Stem Cells International

Review Article

In Vitro Weight-Loaded Cell Models for Understanding Mechanodependent Molecular Pathways Involved in Orthodontic Tooth Movement: A Systematic Review

Table 1

Top four examined genes or substances in studies applying 2D or 3D in vitro WAB loading model on hPDLCs. For each gene or substance, cell culture type and literature reference are given. Additionally, examined force durations and magnitudes are summarized. Force effect on gene expression/substance secretion was evaluated (increase/decrease/no change). In all cases, the most prominent changes (increase/decrease) or “no change” are noted. For each change of expression/secretion, the corresponding maximum (increase/decrease) of force duration and magnitudes are additionally provided.


Gene symbol or metabolite	Cell culture	Reference	Examined force applied		Gene expression			Substance secretion
Gene symbol or metabolite	Cell culture	Reference	Duration (h)	Magnitude (g/cm²)	Increase/decrease/no change	Change in relation to force duration (h)	Change in relation to force magnitude (g/cm²)	Increase/decrease/no change	Change in relation to force duration (h)	Change in relation to force magnitude (g/cm²)

PGE₂	2D	Benjakul et al. in press [17]	48	1.5	na			Increase (qPCR: GAPDH)	48	1.5
		Jin et al. 2015 [18]	0; 0.5; 3; 6; 12	2.0	na			Increase (ELISA)	12	2.0
		Kang et al. 2010 [19]	0.5; 2; 6; 24; 48	2.0	na			Increase (ELISA)	48	2.0
		Kanzaki et al. 2002 [20]	0.5; 1.5; 6; 24; 48 (+ELISA: 60)	0.5; 1.0; 2.0; 3.0; 4.0 (ELISA: 2.0)	na			Increase (ELISA)	60	2.0
		Kirschneck et al. 2015 [21]	24	2.0	na			Not explicitly stated (ELISA)
		Liu et al. 2006 [22]	48	2.0	na			Increase (ELISA)	48	2.0
		Mayahara et al. 2007 [23]	3; 6; 12; 24; 48	2	na			Increase (ELISA)	48	2
		Premaraj et al. 2013 [24]	0.5; 1; 3; 6	5.0	na			Increase (ELISA)	1	5.0
		Proff et al. 2014 [9]	24	2	na			Increase (ELISA)	24	2
		Römer et al. 2013 [25]	24	2	na			Increase (ELISA)	24	2
	3D (Coll. gel)	de Araujo et al. 2007 [26]	3; 12; 24; 48; 72	6.0				Increase (EIA)	72	6.0
	3D (PLGA)	Li et al. 2016 [27]	6; 24; 72	5.0; 15.0; 25.0	na			Increase (ELISA)	24	15.0…25.0
	3D (PLGA)	Yi et al. 2016 [28]	24	25.0				Increase (ELISA)	24	25.0

PTGS2	2D	Jin et al. 2015 [18]	0; 0.5; 3; 6; 12	2.0	Increase (qPCR: GAPDH)	12	2.0
		Kang et al. 2010 [19]	0.5; 2; 6; 24; 48	2.0	Increase (qPCR: GAPDH)	48
		Kanzaki et al. 2002 [20]	0.5; 1.5; 6; 24; 48	0.5; 1.0; 2.0; 3.0; 4.0	Increase (sqPCR: ACTNB)	6	2.0
		Kirschneck et al. 2015 [21]	24	2.0	Increase (qPCR: POL2RA)	24	2.0
		Liu et al. 2006 [22]	48	2.0	Increase (sqPCR: ACTNB)	48	2.0
		Mayahara et al. 2007 [23]	3; 6; 12; 24; 48	2	Increase (qPCR: GAPDH)	48	2
		Mayahara et al. 2010 [29]	3; 6; 12; 24; 48	2.0	Increase (qPCR: GAPDH)	48	2
		Premaraj et al. 2013 [24]	6	0.2; 2.2; 5.0	nd			Increase (WB)	6	5.0
		Proff et al. 2014 [9]	24	2	Increase (qPCR: POL2RA)	24	2	Increase (WB)	24	2
		Römer et al. 2013 [25]	24	2	Increase (qPCR: POL2RA)	24	2
		Wongkhantee et al. 2007 [30]	24	0; 1.25; 2.5	Increase (sqPCR: GAPDH)	24	2.5
	3D (Coll. gel)	de Araujo et al. 2007 [26]	1; 3; 6; 12; 24; 48; 72	3.6; 6.0; 7.1; 9.5	Increase (sqPCR: GAPDH)	6	7.1
	3D (PLGA)	Li et al. 2016 [31]	6; 24; 72	25.0	Increase (qPCR: GAPDH)	6	25.0
		Li et al. 2013 [32]	6; 24; 72	25.0	Increase (qPCR: GAPDH)	6	25.0
		Li et al. 2016 [27]	6; 24; 72	5.0; 15.0; 25.0	Increase (qPCR: GAPDH)	6	25.0
		Li et al. 2011 [33]	6	5; 15; 25; 35	Increase (qPCR: GAPDH)	6	35.0
		Yi et al. 2016 [28]	24	25.0	Increase (qPCR: GAPDH)	24	25.0	Increase (WB)	24	25.0

TNFRSF11B	2D	Benjakul et al. in press [17]	48	1.5	No change (qPCR: GAPDH)			No change
		Jin et al. 2015 [18]	0; 0.5; 3; 6; 12	2.0	No change (qPCR: GAPDH)
		Kanzaki et al. 2002 [20]	0.5; 1.5; 6; 24; 48	0.5; 1.0; 2.0; 3.0; 4.0	No change (sqPCR: ACTNB)
		Kim et al. 2013 [8]	0.5; 2; 6; 24; 48	2.0	Transitory downregulated. (qPCR: GAPDH)	6	2.0	Transitory downregulation (ELISA)	6	2.0
		Kirschneck et al. 2015 [21]	24	2.0	No change (qPCR: POL2RA)
		Lee et al. 2015 [34]	0; 2; 4; 8; 24; 48	2.5	No change (qPCR: ACTNB)
		Liu et al. 2017 [35]	6; 12; 24	0.5; 1.0; 1.5	nd			Decrease (WB)	n. g.	1.5
		Luckprom et al. 2011 [36]	2; 4	2.5	No change (sqPCR: GAPDH)
		Mitsuhashi et al. 2011 [37]	1; 3; 6; 9; 12; 24	4.0	No change (qPCR: ACTNB)
		Nakajima et al. 2008 [38]	0; 1; 3; 6; 9; 12; 24	0.5; 1.0; 2.0; 3.0; 4.0	nd			Increase (ELISA)	24	0.5
		Nishijima et al. 2006 [39]	48	0; 0.5; 1.0; 2.0; 3.0	nd			Decrease (ELISA)	48	2.0
		Römer et al. 2013 [25]	24	2	No change (qPCR: RNA-polymerase-2-polypeptide A)
		Yamada et al. 2013 [40]	12	4.0	Decrease (qPCR: GAPDH)	12	4.0	Decrease (ELISA)	12	4.0
		Yamaguchi et al. 2006 [41]	0; 3; 6; 9; 12; 24; 48	0.5; 1.0; 2.0; 3.0	n. d.			Decrease (ELISA)	12…48	2.0
	3D (Coll. gel)	Kaku et al. 2016 [42]	12; 24	0.5; 1.0; 2.0	Increase (qPCR: GAPDH)	12	1.0
	3D (PLLA modif.)	Liao et al. 2016 [14]	1 d; 3 d; 7 d; 14 d	5.0; 15.0; 25.0; 35.0	No change (qPCR: GAPDH)
	3D (PLGA)	Jianru et al. 2015 [43]	3; 6; 12 (WB: 12)	25.0	Decrease followed by increase (qPCR: GAPDH)	3 (decrease) 12 (increase)	25.0	Increase (WB)	12	25.0
		Li et al. 2016 [31]	6; 24; 72	25.0	Decrease followed by Increase (qPCR: GAPDH)	6 (decrease) 72 (increase)	25.0
		Li et al. 2016 [27]	6; 24; 72	5.0; 15.0; 25.0	Decrease followed by increase (qPCR: GAPDH)	6 (decrease) 72 (increase)	15.0 (decrease) 25.0 (increase)	Decrease followed by Increase (qPCR: GAPDH)	6 (decrease) 72 (increase)	25.0 (decrease) 25.0 (increase)
		Li et al. 2011 [33]	6; 24; 72	25	Decrease followed by increase (qPCR: GAPDH)	6 (decrease) 72 (increase)	25.0
		Yi et al. 2016 [28]	24	25.0	Decrease (qPCR: GAPDH)	24	25.0	No change (WB)

TNFSF11	2D	Benjakul et al. in press [17]	48	1.5	Increase (qPCR: GAPDH)	48	1.5	Increase (qPCR: GAPDH)	48	1.5
		Jin et al. 2015 [18]	0; 0.5; 3; 6; 12	2.0	Increase (qPCR: GAPDH)	12	2.0
		Kang et al. 2013 [44]	2; 48	2.0	Increase (qPCR: GAPDH)	48	2.0
		Kanzaki et al. 2002 [20]	0.5; 1.5; 6; 24; 48	0.5; 1.0; 2.0; 3.0; 4.0	Increase (sqPCR: ACTNB)	48	2.0	Increase (WB): 40-kDa+ 55-kDa	48	2.0
		Kikuta et al. 2015 [45]	1; 3; 6; 9; 12; 24 (+ELISA: 48)	4.0	Increase (qPCR: GAPDH)	12	4.0	Increase (ELISA)	24	4.0
		Kim et al. 2013 [8]	0.5; 2; 6; 24; 48	2.0 ++	Increase (qPCR: GAPDH)	24	2.0	Increase (ELISA)	48	2.0
		Kirschneck et al. 2015 [21]	24	2.0	Increase (qPCR: POL2RA)	24	2.0
		Lee et al. 2015 [34]	0; 2; 4; 8; 24; 48	2.5	Increase (qPCR: ACTNB)	24	2.5
		Liu et al. 2017 [35]	6, 12, 24	0.5; 1.0; 1.5	nd			Increase (WB: GAPDH)	ng	1.5
		Liu et al. 2006 [22]	48	2.0	Increase (sqPCR: ACTNB)	48	2.0
		Luckprom et al. 2011 [36]	2; 4	2.5	Increase (sqPCR: GAPDH)	2	2.5	Increase (WB)	4	2.5
		Mitsuhashi et al. 2011 [37]	1; 3; 6; 9; 12; 24	4.0	Temporary increase (qPCR: ACTNB)	6…9	4.0
		Nakajima et al. 2008 [38]	0; 1; 3; 6; 9; 12; 24	0.5; 1.0; 2.0; 3.0; 4.0	nd			Increase (ELISA)	24	4.0
		Nishijima et al. 2006 [39]	48	0; 0.5; 1.0; 2.0; 3.0	nd			Increase (ELISA)	12…48	2.0
		Römer et al. 2013 [25]	24	2	Increase (qPCR: RNA-polymerase-2-polypeptide A)	24	2
		Wongkhantee et al. 2007 [30]	24	0; 1.25; 2.5	Increase (sqPCR: GAPDH)	24	2.5	Increase (WB; ACTNB)	24	2.5
		Yamada et al. 2013 [40]	12	4.0	Increase (qPCR: GAPDH)	12	4.0	Increase (ELISA)	12	4.0
		Yamaguchi et al. 2006 [41]	0; 3; 6; 9; 12; 24; 48	0.5; 1.0; 2.0; 3.0	nd			Increase (ELISA): sRANKL Increase (WB)	12…48 12	2.0 2.0
	3D (Coll. gel)	Kang et al. 2013 [44]	2; 48	2.0	Increase (qPCR: GAPDH)	2	2.0
	3D (PLLA modif.)	Liao et al. 2016 [14]	1 d; 3 d; 7 d; 14 d	5.0; 15.0; 25.0; 35.0	Increase (qPCR: GAPDH)	Day 14	35.0
	3D (PLGA)	Jianru et al. 2015 [43]	3; 6; 12 (WB: 12)	25.0	Increase (qPCR: GAPDH)	6	25.0	Increase (WB)	12	25.0
		Li et al. 2016 [31]	6; 24; 72	25.0	Increase (qPCR: GAPDH)	6	25.0
		Li et al. 2016 [27]	6; 24; 72	5.0; 15.0; 25.0	Increase (qPCR: GAPDH)	6	25.0	Decrease (ELISA)	72	25.0
		Li et al. 2011 [33]	6; 24; 72	5; 15; 25; 35	Increase (qPCR: GAPDH) Increase followed by no change (qPCR: GAPDH)	6 6 (increase) 72 (no change)	25…35.0 25 25
		Yi et al. 2016 [28]	24	25.0	Increase (qPCR: GAPDH)	24	25.0	Increase (WB)	24	25.0

2D: two-dimensional cell culture; 3D (Coll. gel): three-dimensional cell culture, collagen gel; 3D (PLGA): three-dimensional cell culture using PLGA scaffolds; 3D (PLLA modif.): three-dimensional cell culture, hydrophilically modified PLLA scaffolds; qPCR: quantitative polymerase chain reaction (e.g., real-time PCR); sqPCR: semiquantitative polymerase chain reaction, followed by reference gene used; nr: not reported; na: not applicable; ELISA: enzyme-linked immune absorbent assay; WB: Western blot; IF: immunofluorescence; FLM: fluorescence microscopy; EIA: enzyme immunoassay.