Table 3: Summary of all discussed nanoparticle formulations used for colorectal cancer therapy, for each formulation, size, zeta potential, targeting strategy, administration route, payload, in vitro and in vivo models they have been tested on, the relative results, and references.

NanovectorSize (AVG)ChargeTargeting strategyPayloadAdministration routeModelResultsReferences

Anti-CEA antibody
PaclitaxelIVIn vitro: SW480 (CEA–) and Caco-2 (CEA+) cell cultures.In vitro: CEA dependent uptake and cytotoxicity[26]

Doxorubicin-loaded ONT-conjugated AuNPs23 nmEPR
(tissue retention)
DoxorubicinIntratumouralIn vitro: SW480 cell cultures
In vivo: GFP-transfected SW480 subcutaneously inoculated in BALB/c female mice
In vitro: enhanced DOX cytotoxicity and delivery to the nucleus
In vivo: slower tumor growth compared to control

AuNPs15 nm–21 mVEPRIVIn vitro: SW629 cell cultures
In vivo: SW620 subcutaneously inoculated in female BLAB/c mice
In vitro: reduction in fibrosis-inducing factors secretion
In vivo: decrease in solid stress and improved perfusion; improved cisplatin action

Cu-CiEPRRadiotherapy enhancement
In vitro: SW620 cell culturesIn vitro: nanoparticle dose and radiation dose dependence of cell death by autophagy and apoptosis.[37]

CP-NIC NPs74 nm (length)
13 nm (width)
EPRNicosamideIVIn vitro: HCT116 cell line
In vivo: HCT-116 subcutaneously injected in male nude mice
In vitro: HCT116 cell toxicity similar to free drug, Wnt inhibition
In vivo: tumor growth delay and increase in survival

MSNs-DM1@PDA-PEG-APt170 nm–11 mVEPR
Aptamer against EpCAM
MaytansineIVIn vitro: SW480 and NCM460 cell cultures.

In vivo: SW480 subcutaneously injected in BALB/c mice
In vitro: EpCAM dependent cytostatic and apoptotic effect of targeted NPS
In vivo: reduction of tumor growth rate

FRα targeting
pH-dependent release
DoxorubicinIn vitro: HT-29, Caco-2 and CCD-112 cell linesIn vitro: FR and FRα dependent uptake and cytotoxicity, cytosolic drug delivery.[54]
Dex-SA-DOX-CDDP40 nm—16 mVEPRDoxorubicin
IVIn vitro: CT26 cell lines
In vivo: CT26 cells subcutaneously injected in male BALB/c mice; DMH intraperitoneal injection in BALB/c nude mice.
In vitro: dose-dependent cell toxicity
In vivo: reduction of tumor growth, induction apoptosis, reduced hepatotoxicity and cardiotoxicity

RBC-coated PLGA nanoparticles150 nmEPRGambogic acid (GA)IVIn vitro: SW480
In vivo: Ectopic SW480 bearing BLAB/c bearing mice
In vitro: biocompatible, 48h cytotoxicity similar to free drug
In vivo: Reduction in tumor size and increased survival

anti-EGFR-iRGD-RBCms- PLGA NPs153 nmEPR
Anti-integrin rec. and anti-EGFR targeting
Gambogic acidIVIn vitro: Caco-2, HT-29 (EGFFR+) and SW-480 cell cultures.
In vivo: Caco-2 cells subcutaneously injected in BALB/c mice. Reduced tumor growth and increased survival
In vitro: enhanced HT-29 spheroids penetration, similar GA cytotoxicity compared with the free drug
In vivo: enhanced tumor targeting capability compared to nonfunctionalized NPs

Anti CD113 MoAB – pPEG-PCL/malPEG-PCL167 nm–28 mVEPR
Active targeting against CD113
SN-38IVIn vitro: HT-29, SW620, HCT116 cell cultures.
In vivo: HCT116 subcutaneously injected in BALB/c female mice
In vitro: selective toxicity on CD113 high cells only for actively targeted NPS
In vivo: reduction in CD113 high tumor and histological reduction of CD113 + cells

PPDC nanoparticles105 nmEPRSorafenib CPTIVIn vitro: HT-29 cell cultures (2D and spheroids)
In vivo: HT-29 subcutaneously injected in BALB/c nude mice
In vitro: MMP-espression-dependent drug release and cytotoxic effect
In vivo: Inhibition of tumor growth, reduced vascularization, and tumor necrosis
NKG2D-IL-21 dextran NPs200–400 nm40 mVEPRdsNKG2D-IL-21 plasmidsIVIn vitro: CT-26, NIH-3T3 and RAW264.7 cell lines
Ex vivo: co-culture of above mentioned cell lines with spleen-derived mononuclear cells
In vivo: CT-26 cells subcutaneously injected in BALB/c mice.
In vitro and ex vivo: efficient transfection, induction of immune cells activation.
In vivo: Accumulation in tumor tissue, efficient cells transfection, and induction of immune response leading to tumor growth suppression.

RRHPC/PF33/pDNA127 nm–23 mVActive targeting against CD44 and integrin αvβ3TRAIL pDNAIntraperitonealIn vitro: SW480 tumor cell line
In vivo: SW480 cell intraperitoneally injected in BALB/c female nude mice.
In vitro: Efficient targeting, transfection and apoptosis induction of tumor cell lines.
In vivo: reduced tumor weight and reduced number of tumor nodules.

MSNs-anti-miR-155-PDA-AS1411170 nm–15 mNEPR
Anti-nuclein aptamer
Anti-miR 155 ONTIVIn vitro: SW480, HT-29, SW-620, LoVo and Caco-2 cell lines.
In vitro: BALB/c nude mice subcutaneously injected with SW480 cells
In vitro: aptamer-dependent uptake and anti-miR155 ONT -dependent toxicity, inhibition of colony formation
In vivo: tumor targeting and growth inhibition; synergistic effect with 5-FU

miRNA-139-NPs50–200 nmEPRmiRNA-139IVIn vitro: HCT-116 and LoVo cell lines.
In vivo: HCT-116 subcutaneously injected in nude BALB/c mice; HCT-116 tumours implanted in nude mice colon
In vitro: miRNA-139 dependent toxicity
In vivo: decrease in tumor growth and metastasis; increase in survival

SP-OA-CS143 nm–33 mVEPR
Mannose rec. targeting Hyaluronan rec. targeting
miR-20IVIn vitro: LSECs isolated from Balb/c mice activated with tumor conditioned medium
In vivo: C26 CRC cell administered by intra-splenic injection to produce CRC liver metastasis.
In vitro: efficient LSECs transfection, reduced expression of target proteins regulated by miR-20, reduced LSECs migration.
In vivo:efficient LSEC targeting, reduced number of metastatic foci and reduced LSECs infiltration.

pRLN/pPD-L1 trap LCPs186 nm–6.8 mVActive targeting against Sig-1RpRLNpPD-L1 trapIVIn vitro: CT26-F3 cell cultures.
In vivo: CT26-F3 intrasplenically injected in male or female BALB/c and C57/BL6 mice
In vitro: efficient targeting and transfection.
In vivo: Efficient targeting, transfection and modulation of the metastasis microenvironment towards a pro-inflammatory anticancer phenotype. Synergistic action with checkpoint inhibition. Decreased tumor growth and increased survival.

Gly@Cy7-Si-DOX NPs80–120 nm–30 mV to –80 mVEPRCy7
IVIn vitro: bone marrow and HT29 cell cultures.
In vivo: HT-29 cells subcutaneously injected in BALB/c athymic nude mice.
In vitro: NIR and pH dependent cytotoxicity.
In vivo: tumor accumulation, NIR dependent therapeutic effect, tumor destruction, and improved survival

UCNPs-Ce6-R83780 nm–13 mVEPR (tissue retention)R837
Ce6 PDT-photosensitizers
IntratumouralIn vitro: CT26 cell lines alone on in trans-well co-culture with DCs.
In vivo: CT26 subcutaneous implantation in BALB/c female mice
In vitro: NIR-dependent cell toxicity and DC activation
In vivo: NIR-dependent primary tumor ablation, increased distal tumours reduction, and antitumor immune memory development
HMRu@RBT–SS–Fc150 nm+15 mVEPR
Active targeting against CEA and CD16
RBTIVIn vitro: CT26, Caco-2, SW480, HCT116, CT-26 transfected with CEA cell lines and spheroids.
In vivo: CT-26 cells transfected with CEA subcutaneously injected in BALB/c female mice
In vitro: Efficient active targeting and endocytosis by CEA expressing cells, uptake and photothermal dependent cytotoxicity
In vivo: efficient tumor targeting and killing upon photothermal treatment; elicitation of immune reaction against secondary nonirradiated tumor.

Ox Pt-bp/chol-DHA70–100 nm–21 to –13 mVEPROxPtDHAIntraperitonealIn vitro: CT26 and MC38 cell cultures
In vivo: SD/CD rats, CT26 subcutaneously injected in BALB/c mice and MC38 subcutaneously injected in C57Bl/6 mice
In vitro: Dose-dependent cell death, induction of apoptosis and release of DAMPs.
In vivo: RES avoidance, tumor growth inhibition (especially with PD-L1) blockade and acquisition of antigen-specific immune memory.

EGFR-CPIG80–100 nm40 mVEPR
Active targeting against EGFR
Porphyrin (PDT)
IRDye800CW (MFI)
IVIn vitro: CT26 tumor cell lines.
In vivo: CT26 tumor cells subcutaneously injected in BALB/c male mice.
In vitro: laser intensity and irradiation time-dependent toxicity.
In vivo: tumor growth suppression and eradication, immune cells activation after PDT treatment.

TCL-PDA NPs245 nm–24 mVLNs tropismTCL (providing tumor-associated antigens)SCIn vitro: BMDCs cells isolated from C57BL/6 mice.
In vivo: MC38 cells subcutaneously administerd in C57BL/6 mice
In vitro: BMDCs activation, antigen presentation, pro-inflammatory cytokine secretion.
In vivo: Adaptive immune antitumor response, antitumor immune memory development. Substantial quenching of tumor growth.
aMG1-PEG-HNPs50 nm ca.EOR
Active targeting against MG1
IVIn vitro: CC-531 cell cultures
In vivo: Surgical inoculation of tumours in the liver of Wistar rats
In vitro: MG1-dependent uptake, light-dependent cytotoxicity and increased MRI signal contrast.
In vivo: tumor accumulation and detection through MRI, PTT tumor ablation upon 800 nm laser irradiation

Abbreviations: Avg: average; CRC: colorectal cancer; DOX: doxorubicin; EPR: enhanced permeability and retention effect; IV: intravenous; SC: subcutaneous; LN: lymph node; PDT: Photodynamic treatment; PTT: photothermal treatment.