Osteogenic Potential of Dental Mesenchymal Stem Cells in Preclinical Studies: A Systematic Review Using Modified ARRIVE and CONSORT Guidelines

Table 5

Study characteristics of in vitro experiments with the application of dental stem cells on bone regeneration/osteogenesis potential

(a) Stem cells from apical papilla (SCAPs)

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Abe et al. 2008 [61]	Human	OIM	HA	ALP assay; Staining, SEM	Time dependent ALP activity seen.
Park et al. 2009 [111]	Human	OIM	None	Histochemical staining	Osteoblast differentiation and mineralized nodule formation seen.
Abe et al. 2012 [62]	Human	OIM	None	Histochemical staining	SCAPs differentiate into osteoblasts, adipocytes, chondrocytes, and smooth muscle.
Wang et al. 2012 [35]	Human	OIM	IGF-1	ALP assay; Histochemical staining	IGF-1 enhances osteogenic differentiation but weakens odontogenic differentiation of SCAPs.
Wu et al. 2012 [36]	Human	OIM	bFGF	ALP assay; Histochemical staining	SCAP cultured with bFGF shows decreased mineralized nodule formation and ALP activity, but if pretreated with bFGF increased mineralized nodule formation is seen.
Wang et al. 2013 [63]	Human	OIM	None	ALP assay; Histochemical staining	High ALP activity and RUNX2 upregulation seen.
Qu et al. 2014 [64]	Human	OIM	None	ALP assay; Histochemical staining	Significant mineralization observed and enhanced osteogenesis is linked to DLX2.

(b) Dental papilla stem cells

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Ikeda et al. 2006 [112]	Human	OIM	HA	ALP assay	In vitro osteogenic differentiation observed if cultured in presence of OIM.

(c) Dental follicular stem cells (DFCSs)

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Tsuchiya et al. 2010 [38]	Porcine	OIM	None	ALP assay; Histochemical staining	DFCS has osteogenic potential.
Honda et al. 2011 [39]	Human	GCM	None	ALP assay; Histochemical staining	3 distinct cell populations were identified with DFCS. Among the three, two of them showed strong calcium accumulation.
Viale-Bouroncle et al. 2011 [113]	Human	OIM	Polydimethylsiloxane; Fibronectin	ALP assay	Soft surface improved the induction of osteogenesis differentiation of DFSC compared to higher stiffness.
Aonuma et al. 2012 [114]	Human	OIM	None	ALP assay; Histochemical staining	ALP activity higher than hMSC.
Li et al. 2012 [115]	Rat	OIM	Ad-BMP9; Ad-GFP	Histological staining	BMP 9 enhances osteogenesis of DFCS.
Park et al. 2012 [65]	Human	OIM	None	Histochemical staining	DFSC able to undergo osteogenic differentiation.
Mori et al. 2012 [116]	Human	OIM	None	ALP assay; Histochemical staining	High level of ALP expression, osteogenic potential, and mineralized nodule formation seen.
Rezai Rad et al. 2013 [40]	Rat	OIM	None	ALP assay; Histochemical staining	Osteogenesis of DFSC increased with temperature from 37°C to 40°C but lost its potential at 41°C.
Takahashi et al. 2013 [117]	Human	OIM	None	ALP assay	DFSC can undergo osteogenic differentiation in the absence of dexamethasone and BMP 6 is the key gene in osteogenic differentiation of DFSC.
Yao et al. 2013 [118]	Rat	OIM	hr-BMP6	ALP assay	DFSC lost its osteogenesis during in vitro expansion; addition of BMP-6 dramatically enhances osteogenesis of late passage.

(d) Gingival mesenchymal stem cells (GMSCs)

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Yu et al. 2014 [67]	Human	OIM	None	ALP assay; Histochemical staining	Mineralized nodule formed in the experimental group.

(e) Dental neural crest stem cells

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Degistirici et al. 2010 [119]	Human	OIM	None	ALP assay; Histology	Bone like matrix formation seen.

(f) Stem cells from human exfoliated dentition (SHEDs)

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Miura et al. 2003 [69]	Human	OIM	rhBMP 4	Histochemical staining	Osteogenic differentiation observed.
Vakhrushev et al. 2010 [120]	Human	Serum-free OIM	3D polylactide matrix	Histochemical staining	SHED and BMMSC have similar phenotype and identical osteogenic potential.
Li et al. 2012 [72]	Human	OIM	bFGF	Histochemical staining	bFGF inhibits osteogenic induction.
Viale-Bouroncle et al. 2012 [121]	Human	OIM	PDMS; Fibronectin	ALP assay; Histochemical staining	Rigid scaffold supports proliferation and osteogenesis of SHED.
Vakhrushev et al. 2013 [122]	Human	Serum-free OIM	TCP	Histochemical staining	TCP increases osteogenic differentiation, ossification foci and enhances ECM production by SHED.
Karadzic et al. 2014 [123]	Human	OIM	3D HAP, PLGA, alginate, EVA/EVV	ALP assay; Histology	All four are suitable carrier for SHED. Low level of osteoblastic differentiation is demonstrated in EVA/EVV.
Yu et al. 2014 [124]	Human	OIM	None	ALP assay; Histochemical staining	ALP activity and in vitro mineralization were not different between SCID and SHED. However more TNF-α is seen with SCID.

(g) Dental pulp derived stem cells (DPSCSs) from deciduous/permanent teeth

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Gronthos et al. 2000 [33]	Human	OIM	None	ALP assay	DPSC shows osteogenic potential (formed condensed nodule with high level of calcium) and forms more CFU than BMMSC.
Laino et al. 2005 [45]	Human	OIM	None	ALP assay; Histochemical staining	DPSC able to generate living autologous fibrous bone tissue (LAB).
Laino et al. 2006 [75]	Human	OIM	None	Calcium staining	Demonstrated pluripotency. Able to differentiate into osteoblast.
d’Aquino et al. 2007 [125]	Human	OIM	None	ALP assay; Histochemical staining	DPSC able to form woven bone in vitro.
Cheng et al. 2008 [126]	Chimpanzee	OIM	None	Histochemical staining	Osteogenic capacity of cDPSC was comparable to human BMMSC, DPSC, and rBMSC.
Graziano et al. 2008 [127]	Human	OIM; Rotating culture	HA, Ti, PLGA	ALP assay; Histochemical staining	PLGA shows better scaffold suitability for DPSC (1 mm bone tissue on PLGA, 0.3 mm in Ti, and no bone tissue formation seen in titanium covered with HA).
Morito et al. 2009 [78]	Human	OIM	PLGA; bFGF	ALP assay; Histochemical staining	Membrane bone like tissue formed around PLGA.
Alge et al. 2010 [128]	Rat	OIM	None	ALP assay; Histochemical staining	Significantly higher ALP activity than control group.
Han et al. 2010 [129]	Human	OIM; Mechanical bioreactor	None	ALP assay; Histochemical staining	Mechanical stimulation promotes osteogenic differentiation and osteogenesis of DPSC.
Mangano et al. 2010 [130]	Human	OIM	LST Ti	Histology; SEM	More osteoblast and bone formation seen with laser treated titanium surface.
Mori et al. 2010 [131]	Human	OIM	None	ALP assay	DPSC formed mineralized matrix nodules showing osteoblast features.
Spath et al. 2010 [132]	Human	OIM	Lenti virus vector expressing β galactoside	ALP assay; Histochemical staining	DPSC by explant culture method exhibits elevated proliferation and osteogenic potential.
Chan et al. 2011 [81]	Human	OIM	SAPN	Histochemical staining	DPSC survives encapsulation by SAPN and calcium salt deposition seen.
Galli et al. 2011 [133]	Human	OIM	3DTi	ALP assay; Histochemical staining	Increased expression of ALP genes and BMP 2 genes and increased osteogenic differentiation.
D’Alimonte et al. 2011 [134]	Human	OIM	VEGF-A165 peptide	ALP assay; Histochemical staining	VEGF enhances differentiation of DPSC towards osteoblast and DPSC showed negative hematopoietic marker.
Li et al. 2011 [83]	Human	OIM	3D gelatin	ALP assay; Histochemical staining	Increased ALP activity and osteoblast compared to control group.
Mangano et al. 2011 [135]	Human	OIM	Biocoral	Histology; SEM	Diffuse bone formation seen in the scaffold.
Struys et al. 2011 [136]	Human	OIM	None	TEM; Staining; Image analysis	Presence of multiple mineralization nuclei.
Huang et al. 2012 [137]	Rat	OIM	Flavanoid	ALP assay; Histochemical staining	Flavonoid increases DPSCs ALP activity by 1.47-fold and upregulation of RUNX2by 2.5-fold.
Huang et al. 2012 [138]	Rat	OIM	MAO Ti	ALP assay	Osteogenic potential of DPSC similar to BMMSC.
Khann-Jain et al. 2012 [139]	Human	Human serum (serum-free OIM)	βTCP	ALP assay; Histochemical staining	Matrix mineralization seen. Human serum can be substituted for FBS which facilitates translating from in vitro to clinical trials.
Pisciotta et al. 2012 [85]	Human	Human serum; OIM	Collagen sponge	ALP assay; Histochemical staining	High proliferation rate and osteogenic differentiation of DPSC in human serum compared to FCS.
Taşli et al. 2014 [140]	Human	OIM	BMP2,7; Plasmids, GFP	ALP assay; Histochemical staining	Transfection of human tooth germ cells with BMP2,7, induced osteogenic, and odontogenic differentiation.
Palumbo et al. 2013 [141]	Human	OIM	3D scaffold matrigel; Titanium	SEM; Confocal; TEM	Human osteoblasts from bone biopsies are appropriate compared to DPSCs.
Zavatti et al. 2013 [142]	Human	Ferutinin; OIM	None	Staining	Ferutinin enhances osteoblastic differentiation of DPSC.
Akkouch et al. 2014 [143]	Human	OIM	3D Col/HA/PLCL	Micro-CT; ALP assay; Histochemical staining	30% increase in bone nodule formation and tissue mineralization seen on surface as well inside the scaffold.
Amir et al. 2014 [144]	Macaque Nemestrima	Chitosan; OIM	None	ALP assay; Histochemical staining	Chitosan stimulates proliferation and early osteogenic differentiation of DPSC compared to dexamethasone.
Guo et al. 2014 [145]	Human	OIM	Fluorapatite; PCL	ALP assay; Histochemical staining	Scaffolds provided favorable ECM microenvironment for proliferation and osteogenic differentiation.
Huang et al. 2014 [146]	Human	OIM	Lenti virus; Cloned human OCT4, Nanog	ALP assay; Histochemical staining	OCT 4 and Nanog act as a major regulator in maintaining mesenchymal properties in DPSC.
Jensen et al. 2014 [147]	Human	OIM	NSP-PCL; HT-PCL	ALP assay; Histochemical staining	Both scaffolds promote calcium deposition, but HT-PCL supports only cell proliferation and migration.
Ji et al. 2014 [148]	Human	OIM; Biomimetic bioreactor	3D agarose gel	ALP assay; Histochemical staining	Mechanical loading enhances osteogenesis and bone formation
Kanafi et al. 2014 [149]	Human	OIM	Alginate hydrogel	Calcium quantification assay; Staining	DPSC immobilized in alginate hydrogel exhibits enhanced osteogenic potential
Niu et al. 2014 [91]	Human	OIM cocultured with silicic acid	Collagen	ALP assay; Histochemical staining	ISCS promotes proliferation, osteogenic differentiation, and mineralization compared with NCS.
Taşli et al. 2013 [150]	Human	OIM	NaB	ALP assay; Histochemical staining	NaB significantly increases level of ALP activity and mineralization with higher expression of osteogenic and odontogenic genes.
Woloszyk et al. 2014 [151]	Human	OIM; Spinner flask bioreactor	Silk fibroin	Micro-CT; Histology; ALP assay	DPSCs have the potential to form mineralized matrix when grown on 3D scaffold enhanced by mechanical loading.

(h) Periodontal ligament derived stem cells (PDLSCs)

Reference	Cell source	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Gay et al. 2007 [152]	Human	OIM	None	Histochemical staining	PDLSC has osteogenic differentiation and mineralization potential.
Trubiani et al. 2007 [153]	Human	OIM	Xenogenic Porcine substitute	ALP assay; Histochemical staining	Scaffold able to support PDLSC and demonstrated osteogenic potential.
Zhou et al. 2008 [154]	Human	OIM	None	ALP assay; Histochemical staining	Time dependent increase in matrix calcification observed with PDLSC.
Orciani et al. 2009 [155]	Human	OIM	None	TEM; SEM; ALP assay	NO involved in osteogenesis of PDLSC. In vitro osteogenesis of PDLSC resulted in osteoblast like cells with calcium deposits.
He et al. 2011 [97]	Dog	OIM	Porous n HAC/PLA	ALP assay	Osteogenic differentiation seen on the scaffolds.
Silvério et al. 2010 [51]	Human	OIM	None	Histochemical staining	Deciduous periodontal ligament derived cells promoted 100% mineral nodule formation, while permanent showed 60%.
Zhang et al. 2011 [156]	Rats	OIM	None	Histochemical staining	Decreased osteogenic differentiation seen in PDLSC derived from ovariectomised rats.
Zhou et al. 2011 [49]	Human	OIM	Ibandronate	qRT-PCR	Ibandronate promoted osteoblastic differentiation of PDLSC.
Ge et al. 2012 [157]	Human	OIM	IHGCCS	ALP assay; Histochemical staining	HGCS showed higher ALP activity.
Lee et al. 2012 [47]	Human	OIM	VEGF2; FGF2	ALP assay; Histochemical staining	VEGF has positive effect on osteogenic differentiation. FGF has positive effect on proliferation rate.
Sununliganon and Singhatanadgit 2012 [158]	Human	OIM	None	Staining	PDLSC able to form mineralized mass.
Yu et al. 2012 [48]	Human	OIM	IGF-1	ALP assay; Histochemical staining	IGF-1 stimulates osteogenic potential of PDLSC.
Zhang et al. 2012 [50]	Human	OIM; LMHF	None	ALP assay; Histochemical staining	LMHF promoted osteogenic potential of PDLSC.
Gao et al. 2013 [101]	Human	OIM	None	ALP assay; Histochemical staining	PDLSC able to form mineralized nodule.
Ge et al. 2013 [102]	Human	OIM	HAp; PADM	ALP assay; Histochemical staining	Higher ALP activity and osteogenic differentiation seen in Hap-PADM than pure PADM.
Houshmand et al. 2013 [159]	Human	OIM	EMD	Histochemical staining	EMD has no effect on osteoblastic differentiation of BMMSC or PDLSC.
Kato et al. 2013 [160]	Human	OIM	Synthetic peptide	ALP assay	More number of calcified nodules seen in culture with synthetic peptide.
Kim et al. 2013 [161]	Human	Hesperetin; OIM	None	ALP assay	Significant increase in ALP activity.
Kong et al. 2013 [162]	Human	OIM	None	ALP assay	Periodontal disease derived PDLSC displayed impaired osteogenesis compared to healthy PDLSC.
Singhatanadgit and Varodomrujiranon 2013 [163]	Human	OIM; spheroid culture	Conical polypropylene tube	Staining	Bone like deposit seen. PDLSC may undergo osteogenic differentiation in an osteogenic scaffold-free 3D spheroidal culture.
Yu et al. 2013 [164]	Human	OIM	None	ALP assay; Histochemical staining	Osteogenic differentiation of PDLSC far superior to WJCMSC.
Hakki et al. 2014 [165]	Human	OIM	Type I collagen; BMP6	Histochemical staining	BMP application stimulated mineralized nodule formation.
Jung et al. 2014 [106]	Human	OIM	rAd-EGFP, BMP2	Histochemical staining	Mineralized nodule formation seen. BMP 2 effectively promoted osteogenesis.
Tang et al. 2014 [166]	Human	OIM	None	ALP assay; Histochemical staining	PDLSCs have osteogenic potential and low immunogenicity.
Ye et al. 2014 [167]	Human	OIM	Ad-BMP9	ALP assay; Histochemical staining	BMP 9 promoted matrix mineralization.

(i) Multiple dental stem cells

Reference	Cell source	Comparison	Medium	Scaffold/carriers/cues/markers	Evaluation methods	Observation
Koyama et al. 2009 [168]	Human	DPSC; SHED	OIM	BMP2	ALP assay; Histochemical staining	No difference observed between DPSC and SHED for osteogenic potential.
Chadipiralla et al. 2010 [169]	Human	SHED; PDLSC	Serum-free OIM	Retinoic acid; ITS	ALP assay; Histochemical staining	High proliferation rate seen in PDLSC makes it a better osteogenic cell source. However SHED is more responsive to retinoic acid.
Bakopoulou et al. 2011 [170]	Human	DPSC; SCAP	OIM	None	ALP assay; Histochemical staining	DPSC and SCAP positive for markers of both osteogenic and odontogenic differentiation.
Lee et al. 2011 [171]	Human	DPSC; PDLSC	PRP; OIM	None	ALP assay; Histochemical staining	PRP induces osteogenic and odontogenic differentiation.
Atari et al. 2012 [172]	Human	DPSC; DPMSC	OIM	3D glass scaffold	ALP assay; Histochemical staining	DPPSCs have higher expression of bone markers than DPMSC.
Moshaverinia et al. 2012 [173]	Human	PDLSC; GMSC	OIM	Alginate hydrogel	SEM; XRD; Staining	Osteogenic potential is observed higher for BMMSC followed by PDLSC and lowest in GMSC.
Yang et al. 2013 [56]	Human	PDLSC; GMSC	OIM	None	ALP assay; Histochemical staining	PDLSC showed more effective osteogenic differentiation than GMSC
Davies et al. 2014 [174]	Human	DPSC; ADSC; BMSC	OIM	None	Micro-CT; Histochemical staining; SEM	High volume of mineralized matrix seen in DPSC group but diffused layer of low density seen in SEM.
Moshaverinia et al. 2014 [55]	Human	PDLSC; GMSC	OIM	RGD coupled alginate microsphere	Western blot; Fluorescent image analysis	Osteogenic potential of BMMSC is greater than PDLSC. However PDLSC shows better osteogenic potential than GMSC. Stem cells encapsulated in RGD showed enhanced osteogenesis.