In vivo evaluation of mixtures of uncultured freshly isolated adipose-derived stem cells and demineralized bone matrix for bone regeneration in a rat critically sized calvarial defect model.
Although many studies have suggested that human adipose tissue contains pluripotent stem cells, a few reports are available on stromal vascular fraction (SVF). In the present study, we evaluated the bone formation capacities of SVF. We implanted uncultured freshly isolated adipose-derived stem cells combined with demineralized bone matrix (DBM) to induce bone regeneration in a critically sized rat calvarial defect model. We used DBM (DBX(®)) and/or poly(70L-lactide-co-30DL-lactide) copolymer PLA as a scaffold. Fifty white rats were randomized to 5 different groups (n=10): (1) control, (2) DBM, (3) DBM + SVF, (4) DBM + PLA, and (5) DBM + PLA + SVF groups. After acquiring SVF, an 8-mm critically sized calvarial defect was made in each rat. Specimens were harvested at 8 weeks postimplantation and evaluated radiographically and histologically. New bone formation was qualified by hematoxylin and eosin staining and anti-osteocalcin antibody (OC4-30) immunostaining of calvarial sections. Amounts of mineralization were determined by radiodensitometric analysis. In gross appearance, the DBM + SVF and DBM + PLA + SVF groups showed more abundant bone formation than the other groups. Radiodensitometric evaluations revealed that significant intergroup differences were observed according to the Kruskal-Wallis (rank) test (P=0.030<0.05). The 5 groups show different amounts of filling of bone defects (control: 13.48%; DBM: 39.94%; DBM + SVF: 57.69%; DBM + PLA: 24.86%; DBM + PLA + SVF: 42.75%). Histological evaluation revealed that there was abundant new bone formation in the DBM + SVF and DBM + PLA + SVF groups. It was found that undifferentiated adipose-derived stem cells in the form of SVF induced new bone formation in rat calvarial defects. Accordingly, SVF offers a practical, promising candidate for regenerative tissue engineering or cell-based therapy.
Department of Plastic and Reconstructive Surgery, Inje University Ilsan Paik Hospital, Goyang City, Gyeonggi-do, Republic of Korea.
Seung Chul
SC
Rhee
Yi-hwa
YH
Ji
Nareg A
NA
Gharibjanian
Eun Sang
ES
Dhong
Seung Ha
SH
Park
Eul-Sik
ES
Yoon
D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D000943:Antigens, Differentiation / Q000378:metabolism; D018775:Bone Demineralization Technique; D015519:Bone Density; D001857:Bone Matrix / Q000637:transplantation*; D001861:Bone Regeneration; D002454:Cell Differentiation; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D020007:Implants, Experimental; D051379:Mice; D008819:Mice, Nude; D010006:Osteoblasts / Q000473:pathology; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D012886:Skull / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000166:cytology; D054457:Tissue Scaffolds; D014704:Veratrum Alkaloids / Q000494:pharmacology
D015703:Antigens, CD; D000943:Antigens, Differentiation; D053823:Hedgehog Proteins; D014704:Veratrum Alkaloids; C000541:cyclopamine