自撑式石墨烯水凝胶诱导人脂肪干细胞成骨分化的体外研究

In vitro effects of self-sustaining graphene hydrogel film on the osteogenic differentiation of human adipose-derived stem cells

目的通过体外实验评价本课题组制备的自撑式石墨烯水凝胶(self-sustaining graphene hydrogel,SGH)的骨诱导性能。方法 (1)将人脂肪干细胞(hADSC)接种于SGH和玻片表面,采用荧光显微镜图像计数检测ADSC在材料表面粘附能力;通过扫描电镜观察材料本身的表征及细胞在SGH表面生长的形态;(2)SGH组加入普通培养液作为实验组,玻片组加入成骨培养基作为诱导组,玻片组加入普通培养液作为对照;通过实时定量荧光PCR、ALP活性检测和茜素红半定量测定钙沉积评价各组hADSC的分化及成骨能力。结果 hADSC能以等同于在玻片表面的粘附效率和正常的形态在SGH表面生长。实验组细胞在不同时间点成骨相关基因的表达水平、ALP活性以及钙盐沉积量均显著高于对照组而略低于诱导组(P<0.05)。结论自撑式石墨烯水凝胶作为一种生物相容性材料,具有较强的诱导人脂肪干细胞成骨分化的能力。

Objective To assess in vitro assessment of the excellent osteoinductive ability of a self-sustaining graphene hydrogel on human adipose-derived stem cells （hADSCs）. Methods （1）The hADSCs were cultured on both SGH films and control glass,and cell adhesion and morphology were examined by scanning electron microscopy （SEM） and immunofluorescence analysis;（2）The in vitro experiments were divided into three groups ,SGH films group ,glass group （osteogenic medium） and control glass group （normal growth medium）. In addition,in vitro experiments for osteogenic potential by hADSCs were performed for comparison among the three sample groups. Results The hADSCs, as on control glass ,Nere also attached tightly on the surface of the SGH film and appeared normal cell morphology. The in vitro study showed better osteogenic differentiation of hADSCs on self-sustaining graphene hydrogel group scaffolds than in the control group （P 〈 0.05）, due to the higher ALP activity,osteocalcin mRNA expression and mineralization,while its osteogenic ability was lower than in an osteogenic medium. Conclusions This self-sustaining graphene hydrogel film appears to be highly biocompatible and osteoinductive for hADSCs.