Chondrogenic differentiation and osteogenesis gene expression following a short-time induction of adipose-derived stem cells with bone morphogenetic proteins / 中国组织工程研究

ABSTRACT

OBJECTIVE:To investigate osteogenic and chondrogenic differentiation of adipose-derived stem cells following a short time induction with bone morphogenetic protein 2(BMP-2) or BMP-7.METHODS:The subcutaneous adipose tissue was obtained from adult New Zealand rabbits under aseptic condition,and cultured in vitro with collagenase digestion.All cells were divided into 3 groupsin the BMP-2 group,cells were cultured with medium containing 0.1 g/L vitamin C,10 mmol/L?-sodium glycerophosphate and 10?g/L BMP-2 for 10 minutes,followed by 4-14 days inoculation with density of 18?104 cells per pore.In the BMP-7 group,cells were cultured with BMP-7 with the same methods as BMP-2 group.The cells were cultured with simple culture medium in the control group.RESULTS:Compared to the control group,the number of adipose-derived stem cells,protein level and ALalkaline phosphatase(ALP) activity of BMP-2 and BMP-7 groups was up-regulated 1.78-1.79,1.15-1.95,and 32-40 times,respectively.At days 4 after a 15 minutes induction,the runx-2 gene expression,osteopontin gene,and biglycan gene were increased 1.9,2.2 and 1.3-1.7 folds than that of the control group.Meantime,only the biglycan gene expression was increased 1.3-1.7 folds in the BMP-7 group,the runx-2 gene expression,osteopontin gene was not changed.At day 14 after a 15 minutes induction,there was no alter of runx-2,osteopontin,biglycan,as well as aggrecan gene expression in the BMP-2 group;while down-regulated runx-2,osteopontin,biglycan 1.8,5.0 and 1.7 folds,with increased aggrecan gene in the BMP-7 group.CONCLUSION:Following a short time induction,BMP-2 can stimulate runx-2 and osteogenic expression at 4 days after re-culture,whereas BMP-7 down-regulate genes expression at days 14,yet down-regulate aggrecan mRNA expression.