epidermis-derived JB6 Cl 41-5a cells, by increasing cyclins D1 and A and reducing p27(Kip1) protein levels [84]. Examining these effects in vivo, B6C3F1 mice exhibited epidermal hyperplasia and focal necrosis following topical administration of TCS. Moreover, the pluripotency markers of mouse embryonic stem cells were analyzed following TCS exposure [85]. Alkaline phosphatase (Alp), Sox2, Oct4, and Nanog were all reduced, while miRNA-134 was elevated.

Unlike human and rodent cells, in vivo and in vitro studies on aquatic organisms uniformly agree that TCS is solely proapoptotic in these animals. Pyknotic apoptosis in the central nervous system of zebrafish D. rerio was observed following treatment with either TCS alone or TCS combined with derivatives 2,4,6-trichlorophenol (2,4,6-TCP) and 2,4-dichlorophenol (2,4-DCP) [86, 87]. The TCS-derivative mixture caused pronounced deformities and behavioral abnormalities and perturbed the expression of a panel of neurodevelopmental and apoptotic genes (Table 2). Also, TCS, following both in vivo and in vitro exposure, induced a dose- and time-dependent increase in apoptotic hemocytes of D. polymorpha [88, 89]. Likewise, when the saltwater clam Ruditapes philippinarum was treated with TCS, hemocytes exhibited significant cell death, blunted proliferation, reduced size, and promiment apoptotic DNA fragmentation [65]. TCS-induced apoptosis, or apoptosis-like cell death, was also detected in unicellular organisms, such as the green alga Chlamydomonas reinhardtii and the pathogenic fungus Cryptococcus neoformans [90, 91].


ModelTargetResponse
Gene/proteinMolecular identity

JEG-3 cellsEstradiolMajor female sex hormonesUpregulated by TCS
ProgesteroneUpregulated by TCS
β-hCGMaintenance of pregnancyDownregulated by TCS
Caspase-3Apoptosis regulator; proapoptoticUpregulated by TCS

BG-1 cellsCyclin D1Cell cycle regulatorsUpregulated by TCS
p21Downregulated by TCS
BaxApoptosis regulator; proapoptoticDownregulated by TCS

MCF-7 cellsCyclin D1Cell cycle regulatorsUpregulated by TCS
Cyclin EUpregulated by TCS
p21Downregulated by TCS
BaxApoptosis regulator; proapoptoticDownregulated by TCS
Cathepsin BMetastasis markersUpregulated by TCS
Cathepsin DUpregulated by TCS
MMP-9Upregulated by TCS
MMP-2Upregulated by TCS
CXCR4Upregulated by TCS
SnailMesenchymal markersUpregulated by TCS
SlugUpregulated by TCS

LNCaPp21Cell cycle regulatorDownregulated by TCS

Primary human syncytiotrophoblasts11β-HSD2Fetal development; anticortisolDownregulated by TCS
Caspase-3Apoptosis regulators; proapoptoticUpregulated by TCS
BaxUpregulated by TCS
Bcl-2Apoptosis regulator; antiapoptoticDownregulated by TCS

Rat neural stem cellsCaspase-3Apoptosis regulators; proapoptoticUpregulated by TCS
BaxUpregulated by TCS
Bcl-2Apoptosis regulator; antiapoptoticDownregulated by TCS

Mouse neocortical neuronsGluN1Ionotropic glutamate receptors; neurotransmissionDownregulated by TCS
GluN1Downregulated by TCS
GluN2ADownregulated by TCS
GluN2ADownregulated by TCS
GluN2BUpregulated by TCS
GluN2BDownregulated by TCS
FasRApoptosis regulators; proapoptoticUpregulated by TCS
Caspase-8Upregulated by TCS
Caspase-9Upregulated by TCS
Caspase-3Upregulated by TCS
AhRLigand-activated receptor; detoxificationUpregulated by TCS

JB6 Cl 41-5a cellsCyclin D1Cell cycle regulatorsUpregulated by TCS
Cyclin AUpregulated by TCS
p27Downregulated by TCS

B6C3F1 miceAlpPluripotency markers; stem cell self-renewal and differentiation regulatorsDownregulated by TCS
Oct4Downregulated by TCS
NanogDownregulated by TCS
ALPDownregulated by TCS
Oct 4Downregulated by TCS
NanogDownregulated by TCS
Sox 2Downregulated by TCS
miRNA-134Transcriptional regulator of pluripotency markersUpregulated by TCS

D. rerioOct4Pluripotency markersDownregulated by TCS
NanogDownregulated by TCS
Sox2Upregulated by TCS
p53Cell cycle regulator; tumor suppressorUpregulated by TCS
Casp3Apoptosis regulators; proapoptoticUpregulated by TCS
Casp8Upregulated by TCS
ShhaEarly neurogenesisSensitive to TCS sensitive to TCS
Ngn1Upregulated by TCS
NrdUpregulated by TCS
Elavl3Upregulated by TCS
α1-tubulinNeural maturationUpregulated by TCS
Gap43Upregulated by TCS
GfapDownregulated by TCS
MbpDownregulated by TCS

Abbreviation: Shha: sonic hedgehog a; Ngn1: neurogenin 1; Nrd: NeuroD; Elavl3: ELAV-like, neuron-specific RNA-binding protein 3; Gap43: growth-associated protein 43; Gfap: glial fibrillary acidic protein; Mbp: myelin basic protein.

Collectively, studies on TCS influence on cell fate indicate estrogenic, proliferative, and apoptotic activities. Genes and proteins governing the regulation of cell cycle and apoptosis are particularly sensitive to TCS modulation. The disparity in ultimate cell fate seems to point at an interspecies variation and a dose-specific response, among other experimental details such as cell type and duration of exposure. Elucidating the existence and the identity of a specific molecular “switch” that may tip the scales in favor of either cell death or survival could be an important inquiry for future investigations.

4. Oxidative Stress

Overwhelming evidence has recently accumulated in support of the prooxidative action of TCS. It is prudent to provide an overview of human-based studies first before summarizing notable findings obtained from other model organisms.

In Puerto Rican pregnant women, a correlation between exposure to TCS during pregnancy and oxidative damage, as measured by urinary 8-hydroxyguanosine (8-OHdG), and inflammation was suggested [92]. Similar observations were also mirrored in Chinese and Brazilian children [93, 94]. Conversely, in a global effort comprising nine countries from Asia, Europe, and North America, no relation between urinary TCS and 8-OHdG was established [95].

In vitro studies on human cells have also shed some light on the oxidative potential of TCS. In peripheral blood mononuclear cells (PBMC), 2,4-dichlorophenol (2,4-DCP)—a product of TCS transformation—promoted reactive oxygen species (ROS) generation, with subsequent lipid peroxidation and protein carbonylation [96]. Similarly, TCS caused elevated ROS in Nthy-ori 3-1 human follicular thyroid cells [97] and lipid peroxidation in retinoblastoma (Y79 RB) cells [98]. Our recent investigations on mesenchymal stem cells also showed TCS interference with the activation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2), the “master regulator” of detoxification, and its downstream targets, heme oxygenase 1 (HO-1) and NAD(P)H dehydrogenase [quinone 1] (NQO-1) [66]. Consistently, TCS incorporated in mouthrinse did not exhibit antioxidant activity on fibroblasts [99]. In contrast, TCS reduced ROS levels in VM7Luc4E2 cells, which contributed to its antiapoptotic activity in these malignant breast cells [76].

Mitochondrial damage was also evident in multiple mammalian cells including human PBMC and keratinocytes, exposed to 3.5-350 μM TCS [100]. At concentrations up to 100 μM, TCS caused depolarization of mitochondrial membrane, reduced oxidative phosphorylation, and suppressed ATP synthesis. Weatherly et al. [101] utilized human HMC-1.2 mast cells and primary keratinocytes to show that TCS is a proton ionophore uncoupler and interferes with ATP production.

Animal studies conducted on mice and rats have revealed a profound response in the cellular antioxidant machinery upon TCS treatment. In rat thymocytes, superoxide anions were found to be elevated following TCS treatment [102] which, as Yueh et al. [103] showed, was met with increased expression of key antioxidant enzymes including HO-1, NQO-1, and glutathione S-transferase (GST) in mouse liver. Evidence for testicular DNA damage, elevated malondialdehyde (MDA), and superoxide dismutase (SOD), in addition to diminished catalase (CAT), was related to TCS treatment in weanling rats [104]. Similarly, in lung homogenates of female albino rats, TCS was found to induce lipid peroxidation and severely deplete the levels of other crucial antioxidants: SOD, CAT, and glutathione (GSH) [105]. Increased expression of glutathione peroxidase 1 (Gpx1) and aldehyde oxidase 1 (Aox1) was also observed as a consequence to TCS exposure in C57BL/6 mice [106]. Most recently, Zhang et al. [97] showed downregulation of antioxidant enzymes, Gpx3, Cat, and Sod2, along with elevated MDA, in the hypothalamus of Sprague-Dawley rats. Moreover, it was found that TCS treatment leads to increased ROS and reduced GSH activity in rat neural stem cells [79]. TCS also increased ROS levels in mouse neocortical neurons, along with perturbed regulation of cytochrome P450 family 1, subfamily a, member 1 (CYP1a1) and CYP1b1 [81, 82]. Effects of TCS on cytochromes and hepatic detoxification were also demonstrated in Sprague-Dawley rats, showing increased levels of UDP-glucuronosyltransferase 1-1 (Ugt1a), Ugt2b1, CYP1a1, CYP1a2, CYP2b1, CYP3a1, and sulfotransferase family 1E member 1 (Sult1e1) [97].

Several terrestrial organisms have been employed in the study of TCS toxicology. Caenorhabditis elegans is among the best-studied animal models due to its ease of maintenance and high genetic homology to humans. We have recently shown that TCS leads to overproduction of ROS, inhibition of nuclear translocation of protein skinhead-1 (SKN-1) antioxidant transcription factor, and downregulation of gamma-glutamyl cysteine synthetase (Gcs1) [66]. In a subsequent report, Skn1 expression was found to be upregulated by TCS along with Sod1, Sod4, heat shock proteins (Hsp)-3, -4, -16.2, and -70; and cytochromes Cyp29A2 and Cyp34A9 (https://app.dimensions.ai/details/publication/pub.1103154992#readcube-epdf). TCS also enhanced nuclear translocation of stress-related factor DAF-16, suggesting the occurrence of oxidative stress [107]. In the Earthworm Eisenia fetida, oxidative damage by TCS was manifested as a transient elevation in CAT and GST enzymes, increased MDA, and DNA damage [108]. In a follow-up study by the same group, SOD was also increased and decreased by TCS depending on the concentration used [109], a response mirrored by CAT in the snail Achatina fulica [110]. In that study, TCS caused diminished levels of SOD and peroxidase (POD), along with elevated MDA, among other morphological anomalies.

The ubiquity of TCS in aquatic environments has made animal models from that habitat the subject of extensive investigations on TCS toxicity. Perhaps the most relevant aquatic organism is the zebrafish D. rerio, owing to a strong structural and molecular resemblance to humans. Elucidating the interaction between TCS and the antioxidant system in ZFL liver cells, Zhou et al. [111] showed evidence of induced CYP1A activity along with a general trend of suppression in phase I and II detoxification enzymes. Elevated MDA, along with perturbed homeostasis of GSH, peroxiredoxin-2 (PRD-2), and HSPs, were observed in zebrafish larvae grown in the presence of TCS ([67, 87]).

TCS has been shown to induce MDA and cause oscillations in CAT, ethoxyresorufin-O-deethylase (EROD), erythromycin N-demethylase (ERND), and aminopyrine N-demethylase (APND) in Daphnia magna [112]. Moreover, elevated amino acids, including glutamine, glutamate, and proline, have been attributed to a general oxidative stress state in daphnids [113]. Also, stress-related proteins, including glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and hsp-70, were modulated by TCS in D. polymorpha, in addition to lipid peroxidation [68]. TCS exposure demonstrated reduced oxyradicals and lipofuscin and elevated oxidized glutathione (GSSG) in the digestive gland of swollen river mussels Unio tumidus [114]. In Tigriopus japonicus copepods treated with TCS, increased ROS, SOD, GST, GPx, and GSH content was noted [115]. TCS also caused perturbations in expressional profiles of Cyps, Sod, Gst, and Cat proteins (Table 3) [115].


ModelTargetResponse
BiomarkerMolecular identity

Humans (pregnant women; children)Urinary 8-OHdGOxidized deoxyguanosine; DNA damageUpregulated by TCS

Nthy-ori 3-1 cellsROSMetabolic oxygen by-productsUpregulated by TCS
PBMCROSUpregulated by TCS
Lipid peroxidationOxidized lipidsUpregulated by TCS
Protein carbonylationOxidized proteinsUpregulated by TCS

Y79 RB cellsLipid peroxidationOxidized lipidsUpregulated by TCS

Human bone marrow-derived mesenchymal stem cellsNrf2Antioxidant regulatorDownregulated by TCS
Ho-1Antioxidant enzymesDownregulated by TCS
Nqo-1Downregulated by TCS

VM7Luc4E2 cellsROSMetabolic oxygen by-productsUpregulated by TCS

Mouse liverO2Antioxidant enzymesUpregulated by TCS
HO-1Upregulated by TCS
NQO-1Upregulated by TCS
GSTUpregulated by TCS

Weanling ratsMDAOxidized lipid markerUpregulated by TCS
SODAntioxidant enzymesUpregulated by TCS
CATDownregulated by TCS

Female albino rat lung homogenatesLipid peroxidationOxidized lipidsUpregulated by TCS
SODAntioxidantsDownregulated by TCS
CATDownregulated by TCS
GSHDownregulated by TCS

C57BL/6 mice liverGpx1Antioxidant enzyme; glutathione homeostasisUpregulated by TCS
Aox1Superoxide and hydrogen peroxide formationUpregulated by TCS

Sprague-Dawley rat hypothalamusMDAOxidized lipid markerUpregulated by TCS
Gpx3Antioxidant enzyme; glutathione homeostasisDownregulated by TCS
CatAntioxidant enzymesDownregulated by TCS
Sod2Downregulated by TCS

Rat neural stem cellsROSMetabolic oxygen by-productsUpregulated by TCS
GSHAntioxidantDownregulated by TCS

Mouse neocortical neuronsROSMetabolic oxygen by-productsUpregulated by TCS
Cyp1a1Cytochrome family enzymes; detoxificationDownregulated by TCS
CYP1a1Downregulated by TCS
Cyp1b1Downregulated by TCS
Cyp1b1Upregulated by TCS
Sprague-Dawley rat liverCyp1a1Upregulated by TCS
Cyp1a2Upregulated by TCS
Cyp2b1Upregulated by TCS
CYP2b1Upregulated by TCS
Cyp3a1Upregulated by TCS
Ugt2b1Glucuronidation enzymes; detoxificationUpregulated by TCS
Ugt2b1Upregulated by TCS
Sult1e1Sulfation enzyme; detoxificationUpregulated by TCS
Sult1e1Upregulated by TCS

C. elegansROSMetabolic oxygen by-productsUpregulated by TCS
Skn1Stress response regulatorUpregulated by TCS
SKN-1Downregulated by TCS
Gcs1Antioxidant enzymesDownregulated by TCS
Sod1Upregulated by TCS
Sod4Upregulated by TCS
Hsp-3Stress response; protein stabilizationUpregulated by TCS
Hsp-4Upregulated by TCS
Hsp-16.2Upregulated by TCS
Hsp-70Upregulated by TCS
Cyp29A2Cytochrome family enzymes; detoxificationUpregulated by TCS
Cyp34A9Upregulated by TCS
DAF-16Stress responseUpregulated by TCS

E. fetidaMDAOxidized lipid markerUpregulated by TCS
CATAntioxidant enzymesUpregulated by TCS
GSTUpregulated by TCS
SODSensitive to TCS to TCS

A. fulicaMDAOxidized lipid markerUpregulated by TCS
CATAntioxidant enzymesSensitive to TCS to TCS
SODDownregulated by TCS
PODDownregulated by TCS

ZFL liver cellsCYP1ACytochrome family enzyme; detoxificationUpregulated by TCS

D. rerio larvaeGPxAntioxidant enzymes; glutathione homeostasisUpregulated by TCS
GRDownregulated by TCS
PRD-2Antioxidant enzymeDownregulated by TCS
Hsp-5Stress response; protein stabilizationUpregulated by TCS
Hsp-90 βUpregulated by TCS

D. magnaMDAOxidized lipid markerUpregulated by TCS
CATAntioxidant enzymesSensitive to TCS to TCS
ERODDetoxification enzymesSensitive to TCS to TCS
ERNDSensitive to TCS to TCS
APNDSensitive to TCS to TCS
GlutamineAmino acids; markers of protein oxidation/breakdownUpregulated by TCS
GlutamateUpregulated by TCS
ProlineUpregulated by TCS

D. polymorpha gillsHsp-70Stress response; protein stabilizationSensitive to TCS to TCS

U. tumidus digestive glandGAPDHOxidoreductase; glucose metabolismSensitive to TCS to TCS
GSSGOxidized glutathione; antioxidantUpregulated by TCS
OxyradicalsOxygen-containing radicals; prooxidantsDownregulated by TCS
LipofuscinLysosomal pigment granules; toxicity markerDownregulated by TCS

T. japonicusROSMetabolic oxygen by-productsUpregulated by TCS
SodAntioxidant enzymesSensitive to TCS to TCS
SODUpregulated by TCS
CatSensitive to TCS to TCS
Gst variantsAntioxidants; glutathione homeostasisSensitive to TCS to TCS
GSTUpregulated by TCS
GPxUpregulated by TCS
GSHUpregulated by TCS
Cyp3026a3Cytochrome family enzymes; detoxificationUpregulated by TCS
Cyp3037a1Upregulated by TCS

P. fulvidracoMDAOxidized lipid markerSensitive to TCS to TCS
CATAntioxidant enzymeUpregulated by TCS
GstAntioxidant enzyme; glutathione homeostasisSensitive to TCS to TCS
ERODDetoxification enzymesUpregulated by TCS
ERNDUpregulated by TCS
APNDUpregulated by TCS
Cyp1aCytochrome family enzymes; detoxificationSensitive to TCS to TCS
Cyp3aSensitive to TCS to TCS

H. fossilisCATAntioxidant enzymesUpregulated by TCS
SODUpregulated by TCS
GSHAntioxidants; glutathione homeostasisDownregulated by TCS
GSTDownregulated by TCS
GPxDownregulated by TCS

C. auratusMDAOxidized lipid markerUpregulated by TCS
CATAntioxidant enzymesUpregulated by TCS
SODDownregulated by TCS
GSHAntioxidant; glutathione homeostasisUpregulated by TCS

Brachionus koreanusROSMetabolic oxygen by-productsUpregulated by TCS
Gst variantsAntioxidant enzyme; glutathione homeostasisSensitive to TCS to TCS
GpxSensitive to TCS to TCS
GSTUpregulated by TCS
SodAntioxidant enzymesSensitive to TCS to TCS
CatSensitive to TCS to TCS
Cyp3042a1Cytochrome family enzymes; detoxificationSensitive to TCS to TCS
Cyp43a1Sensitive to TCS to TCS
Hsp10Stress response; protein stabilizationSensitive to TCS to TCS
Hsp21Upregulated by TCS
Hsp27Upregulated by TCS
Hsp30Sensitive to TCS to TCS
Hsp40Sensitive to TCS to TCS
Hsp40hSensitive to TCS to TCS
Hsp60Sensitive to TCS to TCS
Hsp70Upregulated by TCS
Hsc70Upregulated by TCS
Hsp90α1Sensitive to TCS to TCS
Hsp90α2Sensitive to TCS to TCS
Hsp90βSensitive to TCS to TCS

B. gargarizans liverSodAntioxidant enzymeDownregulated by TCS
PhgpxAntioxidant enzyme; glutathione homeostasisDownregulated by TCS
P. perezi larvaeGSTUpregulated by TCS

R. philippinarum digestive glandMDAOxidized lipid markerUpregulated by TCS
CATAntioxidant enzymesSensitive to TCS to TCS
SODSensitive to TCS to TCS
GPx variantsAntioxidant enzymes; glutathione homeostasisSensitive to TCS to TCS
GSTSensitive to TCS to TCS
GRSensitive to TCS to TCS
ERODDetoxification enzymeSensitive to TCS to TCS

O. mykiss liver and kidneyCatAntioxidant enzymesDownregulated by TCS
SodUpregulated by TCS
Gpx variantsAntioxidant enzymes; glutathione homeostasisUpregulated by TCS
GstaUpregulated by TCS
Hsp90bbStress response; protein stabilizationUpregulated by TCS
Hsp90baUpregulated by TCS
Hsc70aUpregulated by TCS

C. reinhardtiiROSMetabolic oxygen by-productsUpregulated by TCS
MDAOxidized lipid markerUpregulated by TCS
SodAntioxidant enzymeUpregulated by TCS
GpxAntioxidant enzyme; glutathione homeostasisUpregulated by TCS
R. rubrum S1HGpxUpregulated by TCS
GrxCAntioxidant enzymes; glutathione homeostasisUpregulated by TCS
TrxBUpregulated by TCS
OsmCAntioxidant enzymeUpregulated by TCS
DnaJHeat shock protein; general stress markerUpregulated by TCS
RpoNRNA polymerase factor sigma-54; general stress markerDownregulated by TCS
TerATellurite resistance protein A; general stress markerUpregulated by TCS
Psp variantsPhage shock proteins; general stress markersSensitive to TCS to TCS
ClpPATP-dependent protease, proteolytic subunit; general stress markerUpregulated by TCS
HrcAHeat-inducible transcription suppressor; general stress markerUpregulated by TCS

E. coli K12, MG1655OxyRROS sensor proteinsUpregulated by TCS
GrxAntioxidant enzymes; glutathione homeostasisUpregulated by TCS
Sod variantsAntioxidant enzymesUpregulated by TCS
Cat variantsUpregulated by TCS
Ahp variantsAntioxidant enzymesUpregulated by TCS

E. coliROSMetabolic oxygen by-productsUpregulated by TCS
YgiWAntioxidant proteinsDownregulated by TCS
SoxSDownregulated by TCS
YhcNDownregulated by TCS

Abbreviation: TrxB: thioredoxin; OsmC: peroxiredoxin osmotically inducible protein C-like. Effects of 2,4-DCP, a by-product of TCS degradation.

TCS treatment in the yellow catfish Pelteobagrus fulvidraco revealed induced CAT, EROD, ERND, and APND [116]. Expressional profiling of Cyp1a, Cyp3a, and Gst showed both up- and downregulation depending on TCS concentration and length of exposure, a pattern that was also seen with MDA formation. When another catfish, Heteropneustes fossilis, was treated with a cosmetic effluent rich in TCS, increased SOD and CAT activities and reduced GSH, GST, and GPx were noted [117].

Oxidative damage by TCS was also evident in the goldfish Carassius auratus, as MDA, CAT, and GSH were elevated in addition to a reduced total antioxidant capacity [118]. Variable responses by antioxidant enzymes and in MDA levels were recorded in the goldfish’s liver after TCS treatment under a pH range of 6 to 9 [119]. The oxidative potential of TCS was also evident in the rotifer Brachionus koreanus, detected as ROS overproduction and enhanced GST activity, in addition to transcriptional modulation of cytochromes, antioxidant genes Gst, Gpx, Sod, and Cat and chaperons (Table 3) [120]. Moreover, TCS inhibited Sod and phospholipid hydroperoxide glutathione peroxidase (Phgpx) expression in the liver of Bufo gargarizans tadpoles [121] and induced GST in Pelophylax perezi frog larvae [122].

Sendra et al. [123] studied the combined effect of titanium dioxide (TiO2) and a heterogeneous mixture of organic compounds including TCS using the clam Ruditapes philippinarum. Modulations in EROD, SOD, CAT, GPx, GST, and GR enzyme activities were noted in the clam’s digestive gland, in parallel with increased lipid peroxidation. TCS exposure caused alterations in Cat, Sod, Gpx1, Gpx2, Gsta, Hsp90bb, Hsp90ba, and Hsc70a genes in rainbow trout Oncorhynchus mykiss [124]. Although in one report TCS failed to elicit oxidative stress in the green algae Chlamydomonas reinhardtii [125], another report detected ROS formation following TCS exposure [90], which was also most recently confirmed by significantly increased MDA, downregulated Gpx, and upregulated Sod expression [126].

The antimicrobial nature of TCS makes bacteria an appropriate target for mechanistic studies. Using Rhodospirillum rubrum S1H, Pycke et al. [57] detected upregulation in a host of TCS-induced oxidative response genes, most notably Gpx. In E. coli K12, MG1655, the electro-Fenton transformation of TCS caused activation of genes related to ROS sensing, along with reduced glutaredoxin (Grx), Sod, Cat, and alkyl hydroperoxide reductase (Ahpr) [55]. Very recently, ROS formation by TCS was associated with diminished expression of antioxidants in E. coli (Table 3), an event that preceded mutagenesis and enhanced drug resistance in that species [56]. TCS was also recently used to validate novel self-luminescent bioreporter strains of Nostoc sp. PCC 7120 using Sod promoters [127].

Collectively, monumental evidence demonstrates the prooxidant properties of TCS evident as both overproduction of ROS and interference with the cellular antioxidant defensome. TCS is toxic in part by inducing oxidative damage in a wide range of organisms and by targeting a defined cluster of proteins in a fashion that is conserved among diverse species. Nonetheless, the vast majority of data are collected from non-human models, and, as is the case with other toxicological reports of TCS, studies conducted on man or human-derived tissues are severely lacking.

5. Immunity and Inflammation

TCS has, for a long time, been recognized as an effective therapy for infectious dermatitis [2931], and the observed curative capacity of the compound was solely attributed to its antimicrobial activity. It was not until the end of last century that associations between TCS exposure and remission of noninfectious inflammation were made [33, 35, 36], and the use of antibacterials as anti-inflammatory agents has gained deserved attention during the past two decades. For example, an appreciable number of antibiotics, including macrolides and quinolones, have been shown to possess anti-inflammatory activity [128132]. Follow-up efforts have successfully provided solid evidence for the direct interaction of TCS with inflammatory pathways.

Gaffar et al. [133] reported that TCS inhibits cyclooxygenase-1 (COX-1) and COX-2, 5-lipoxygenase and (LPO), 15-LPO, and interleukin- (IL-) 1β-induced prostaglandin E2 (PGE2) in gingival cells. TCS was also shown to suppress a wider range of inflammatory mediators including IL-1β-induced prostaglandin I2 (PGI2) and arachidonic acid, tumor necrosis factor (TNF)α-induced PGE2, phospholipase A2 (PLA2), and COX [134]. Moreover, in a double-blind crossover study, participants who used a mouthrinse with added 0.15% TCS developed significantly less oral erythematous lesions than those who used a TCS-free mouthrinse [135]. By then, the anti-inflammatory properties of TCS were established and were widely accepted within the scientific and medical communities.

TCS in prosthetic devices was found to have no influence on the acute phase response [136], and only modest differences were seen between TCS and stannous fluoride dentrifice [137]. Nevertheless, TCS, when applied intracrevicularly, improved clinical parameters of gingivitis [138]. In a recent double-blind, randomized, crossover study, it was concluded that TCS-containing toothpaste inhibits inflammation in peri-implant tissue [139].

To date, elaborations on the anti-inflammatory nature of TCS have been the focus of subsequent studies. Mustafa et al. [140142] identified IL-1β, interferon (IFN)γ, major histocompatibility complex (MHC) class II, and PGE synthase-1, as targets of TCS in human gingival fibroblasts. Of note, studies to discern the subcellular localization of TCS show preference for nuclear, as opposed to cytosolic, accumulation. Although initial uptake was considerably higher in the cytoplasm, a great proportion of cytosolic TCS was eliminated after repeated washing, while nuclear retention was observed [143]. This may explain the perturbed inflammatory signaling associated with TCS. Moreover, in primary human oral epithelial cells, TCS attenuated LPS-induced cytokine response including IL-8, IL-1α, and TNFα and aggravated the antimicrobial response, which was mediated through microRNA (miRNA) regulation of the toll-like receptor (TLR) pathway [144]. The findings were also reciprocated in cells derived from diabetic patients, with an exaggerated TLR response [145]. It was revealed that TCS, nevertheless, abrogated LPS-induced TLR response, again, through regulating miRNAs (stimulating miR146a and inhibiting miR155s).

In skin and leukocytes of mice topically treated with TCS, alterations in inflammatory responses were mediated through TLR4 [146]. Likewise, TCS downregulated parathyroid hormone- (PTH-) or PGE2-stimulated matrix metalloproteinase-13 (MMP-13) expression in rat osteoblastic osteosarcoma cells [147]. Since hyperactive MMP-13 is implicated in periodontal disease, it was suggested that TCS might have a protective role against oral inflammatory conditions through its action on that enzyme, among others [148].

Interestingly, favorable results have been observed for TCS against other inflammatory conditions including cardiovascular disease and hidradenitis suppurativa [149, 150]. Moreover, the use of TCS-impregnated ureteral stents seems to be a promising approach to combat urinary tract infections (UTI) and associated inflammation [151, 152]. Along those lines, an increased urinary TCS was related to increased serum IL-6 in pregnant women [92], pointing at a possible pro- or anti-inflammatory role.

In a unique effort by Barros et al. [153], TCS modulation of the inflammatory response in an ex vivo whole blood stimulation assay was investigated. In that study, TCS inhibited multiple inflammatory mediators induced by LPS, including interleukins, most notably IL-1 & IL-6, IFNs, and colony-stimulating factor (CSF) 2. Activation of type 1 T helper lymphocytes was interrupted through the action of TCS on CD70. In a related report, TCS also reduced the capacity of natural killer (NK) lymphocytes to lyse chronic myelogenous leukemia K562 cells [154]. Recently, chitosan-TCS particles reduced the expression of IL-1β-induced Cox2 and Il6, among other immune molecules in gingival fibroblasts (Table 4) [155], showcasing the vast amenability of this antimicrobial to nanoparticle manipulation.


ModelTargetResponse
BiomarkerMolecular identity

Human gingival fibroblastsCOX-1/2Inflammatory mediatorsDownregulated by TCS
5/15-LPODownregulated by TCS
PGE2Downregulated by TCS
PGI2Downregulated by TCS
Arachidonic acidDownregulated by TCS
PLA2Downregulated by TCS
PGE synthase-1Downregulated by TCS
IFNγImmune/inflammatory cytokinesDownregulated by TCS
IL-1βDownregulated by TCS
MHC IICell surface proteins; adaptive immunity regulatorsDownregulated by TCS
Cox2Inflammatory mediatorDownregulated by TCS
Il6Immune/inflammatory cytokinesDownregulated by TCS
Il1bDownregulated by TCS
Tlr6Innate immunity receptorUpregulated by TCS

Human primary oral epithelial cellsIL-8Immune/inflammatory cytokinesDownregulated by TCS
IL-1αDownregulated by TCS
TNFαDownregulated by TCS
miR146aTranscriptional regulators of TLR responseUpregulated by TCS
miR155sDownregulated by TCS

Mouse skin and leukocytesS100A8/A9Inflammatory modulator; Ca2+-binding proteinUpregulated by TCS
Tlr4Innate immunity receptorsUpregulated by TCS
TLR4Upregulated by TCS
Tlr1Upregulated by TCS
Tlr2Upregulated by TCS
Tlr6Upregulated by TCS

Rat osteoblastic osteosarcoma cellsMMP-13Endopeptidase; collagen degradationDownregulated by TCS

Human oral fluidsIL-1αImmune/inflammatory cytokinesDownregulated by TCS
IL-1βSensitive to TCS
IL-8Sensitive to TCS
MCP-1Sensitive to TCS
TIMP-2MMP regulator proteinsSensitive to TCS
TIMP-1Downregulated by TCS
MMP-8/9Endopeptidases; extracellular matrix degradationDownregulated by TCS

Human urineIL-6Immune/inflammatory cytokinesUpregulated by TCS
Sprague-Dawley ratsTNFαUpregulated by TCS
IL-6Upregulated by TCS

Human whole blood leukocytesCsf2Hematopoietic stem cell growth and maintenanceDownregulated by TCS
Ifna1Immune/inflammatory cytokinesDownregulated by TCS
Ifna2Downregulated by TCS
Ifna4Downregulated by TCS
Ifna8Downregulated by TCS
Il-1f10Downregulated by TCS
Il-1f5Downregulated by TCS
Il-1f7Downregulated by TCS
Il-1f8Downregulated by TCS
Il-1f9Downregulated by TCS
Il-6Downregulated by TCS
Il-11Downregulated by TCS
Il-13Downregulated by TCS
Il-25Downregulated by TCS
Il-19Downregulated by TCS
Il-21Downregulated by TCS
Il-9Downregulated by TCS
Cd70Cell surface receptor/ligand; activated lymphocytesDownregulated by TCS
Bmp2Growth factors; bone and cartilage developmentUpregulated by TCS
Bmp6Upregulated by TCS
Tnfrsf11bTNFSF11 receptorDownregulated by TCS
Gdf3Growth/differentiation factorsDownregulated by TCS
Gdf2Downregulated by TCS
Gdf5Downregulated by TCS
Gdf9Downregulated by TCS
InhbaHypothalamus-pituitary axis regulatorDownregulated by TCS
Lefty2Left-right determination factor 2; left-right asymmetry of organsDownregulated by TCS

Sprague-Dawley ratsTNFαImmune/inflammatory cytokineUpregulated by TCS
IL-6Upregulated by TCS

Abbreviation: MCP: monocyte chemoattractant protein; TIMP: tissue inhibitor of metalloproteinase; Bmp: bone morphogenetic protein; Gdf: growth differentiation factor; Inhba: inhibin beta A chain.

ModelEffectClassification

HepG2 cellsGlobal DNA hypomethylationLimited evidence of carcinogenicity
V79 cellsChromosomal aberrations
MouseSomatic mutation (positive spot test)
Increased incidence of liver tumors
Aggravated hepatocellular carcinoma
Exacerbated colon tumorigenesis

D. polymorphaDNA damage (positive comet assay)N/A
U. tumidusDNA strand breaks (Hoescht 33342 fluorescence)
E. FetidaDNA damage (positive Comet assay)
D. magnaDNA damage (positive Comet assay)
A. salinaDNA damage (positive Comet assay)
H. fossilisDNA damage (positive Comet assay)
C. auratusDNA damage (positive Comet assay)
O. mykissDNA damage (positive Comet assay)
T. thermophilaDNA damage (positive Comet assay)
A. cepaChromosomal stickiness, reduced mitotic activity, and ana-telophase bridges (positive Feulgen reaction)

N/A = data from non-mammalian animals are not considered for ECHA mutagenicity/carcinogenicity classification.

Other in vivo studies on rodents and marine organisms clarified further the immunomodulatory properties of TCS. For instance, in mice subjected to an acute, systemic E. coli infection, Sharma et al. [156] demonstrated that cotreatment with TCS significantly reversed the damage caused by the bacteria. Specifically, TCS prolonged survival; lessened hepatic congestion, hemorrhage, and fatty changes; and reduced blood liver enzymes, serum TNFα, and the severity of bacteremia. In accordance with published data, TCS was similarly immunosuppressive in aquatic mussels (M. galloprovincialis) and clams (R. philippinarum) [64, 65].

Contrary to the overwhelming evidence of the anti-inflammatory function of TCS, a number of studies have nonetheless identified a proinflammatory role by the antiseptic. For example, upon intratracheal instillation of TCS in Sprague-Dawley rats, elevated total cell (TC) count, polymorphonuclear leukocytes (PMNs), total protein (TP), LDH, TNFα, and IL-6 were observed in bronchoalveolar lavage (BAL) fluid [83], which, except for TP, returned to baseline levels 14 days after exposure. Consonantly, it has also been demonstrated that TCS exacerbates diethylnitrosamine-induced hepatocellular carcinoma in C57BL/6 mice [103]. Likewise, TCS was very recently found to increase Tlr4 expression to promote colitis and aggravate colitis-related cancer in C57BL/6 mice [157].

It is evident from the wealth of information present that TCS is a modulator of immune and inflammatory reactions. The sum of data from in vitro and in vivo studies indicates that TCS, on its own, is immunosuppressive. Nevertheless, increasing evidence seems to suggest that in the presence of an existing adverse condition, such as inflammation or tumor, TCS further potentiates and worsens the eventual outcome. Investigations into the molecular basis behind this unique behavior are particularly warranted.

6. Genotoxicity and Carcinogenicity

Among the most important aspects of toxicological profiling of compounds is their interaction with the molecule of life—the DNA. Early efforts [42, 158] point at a possible role for TCS in somatic mutations observed in mice. TCS also caused a significant reduction in global DNA methylation in human hepatocellular carcinoma HepG2 cells, a finding associated with liver tumor [159]. Similarly, TCS caused a dose-responsive increase in chromosomal aberrations in lung fibroblast V79 cells, but not in ovary CHO cells, of the Chinese hamster Cricetulus griseus [12]. In a comparative study on Drosophila melanogaster using three mouthwashes, namely, Cepacol® (0.05% cetylpyridinium chloride), Periogard® (0.12% chlorhexidine digluconate), and Plax® (0.03% TCS), it was concluded that only the ethanol content in Cepacol®, but not other active ingredients, caused mitotic recombination between homologous chromosomes [160]. On the other hand, TCS induced dose-responsive DNA damage in hemocytes of the zebra mussel D. polymorpha [88], and strand breaks in the digestive gland of U. tumidus mussels [114]. A similar dose-dependent DNA damage was also observed in the earthworm E. fetida [108, 109], but not in E. andrei [161].

Comparing TCS to other toxicants in the larvae of freshwater insect Chironomus riparius, Martinez-Paz et al. [162] found TCS, along with nonylphenol, to be the most potent in causing DNA breakage. It was also noted that TCS, either alone or in combination with carbendazim, induced DNA damage in D. magna [163]. Using the brine shrimp Artemia salina, a time-dependent pattern of TCS-induced genotoxicity was identified [164]. Moreover, TCS was genotoxic in the catfish Heteropneustes fossilis, goldfish C. auratus, and rainbow trout O. mykiss [117, 118, 124]. Importantly, when TCS at an environmentally relevant concentration (3 nM) was treated to the freshwater protozoan Tetrahymena thermophila, notable DNA damage, without significant perturbation in growth or cell viability, was evident [165]. In a more detailed study on E. coli, Gou et al. [55] revealed that the electro-Fenton transformation of TCS caused upregulation of a host of genes involved in the DNA repair machinery, indicative of DNA stress. These genes belong to base excision repair (mutT and nfo), nucleotide excision repair (uvrA and uvrD), mismatch repair (uvrD and ssb), and double-strand break repair (ssb and recN). Chromosomal stickiness, reduced mitotic activity, and ana-telophase bridges were also noticeable in the bulb onion Allium cepa following TCS treatment [166].

In a recent proof-of-concept study, the promising potential of a toxicogenomic approach as a follow-up to positive in vitro genotoxicity data was evaluated. Using TCS as a testing compound, it was shown that the antimicrobial is non-DNA reactive and that it is genotoxic solely in vitro as opposed to in vivo [167].

Ambiguity surrounding the carcinogenicity of TCS still remains today. Investigators have generally been able to provide evidence for carcinogenic effects in animal models but not in humans. Of the earliest studies in this regard was a report by Lyman and Furia [38] identifying TCS as a carcinogen in mice. Other studies on mice have been in agreement with that conclusion. For example, it was noted that chronic TCS exposure increased the incidence of liver neoplasms [12] and aggravated hepatocellular carcinoma [103]. Furthermore, TCS caused colonic inflammation and worsened colitis or tumorigenesis induced by dextran sodium sulfate [168]. These findings, were, however, not paralleled in rats, hamsters, or baboons [12, 169]. More importantly, in vivo human studies of TCS are scarce, and aspects related to TCS-induced oncogenesis are lacking. Consequently, whether TCS poses a carcinogenic hazard to humans is unknown and requires further investigation. Nonetheless, the interaction of TCS with human-derived cancer cells in vitro has recently gained considerable attention (reviewed under Therapeutic Proposals).

In light of available data (Table 5), TCS demonstrates carcinogenicity solely in mice and within a narrow range of tissues (the liver and colon), which constitutes limited evidence of carcinogenicity according to ECHA (https://echa.europa.eu/documents/10162/23036412/clp_en.pdf/58b5dc6d-ac2a-4910-9702-e9e1f5051cc5). Hence, TCS is not classifiable as a carcinogen (http://ec.europa.eu/health/scientific_committees/consumer_safety/docs/sccs_o_054.pdf). It must be noted that in case future assessment conclusively rules out TCS as a human carcinogen, caution with its use must still be exercised given the established carcinogenicity of its transformation products—dioxins, chloroform, and anilines [170].

7. Cellular Signaling

Adaptations to the ever-changing intracellular and surrounding environments are achieved, in large part, by effective communication. Transmission of information that carries specific instructions is executed by messengers that function in tandem within a defined pathway. Tasks, however, are usually accomplished through the sequential transduction of multiple messages along a complex, intertwining network that involves a wide assortment of mediators [171]. Hence, the participation of cell signaling cascades in the response to xenobiotics cannot be overlooked.

The use of human cell lines has provided a wealth of information particularly regarding the study of signaling molecules responsive to stressors and xenobiotics, including TCS. In H460 lung cancer cells, TCS promoted migration and invasion through focal adhesion kinase/ATP-dependent tyrosine kinase (FAK/Akt) and Ras-related C3 botulinum toxin substrate 1 (Rac1) [72]. Evidence similarly exists for the classical mitogen-activated protein kinases (MAPK) as targets of TCS. For example, proliferation of JB6 Cl 41-5a cells as induced by TCS was accompanied by activation of extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinases (JNK), and p38 MAPKs, in addition to Akt [84]. Importantly, blocking either MEK1/2 or phosphoinositide 3-kinase (PI3K) significantly attenuated TCS-induced proliferation. In another study on rat neural stem cells, TCS-induced cytotoxicity and apoptosis were accompanied by activation of p38 and JNK and suppression of ERK, Akt, and PI3K [79]. This points at the involvement of these proteins in both cellular survival and death as brought about by TCS. Recently, TCS was shown to activate p38 and JNK in vivo as detected in the hypothalamus of Sprague-Dawley rats and in vitro utilizing human Nthy-ori 3-1 thyroid follicular cells [97]. In that study, TCS stimulated the thyrotropin-releasing hormone receptor through p38 MAPK, which, in turn, influenced the thyroid peroxidase (TPO) level.

In suppressing TLR signaling in whole blood leukocytes, TCS downregulated the expression of several signaling mediators, most notably, NF-κB-inducing kinase (Nik) and C-jun, which accounted for the overall blunted inflammatory response to LPS in these cells [153]. Furthermore, suppression of Mmp-13 expression in mouse osteoblastic osteocarcinoma cells by TCS was possibly related to its inhibition of Fos/Jun and AP-1 sequence binding in both the Mmp-13 and C-fos promoters [147].

The endocrine-disrupting activity of TCS, specifically its estrogenicity, has been of great interest to researchers. Kim et al. [73] utilized BG-1 ovarian cancer cells to show that the proliferative effects of TCS were mediated through ERα. Confirming the ER’s role, the use of ICI 182,780 reversed the proliferative properties of TCS along with associated perturbations in cyclin D1, p21, and Bax expression and protein levels. Likewise, the ER is implicated in TCS-induced proliferation of MCF-7 cells and increased breast tumor mass in mice [74, 75, 172]. This was similarly indicated by TCS inhibition with ICI 182,780 or kaempferol and the stimulation of insulin-like growth factor (IGF) signaling, namely, phosphorylated insulin receptor substrate (pIRS-1), pAkt, pMEK1/2, and pERK1/2 [75]. Notably, kaempferol also inhibited TCS-induced VM7Luc4E2 cell growth [76]. These observations are in congruence with an earlier report by Huang et al. [173] describing the estrogenic activities of nanomolar concentrations of TCS in the same cells. Investigating ER-responsive genes on the transcriptional and translational levels, it was shown that TCS induced pS2 but blunted ERα mRNA and protein levels, the latter of which was related to elevated miR-22, miR-206, and miR-193b miRNAs.

Recent studies have also argued for the dual effect of TCS on ER signaling. For example, Henry and Fair [174] demonstrated that, when administered alone to MCF7 cells, TCS at 7 nM to 700 μM exhibits estrogenic activity but becomes antiestrogenic in the presence of E2. Along those lines, it was shown that TCS, on its own, lacked any effect on rat uterine growth, but could still potentiate the effect of ethinylestradiol (EE) [175]. In a follow-up investigation, it was reported that TCS promotes EE-induced inhibition of ERα and ERβ expression and when given alone does not activate ER at concentrations from 30 nM to 100 μM [176]. Furthermore, TCS diminished E2 and estrogen sulfotransferase in sheep placenta [177]. This is in contrast to the increased activity of ERβ but not ERα caused by a TCS-derivative mixture, which led to neurological and behavioral abnormalities in zebrafish [87]. Also, Sprague-Dawley rats given TCS showed increased uterine weight and Calbindin-d(9k) (CaBP-9k) expression, which was also reciprocated in pituitary GH3 cells [178]. Reversal of both anomalies by ICI 182,780 and RU 486 points at a possible estrogenic role of the antimicrobial.

Very recently, Serra et al. [179] challenged accumulating evidence of TCS estrogenicity by showing the lack of agonistic or antagonistic effect in vivo and in vitro. While up to 0.3 μM TCS did not modulate ER-dependent brain aromatase in zebrafish embryos, interference with the enzyme’s activity, and with E2 activation of the enzyme observed at 1 μM, was not attributed to TCS-ER interaction. Moreover, up to 10 μM TCS lacked estrogenic effects in ER-expressing zebrafish liver cells as well as in MCF-7 cells [179]. Additionally, in a screening study of the estrogenicity of a group of endocrine-disrupting chemicals on fish species, TCS failed to significantly elicit a response in an in vitro ERα reporter gene assay [180].

In light of available evidence, the general consensus seems to indicate that the estrogenicity of TCS is contingent upon multiple factors, including concentration, species, duration of exposure, and whether TCS is administered alone or in combination with other molecules.

With regard to the androgenic properties of TCS, it was revealed that TCS interferes with testosterone- (TSN-) related transcription but promotes that dependent on androgen [181, 182]. In a recent in vivo study on weanling male rats, Riad et al. [104] reported that TCS, either alone or combined with butylparaben, reduced TSN, leutinizing hormone (LH), and follicle-stimulating hormone (FSH), while increased E2 was observed upon single TCS administration Also, TCS-induced proliferation and migration of LNCaP cells were significantly reduced in presence of bicalutamide, an androgen receptor (AR) antagonist [77]. These findings support a previous report by Ahn et al. [183] in which 1 μM TCS reduced E2-induced ER activation by 50% and AR in human BG1Luc4E2 ovarian adenocarcinoma cells and T47D-ARE breast cancer cells, respectively. Evidence for TCS estrogenicity was detected in MCF7 cells when [(3)H]estradiol was successfully displaced from the ER by the antimicrobial [184]. Furthermore, 10 μM TCS attenuated E2-dependent ERE-CAT reporter gene induction, while 0.1 and 1 μM TCS inhibited TSN-stimulated LTR-CAT reporter gene in both T47D cells and S115 mouse mammary tumor cells [184]. TCS was also determined to have a weak effect on AhR in recombinant rat hepatoma (H4L1.1c4) cells. Finally, Forgacs et al. [185] showed that TCS interferes with recombinant hCG stimulation of TSN in a novel BLTK1 murine Leydig cell model. Most recently, however, no significant influence on androgen synthesis or activity by TCS was observed in Wistar rats [186].

Controversy surrounding the interaction between TCS and members of the peroxisome proliferator-activated receptors (PPARs) has gained considerable attention as of late. This has essentially stemmed from the apparent discrepancy between data obtained from humans and those from rodents. In comparing the differential modulation of TCS on PPARα in HepG2 cells and mouse hepatoma Hepa1c1c7 cells, distinct responses were observed by Wu et al. [187]. Protein levels of PPARα downstream target, acyl-coenzyme A oxidase, were decreased in HepG2 cells but were increased in Hepa1c1c7, which also showed higher DNA synthesis and blunted apoptosis through transforming growth factor (TGF-β). PPAR signaling was similarly identified as a target of TCS through genome-wide CRISPR-Cas9 screening in HepG2 cells [188], zebrafish [189], and Gallus gallus chicken embryos [190]. In the latter model, PPAR signaling members Cyp7a1, fatty acid-binding protein 1 (Fabp1), acyl-CoA synthetase long-chain family member 5 (Acsl5), acyl-CoA oxidase 2 (Acox2), and perilipin 1 (Plin1) were upregulated, whereas angiopoietin-like 4 (Angptl) was downregulated.

TCS administered to pregnant mice caused insulin resistance, hypothyroidism, diminished glucose transporter 4 (GLUT4) expression, and inhibition of Akt and mTOR phosphorylation [191, 192]. While thyroxine corrected these adversaries, PPARγ activator, rosiglitazone, solely reversed the decrease in Akt phosphorylation in adipose tissue and in muscle [192]. PPARγ is known to ameliorate mTOR suppression-induced glucose intolerance in rats [193], further underlining the far-reaching effects of TCS action.

Although TCS has been reported to promote hepatocyte proliferation in mice through PPAR [12], Yueh et al. [103] found no appreciable induction of PPARα following TCS treatment. Importantly, the authors also identified constitutive androstane receptor (CAR) as a possible aggravator of TCS-induced tumorigenesis, given the halved tumor number in Car–/− mice compared to their Car+/− counterparts. TCS, as is the case with PPARs, is reported to exhibit varying affinities for CAR and pregnane X receptor (PXR) in humans and rodents. A weak agonist for human CAR, TCS was found to be a reverse agonist for rodent CAR, an agonist for human PXR, and had no effect on rodent PXR [194].

Calcium concentration within cells influences protein conformation and dynamics. Protein binding of Ca2+, on the other hand, maintains the ion’s content within a physiological range and sets forth diverse cellular activities related to gene expression, motility, secretion, and survival [195]. Beside proteins, intracellular Ca2+ levels are modulated by a variety of stimuli, including xenobiotic exposure. Through the Ca2+ channel ryanodine (Ry) receptor type 1 (RyR1), TCS increased cytosolic Ca2+ dose-dependently in primary skeletal myotubes irrespective of extracellular Ca2+ [183]. Accordingly, muscle contractility was compromised upon TCS exposure in vitro and in vivo [196]. Results from this study indicate that TCS impaired excitation-contraction coupling (ECC) in cardiac and skeletal muscles and enhanced electrically induced Ca2+ transients in myotubes without depleting intracellular Ca2+ and notwithstanding RyR1 blockage. TCS also efficiently blocked excitation-coupled Ca2+ entry and interfered with the bidirectional signaling between RyR1 channels and Ca2+ ions. Likewise, TCS compromised ECC in larval fathead minnows Pimephales promelas, as evidenced by altered RyR and dihydropyridine receptor (DHPR) mRNA and protein levels and weakened ligand binding to both receptors in adult muscle homogenates [197].

In rat thymocytes, TCS elevated intracellular Ca2+ levels and opened Ca2+-responsive K+ channels, eventually leading to membrane hyperpolarization [198]. Also, TCS prevented Ca2+-induced mitochondrial swelling in rat liver [199]. A more in-depth analysis of TCS modulation of Ca2+ homeostasis was conducted on rat basophilic leukemia (RBL) mast cells [24]. In this cell type, TCS caused mitochondrial fission and diminished membrane potential and translocation, with compromised ATP production and elevated ROS. These changes were associated with perturbed mitochondrial and endoplasmic reticulum Ca2+ and depleted cytosolic Ca2+ levels following antigen stimulation. Accordingly, TCS-induced degranulation of mast cell may at least in part be attributed to Ca2+ mobilization.

Calcium modulation by TCS has also been investigated in other organisms. In C. reinhardtii exposed to 14 μM TCS, increased Ca2+ levels with oxidative stress, cell and mitochondrial membrane depolarization, compromised photosynthesis, and caspase activation were noted [90]. Importantly, chelation of intracellular Ca2+ ions by BAPTA-AM protected the algae from TCS-induced Ca2+ dysregulation. These observations strongly implicate Ca2+ as a mediator of a wide array of toxic anomalies attributed to TCS.

Literature concerning the xenobiotic response to TCS has revealed important signaling pathways activated or suppressed by TCS (Table 6). Distinct outcomes exist among species and even within the same species based on experimental conditions and model under investigation. Although important milestones in TCS signaling have been achieved so far, there remains a lot to be discovered, especially in human-based systems, about the modulatory effects of TCS on cellular physiology. In particular, the response of many human cell types and tissues to TCS treatment is unknown, and identification of signaling pathways and their roles in cellular growth, metabolism, and overall function is therefore advised.


ModelTargetResponse
PathwaysTCS role

H460 cellsFAK/AktCellular migration and invasionUpregulated by TCS
Rac1Upregulated by TCS

JB6 Cl 41-5a cellsERK1/2Cell proliferationUpregulated by TCS
JNKUpregulated by TCS
p38Upregulated by TCS
AktUpregulated by TCS
PI3KUpregulated by TCS

Rat neural stem cellsJNKCytotoxicity and apoptosisUpregulated by TCS
p38
Upregulated by TCS
ERK
AktDownregulated by TCS
PI3KDownregulated by TCS
Downregulated by TCS

Sprague-Dawley rats hypothalamus and Nthy-ori 3-1 cellsJNKReduced TPO; hypothyroidismUpregulated by TCS
p38Upregulated by TCS

Whole blood leukocytesNikAnti-inflammatory responseDownregulated by TCS
CjunDownregulated by TCS
Mouse osteoblastic osteocarcinomaFosDownregulated by TCS
JunDownregulated by TCS
Ap1Downregulated by TCS

BG-1ERαCell proliferationUpregulated by TCS
MCF-7 cellsERαSensitive to TCS
pIRS-1Upregulated by TCS
pAKTUpregulated by TCS
pMEK1/2Upregulated by TCS
pERK1/2Upregulated by TCS
VM7Luc4E2 cellsErαDownregulated by TCS
Ps2Upregulated by TCS
ERαDownregulated by TCS
pS2Upregulated by TCS
miR-22Upregulated by TCS
miR-206Upregulated by TCS
miR-193bUpregulated by TCS

Sheep placentaE2Anti-estrogenicityDownregulated by TCS
Estrogen sulfotransferaseDownregulated by TCS
BG1Luc4E2 cellsERDownregulated by TCS

Sprague-Dawley rats and GH3 cellsCaBP-9 kEstrogenicityUpregulated by TCS

LNCaPARAndrogenicity; cell proliferation, and migrationUpregulated by TCS

T47D-ARE cellsARAnti-androgenicityDownregulated by TCS

H4L1.1c4 cellsARPro(anti)-androgenicitySensitive to TCS

HepG2 cellsAcyl-coenzyme A oxidaseBlunted lipid metabolismDownregulated by TCS

Hepa1c1c7 cellsAcyl-coenzyme A oxidaseEnhanced lipid metabolism and DNA synthesisUpregulated by TCS
TGF-βAntiapoptosisDownregulated by TCS

D. rerioPPARαEnhanced lipid metabolismUpregulated by TCS
PPARγUpregulated by TCS
G. gallus embryo liversPPARαUpregulated by TCS

ICR miceAktImpaired glucose metabolismDownregulated by TCS
mTORDownregulated by TCS

C57BL/6 miceCARTumorigenesisUpregulated by TCS

HepG2 cellsCAREnhanced hepatic catabolismUpregulated by TCS
PXRUpregulated by TCS

Rodent FAO hepatoma cellsCARReduced hepatic catabolismDownregulated by TCS

Primary skeletal myotubesCa2+Diminished muscle contractilityUpregulated by TCS
RyR1Upregulated by TCS
P. promelas muscle homogenatesRyr2Sensitive to TCS
Ryr3Downregulated by TCS
RyRDownregulated by TCS

Rat thymocytesCa2+Cell membrane hyperpolarizationUpregulated by TCS

RBL cellsCa2+Mast cell degranulationDownregulated by TCS

C. reinhardtiiCa2+Dampened photosynthesisUpregulated by TCS

TCS is anti-estrogenic in the presence of E2.

8. Therapeutic Proposals

The first specific action mechanism of TCS in prokaryotes was only demonstrated 20 years ago, when inhibition of fatty acid synthesis in Escherichia coli was noted following exposure to TCS [200, 201]. TCS irreversibly inhibited the fatty acid biosynthesis enzyme, enoyl–acyl carrier protein reductase (ACP), by mimicking its natural substrate in vivo. Further, a mutated or overexpressed ACP, encoded by fabI, was shown to confer TCS resistance in the bacterium. These findings established ACP as a specific, subcellular TCS target. Efforts have thus far revealed the susceptibility of a host of other pathogens to inhibition of fatty acid synthesis by TCS. These include Staphylococcus aureus, M. tuberculosis, Helicobacter pylori, Haemophilus influenzae, Plasmodium falciparum, Toxoplasma gondii, Leishmania spp., and Trypanosoma spp. [52, 202208]. In humans, fatty acid synthase (FAS) is the only multienzyme complex that is responsible for the endogenous synthesis of saturated fatty acids from acetyl-CoA and malonyl-CoA [209, 210]. Although a BLAST analysis of E. coli FabI protein and FAS showed no homology, appreciable sequence similarities were nevertheless found with polyketide synthase and type I FAS of M. tuberculosis [211].

The success of cerulenin, a mycotoxin with fatty acid inhibitory action, in suppressing tumor progression in vivo has spawned several reports in support of fatty acid synthesis inhibition as an emerging target for chemotherapy [212]. The earliest study in this regard investigated the cytotoxicity of TCS in MCF-7 and SKBr-3 breast cancer cells [211]. It was revealed that TCS at 10-50 μM is cytotoxic and antiproliferative, induces morphological alterations, and inhibits FAS. These findings corroborate an earlier observation linking FAS inhibition with apoptotic death of breast cancer cells [211, 213, 214]. TCS was similarly found to inhibit the development of methylnitrosourea-induced breast cancer in Sprague-Dawley rats [209]. In human A-375 melanoma cells, TCS inhibited growth at 40 μM [215]. TCS was similarly found to be dose-dependently proapoptotic in prostate cancer cells, with IC50 values as low as 4.5-7.8 μM [216]. Whereas no cytotoxicity was observed in NIH3T3 fibroblasts at concentrations up to 60 μM, values of IC50 ranging from 0.74 to 62 μM were nonetheless observed in nonmalignant prostate cells. This suggests two things; first, that prostate cells are relatively more sensitive to TCS toxicity than fibroblasts and presumably other nonmalignant cell types, and second, that malignant prostate cells exhibit higher chemosensitivity compared to their nonmalignant counterparts. This differential susceptibility could be due to overexpressed FAS in malignant cells. However, in contrast to these reports, at concentrations up to 345 μM, TCS was found to be preferentially cytotoxic to Y79 RB cells over mouse 3T3 fibroblasts and human MIO-M1 Müller glial cells as indicated by IC50 values, creating a large therapeutic index of 7.1 and 5.3, respectively [217]. FAS suppression, depleted fatty acid content, lipid peroxidation, and apoptotic death were noted in Y79 RB cells at the same TCS concentration range [98]. Recently, TCS at 40 μM was also shown to be effective against MiaPaCa-2 and AsPC-1 pancreatic cancer cells suppressing proliferation and eliciting apoptotic death [218]. Of note, in a related study, TCS impeded mouse preadipocyte differentiation [219]. Given the regulation of food intake by FAS, and the susceptibility of adipocyte development to TCS inhibition, it was suggested that TCS may possess anti-obesogenic properties.

The differential expression and activity of FAS in healthy and malignant tissues, where it is upregulated in the latter [220, 221], indicate a possibly high therapeutic index. The long history of human use, and the ubiquity of TCS in consumer products, coupled with encouraging in vivo results, cements the antimicrobial as a promising candidate for chemotherapy. As noted earlier, it must be stressed that variations in the final outcome of TCS treatment largely depend on experimental setup. Moreover, limited data from animal studies suggest that in the presence of a preexisting tumor, TCS administration seems to exacerbate the condition. This observation is concerning and indeed warrants further investigation before TCS can be invested in for clinical trials.

9. Conclusion

TCS is a synthetic antimicrobial with a long history of human use. At concentrations well below those present in commercial products, data from in vitro and in vivo studies have provided evidence of adverse effects on diverse molecular pathways. Most alarmingly is TCS enhancement of malignant cell proliferation in vitro and tumor growth in vivo. On the other hand, TCS has also been shown to be protective against malignant cell growth and proliferation, possibly opening the door for its use in chemotherapy. Clearly, dose and time dependence is an important factor in determining the eventual denouement of the chemical. In spite of the numerous publications dissecting the signaling pathways responsive to TCS, it is evident that a severe paucity surrounding human-based in vivo and in vitro studies still remains today. Future studies, thus, should focus on identifying signaling molecules differentially regulated by TCS and characterize their roles in toxic or protective effects in different cell types. Insights gained from such revelations will be invaluable to possibly validate targets for drug development or devise possible TCS adjuvants or inhibitors.

Conflicts of Interest

The authors declare that there are no conflicts of interest regarding the publication of this paper.

Acknowledgments

We thank the members of the Lee Laboratory for helpful advice and discussion during this work. This work was supported in part by the Brody Brothers Grant (BBE216102), NIH (1R15AG060373-01) to M-H.L., and the Saudi Government Graduate Scholarship (through King Saud University) to M.A.A.

References

  1. R. Montville and D. W. Schaffner, “A meta-analysis of the published literature on the effectiveness of antimicrobial soaps,” Journal of Food Protection, vol. 74, no. 11, pp. 1875–1882, 2011. View at: Publisher Site | Google Scholar
  2. V. Kjaerheim, A. Skaare, P. Barkvoll, and G. Rolla, “Antiplaque, antibacterial, and anti-inflammatory properties of triclosan mouthrinses in combination with zinc citrate or polyvinylmethylether maleic acid (PVM-MA) copolymer,” European Journal of Oral Sciences, vol. 104, no. 5-6, pp. 529–534, 1996. View at: Publisher Site | Google Scholar
  3. V. Kjaerheim, S. M. Waaler, and G. Rolla, “Organic solvents and oils as vehicles for triclosan in mouthrinses: a clinical study,” Scandinavian Journal of Dental Research, vol. 102, no. 5, pp. 306–308, 1994. View at: Google Scholar
  4. A. B. Skaare, V. Kjaerheim, P. Barkvoll, and G. Rolla, “Does the nature of the solvent affect the anti-inflammatory capacity of triclosan? An experimental study,” Journal of Clinical Periodontology, vol. 24, no. 2, pp. 124–128, 1997. View at: Publisher Site | Google Scholar
  5. V. Kjaerheim, S. M. Waaler, and G. Rolla, “Significance of choice of solvents for the clinical effect of triclosan-containing mouthrinses,” Scandinavian Journal of Dental Research, vol. 102, no. 4, pp. 202–205, 1994. View at: Google Scholar
  6. M. A. Alfhili, D. S. Yoon, T. A. Faten et al., “Non-ionic surfactants antagonize toxicity of potential phenolic endocrine-disrupting chemicals, including Triclosan in Caenorhabditis elegans,” Molecules and Cells, vol. 41, no. 12, pp. 1052–1060, 2018. View at: Publisher Site | Google Scholar
  7. S. M. Waaler, G. Rölla, K. K. Skjörland, and B. Ögaard, “Effects of oral rinsing with triclosan and sodium lauryl sulfate on dental plaque formation: a pilot study,” Eropean Journal of Oral Sciences, vol. 101, no. 4, pp. 192–195, 1993. View at: Publisher Site | Google Scholar
  8. K. Bellamy, R. Alcock, J. R. Babb, J. G. Davies, and G. A. J. Ayliffe, “A test for the assessment of ‘hygienic’ hand disinfection using rotavirus,” The Journal of Hospital Infection, vol. 24, no. 3, pp. 201–210, 1993. View at: Publisher Site | Google Scholar
  9. J. Regos and H. R. Hitz, “Investigations on the mode of action of Triclosan, a broad spectrum antimicrobial agent,” Zentralblatt fur Bakteriologie, Parasitenkunde, Infektionskramkheiten und hygiene, Erste Abteilung Originale, Reihe A: Medizinische Mikrobiologie und Parasitologie, vol. 226, no. 3, pp. 390–401, 1974. View at: Google Scholar
  10. J. Regös, O. Zak, R. Solf, W. A. Vischer, and E. G. Weirich, “Antimicrobial spectrum of triclosan, a broad-spectrum antimicrobial agent for topical application. II. Comparison with some other antimicrobial agents,” Dermatologica, vol. 158, no. 1, pp. 72–79, 1979. View at: Publisher Site | Google Scholar
  11. R. C. Petersen, “Triclosan antimicrobial polymers,” AIMS Molecular Science, vol. 3, no. 1, pp. 88–103, 2016. View at: Publisher Site | Google Scholar
  12. J. V. Rodricks, J. A. Swenberg, J. F. Borzelleca, R. R. Maronpot, and A. M. Shipp, “Triclosan: a critical review of the experimental data and development of margins of safety for consumer products,” Critical Reviews in Toxicology, vol. 40, no. 5, pp. 422–484, 2010. View at: Publisher Site | Google Scholar
  13. X. Chen, J. L. Nielsen, K. Furgal, Y. Liu, I. B. Lolas, and K. Bester, “Biodegradation of triclosan and formation of methyl-triclosan in activated sludge under aerobic conditions,” Chemosphere, vol. 84, no. 4, pp. 452–456, 2011. View at: Publisher Site | Google Scholar
  14. M. Adolfsson-Erici, M. Pettersson, J. Parkkonen, and J. Sturve, “Triclosan, a commonly used bactericide found in human milk and in the aquatic environment in Sweden,” Chemosphere, vol. 46, no. 9-10, pp. 1485–1489, 2002. View at: Publisher Site | Google Scholar
  15. T. Geens, H. Neels, and A. Covaci, “Distribution of bisphenol-A, triclosan and n-nonylphenol in human adipose tissue, liver and brain,” Chemosphere, vol. 87, no. 7, pp. 796–802, 2012. View at: Publisher Site | Google Scholar
  16. L. Hovander, T. Malmberg, M. Athanasiadou et al., “Identification of hydroxylated PCB metabolites and other phenolic halogenated pollutants in human blood plasma,” Archives of Environmental Contamination and Toxicology, vol. 42, no. 1, pp. 105–117, 2002. View at: Publisher Site | Google Scholar
  17. L. W. B. Olaniyan, N. Mkwetshana, and A. I. Okoh, “Triclosan in water, implications for human and environmental health,” Springerplus, vol. 5, no. 1, p. 1639, 2016. View at: Publisher Site | Google Scholar
  18. J. Schulze, F. H. Marquardt, F. Lyman, and C. Spitzer, “Determination of free and conjugated triclosan-1 in blood by electron capture gas liquid chromatography-2,” Journal of the American Oil Chemists' Society, vol. 52, no. 7, pp. 215–218, 1975. View at: Publisher Site | Google Scholar
  19. L. M. Weatherly and J. A. Gosse, “Triclosan exposure, transformation, and human health effects,” Journal of Toxicology and Environmental Health Part B, Critical Reviews, vol. 20, no. 8, pp. 447–469, 2017. View at: Publisher Site | Google Scholar
  20. Q. Wu, H. Shi, C. D. Adams, T. Timmons, and Y. Ma, “Oxidative removal of selected endocrine-disruptors and pharmaceuticals in drinking water treatment systems, and identification of degradation products of triclosan,” Science of The Total Environment, vol. 439, pp. 18–25, 2012. View at: Publisher Site | Google Scholar
  21. C. Queckenberg, J. Meins, B. Wachall et al., “Absorption, pharmacokinetics, and safety of triclosan after dermal administration,” Antimicrobial Agents and Chemotherapy, vol. 54, no. 1, pp. 570–572, 2010. View at: Publisher Site | Google Scholar
  22. G. Sandborgh-Englund, M. Adolfsson-Erici, G. Odham, and J. Ekstrand, “Pharmacokinetics of triclosan following oral ingestion in humans,” Journal of Toxicology and Environmental Health Part A, vol. 69, no. 20, pp. 1861–1873, 2006. View at: Publisher Site | Google Scholar
  23. S. B. Levy, “Antibacterial household products: cause for concern,” Emerging Infectious Diseases, vol. 7, Supplement 3, pp. 512–515, 2001. View at: Publisher Site | Google Scholar
  24. L. M. Weatherly, A. J. Nelson, J. Shim et al., “Antimicrobial agent triclosan disrupts mitochondrial structure, revealed by super-resolution microscopy, and inhibits mast cell signaling via calcium modulation,” Toxicology and Applied Pharmacology, vol. 349, pp. 39–54, 2018. View at: Publisher Site | Google Scholar
  25. J. L. Fang, M. Vanlandingham, G. G. da Costa, and F. A. Beland, “Absorption and metabolism of triclosan after application to the skin of B6C3F1 mice,” Environmental Toxicology, vol. 31, no. 5, pp. 609–623, 2016. View at: Publisher Site | Google Scholar
  26. T. Moss, D. Howes, and F. M. Williams, “Percutaneous penetration and dermal metabolism of triclosan (2,4, 4-trichloro-2-hydroxydiphenyl ether),” Food and Chemical Toxicology, vol. 38, no. 4, pp. 361–370, 2000. View at: Publisher Site | Google Scholar
  27. E. Franz and S. Weidner-Strahl, “The effectiveness of topical antibacterials in acne: a double-blind clinical study,” The Journal of International Medical Research, vol. 6, no. 1, pp. 72–77, 1978. View at: Publisher Site | Google Scholar
  28. T. W. Lee, J. C. Kim, and S. J. Hwang, “Hydrogel patches containing triclosan for acne treatment,” European Journal of Pharmaceutics and Biopharmaceutics, vol. 56, no. 3, pp. 407–412, 2003. View at: Publisher Site | Google Scholar
  29. P. Kalliomaki and K. Kuokkanen, “Comparative study on the efficacy and tolerance of the ointments CGP433 and GP41353 in the treatment of infectious dermatitis,” Zeitschrift für Hautkrankheiten, vol. 54, no. 14, pp. 668–670, 1979. View at: Google Scholar
  30. H. Weitgasser, C. Schindléry, and V. Macarol, “A comparative multicentre trial of halometasone/triclosan cream and betamethasone dipropionate/gentamicin sulphate cream in the treatment of infected acute eczematous dermatitis,” The Journal of International Medical Research, vol. 11, Supplment 1, pp. 43–47, 1983. View at: Google Scholar
  31. A. Aliaga, A. Castells, D. Kriznik et al., “An overview of two comparative multicentre trials with halometasone/triclosan cream in acute superficial bacterial skin infections,” The Journal of International Medical Research, vol. 11, Supplement 1, pp. 53–57, 1983. View at: Google Scholar
  32. L. Huber, “Role of Klion ointment in the treatment of crural ulcer,” Therapia Hungarica, vol. 39, no. 3, pp. 148–150, 1991. View at: Google Scholar
  33. P. Barkvoll and G. Rolla, “Triclosan protects the skin against dermatitis caused by sodium lauryl sulphate exposure,” Journal of Clinical Periodontology, vol. 21, no. 10, pp. 717–719, 1994. View at: Publisher Site | Google Scholar
  34. A. Skaare, G. Eide, B. Herlofson, and P. Barkvoll, “The effect of toothpaste containing triclosan on oral mucosal desquamation. A model study,” Journal of Clinical Periodontology, vol. 23, no. 12, pp. 1100–1103, 1996. View at: Publisher Site | Google Scholar
  35. V. Kjaerheim, P. Barkvoll, S. M. Waaler, and G. Rolla, “Triclosan inhibits histamine-induced inflammation in human skin,” Journal of Clinical Periodontology, vol. 22, no. 6, pp. 423–426, 1995. View at: Publisher Site | Google Scholar
  36. P. Barkvoll and G. Rolla, “Triclosan reduces the clinical symptoms of the allergic patch test reaction (APR) elicited with 1% nickel sulphate in sensitised patients,” Journal of Clinical Periodontology, vol. 22, no. 6, pp. 485–487, 1995. View at: Publisher Site | Google Scholar
  37. V. Kjaerheim, A. Roed, P. Brodin, and G. Rolla, “Effects of triclosan on the rat phrenic nerve-diaphragm preparation,” Journal of Clinical Periodontology, vol. 22, no. 6, pp. 488–493, 1995. View at: Publisher Site | Google Scholar
  38. F. L. Lyman and T. Furia, “Toxicology of 2, 4, 4-trichloro-2-hydroxy-diphenyl ether,” IMS, Industrial Medicine and Surgery, vol. 38, no. 2, pp. 64–71, 1969. View at: Google Scholar
  39. F. L. Lyman and T. E. Furia, “Toxicology of 2,4,4-trichloro-2-hydroxyphenyl ether,” IMS, Industrial Medicine and Surgery, vol. 37, no. 7, p. 546, 1968. View at: Google Scholar
  40. E. Arrhenius, L. Renberg, L. Johansson, and M. A. Zetterqvist, “Disturbance of microsomal detoxication mechanisms in liver by chlorophenol pesticides,” Chemico-Biological Interactions, vol. 18, no. 1, pp. 35–46, 1977. View at: Publisher Site | Google Scholar
  41. A. Y. K. Chow, G. H. Hirsch, and H. S. Buttar, “Nephrotoxic and hepatotoxic effects of triclosan and chlorhexidine in rats,” Toxicology and Applied Pharmacology, vol. 42, no. 1, pp. 1–10, 1977. View at: Publisher Site | Google Scholar
  42. L. B. Russell and C. S. Montgomery, “Use of the mouse spot test to investigate the mutagenic potential of triclosan (Irgasan DP300),” Mutation Research, vol. 79, no. 1, pp. 7–12, 1980. View at: Publisher Site | Google Scholar
  43. T. L. Miller, D. J. Lorusso, and M. L. Deinzer, “The acute toxicity of nonachloropredioxin and 3- and 4-hydroxynonachlorodiphenyl ether in mice,” Journal of Toxicology and Environmental Health, vol. 10, no. 4-5, pp. 699–707, 1982. View at: Publisher Site | Google Scholar
  44. T. L. Miller, D. J. Lorusso, M. L. Walsh, and M. L. Deinzer, “The acute toxicity of penta-, hexa-, and heptachlorohydroxydiphenyl ethers in mice,” Journal of Toxicology and Environmental Health, vol. 12, no. 2-3, pp. 245–253, 1983. View at: Publisher Site | Google Scholar
  45. J. Roed-Petersen, G. Auken, and N. Hjorth, “Contact sensitivity to Irgasan DP 300,” Contact Dermatitis, vol. 1, no. 5, pp. 293-294, 1975. View at: Publisher Site | Google Scholar
  46. B. Steinkjer and L. R. Braathen, “Contact dermatitis from triclosan (Irgasan DP 300),” Contact Dermatitis, vol. 18, no. 4, pp. 243-244, 1988. View at: Publisher Site | Google Scholar
  47. E. Storer, K. J. Koh, and L. Warren, “Severe contact dermatitis as a result of an antiseptic bath oil,” The Australasian Journal of Dermatology, vol. 45, no. 1, pp. 73–75, 2004. View at: Publisher Site | Google Scholar
  48. S. Veronesi, S. M. P. de Padova, D. Vanni, and M. Melino, “Contact dermatitis to triclosan,” Contact Dermatitis, vol. 15, no. 4, pp. 257-258, 1986. View at: Publisher Site | Google Scholar
  49. C. S. M. Wong and M. H. Beck, “Allergic contact dermatitis from triclosan in antibacterial handwashes,” Contact Dermatitis, vol. 45, no. 5, p. 307, 2001. View at: Publisher Site | Google Scholar
  50. W. A. Vischer and J. Regös, “Antimicrobial spectrum of Triclosan, a broad-spectrum antimicrobial agent for topical application,” Zentralblatt fur Bakteriologie, Parasitenkunde, Infektionskramkheiten und hygiene, Erste Abteilung Originale, Reihe A: Medizinische Mikrobiologie und Parasitologie, vol. 226, no. 3, pp. 376–389, 1974. View at: Google Scholar
  51. P. Gilbert and A. J. McBain, “Literature-based evaluation of the potential risks associated with impregnation of medical devices and implants with triclosan,” Surgical Infections, vol. 3, Supplement 1, pp. S55–S63, 2002. View at: Publisher Site | Google Scholar
  52. R. J. Heath, J. Li, G. E. Roland, and C. O. Rock, “Inhibition of the Staphylococcus aureus NADPH-dependent enoyl-acyl carrier protein reductase by triclosan and hexachlorophene,” The Journal of Biological Chemistry, vol. 275, no. 7, pp. 4654–4659, 2000. View at: Publisher Site | Google Scholar
  53. J. C. Betts, A. McLaren, M. G. Lennon et al., “Signature gene expression profiles discriminate between isoniazid-, thiolactomycin-, and triclosan-treated Mycobacterium tuberculosis,” Antimicrobial Agents and Chemotherapy, vol. 47, no. 9, pp. 2903–2913, 2003. View at: Publisher Site | Google Scholar
  54. V. B. Srinivasan, B. B. Singh, N. Priyadarshi, N. K. Chauhan, and G. Rajamohan, “Role of novel multidrug efflux pump involved in drug resistance in Klebsiella pneumoniae,” PLoS One, vol. 9, no. 5, article e96288, 2014. View at: Publisher Site | Google Scholar
  55. N. Gou, S. Yuan, J. Lan, C. Gao, A. N. Alshawabkeh, and A. Z. Gu, “A quantitative toxicogenomics assay reveals the evolution and nature of toxicity during the transformation of environmental pollutants,” Environmental Science & Technology, vol. 48, no. 15, pp. 8855–8863, 2014. View at: Publisher Site | Google Scholar
  56. J. Lu, M. Jin, S. H. Nguyen et al., “Non-antibiotic antimicrobial triclosan induces multiple antibiotic resistance through genetic mutation,” Environment International, vol. 118, pp. 257–265, 2018. View at: Publisher Site | Google Scholar
  57. B. F. G. Pycke, G. Vanermen, P. Monsieurs et al., “Toxicogenomic response of Rhodospirillum rubrum S1H to the micropollutant triclosan,” Applied and Environmental Microbiology, vol. 76, no. 11, pp. 3503–3513, 2010. View at: Publisher Site | Google Scholar
  58. L. M. McMurry, M. Oethinger, and S. B. Levy, “Overexpression of marA, soxS, or acrAB produces resistance to triclosan in laboratory and clinical strains of Escherichia coli,” FEMS Microbiology Letters, vol. 166, no. 2, pp. 305–309, 1998. View at: Publisher Site | Google Scholar
  59. P. Nuonming, S. Khemthong, T. Dokpikul, R. Sukchawalit, and S. Mongkolsuk, “Characterization and regulation of AcrABR, a RND-type multidrug efflux system, in Agrobacterium tumefaciens C58,” Microbiological Research, vol. 214, pp. 146–155, 2018. View at: Publisher Site | Google Scholar
  60. T. L. Miller and M. L. Deinzer, “Effects of nonachloropredioxin and other hydroxychlorodiphenyl ethers on biological membranes,” Journal of Toxicology and Environmental Health, vol. 6, no. 1, pp. 11–25, 1980. View at: Publisher Site | Google Scholar
  61. D. J. Lorusso, T. L. Miller, and M. L. Deinzer, “Effect of hydroxychlorodiphenyl ethers (chlorinated pre-and isopredioxins) on erythrocyte membrane adenosinetriphosphatase activity,” Journal of Toxicology and Environmental Health, vol. 8, no. 1-2, pp. 215–223, 1981. View at: Publisher Site | Google Scholar
  62. J. Villalain, C. R. Mateo, F. J. Aranda, S. Shapiro, and V. Micol, “Membranotropic effects of the antibacterial agent Triclosan,” Archives of Biochemistry and Biophysics, vol. 390, no. 1, pp. 128–136, 2001. View at: Publisher Site | Google Scholar
  63. J. Guillén, A. Bernabeu, S. Shapiro, and J. Villalaín, “Location and orientation of Triclosan in phospholipid model membranes,” European Biophysics Journal, vol. 33, no. 5, pp. 448–453, 2004. View at: Publisher Site | Google Scholar
  64. L. Canesi, C. Ciacci, L. C. Lorusso et al., “Effects of Triclosan on Mytilus galloprovincialis hemocyte function and digestive gland enzyme activities: possible modes of action on non target organisms,” Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, vol. 145, no. 3, pp. 464–472, 2007. View at: Publisher Site | Google Scholar
  65. V. Matozzo, A. Costa Devoti, and M. G. Marin, “Immunotoxic effects of triclosan in the clam Ruditapes philippinarum,” Ecotoxicology, vol. 21, no. 1, pp. 66–74, 2012. View at: Publisher Site | Google Scholar
  66. D. S. Yoon, Y. Choi, D. S. Cha et al., “Triclosan disrupts SKN-1/Nrf2-mediated oxidative stress response in C. elegans and human mesenchymal stem cells,” Scientific Reports, vol. 7, no. 1, article 12592, 2017. View at: Publisher Site | Google Scholar
  67. E. Falisse, A. S. Voisin, and F. Silvestre, “Impacts of triclosan exposure on zebrafish early-life stage: toxicity and acclimation mechanisms,” Aquatic Toxicology, vol. 189, pp. 97–107, 2017. View at: Publisher Site | Google Scholar
  68. C. Riva, S. Cristoni, and A. Binelli, “Effects of triclosan in the freshwater mussel Dreissena polymorpha: a proteomic investigation,” Aquatic Toxicology, vol. 118-119, pp. 62–71, 2012. View at: Publisher Site | Google Scholar
  69. H. Babich and J. P. Babich, “Sodium lauryl sulfate and triclosan: in vitro cytotoxicity studies with gingival cells,” Toxicology Letters, vol. 91, no. 3, pp. 189–196, 1997. View at: Publisher Site | Google Scholar
  70. H. L. Zuckerbraun, H. Babich, R. May, and M. C. Sinensky, “Triclosan: cytotoxicity, mode of action, and induction of apoptosis in human gingival cells in vitro,” European Journal of Oral Sciences, vol. 106, Part 1, no. 2, pp. 628–636, 1998. View at: Publisher Site | Google Scholar
  71. E. Honkisz, D. Zieba-Przybylska, and A. K. Wojtowicz, “The effect of triclosan on hormone secretion and viability of human choriocarcinoma JEG-3 cells,” Reproductive Toxicology, vol. 34, no. 3, pp. 385–392, 2012. View at: Publisher Site | Google Scholar
  72. T. Winitthana, S. Lawanprasert, and P. Chanvorachote, “Triclosan potentiates epithelial-to-mesenchymal transition in anoikis-resistant human lung cancer cells,” PLoS One, vol. 9, no. 10, article e110851, 2014. View at: Publisher Site | Google Scholar
  73. Y. S. Kim, H. W. Seo, M. H. Lee, D. K. Kim, H. Jeon, and D. S. Cha, “Protocatechuic acid extends lifespan and increases stress resistance in Caenorhabditis elegans,” Archives of Pharmacal Research, vol. 37, no. 2, pp. 245–252, 2014. View at: Publisher Site | Google Scholar
  74. H. R. Lee, K. A. Hwang, K. H. Nam, H. C. Kim, and K. C. Choi, “Progression of breast cancer cells was enhanced by endocrine-disrupting chemicals, triclosan and octylphenol, via an estrogen receptor-dependent signaling pathway in cellular and mouse xenograft models,” Chemical Research in Toxicology, vol. 27, no. 5, pp. 834–842, 2014. View at: Publisher Site | Google Scholar
  75. S. H. Kim, K. A. Hwang, and K. C. Choi, “Treatment with kaempferol suppresses breast cancer cell growth caused by estrogen and triclosan in cellular and xenograft breast cancer models,” The Journal of Nutritional Biochemistry, vol. 28, pp. 70–82, 2016. View at: Publisher Site | Google Scholar
  76. G. A. Lee, K. C. Choi, and K. A. Hwang, “Treatment with phytoestrogens reversed triclosan and bisphenol A-induced anti-apoptosis in breast cancer cells,” Biomolecules & Therapeutics, vol. 26, no. 5, pp. 503–511, 2018. View at: Publisher Site | Google Scholar
  77. S. H. Kim, K. A. Hwang, S. M. Shim, and K. C. Choi, “Growth and migration of LNCaP prostate cancer cells are promoted by triclosan and benzophenone-1 via an androgen receptor signaling pathway,” Environmental Toxicology and Pharmacology, vol. 39, no. 2, pp. 568–576, 2015. View at: Publisher Site | Google Scholar
  78. N. Zhang, W. Wang, W. Li et al., “Inhibition of 11β-HSD2 expression by triclosan via induction of apoptosis in human placental syncytiotrophoblasts,” The Journal of Clinical Endocrinology and Metabolism, vol. 100, no. 4, pp. E542–E549, 2015. View at: Publisher Site | Google Scholar
  79. B. K. Park, E. L. T. Gonzales, S. M. Yang, M. Bang, C. S. Choi, and C. Y. Shin, “Effects of triclosan on neural stem cell viability and survival,” Biomolecules & Therapeutics, vol. 24, no. 1, pp. 99–107, 2016. View at: Publisher Site | Google Scholar
  80. K. A. Szychowski, A. M. Sitarz, and A. K. Wojtowicz, “Triclosan induces Fas receptor-dependent apoptosis in mouse neocortical neurons in vitro,” Neuroscience, vol. 284, pp. 192–201, 2015. View at: Publisher Site | Google Scholar
  81. K. A. Szychowski, A. Wnuk, M. Kajta, and A. K. Wojtowicz, “Triclosan activates aryl hydrocarbon receptor (AhR)-dependent apoptosis and affects Cyp1a1 and Cyp1b1 expression in mouse neocortical neurons,” Environmental Research, vol. 151, pp. 106–114, 2016. View at: Publisher Site | Google Scholar
  82. K. A. Szychowski, A. Wnuk, J. Rzemieniec, M. Kajta, T. Leszczyńska, and A. K. Wójtowicz, “Triclosan-evoked neurotoxicity involves NMDAR subunits with the specific role of GluN2A in caspase-3-dependent apoptosis,” Molecular Neurobiology, vol. 56, no. 1, pp. 1–12, 2019. View at: Publisher Site | Google Scholar
  83. J. T. Kwon, Y. S. Yang, M. S. Kang et al., “Pulmonary toxicity screening of triclosan in rats after intratracheal instillation,” The Journal of Toxicological Sciences, vol. 38, no. 3, pp. 471–475, 2013. View at: Publisher Site | Google Scholar
  84. Y. Wu, F. A. Beland, S. Chen, and J. L. Fang, “Extracellular signal-regulated kinases 1/2 and Akt contribute to triclosan-stimulated proliferation of JB6 Cl 41-5a cells,” Archives of Toxicology, vol. 89, no. 8, pp. 1297–1311, 2015. View at: Publisher Site | Google Scholar
  85. X. Chen, B. Xu, X. Han et al., “The effects of triclosan on pluripotency factors and development of mouse embryonic stem cells and zebrafish,” Archives of Toxicology, vol. 89, no. 4, pp. 635–646, 2015. View at: Publisher Site | Google Scholar
  86. J. Kim, H. Oh, B. Ryu et al., “Triclosan affects axon formation in the neural development stages of zebrafish embryos (Danio rerio),” Environmental Pollution, vol. 236, pp. 304–312, 2018. View at: Publisher Site | Google Scholar
  87. J. Liu, L. Sun, H. Zhang et al., “Response mechanisms to joint exposure of triclosan and its chlorinated derivatives on zebrafish (Danio rerio) behavior,” Chemosphere, vol. 193, pp. 820–832, 2018. View at: Publisher Site | Google Scholar
  88. A. Binelli, D. Cogni, M. Parolini, C. Riva, and A. Provini, “Cytotoxic and genotoxic effects of in vitro exposure to triclosan and trimethoprim on zebra mussel (Dreissena polymorpha) hemocytes,” Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, vol. 150, no. 1, pp. 50–56, 2009. View at: Publisher Site | Google Scholar
  89. A. Binelli, D. Cogni, M. Parolini, C. Riva, and A. Provini, “In vivo experiments for the evaluation of genotoxic and cytotoxic effects of Triclosan in Zebra mussel hemocytes,” Aquatic Toxicology, vol. 91, no. 3, pp. 238–244, 2009. View at: Publisher Site | Google Scholar
  90. M. Gonzalez-Pleiter, C. Rioboo, M. Reguera et al., “Calcium mediates the cellular response of Chlamydomonas reinhardtii to the emerging aquatic pollutant Triclosan,” Aquatic Toxicology, vol. 186, pp. 50–66, 2017. View at: Publisher Site | Google Scholar
  91. E. Movahed, G. M. Y. Tan, K. Munusamy et al., “Triclosan demonstrates synergic effect with amphotericin B and fluconazole and induces apoptosis-like cell death in Cryptococcus neoformans,” Frontiers in Microbiology, vol. 7, p. 360, 2016. View at: Publisher Site | Google Scholar
  92. D. J. Watkins, K. K. Ferguson, L. V. Anzalota Del Toro, A. N. Alshawabkeh, J. F. Cordero, and J. D. Meeker, “Associations between urinary phenol and paraben concentrations and markers of oxidative stress and inflammation among pregnant women in Puerto Rico,” International Journal of Hygiene and Environmental Health, vol. 218, no. 2, pp. 212–219, 2015. View at: Publisher Site | Google Scholar
  93. Y. Lv, C. Rui, Y. Dai et al., “Exposure of children to BPA through dust and the association of urinary BPA and triclosan with oxidative stress in Guangzhou, China,” Environmental Science: Processes & Impacts, vol. 18, no. 12, pp. 1492–1499, 2016. View at: Publisher Site | Google Scholar
  94. B. A. Rocha, A. G. Asimakopoulos, M. Honda et al., “Advanced data mining approaches in the assessment of urinary concentrations of bisphenols, chlorophenols, parabens and benzophenones in Brazilian children and their association to DNA damage,” Environment International, vol. 116, pp. 269–277, 2018. View at: Publisher Site | Google Scholar
  95. A. P. Iyer, J. Xue, M. Honda et al., “Urinary levels of triclosan and triclocarban in several Asian countries, Greece and the USA: association with oxidative stress,” Environmental Research, vol. 160, pp. 91–96, 2018. View at: Publisher Site | Google Scholar
  96. B. Bukowska, P. Wieteska, M. Kwiatkowska, P. Sicinska, and J. Michalowicz, “Evaluation of the effect of 2,4-dichlorophenol on oxidative parameters and viability of human blood mononuclear cells (in vitro),” Human & Experimental Toxicology, vol. 35, no. 7, pp. 775–784, 2016. View at: Publisher Site | Google Scholar
  97. P. Zhang, M. Yang, L. Zeng, and C. Liu, “P38/TRHr-dependent regulation of TPO in thyroid cells contributes to the hypothyroidism of triclosan-treated rats,” Cellular Physiology and Biochemistry, vol. 45, no. 4, pp. 1303–1315, 2018. View at: Publisher Site | Google Scholar
  98. S. Vandhana, K. Coral, U. Jayanthi, P. R. Deepa, and S. Krishnakumar, “Biochemical changes accompanying apoptotic cell death in retinoblastoma cancer cells treated with lipogenic enzyme inhibitors,” Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, vol. 1831, no. 9, pp. 1458–1466, 2013. View at: Publisher Site | Google Scholar
  99. M. Battino, M. S. Ferreiro, D. Fattorini, and P. Bullon, “In vitro antioxidant activities of mouthrinses and their components,” Journal of Clinical Periodontology, vol. 29, no. 5, pp. 462–467, 2002. View at: Publisher Site | Google Scholar
  100. C. Ajao, M. A. Andersson, V. V. Teplova et al., “Mitochondrial toxicity of triclosan on mammalian cells,” Toxicology Reports, vol. 2, pp. 624–637, 2015. View at: Publisher Site | Google Scholar
  101. L. M. Weatherly, J. Shim, H. N. Hashmi, R. H. Kennedy, S. T. Hess, and J. A. Gosse, “Antimicrobial agent triclosan is a proton ionophore uncoupler of mitochondria in living rat and human mast cells and in primary human keratinocytes,” Journal of Applied Toxicology, vol. 36, no. 6, pp. 777–789, 2016. View at: Publisher Site | Google Scholar
  102. I. Tamura, Y. Kanbara, M. Saito et al., “Triclosan, an antibacterial agent, increases intracellular Zn2+ concentration in rat thymocytes: its relation to oxidative stress,” Chemosphere, vol. 86, no. 1, pp. 70–75, 2012. View at: Publisher Site | Google Scholar
  103. M. F. Yueh, K. Taniguchi, S. Chen et al., “The commonly used antimicrobial additive triclosan is a liver tumor promoter,” Proceedings of the National Academy of Sciences of the United States of America, vol. 111, no. 48, pp. 17200–17205, 2014. View at: Publisher Site | Google Scholar
  104. M. A. Riad, M. M. Abd-Rabo, S. A. Abd El Aziz, A. M. El Behairy, and M. M. Badawy, “Reproductive toxic impact of subchronic treatment with combined butylparaben and triclosan in weanling male rats,” Journal of Biochemical and Molecular Toxicology, vol. 32, no. 3, article e22037, 2018. View at: Publisher Site | Google Scholar
  105. A. T. Mohammed, A. A.-R. Mohamed, and H. Ali, “Pulmonary apoptotic and oxidative damaging effects of Triclosan alone or in combination with fluoride in Sprague Dawley rats,” Acta Histochemica, vol. 119, no. 4, pp. 357–363, 2017. View at: Publisher Site | Google Scholar
  106. Z. Wang, X. Li, and J. E. Klaunig, “Investigation of the mechanism of triclosan induced mouse liver tumors,” Regulatory Toxicology and Pharmacology, vol. 86, pp. 137–147, 2017. View at: Publisher Site | Google Scholar
  107. K. A. Lenz, C. Pattison, and H. Ma, “Triclosan (TCS) and triclocarban (TCC) induce systemic toxic effects in a model organism the nematode Caenorhabditis elegans,” Environmental Pollution, vol. 231, Part 1, pp. 462–470, 2017. View at: Publisher Site | Google Scholar
  108. D. Lin, Q. Zhou, X. Xie, and Y. Liu, “Potential biochemical and genetic toxicity of triclosan as an emerging pollutant on earthworms (Eisenia fetida),” Chemosphere, vol. 81, no. 10, pp. 1328–1333, 2010. View at: Publisher Site | Google Scholar
  109. D. Lin, X. Xie, Q. Zhou, and Y. Liu, “Biochemical and genotoxic effect of triclosan on earthworms (Eisenia fetida) using contact and soil tests,” Environmental Toxicology, vol. 27, no. 7, pp. 385–392, 2012. View at: Publisher Site | Google Scholar
  110. X. Wang, Z. Liu, W. Wang et al., “Assessment of toxic effects of triclosan on the terrestrial snail (Achatina fulica),” Chemosphere, vol. 108, pp. 225–230, 2014. View at: Publisher Site | Google Scholar
  111. Z. Zhou, J. Yang, and K. M. Chan, “Toxic effects of triclosan on a zebrafish (Danio rerio) liver cell line, ZFL,” Aquatic Toxicology, vol. 191, pp. 175–188, 2017. View at: Publisher Site | Google Scholar
  112. Y. Peng, Y. Luo, X. P. Nie, W. Liao, Y. F. Yang, and G. G. Ying, “Toxic effects of triclosan on the detoxification system and breeding of Daphnia magna,” Ecotoxicology, vol. 22, no. 9, pp. 1384–1394, 2013. View at: Publisher Site | Google Scholar
  113. V. Kovacevic, A. J. Simpson, and M. J. Simpson, “(1)H NMR-based metabolomics of Daphnia magna responses after sub-lethal exposure to triclosan, carbamazepine and ibuprofen,” Comparative Biochemistry and Physiology Part D, Genomics & Proteomics, vol. 19, pp. 199–210, 2016. View at: Publisher Site | Google Scholar
  114. H. I. Falfushynska, L. L. Gnatyshyna, O. Y. Osadchuk et al., “Diversity of the molecular responses to separate wastewater effluents in freshwater mussels,” Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, vol. 164, pp. 51–58, 2014. View at: Publisher Site | Google Scholar
  115. J. C. Park, J. Han, M. C. Lee, J. S. Seo, and J. S. Lee, “Effects of triclosan (TCS) on fecundity, the antioxidant system, and oxidative stress-mediated gene expression in the copepod Tigriopus japonicus,” Aquatic Toxicology, vol. 189, pp. 16–24, 2017. View at: Publisher Site | Google Scholar
  116. P. Ku, X. Wu, X. Nie et al., “Effects of triclosan on the detoxification system in the yellow catfish (Pelteobagrus fulvidraco): expressions of CYP and GST genes and corresponding enzyme activity in phase I, II and antioxidant system,” Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, vol. 166, pp. 105–114, 2014. View at: Publisher Site | Google Scholar
  117. P. Banerjee, T. K. Dey, S. Sarkar, S. Swarnakar, A. Mukhopadhyay, and S. Ghosh, “Treatment of cosmetic effluent in different configurations of ceramic UF membrane based bioreactor: toxicity evaluation of the untreated and treated wastewater using catfish (Heteropneustes fossilis),” Chemosphere, vol. 146, pp. 133–144, 2016. View at: Publisher Site | Google Scholar
  118. F. Wang, R. Xu, F. Zheng, and H. Liu, “Effects of triclosan on acute toxicity, genetic toxicity and oxidative stress in goldfish (Carassius auratus),” Experimental Animals, vol. 67, no. 2, pp. 219–227, 2018. View at: Publisher Site | Google Scholar
  119. C. Li, R. Qu, J. Chen et al., “The pH-dependent toxicity of triclosan to five aquatic organisms (Daphnia magna, Photobacterium phosphoreum, Danio rerio, Limnodrilus hoffmeisteri, and Carassius auratus),” Environmental Science and Pollution Research International, vol. 25, no. 10, pp. 9636–9646, 2018. View at: Publisher Site | Google Scholar
  120. J. Han, E. J. Won, U. K. Hwang, I. C. Kim, J. H. Yim, and J. S. Lee, “Triclosan (TCS) and Triclocarban (TCC) cause lifespan reduction and reproductive impairment through oxidative stress-mediated expression of the defensome in the monogonont rotifer (Brachionus koreanus),” Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, vol. 185-186, pp. 131–137, 2016. View at: Publisher Site | Google Scholar
  121. L. Chai, A. Chen, P. Luo, H. Zhao, and H. Wang, “Histopathological changes and lipid metabolism in the liver of Bufo gargarizans tadpoles exposed to Triclosan,” Chemosphere, vol. 182, pp. 255–266, 2017. View at: Publisher Site | Google Scholar
  122. D. Martins, M. S. Monteiro, A. M. V. M. Soares, and C. Quintaneiro, “Effects of 4-MBC and triclosan in embryos of the frog Pelophylax perezi,” Chemosphere, vol. 178, pp. 325–332, 2017. View at: Publisher Site | Google Scholar
  123. M. Sendra, M. G. Pintado-Herrera, G. V. Aguirre-Martinez et al., “Are the TiO2 NPs a “Trojan horse” for personal care products (PCPs) in the clam Ruditapes philippinarum?” Chemosphere, vol. 185, pp. 192–204, 2017. View at: Publisher Site | Google Scholar
  124. E. Capkin, T. Ozcelep, S. Kayis, and I. Altinok, “Antimicrobial agents, triclosan, chloroxylenol, methylisothiazolinone and borax, used in cleaning had genotoxic and histopathologic effects on rainbow trout,” Chemosphere, vol. 182, pp. 720–729, 2017. View at: Publisher Site | Google Scholar
  125. A. C. Almeida, T. Gomes, K. Langford, K. V. Thomas, and K. E. Tollefsen, “Oxidative stress in the algae Chlamydomonas reinhardtii exposed to biocides,” Aquatic Toxicology, vol. 189, pp. 50–59, 2017. View at: Publisher Site | Google Scholar
  126. C. G. Pan, F. J. Peng, W. J. Shi, L. X. Hu, X. D. Wei, and G. G. Ying, “Triclosan-induced transcriptional and biochemical alterations in the freshwater green algae Chlamydomonas reinhardtii,” Ecotoxicology and Environmental Safety, vol. 148, pp. 393–401, 2018. View at: Publisher Site | Google Scholar
  127. J. Hurtado-Gallego, K. Martin-Betancor, I. Rodea-Palomares, F. Leganes, R. Rosal, and F. Fernandez-Pinas, “Two novel cyanobacterial bioluminescent whole-cell bioreporters based on superoxide dismutases MnSod and FeSod to detect superoxide anion,” Chemosphere, vol. 201, pp. 772–779, 2018. View at: Publisher Site | Google Scholar
  128. O. Culic, V. Erakovic, and M. J. Parnham, “Anti-inflammatory effects of macrolide antibiotics,” European Journal of Pharmacology, vol. 429, no. 1-3, pp. 209–229, 2001. View at: Publisher Site | Google Scholar
  129. Y. Iino, M. Toriyama, Y. Natori, K. Kudo, and A. Yuo, “Erythromycin inhibition of lipopolysaccharide-stimulated tumor necrosis factor alpha production by human monocytes in vitro,” The Annals of Otology, Rhinology & Laryngology. Supplement, vol. 101, 10 Supplement, pp. 16–20, 1992. View at: Publisher Site | Google Scholar
  130. O. M. Korzeniowski, “Effects of antibiotics on the mammalian immune system,” Infectious Disease Clinics of North America, vol. 3, no. 3, pp. 469–478, 1989. View at: Google Scholar
  131. S. M. Uriarte, R. E. Molestina, R. D. Miller et al., “Effects of fluoroquinolones on the migration of human phagocytes through Chlamydia pneumoniae-infected and tumor necrosis factor alpha-stimulated endothelial cells,” Antimicrobial Agents and Chemotherapy, vol. 48, no. 7, pp. 2538–2543, 2004. View at: Publisher Site | Google Scholar
  132. B. Van Vlem, R. Vanholder, P. De Paepe, S. Ringoir, and D. Vogelaers, “Immunomodulating effects of antibiotics: literature review,” Infection, vol. 24, no. 4, pp. 275–291, 1996. View at: Publisher Site | Google Scholar
  133. A. Gaffar, D. Scherl, J. Afflitto, and E. J. Coleman, “The effect of triclosan on mediators of gingival inflammation,” Journal of Clinical Periodontology, vol. 22, no. 6, pp. 480–484, 1995. View at: Publisher Site | Google Scholar
  134. T. Modéer, A. Bengisson, and G. Rölla, “Triclosan reduces prostaglandin biosynthesis in human gingival fibroblasts challenged with interleukin-1 in vitro,” Journal of Clinical Periodontology, vol. 23, no. 10, pp. 927–933, 1996. View at: Publisher Site | Google Scholar
  135. A. B. Skaare, G. Rolla, and P. Barkvoll, “The influence of triclosan, zinc or propylene glycol on oral mucosa exposed to sodium lauryl sulphate,” European Journal of Oral Sciences, vol. 105, Part 2, no. 5, pp. 527–533, 1997. View at: Publisher Site | Google Scholar
  136. T. M. Hernandez-Richter, M. W. Wichmann, W. Schrodl, M. K. Angele, K. Heinritzi, and F. W. Schildberg, “The acute phase response following implantation of triclosan-bonded vascular prostheses,” Clinical and Experimental Medicine, vol. 1, no. 1, pp. 35–41, 2001. View at: Publisher Site | Google Scholar
  137. S. Kumar, “Little difference between triclosan and stannous fluoride dentifrices on gingival inflammation,” Evidence-Based Dentistry, vol. 16, no. 1, pp. 13-14, 2015. View at: Publisher Site | Google Scholar
  138. D. K. Suresh, K. L. Vandana, and D. S. Mehta, “Intracrevicular application of 0.3% Flurbiprofen gel and 0.3% Triclosan gel as anti inflammatory agent. A comparative clinical study,” Indian Journal of Dental Research, vol. 12, no. 2, pp. 105–112, 2001. View at: Google Scholar
  139. F. V. Ribeiro, M. Z. Casati, R. C. Casarin et al., “Impact of a triclosan-containing toothpaste during the progression of experimental peri-implant mucositis: clinical parameters and local pattern of osteo-immunoinflammatory mediators in peri-implant fluid,” Journal of Periodontology, vol. 89, no. 2, pp. 203–212, 2018. View at: Publisher Site | Google Scholar
  140. M. Mustafa, M. Bakhiet, B. Wondimu, and T. Modeer, “Effect of triclosan on interferon-gamma production and major histocompatibility complex class II expression in human gingival fibroblasts,” Journal of Clinical Periodontology, vol. 27, no. 10, pp. 733–737, 2000. View at: Publisher Site | Google Scholar
  141. M. Mustafa, B. Wondimu, M. Ibrahim, and T. Modeer, “Effect of triclosan on interleukin-1beta production in human gingival fibroblasts challenged with tumor necrosis factor alpha,” European Journal of Oral Sciences, vol. 106, Part 1, no. 2, pp. 637–643, 1998. View at: Publisher Site | Google Scholar
  142. M. Mustafa, B. Wondimu, T. Yucel-Lindberg, A. T. Kats-Hallstrom, A. S. Jonsson, and T. Modeer, “Triclosan reduces microsomal prostaglandin E synthase-1 expression in human gingival fibroblasts,” Journal of Clinical Periodontology, vol. 32, no. 1, pp. 6–11, 2005. View at: Publisher Site | Google Scholar
  143. M. Mustafa, B. Wondimu, K. Hultenby, T. Yucel-Lindberg, and T. Modeer, “Uptake, distribution and release of 14C-triclosan in human gingival fibroblasts,” Journal of Pharmaceutical Sciences, vol. 92, no. 8, pp. 1648–1653, 2003. View at: Publisher Site | Google Scholar
  144. M. A. Wallet, N. L. Calderon, T. R. Alonso et al., “Triclosan alters antimicrobial and inflammatory responses of epithelial cells,” Oral Diseases, vol. 19, no. 3, pp. 296–302, 2013. View at: Publisher Site | Google Scholar
  145. K. G. Neiva, N. L. Calderon, T. R. Alonso, F. Panagakos, and S. M. Wallet, “Type 1 diabetes-associated TLR responsiveness of oral epithelial cells,” Journal of Dental Research, vol. 93, no. 2, pp. 169–174, 2014. View at: Publisher Site | Google Scholar
  146. N. B. Marshall, E. Lukomska, A. P. Nayak, C. M. Long, J. M. Hettick, and S. E. Anderson, “Topical application of the anti-microbial chemical triclosan induces immunomodulatory responses through the S100A8/A9-TLR4 pathway,” Journal of Immunotoxicology, vol. 14, no. 1, pp. 50–59, 2017. View at: Publisher Site | Google Scholar
  147. V. M. Barnes, T. Xu, E. Shimizu et al., “Triclosan blocks MMP-13 expression in hormone-stimulated osteoblasts,” Journal of Periodontology, vol. 84, no. 11, pp. 1–9, 2013. View at: Publisher Site | Google Scholar
  148. B. A. Pancer, D. Kott, J. V. Sugai et al., “Effects of triclosan on host response and microbial biomarkers during experimental gingivitis,” Journal of Clinical Periodontology, vol. 43, no. 5, pp. 435–444, 2016. View at: Publisher Site | Google Scholar
  149. M. P. Cullinan, J. E. Palmer, M. J. Faddy et al., “The influence of triclosan on biomarkers of cardiovascular risk in patients in the Cardiovascular and Periodontal Study (CAPS): a randomized controlled trial,” Journal of Periodontology, vol. 86, no. 7, pp. 847–855, 2015. View at: Publisher Site | Google Scholar
  150. S. Hessam, M. Sand, N. M. Meier, T. Gambichler, L. Scholl, and F. G. Bechara, “Combination of oral zinc gluconate and topical triclosan: an anti-inflammatory treatment modality for initial hidradenitis suppurativa,” Journal of Dermatological Science, vol. 84, no. 2, pp. 197–202, 2016. View at: Publisher Site | Google Scholar
  151. P. A. Cadieux, B. H. Chew, B. E. Knudsen et al., “Triclosan loaded ureteral stents decrease proteus mirabilis 296 infection in a rabbit urinary tract infection model,” The Journal of Urology, vol. 175, no. 6, pp. 2331–2335, 2006. View at: Publisher Site | Google Scholar
  152. C. N. Elwood, B. H. Chew, S. Seney, J. Jass, J. D. Denstedt, and P. A. Cadieux, “Triclosan inhibits uropathogenic Escherichia coli-stimulated tumor necrosis factor-alpha secretion in T24 bladder cells in vitro,” Journal of Endourology, vol. 21, no. 10, pp. 1217–1222, 2007. View at: Publisher Site | Google Scholar
  153. S. P. Barros, S. Wirojchanasak, D. A. Barrow, F. S. Panagakos, W. Devizio, and S. Offenbacher, “Triclosan inhibition of acute and chronic inflammatory gene pathways,” Journal of Clinical Periodontology, vol. 37, no. 5, pp. 412–418, 2010. View at: Publisher Site | Google Scholar
  154. F. Udoji, T. Martin, R. Etherton, and M. M. Whalen, “Immunosuppressive effects of triclosan, nonylphenol, and DDT on human natural killer cells in vitro,” Journal of Immunotoxicology, vol. 7, no. 3, pp. 205–212, 2010. View at: Publisher Site | Google Scholar
  155. L. Pavez, N. Tobar, C. Chacon et al., “Chitosan-triclosan particles modulate inflammatory signaling in gingival fibroblasts,” Journal of Periodontal Research, vol. 53, no. 2, pp. 232–239, 2018. View at: Publisher Site | Google Scholar
  156. S. Sharma, T. N. C. Ramya, A. Surolia, and N. Surolia, “Triclosan as a systemic antibacterial agent in a mouse model of acute bacterial challenge,” Antimicrobial Agents and Chemotherapy, vol. 47, no. 12, pp. 3859–3866, 2003. View at: Publisher Site | Google Scholar
  157. H. Yang, W. Wang, K. A. Romano et al., “A common antimicrobial additive increases colonic inflammation and colitis-associated colon tumorigenesis in mice,” Science Translational Medicine, vol. 10, no. 443, article eaan4116, 2018. View at: Publisher Site | Google Scholar
  158. A. Kanetoshi, E. Katsura, H. Ogawa, T. Ohyama, H. Kaneshima, and T. Miura, “Acute toxicity, percutaneous absorption and effects on hepatic mixed function oxidase activities of 2,4,4-trichloro-2-hydroxydiphenyl ether (Irgasan DP300) and its chlorinated derivatives,” Archives of Environmental Contamination and Toxicology, vol. 23, no. 1, pp. 91–98, 1992. View at: Google Scholar
  159. H. Ma, L. Zheng, Y. Li et al., “Triclosan reduces the levels of global DNA methylation in HepG2 cells,” Chemosphere, vol. 90, no. 3, pp. 1023–1029, 2013. View at: Publisher Site | Google Scholar
  160. F. Rodrigues, M. Lehmann, V. S. do Amaral, M. L. Reguly, and H. H. R. de Andrade, “Genotoxicity of three mouthwash products, Cepacol®, Periogard®, and Plax®, in the Drosophila wing-spot test,” Environmental and Molecular Mutagenesis, vol. 48, no. 8, pp. 644–649, 2007. View at: Publisher Site | Google Scholar
  161. F. Chevillot, M. Guyot, M. Desrosiers et al., “Accumulation and sublethal effects of triclosan and its transformation product methyl-triclosan in the earthworm Eisenia andrei exposed to environmental concentrations in an artificial soil,” Environmental Toxicology and Chemistry, vol. 37, no. 7, pp. 1940–1948, 2018. View at: Publisher Site | Google Scholar
  162. P. Martinez-Paz, M. Morales, J. L. Martinez-Guitarte, and G. Morcillo, “Genotoxic effects of environmental endocrine disruptors on the aquatic insect Chironomus riparius evaluated using the comet assay,” Mutation Research, vol. 758, no. 1-2, pp. 41–47, 2013. View at: Publisher Site | Google Scholar
  163. A. R. R. Silva, D. N. Cardoso, A. Cruz et al., “Ecotoxicity and genotoxicity of a binary combination of triclosan and carbendazim to Daphnia magna,” Ecotoxicology and Environmental Safety, vol. 115, pp. 279–290, 2015. View at: Publisher Site | Google Scholar
  164. X. Xu, Y. Lu, D. Zhang et al., “Toxic assessment of triclosan and triclocarban on Artemia salina,” Bulletin of Environmental Contamination and Toxicology, vol. 95, no. 6, pp. 728–733, 2015. View at: Publisher Site | Google Scholar
  165. L. Gao, T. Yuan, P. Cheng et al., “Effects of triclosan and triclocarban on the growth inhibition, cell viability, genotoxicity and multixenobiotic resistance responses of Tetrahymena thermophila,” Chemosphere, vol. 139, pp. 434–440, 2015. View at: Publisher Site | Google Scholar
  166. O. Herrero, J. M. Perez Martin, P. Fernandez Freire, L. Carvajal Lopez, A. Peropadre, and M. J. Hazen, “Toxicological evaluation of three contaminants of emerging concern by use of the Allium cepa test,” Mutation Research, vol. 743, no. 1-2, pp. 20–24, 2012. View at: Publisher Site | Google Scholar
  167. T. Y. Doktorova, G. Ates, M. Vinken, T. Vanhaecke, and V. Rogiers, “Way forward in case of a false positive in vitro genotoxicity result for a cosmetic substance?” Toxicology In Vitro, vol. 28, no. 1, pp. 54–59, 2014. View at: Publisher Site | Google Scholar
  168. K. Z. Sanidad, H. Xiao, and G. Zhang, “Triclosan, a common antimicrobial ingredient, on gut microbiota and gut health,” Gut Microbes, pp. 1–4, 2018. View at: Publisher Site | Google Scholar
  169. H. N. Bhargava and P. A. Leonard, “Triclosan: applications and safety,” American Journal of Infection Control, vol. 24, no. 3, pp. 209–218, 1996. View at: Publisher Site | Google Scholar
  170. R. U. Halden, A. E. Lindeman, A. E. Aiello et al., “The Florence statement on triclosan and triclocarban,” Environmental Health Perspectives, vol. 125, no. 6, article 064501, 2017. View at: Publisher Site | Google Scholar
  171. I. J. Uings and S. N. Farrow, “Cell receptors and cell signalling,” Molecular Pathology, vol. 53, no. 6, pp. 295–299, 2000. View at: Publisher Site | Google Scholar
  172. G. A. Lee, K. A. Hwang, and K. C. Choi, “Inhibitory effects of 3,3-diindolylmethane on epithelial-mesenchymal transition induced by endocrine disrupting chemicals in cellular and xenograft mouse models of breast cancer,” Food and Chemical Toxicology, vol. 109, Part 1, pp. 284–295, 2017. View at: Publisher Site | Google Scholar
  173. H. Huang, G. Du, W. Zhang et al., “The in vitro estrogenic activities of triclosan and triclocarban,” Journal of Applied Toxicology, vol. 34, no. 9, pp. 1060–1067, 2014. View at: Publisher Site | Google Scholar
  174. N. D. Henry and P. A. Fair, “Comparison of in vitro cytotoxicity, estrogenicity and anti-estrogenicity of triclosan, perfluorooctane sulfonate and perfluorooctanoic acid,” Journal of Applied Toxicology, vol. 33, no. 4, pp. 265–272, 2013. View at: Publisher Site | Google Scholar
  175. T. E. Stoker, E. K. Gibson, and L. M. Zorrilla, “Triclosan exposure modulates estrogen-dependent responses in the female Wistar rat,” Toxicological Sciences, vol. 117, no. 1, pp. 45–53, 2010. View at: Publisher Site | Google Scholar
  176. G. W. Louis, D. R. Hallinger, and T. E. Stoker, “The effect of triclosan on the uterotrophic response to extended doses of ethinyl estradiol in the weanling rat,” Reproductive Toxicology, vol. 36, pp. 71–77, 2013. View at: Publisher Site | Google Scholar
  177. M. O. James, W. Li, D. P. Summerlot, L. Rowland-Faux, and C. E. Wood, “Triclosan is a potent inhibitor of estradiol and estrone sulfonation in sheep placenta,” Environment International, vol. 36, no. 8, pp. 942–949, 2010. View at: Publisher Site | Google Scholar
  178. E. M. Jung, B. S. An, K. C. Choi, and E. B. Jeung, “Potential estrogenic activity of triclosan in the uterus of immature rats and rat pituitary GH3 cells,” Toxicology Letters, vol. 208, no. 2, pp. 142–148, 2012. View at: Publisher Site | Google Scholar
  179. H. Serra, F. Brion, J. M. Porcher, H. Budzinski, and S. Ait-Aissa, “Triclosan lacks anti-estrogenic effects in zebrafish cells but modulates estrogen response in zebrafish embryos,” International Journal of Molecular Sciences, vol. 19, no. 4, p. 1175, 2018. View at: Publisher Site | Google Scholar
  180. S. Miyagawa, A. Lange, I. Hirakawa et al., “Differing species responsiveness of estrogenic contaminants in fish is conferred by the ligand binding domain of the estrogen receptor,” Environmental Science & Technology, vol. 48, no. 9, pp. 5254–5263, 2014. View at: Publisher Site | Google Scholar
  181. J. Chen, K. C. Ahn, N. A. Gee, S. J. Gee, B. D. Hammock, and B. L. Lasley, “Antiandrogenic properties of parabens and other phenolic containing small molecules in personal care products,” Toxicology and Applied Pharmacology, vol. 221, no. 3, pp. 278–284, 2007. View at: Publisher Site | Google Scholar
  182. V. Christen, P. Crettaz, A. Oberli-Schrammli, and K. Fent, “Some flame retardants and the antimicrobials triclosan and triclocarban enhance the androgenic activity in vitro,” Chemosphere, vol. 81, no. 10, pp. 1245–1252, 2010. View at: Publisher Site | Google Scholar
  183. K. C. Ahn, B. Zhao, J. Chen et al., “In vitro biologic activities of the antimicrobials triclocarban, its analogs, and triclosan in bioassay screens: receptor-based bioassay screens,” Environmental Health Perspectives, vol. 116, no. 9, pp. 1203–1210, 2008. View at: Publisher Site | Google Scholar
  184. R. H. Gee, A. Charles, N. Taylor, and P. D. Darbre, “Oestrogenic and androgenic activity of triclosan in breast cancer cells,” Journal of Applied Toxicology, vol. 28, no. 1, pp. 78–91, 2008. View at: Publisher Site | Google Scholar
  185. A. L. Forgacs, Q. Ding, R. G. Jaremba, I. T. Huhtaniemi, N. A. Rahman, and T. R. Zacharewski, “BLTK1 murine Leydig cells: a novel steroidogenic model for evaluating the effects of reproductive and developmental toxicants,” Toxicological Sciences, vol. 127, no. 2, pp. 391–402, 2012. View at: Publisher Site | Google Scholar
  186. W. T. Farmer, G. W. Louis, A. R. Buckalew, D. R. Hallinger, and T. E. Stoker, “Evaluation of triclosan in the Hershberger and H295R steroidogenesis assays,” Toxicology Letters, vol. 291, pp. 194–199, 2018. View at: Publisher Site | Google Scholar
  187. Y. Wu, Q. Wu, F. A. Beland, P. Ge, M. G. Manjanatha, and J. L. Fang, “Differential effects of triclosan on the activation of mouse and human peroxisome proliferator-activated receptor alpha,” Toxicology Letters, vol. 231, no. 1, pp. 17–28, 2014. View at: Publisher Site | Google Scholar
  188. P. Xia, X. Zhang, Y. Xie, M. Guan, D. L. Villeneuve, and H. Yu, “Functional toxicogenomic assessment of triclosan in human HepG2 cells using genome-wide CRISPR-Cas9 screening,” Environmental Science & Technology, vol. 50, no. 19, pp. 10682–10692, 2016. View at: Publisher Site | Google Scholar
  189. D. E. Haggard, P. D. Noyes, K. M. Waters, and R. L. Tanguay, “Phenotypically anchored transcriptome profiling of developmental exposure to the antimicrobial agent, triclosan, reveals hepatotoxicity in embryonic zebrafish,” Toxicology and Applied Pharmacology, vol. 308, pp. 32–45, 2016. View at: Publisher Site | Google Scholar
  190. J. Guo, S. Ito, H. T. Nguyen et al., “Effects of prenatal exposure to triclosan on the liver transcriptome in chicken embryos,” Toxicology and Applied Pharmacology, vol. 347, pp. 23–32, 2018. View at: Publisher Site | Google Scholar
  191. X. Cao, X. Hua, X. Wang, and L. Chen, “Exposure of pregnant mice to triclosan impairs placental development and nutrient transport,” Scientific Reports, vol. 7, no. 1, article 44803, 2017. View at: Publisher Site | Google Scholar
  192. X. Hua, X. Y. Cao, X. L. Wang, P. Sun, and L. Chen, “Exposure of pregnant mice to triclosan causes insulin resistance via thyroxine reduction,” Toxicological Sciences, vol. 160, no. 1, pp. 150–160, 2017. View at: Publisher Site | Google Scholar
  193. W. T. Festuccia, P. G. Blanchard, T. Belchior et al., “PPARγ activation attenuates glucose intolerance induced by mTOR inhibition with rapamycin in rats,” American Journal of Physiology Endocrinology and Metabolism, vol. 306, no. 9, pp. E1046–E1054, 2014. View at: Publisher Site | Google Scholar
  194. K. B. Paul, J. T. Thompson, S. O. Simmons, J. P. Vanden Heuvel, and K. M. Crofton, “Evidence for triclosan-induced activation of human and rodent xenobiotic nuclear receptors,” Toxicology In Vitro, vol. 27, no. 7, pp. 2049–2060, 2013. View at: Publisher Site | Google Scholar
  195. D. E. Clapham, “Calcium signaling,” Cell, vol. 131, no. 6, pp. 1047–1058, 2007. View at: Publisher Site | Google Scholar
  196. G. Cherednichenko, R. Zhang, R. A. Bannister et al., “Triclosan impairs excitation-contraction coupling and Ca2+ dynamics in striated muscle,” Proceedings of the National Academy of Sciences of the United States of America, vol. 109, no. 35, pp. 14158–14163, 2012. View at: Publisher Site | Google Scholar
  197. E. B. Fritsch, R. E. Connon, I. Werner et al., “Triclosan impairs swimming behavior and alters expression of excitation-contraction coupling proteins in fathead minnow (Pimephales promelas),” Environmental Science & Technology, vol. 47, no. 4, pp. 2008–2017, 2013. View at: Publisher Site | Google Scholar
  198. T. Kawanai, “Triclosan, an environmental pollutant from health care products, evokes charybdotoxin-sensitive hyperpolarization in rat thymocytes,” Environmental Toxicology and Pharmacology, vol. 32, no. 3, pp. 417–422, 2011. View at: Publisher Site | Google Scholar
  199. V. V. Teplova, K. N. Belosludtsev, and A. G. Kruglov, “Mechanism of triclosan toxicity: mitochondrial dysfunction including complex II inhibition, superoxide release and uncoupling of oxidative phosphorylation,” Toxicology Letters, vol. 275, pp. 108–117, 2017. View at: Publisher Site | Google Scholar
  200. G. McDonnell and A. D. Russell, “Antiseptics and disinfectants: activity, action, and resistance,” Clinical Microbiology Reviews, vol. 12, no. 1, pp. 147–179, 1999. View at: Publisher Site | Google Scholar
  201. L. M. McMurry, M. Oethinger, and S. B. Levy, “Triclosan targets lipid synthesis,” Nature, vol. 394, no. 6693, pp. 531-532, 1998. View at: Publisher Site | Google Scholar
  202. J. G. Beeson, P. A. Winstanley, G. I. McFadden, and G. V. Brown, “New agents to combat malaria,” Nature Medicine, vol. 7, no. 2, pp. 149-150, 2001. View at: Publisher Site | Google Scholar
  203. H. H. Lee, J. Yun, J. Moon et al., “Crystallization and preliminary X-ray crystallographic analysis of enoyl-acyl carrier protein reductase from Helicobacter pylori,” Acta Crystallographica Section D, Biological Crystallography, vol. 58, no. 6, pp. 1071–1073, 2002. View at: Publisher Site | Google Scholar
  204. J. Marcinkeviciene, W. Jiang, L. M. Kopcho, G. Locke, Y. Luo, and R. A. Copeland, “Enoyl-ACP reductase (FabI) of Haemophilus influenzae: steady-state kinetic mechanism and inhibition by triclosan and hexachlorophene,” Archives of Biochemistry and Biophysics, vol. 390, no. 1, pp. 101–108, 2001. View at: Publisher Site | Google Scholar
  205. R. McLeod, S. P. Muench, J. B. Rafferty et al., “Triclosan inhibits the growth of Plasmodium falciparum and Toxoplasma gondii by inhibition of apicomplexan Fab I,” International Journal for Parasitology, vol. 31, no. 2, pp. 109–113, 2001. View at: Publisher Site | Google Scholar
  206. S. L. Parikh, G. Xiao, and P. J. Tonge, “Inhibition of InhA, the enoyl reductase from Mycobacterium tuberculosis, by triclosan and isoniazid,” Biochemistry, vol. 39, no. 26, pp. 7645–7650, 2000. View at: Publisher Site | Google Scholar
  207. C. W. Roberts, R. McLeod, D. W. Rice, M. Ginger, M. L. Chance, and L. J. Goad, “Fatty acid and sterol metabolism: potential antimicrobial targets in apicomplexan and trypanosomatid parasitic protozoa,” Molecular and Biochemical Parasitology, vol. 126, no. 2, pp. 129–142, 2003. View at: Publisher Site | Google Scholar
  208. N. Surolia and A. Surolia, “Triclosan offers protection against blood stages of malaria by inhibiting enoyl-ACP reductase of Plasmodium falciparum,” Nature Medicine, vol. 7, no. 2, pp. 167–173, 2001. View at: Publisher Site | Google Scholar
  209. S. Lu and M. C. Archer, “Fatty acid synthase is a potential molecular target for the chemoprevention of breast cancer,” Carcinogenesis, vol. 26, no. 1, pp. 153–157, 2005. View at: Publisher Site | Google Scholar
  210. R. Lupu and J. Menendez, “Pharmacological inhibitors of fatty acid synthase (FASN)--catalyzed endogenous fatty acid biogenesis: a new family of anti-cancer agents?” Current Pharmaceutical Biotechnology, vol. 7, no. 6, pp. 483–494, 2006. View at: Publisher Site | Google Scholar
  211. B. Liu, Y. Wang, K. Fillgrove, and V. Anderson, “Triclosan inhibits enoyl-reductase of type I fatty acid synthase in vitro and is cytotoxic to MCF-7 and SKBr-3 breast cancer cells,” Cancer Chemotherapy and Pharmacology, vol. 49, no. 3, pp. 187–193, 2002. View at: Publisher Site | Google Scholar
  212. E. S. Pizer, C. Jackisch, F. D. Wood, G. R. Pasternack, N. E. Davidson, and F. P. Kuhajda, “Inhibition of fatty acid synthesis induces programmed cell death in human breast cancer cells,” Cancer Research, vol. 56, no. 12, pp. 2745–2747, 1996. View at: Google Scholar
  213. F. P. Kuhajda, K. Jenner, F. D. Wood et al., “Fatty acid synthesis: a potential selective target for antineoplastic therapy,” Proceedings of the National Academy of Sciences of the United States of America, vol. 91, no. 14, pp. 6379–6383, 1994. View at: Publisher Site | Google Scholar
  214. F. P. Kuhajda, E. S. Pizer, J. N. Li, N. S. Mani, G. L. Frehywot, and C. A. Townsend, “Synthesis and antitumor activity of an inhibitor of fatty acid synthase,” Proceedings of the National Academy of Sciences of the United States of America, vol. 97, no. 7, pp. 3450–3454, 2000. View at: Publisher Site | Google Scholar
  215. T. S. Ho, Y. P. Ho, W. Y. Wong, L. Chi-Ming Chiu, Y. S. Wong, and V. Eng-Choon Ooi, “Fatty acid synthase inhibitors cerulenin and C75 retard growth and induce caspase-dependent apoptosis in human melanoma A-375 cells,” Biomedicine & Pharmacotherapy, vol. 61, no. 9, pp. 578–587, 2007. View at: Publisher Site | Google Scholar
  216. M. C. Sadowski, R. H. Pouwer, J. H. Gunter, A. A. Lubik, R. J. Quinn, and C. C. Nelson, “The fatty acid synthase inhibitor triclosan: repurposing an anti-microbial agent for targeting prostate cancer,” Oncotarget, vol. 5, no. 19, pp. 9362–9381, 2014. View at: Publisher Site | Google Scholar
  217. P. R. Deepa, S. Vandhana, U. Jayanthi, and S. Krishnakumar, “Therapeutic and toxicologic evaluation of anti-lipogenic agents in cancer cells compared with non-neoplastic cells,” Basic & Clinical Pharmacology & Toxicology, vol. 110, no. 6, pp. 494–503, 2012. View at: Publisher Site | Google Scholar
  218. K. Nishi, K. Suzuki, J. Sawamoto et al., “Inhibition of fatty acid synthesis induces apoptosis of human pancreatic cancer cells,” Anticancer Research, vol. 36, no. 9, pp. 4655–4660, 2016. View at: Publisher Site | Google Scholar
  219. B. Schmid, J. F. Rippmann, M. Tadayyon, and B. S. Hamilton, “Inhibition of fatty acid synthase prevents preadipocyte differentiation,” Biochemical and Biophysical Research Communications, vol. 328, no. 4, pp. 1073–1082, 2005. View at: Publisher Site | Google Scholar
  220. E. S. Pizer, R. J. Kurman, G. R. Pasternack, and F. P. Kuhajda, “Expression of fatty acid synthase is closely linked to proliferation and stromal decidualization in cycling endometrium,” International Journal of Gynecological Pathology, vol. 16, no. 1, pp. 45–51, 1997. View at: Publisher Site | Google Scholar
  221. R. E. Wilentz, L. A. Witters, and E. S. Pizer, “Lipogenic enzymes fatty acid synthase and acetyl-coenzyme A carboxylase are coexpressed with sterol regulatory element binding protein and Ki-67 in fetal tissues,” Pediatric and Developmental Pathology, vol. 3, no. 6, pp. 525–531, 2000. View at: Publisher Site | Google Scholar

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