多肽水凝胶复合支架对大鼠脂肪干细胞体外增殖和分化的影响  

The effects of peptide hydrogel composite scaffold on the proliferation and differentiation of RAT adipose-derived stem cells

作　　者：彭莎莎[1] 姚菁[1] 陈刚[1] 连小洁[2,3] 张晓军 崔菡 宋天喜 夏阳[1] 章非敏[1] 顾宁[4] 崔福斋[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]清华大学材料系,北京100084 [3]北京奥精医药科技有限公司,北京100176 [4]东南大学苏州研究院,江苏苏州215123

出　　处：《南京医科大学学报（自然科学版）》2013年第3期318-322,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家973项目(2011CB606205);国家自然科学基金(50830102);苏州市国际科技合作项目(SH201114);江苏高校优势学科建设工程资助项目江苏省医学领军人才与创新团队;江苏省研究生创新工程(CXLX11_0724)

摘　　要：目的:研究多肽水凝胶复合骼金(nano-hydroxyapatite/collagen,nHAC)对大鼠脂肪干细胞(adipose-derived stemcells,ADSCs)体外增殖和早期骨向分化的作用。方法:用1%的多肽水凝胶与nHAC制备复合支架,未复合水凝胶的nHAC为对照组,使用扫描电子显微镜(field emission scanning electron microscope,FESEM)和共聚焦显微镜(Confocal laser scanningmicroscope,CLSM)进行表征,将ADSCs接种到两种支架材料上,以CCK-8检测1、3、5、7 d的增殖情况,以碱性磷酸酶(ALP)活性为早期骨向分化指标,检测其3、5、7 d的活性。结果:多肽水凝胶在nHAC表面形成涂层,ADSCs在有涂层的nHAC上黏附、铺展得更好,两种支架上的ADSCs在1、3、5、7 d都持续增殖,其中两组间在1、3 d无差异,5、7 d有差异,实验组增殖更快(P<0.05);ALP值在3、5、7 d都持续增高,实验组更高,两组间有差异(P<0.05),但在3、5、7 d时两两比较差异不显著(P>0.05)。结论:多肽水凝胶复合支架材料能显著促进ADSCs的黏附增殖,对细胞早期骨向分化有一定的促进作用,但不如对细胞增殖的促进作用显著。Objective：To study the effects of peptide hydrogel composite scaffold on the proliferation and early osteogenic differentiation of rat adipose-derived stem cells（ADSCs）. Methods： Nano-Hydroxyapatite/Collagen （nHAC） scaffold was coated by 1% peptide hydrogel, nHAC without peptide coating was set as control. They were characterized by field emission scanning electron microscope （FESEM） and confocal laser scanning microscope （CLSM）. ADSCs were chosen to seed onto the nHAC coated and uncoated with peptide hydrogel. CCK-8 kit was used to observe cell proliferation. The alkaline phosphate （ALP） activity was detected the early osteogentic differentiation. Results：Peptide coating was formed on the surface of scaffold. ADSCs adhered to and fully extended on the peptide hydrogel composite scaffold under CLSM. The data of proliferation and ALP continually increased in both groups. There was no difference in cell proliferation at 1 d,3 d（P 〉 0.05）. At 5 d,7 d,ADSCs on the coated nHAC proliferated more significantly （P 〈 0.05）. There was no difference among 3 d,5 d and 7 d in the ALP activity （P 〉 0.05）. Conclusions：The peptide hydrogel composite scaffold could improve adhesion and proliferation of ADSCs, and also can improve early osteogentie differentiation of the cell, although not so obvious as the former two.

关 键 词：多肽水凝胶 纳米羟基磷灰石 胶原 脂肪干细胞 增殖 分化 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]