流体剪切力对人脂肪基质细胞成骨相关基因表达影响的研究

摘要

目的 探讨流体剪切力(FSS)对人脂肪基质细胞(ADSC)成骨相关基因骨钙素(OCN)、转录因子Osterix(Osx)及核心结合因子(Cbf) al/Runt相关转录因子(Runx)2表达的影响.方法 将脂肪抽吸术获取的人体脂肪进行常规培养(A1、A2组)和诱导培养(A3、A4组),然后分别对其加载强度为0.3 Pa的FSS(A1、A3组),未加FSS的为对照组(A2、A4组),通过逆转录-聚合酶链反应检测骨钙素、转录因子Osx及Cbfal/Runx2基因表达的情况.结果 A3、A4组在FSS加载后,其成骨相关基因的表达增强;而A1、A2组未见成骨特异性基因条带.这表明在常规培养条件下,无论是否有FSS的刺激,均无成骨特异性基因表达.结论 脂肪基质细胞在诱导培养条件下,FSS可促使其向成骨细胞分化,可作为骨组织工程的种子细胞.%Objective This study aimed to investigate the influence of fluid shear stress(FSS) on osteogenic genes,code for steocalcin(OCN),Osterix(Osx),and core binding factor(Cbf) al/Runt-related transcription factor(Runx)2,in human adipose-derived stromal cells(ADSCs).Methods Human ADSCs were cultured and proliferated in vitro by common culture(groups A1 and A2) and inducing culture(groups A3 and A4).These groups were then subjected to a FSS of 0.3 Pa(groups A1 and A3),whereas the group without FSS were used as control(groups A2 and A4).After adding FSS,the gene expression levels of OCN,Osx,and Cbfal/Runx2 were examined by reverse transcription polymerase chain reaction.Results The gene expression levels of OCN,Osx,and Cbfal/Runx2 in groups A3 and A4 were enhanced after applying FSS.However,the gene expression levels were undetectable in groups A1 and A2.Therefore,osteogenic gene expression in human ADSCs is not present in common culture conditions even after the application of FSS.Conclusion In inducing culture condition,FSS can enhance the expression levels of the osteogenic genes OCN,Osx,and Cbfal/Runx2 in human ADSCs and promote the differentiation of osteoblasts,which can be used as seed cells for bone tissue engineering.