曲古抑菌素A对大鼠脂肪干细胞在二维及三维培养下早期骨向分化的影响  

Effects of trichostatin A on the early osteogenic differentiation of RAT adipose-derived stem cells cultured in two-dimensional and three-dimensional systems

作　　者：李娜[1] 陈刚[1] 刘珏[1] 章非敏[1] 夏阳[1] 顾宁[2] 

机构地区：[1]南京医科大学口腔医学研究所,江苏南京210029 [2]东南大学苏州研究院,江苏苏州215000

出　　处：《南京医科大学学报（自然科学版）》2014年第4期472-477,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏高校优势学科建设工程资助项目(2011-137);江苏省医学领军人才与创新团队;苏州市国际科技合作项目(SH201114)

摘　　要：目的:研究曲古抑菌素A(trichostatin A,TSA)对大鼠脂肪干细胞(adipose-derived stem cells,ADSCs)在二维培养板(TCP)和三维复合纳米羟基磷灰石/胶原(nano-hydroxyapatite/collagen,nHAC)支架上早期骨向分化的影响。方法:处理组分别在含75 nmol/L TSA的二维培养板和三维nHAC上培养ADSCs(TCPT组和nHACT组),未添加TSA的两组(TCP组和nHAC组)为对照组。使用扫描电子显微镜(field emission scanning electron microscope,FESEM)对接种ADSCs 1、3 d后nHAC和nHACT组细胞的生长情况进行表征,以碱性磷酸酶(ALP)活性为早期骨向分化指标,分别检测3、5、7 d时的ALP活性,用Western blot检测成骨指标Runx2、骨桥蛋白(OPN)、骨形态发生蛋白2(BMP2)的表达。结果:ADSCs在三维培养中均表现出良好的粘附、铺展,其中经TSA诱导的细胞对支架的黏附、铺展更好;ALP值在3、5、7 d持续增高,其中以nHACT组最高,组间差异显著(P<0.05);Western blot结果显示Runx2、OPN、BMP2在nHACT组表达最高,与ALP结果一致。结论:TSA对ADSCs早期骨向分化有促进作用,复合三维支架共培养后促进作用显著。Objective：To study the effects of trichostatin A（TSA） on the osteogenic differentiation of rat adipose-derived stem cells （ADSCs） in two-dimensional tissue cuhure plastic and three-dimensional nano-hydroxyapatite/collagen （nHAC） scaffold. Methods： ADSCs were cultured in two-dimensional tissue culture plastic （TCPT） and three-dimenslonal nHAC （nHACT） which contained 75 nmol/L TSA, and two groups without TSA （the TCP group and the nHAC group） were set as control. One and three days after ADSCs seeded onto the nHAC scaffold,the cells were characterized by field emission scanning electron microscope （FESEM）. The alkaline phosphate （ALP） activity was detected to evaluate the early osteogentic differentiation at 3,5,7 d. Western blot was performed to detect the expression of Runx2, OPN and BMP2 proteins. Results： On nHAC, ADSCs were well adhered to and fully extended on the scaffold with TSA. ADSCs on TSA treated scaffold presented better. The data of ALP was continuously increased at 3,5,7 d. The highest expression of ALP occurred in nHACT had significant difference between groups （P 〈 0.05）. Western blot demonstrated that Runx2, OPN and BMP2 protein in the nHACT groups increased more evidently than the other groups,with the same trends as ALP results. Conclusion：TSA could improve the early osteogentic differentiation of ADSCs,especially in the three-dimensional nHAC scaffold.

关 键 词：曲古抑菌素A 纳米羟基磷灰石 胶原 脂肪干细胞 分化 

分 类 号：Q254[生物学—细胞生物学]