Stem Cells International

Review Article

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

Table 4

Study characteristics of included in vivo experiments with the application of dental stem cells on Bone regeneration.
(a) Stem cells from apical papilla (SCAPs)
ReferenceCell sourceSpeciesGenderAge
Week/months		Weight (mg/kg)Total number of animalsDefect type and locationTransplanted cell numberScaffold/growth factors/cuesPeriodEvaluation methodsObservation
Abe et al. 2008 [61]HumanRatnanananaSC pouch5 × 105HA12 wkHistologyEctopic bone like tissue on the border of the scaffold
Abe et al. 2012 [62]Human MiceM4 wknanaSC pouch5 × 104Porous HA12 wk HistologyEctopic bone like tissue on the border of the scaffold
Wang et al. 2013 [63]Human Micenanana12Renal capsule1 × 106Absorbable gelatin sponge2 wkHistologyCalcified tissue formation
Qu et al. 2014 [64]HumanMiceF10 wknanaSC4 × 106HA/TCP
BMP4		8 wk HistologyDLX2 overexpression enhances mineralized tissue formation.
(b) Dental follicular stem cells (DFCSs)
ReferenceCell sourceSpeciesGenderAge
Week/months		Weight (mg/kg)Total number of animalsDefect type and location Transplanted cell numberScaffold/growth factors/cuesPeriod Evaluation methods Observation
Xu et al. 2009 [37] Rat Mice nanananaSc pouch4 × 1063D-β TCP
BMP 2		8 wkHistologyLacked new bone formation
Tsuchiya et al. 2010 [38]Porcine Ratnanana12CSD calvarium 5 mm 1 × 106None1 wk
4 wk		HistologyNo new bone formation. Apparent bone like structure
Honda et al. 2011 [39]Human Ratnanana24CSD calvarium
8 mm		2 × 106/pelletNone1 wk
4 wk		Histology Bone formation with evidence of vascular invasion similar to intramembranous ossification
Park et al. 2012 [65]Human Micem8 wkna4SC pouch1 × 106DBM
Fibrin glue		4 wkCT
Histology		Trabecular bone generation with vessels
(c) Gingival mesenchymal stem cells (GMSCs)
ReferenceCell sourceSpeciesGenderAge
Week/months		Weight (mg/kg)Total number of animalsDefect type and location Transplanted cell numberScaffold/growth factors/cuesPeriod Evaluation methods Observation
Wang et al. 2011 [66]Human Rat
 Mice		F6–8 wk
 8–10 wk		160–180 g
 na		10
 3		Mandibular body defect (5 × 2 × 1 mm)
SC pouch		na 5 × 106Type 1 collagenHistology8 wk
 6 wk		Bone formation in the defected area
Yu et al. 2014 [67]DogDogMna10-11 kg4Class III furcation defect eGFPHistology 8 wkEnhanced new bone formation GMSC (47.11 ± 7.91%) versus control group ( 37 ± 9.53)
Xu et al. 2014 [68]HumanMiceM7 wkna36Rt mandibular body (1.5 mm diameter) 1 × 106GFP as markerHistology1 wk
2 wk
3 wk		Active bone formation at 3 wk
(d) Stem cells from human exfoliated dentition (SHEDs)
ReferenceCell sourceSpeciesGenderAge
Week/months		Weight (mg/kg)Total number of animalsDefect type and locationTransplanted cell numberScaffold/growth factors/cuesPeriodEvaluation methodsObservation
Miura et al. 2003 [69]HumanMicenanananaSC2 × 106HA/TCP8 wkHistology Induce new bone formation
Seo et al. 2008 [70]HumanMicenanana18Calvaria (2.7 mm)2 × 106HA/TCP6–8 wk
6 month		HistologyRobust bone formation without hematopoietic bone marrow
Zheng et al. 2009 [71]MinipigMinipigF4–6 m20–30 kg16Bilateral parasymphyseal CSD (2.5 × 1.5 × 1.5 cm3)  
1 × 1 × 0.5 cm3  		2 × 107 to
4 × 108		PT67/eGFP
β TCP
HA/TCP		24 wk
2 wk [3]
4 wk [3]		-CT
Histology		Defects restored with new bone at 6 m
Li et al. 2012 [72]HumanMiceF8–12 wknanaSC pouch4 × 106HA/TCP
bFGF		8 wkHistology b FGF downregulated STRO-1, CD146, CD90, and CD73 expression of SHED
Vakhrushev et al. 2012 [73]HumanMicenananananana3D PLGA1 monthDAPI stainingMore intense expression of osteocalcin on scaffolds with SHED
Alkaisi et al. 2013 [74]HumanNew Zealand Rabbitna3–5 months2.7 ± 0.31 kg22Distraction of 6.2 mm between first lower premolar and mental foramen6 × 106None2 wk
4 wk
6 wk		Radiology
Histology		New bone formation with thick cortices and marrow cavity at 6 wk
Behina et al. 2014 [41]Human SHED 5 yr agoDogMna15–25 kg4Mandibular through-through (9 mm diameter)naCollagen12 wkHistology5-year cryopreserved SHED able to proliferate and osteogenesis without immune response. Bone formation is same as control group
(e) Dental pulp derived stem cells (DPSCSs) from deciduous/permanent teeth
ReferenceCell sourceSpeciesGenderAge
Week/months		Weight (mg/kg)Total number of animalsDefect type and location Transplanted cell numberScaffold/growth factors/cuesPeriod Evaluation methods Observation
Laino et al. 2006 [75]Human (deciduous teeth)Ratna10–12 wkna5SCWoven bone obtained by in vitro SHED cultureWoven bone4 wkHistologyWoven bone remodeled to lamellar bone with osteocytes entrapped within the lamella
Otaki et al. 2007 [76]HumanMicena7 wknanaSC2 × 106 to 1.8 × 107HA/TCP7 wk
15 wk		Histology50% bone formation seen
de Mendonça Costa et al. 2008 [77]HumanRatM4 months320–420 gm8Cranium
(5 × 8 mm)		1 × 106Collagen membrane7 d
20 d
30 d
60 d
120 d 		HistologyDefect healed with new bone formation
Zhang et al. 2008 [44]RatMicena10 wkna10SC5 × 106HA/TCP5 wk
10 wk		HistologyNo evidence of bone formation
Morito et al. 2009 [78]HumanMicena10 wknanaSC4 × 105PLGA with Calcium Phosphate5 wk
10 wk		HistologyConfirmed bone and cartilage formation
Yang et al. 2009 [79]RatMicena10 wkna12SC5 × 106AdBMP-2
HA/TCP		1 wk
4 wk
12 wk		HistologyEnhance mineral tissue formation
Kraft et al. 2010 [80]HumanMiceF8 wkna21.5 cm deep pouch5 × 105HA-TCP8 wkHistologyLamellar bone like structure
Chan et al. 2011 [81]HumanMicena6 wkna5SC pouch1 × 105SAPN4 wkHistologyMineralized tissue formed
Ito et al. 2011 [82]DogDogna2 yrna3Hemimandible 10 × 10 mm1 × 107PRP gel8 wkHistologySignificant amount of new bone formation seen in the defect
Li et al. 2011 [83]HumanMicena6 wkna8SCnaNone4 wkHistology
X ray		Bone formation seen.
Liu et al. 2011 [84]RabbitNew Zealand RabbitFNa2.5–3 kg36Segmental
10 × 4 × 3 mm		1 × 108n HAC/PLA
rh-BMP-2
eFG		12 wkHistology
X ray		Bone regenerated in the defect area
Pisciotta et al. 2012 [85]HumanRatM14 wkna105.8 × 1.5 mm cranial1 × 106Collagen sponge6 wkHistologyRegeneration of resected bone
Riccio et al. 2012 [86]HumanRatM12–14 wkna155 × 8 mm parietalnaSilk fibroin4 wkHistologyInduce new bone formation in the critical sized defect
Annibali et al. 2013 [42]HumanMicena50 daysna75Parietal (4 × 1 mm)1 × 106DBB β TCP
Hydrogel-ceramic composite sponge		1 wk
2 wk
4 wk
8 wk		HistologyTE constructs did not significantly improve bone regeneration
Khorsand et al. 2013 [87]DogDogM1-2 yr14–22 kg103 × 5 × 8 mm2 × 107BIO-OSS8 wkHistologyWoven bone formation seen and no significant difference seen between control and experimental group
Maraldi et al. 2013 [88]HumanRatM12–14 wkna30Parietal 5 × 8 mmnaCollagen4 wk
8 wk		HistologyNew bone formation seen in the defect
Wang et al. 2013 [89]RatRatF8 wkna30Ovariectomy
Renal capsule		1 × 106Absorbable gelatin sponge14 daysHistologyEstrogen deficiency inhibits osteogenic potential of DPSCS (downregulated by NF-B pathway)
Annibali et al. 2014 [43]HumanRatna50 daysna8Parietal (5 × 1 mm)naGDPB
β TCP		2 wk
4 wk
8 wk
12 wk		µ-CT
µ-PET		Addition of stem cell did not increase new bone formation
Ling et al. 2014 [90]RabbitNew Zealand Rabbitnana2.5–3 kg6SC1 × 106 n HAC/PLA 
β TCP		8 wkHistologyMature bone formation seen
Niu et al. 2014 [91]HumanMiceM5 wkna6SC5 × 106ISCS
NCS		8 wkHistologyNew bone formation seen.
(f) Periodontal ligament derived stem cells (PDLSCs)
ReferenceCell sourceSpeciesGenderAge
Week/months		Weight (mg/kg)Total no of animalsDefect type and location Transplanted cell numberScaffold/growth factors/cuesPeriod Evaluation methods Observation
Doǧan et al. 2002 [92]DogDognanana1Class II furcation defect2 × 105Blood clot42 daysHistologyPDLSC promote bone regeneration
Seo et al. 2004 [46]HumanRat
Mice		na12–10 wknaRat-6
Mice-12		Rat-2 mm2 periodontal defect
Mice-SC		Rat-2 × 106 
Mice-4 × 106		HA-TCP6–8 wkHistologyNo bone formation seen
Murano et al. 2006 [93]DogDognanana15Class III furcation defectnaNone2 wk
4 wk
8 wk		HistologyBone regeneration with filling of most defect along with cementum formation
Iwata et al. 2009 [94]DogDogMna10 kg43-wall defect (5 × 5 × 4 mm)naPGA6 wkHistology
Micro-CT		Significant new bone formation compared to control group
Kim et al. 2009 [95]DogDogMna12–15 Kg4Mandibular 5 × 10 mm saddle defect1 × 106HA/TCP16 wkHistologyDefect regenerated new bone
Ding et al. 2010 [96]MinipigMinipigM & F6–8 m30–40 kg153 × 7 × 5 mm periodontal defect2-cell sheet/defectHA/TCP0 wk
12 wk		CT-Scan
Histology		PDLSC sheet repair allogeneic bone defect
He et al. 2011 [97]DogDogna2 yearnanaSC pocket2 × 106nHAC/PLA8 wkHistologyNew bone like tissue seen
Grimm et al. 2011 [98]HumanRatsna10 wkna172.5 × 2.5 × 2 mm3 periodontal defect1 × 105Collagen sponge2 wk
6 wk
8 wk		HistologyPDLSC able to regenerate bone
Lee et al. 2012 [47]HumanMiceM6–8 wknanaSCnaHA/TCP
VEGF
FGF-2		8 wkHistologyHard tissue formation seen.
Suaid et al. 2012 [99]DogDogna1.46 ± 0.18 years10–20 kg7Bilateral Class III defect3 × 105Collagen12 wkHistologyNew bone formation seen in the defect
Tour et al. 2012 [100]RatRatMna350 gm24CSD Calvaria
8 mm		2 × 105HA-ECM12 wkHistologyBone regeneration observed in the CSD
Yu et al. 2012 [48]HumanMicenanananaRenal capsule1 × 106Absorbable gelatin sponge
IGF-1		6–8 wkHistologyIGF-1 enhances osteogenic differentiation of PDLSC
Immature bone like structure formed
Gao et al. 2013 [101]HumanMiceM4–6 wkna12SCna Osthole 
HA-TCP		4 wkHistologySignificant bone formation seen
Ge et al. 2013 [102]HumanRatM8 wk180–220 gm18Bilateral parietal defect
5 mm diameter		1 × 107HGCCS
GCF		12 wkHistologyBone formation seen in the defect
Mrozik et al. 2013 [103]SheepSheepna3–5 years63.5–72 kg13Rectangular 0-wall defect (10 mm deep)1 × 107Gelfoam4 wkHistologyNew alveolar bone formation seen, not significant with gelfoam alone group but significant with control group
Yu et al. 2013 [104]RatRatna7 wkna12Bilateral 3 wall bone defect
(2 × 2 × 1.7 mm3)		4 × 106Gelatin sponges6 wkHistologyNew bone formed in the defect
Han et al. 2014 [105]RatRatFna220–250 g36Periodontal defect1 × 106Gel foam1 wk
2 wk
3 wk
4 wk		HistologyComplete bridging of osseous defect with mineralized tissue containing osteocytes
Jung et al. 2014 [106]HumanMicena6 wkna14SCnarAD-EGFP
hBMP2		2 wk
8 wk		HistologyEctopic Bone formation seen
Park et al. 2015 [107]DogDognana10–12 kg6Peri-implantitisnaHA
Ad BMP2		7.5 monthsHistologyNew bone formation and re osseointegration of implants seen
Yu et al. 2014 [108]DogRatM2 m150 g24CSD calvaria (4 mm wide)2 × 106 Bio-oss8 wkMicro-CT
Histology		Defect regenerated new bone
Yu et al. 2014 [109]DogDogM18 m14.5 kg6Maxillary sinus floor augmentation2 × 106 Bio-oss8 wkMicro-CT
Histology		New bone formation seen
Zhao and Liu 2014 [110]HumanMicenanananaSC4 × 106Ceramic bovine bone simvastatin8 wkHistologyBone like hard tissue formation on the scaffold. Larger amount seen in PDLSC and scaffold with simvastatin group
(g) Multiple dental stem cells
ReferenceCell source Type comparedSpeciesGenderAge
Week/months		Weight (mg/kg)Total number of animalsDefect type and location Transplanted cell numberScaffold/growth factors/cuesPeriod Evaluation methods Observation
Yamada et al. 2011 [52]Dogc DPSC
p DTSC		Dogna2 yrnanaThree 10 mm diameter mandibular defectsnaPRP8 wk
16 wk		HistologyWell-formed new bone with vascularity is seen in all groups studied.
Wang et al. 2012 [53]HumanSHED
DPSC		Micena8 wknanaSC2 × 106CBB
Fibrin gel		8 wkHistologyHigher osteogenic differentiation and bone formation seen in SHED compared to DPSC.
Moshaverinia et al. 2013 [54]HumanPDLSC
GMSC		Micena5 monthsnanaSC2 × 106Injectable alginate hydrogel8 wkMicro-CT
Histology		ALP activity as well as mineralized tissue formation of PDLSC is better than GMSC but comparatively less than BMMSC.
Yang et al. 2013 [56]HumanPDLSC
GMSC		MiceM6 wknanaSC2 × 105Artificial bone repair material8 wkHistologySignificant bone formation seen. However GMSC demonstrated better osteogenic potential and bone formation in inflammatory condition compared to PDLSC.
Moshaverinia et al. 2014 [55]HumanPDLSC
GMSC		Micena5 monthsna165 mm diameter calvarial defect4 × 106RGD-coupled alginate8 wkMicro-CT
Histology		Bone regeneration in defect area (greater in BMMSC, moderate in PDLSC, lesser in GMSC groups)