转染Ad-hBMP2的脂肪干细胞与壳聚糖/磷酸三钙复合物支架的相容性  

Compatibility of Adipose-derived Stem Cells Transfected by Ad-hBMP2 Gene and CTCP Scaffold in vitro

作　　者：方忠[1] 杨琴[1] 熊伟[1] 李光辉[1] 廖晖[1] 李锋[1] 肖骏[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《华中科技大学学报（医学版）》2012年第3期291-295,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30772206)

摘　　要：目的探讨转染腺病毒骨形态发生蛋白(Ad-hBMP2)基因的脂肪干细胞(ADSCs)与壳聚糖/磷酸三钙(CTCP)复合物支架的相容性,以期为ADSCs修复骨缺损寻找理想的组织工程骨支架材料。方法将壳聚糖与磷酸三钙进行复合制成CTCP复合物材料,再将其与转染Ad-hBMP2的ADSCs(密度1×106/mL)复合培养,进行细胞复合物支架的一般与超微形态学观察,观察细胞粘附能力、增殖活力,RT-PCR及Western blot测定成骨细胞骨钙素和Ⅰ型胶原水平。结果 CTCP支架孔径200～350μm,孔隙率83%;电镜显示转染Ad-hBMP2的ADSCs与CTCP复合物在体外培养期间支架无塌陷及形变,且其在支架上粘附、增殖良好,并能分泌细胞外基质如骨钙素和Ⅰ型胶原等;随着培养时间延长,骨的组织学特征日益明显。结论 CTCP支架具有适宜的孔结构和良好的生物相容性及骨诱导作用,可以作为骨组织工程较理想的支架材料,且与转染Ad-hBMP2的ADSCs相结合更易形成组织工程骨。Objective To observe the biological behavior of cultured adipose-derived stem cells（ADSCs）transfected by Ad-hBMP2 combined with chitosan/tricalcium phosphate（CTCP）scaffold and investigate the feasibility of the composite for cartilage tissue engineering.Methods The ADSCs（1×106/mL）transfected with Ad-hBMP2 plasmid vector were co-cultured with the CTCP scaffold.The adhesion and proliferation of ADSCs,and the morphological changes were observed.RT-PCR,Western blot and immunohistochemistry were applied to detect the expression of Osteocalcin and collagen Ⅰ in the scaffold.Results The poriferous CTCP scaffold has macro and micro poriferous structures,and the porous rate was 83%.The ADSCs transfected by Ad-hBMP2 have been successfully cultured in vitro.The induced cells adhered to the surface of the scaffold and proliferated well.The RT-PCR,Western blot and immunohistochemistry revealed that the expression of Osteocalcin and collagen Ⅰ was detected after the co-culture.Conclusion The poriferous CTCP scaffold with excellent property should be a good ＂matrix＂ for ADSCs transfected by Ad-hBMP2,and could be used for bone tissue engineering.

关 键 词：Ad-hBMP2基因 支架 脂肪干细胞 组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]