转化生长因子-β基因转染骨髓基质干细胞在多肽修饰的聚乳酸-羟基乙酸上生长情况的研究  

Growth of mesenchymal stem cells transfected by transforming growth factor 15 gene on PLGA scaffold modified with RGD peptide

作　　者：李长文[1] 郑启新[1] 郭晓东[1] 全大萍[2] 赵洁[2] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,武汉430022 [2]中山大学高分子研究所

出　　处：《中华创伤骨科杂志》2008年第1期54-58,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金项目（30170270,30200063,30470483）

摘　　要：目的探讨转转化生长因子-β（TGF-β）基因的骨髓基质干细胞（BMSCs）在RGD多肽表面修饰的聚乳酸-羟基乙酸（PLGA）材料上的增殖、黏附、分化情况，以研究TGF-β基因和RGD多肽在调控种子细胞生物学行为方面的作用。方法将GRGDSPC多肽共价结合到改性PLGA支架材料上，把TGF-β基因转入BMSCs，以单纯PLGA支架材料、单纯BMSCs作为对照，分别将单纯BMSCs和转TGF-β基因BMSCs两组细胞种植于两种材料上，培养1、2、3d后，计数细胞密度以反映细胞增殖情况；接种后4、12h，沉淀法定量检测黏附的细胞数；培养24h后用FITC连接的鬼笔环肽对细胞骨架染色，观察细胞骨架的组织情况；用成骨性培养基培养7、14、21d，检测细胞中碱性磷酸酶（ALP）活性来了解BMSCs分化情况。结果转TGF-β基因细胞密度显著增大，且在各时间点有较高的ALP活性。RGD多肽修饰的PLGA材料上细胞黏附率高，并有较高的荧光素密度及较粗大的细胞骨架，且在第14天时表现出高的ALP活性。结论TGF-β基因和RGD多肽同时应用有利于BMSCs黏附到支架材料上，并能促进其增殖和向成骨细胞方向分化。Objective To explore the effect of transforming growth factor β（TGF-β） gene and RGD peptide on cell biological behavior by investigating the proliferation, adhesion and differentiation of BMSCs transfected by TGF-β1 gene on the PLGA scaffold modified by RGD peptide. Methods Peptide Gly-Arg-Gly-Asp-Ser-Pro-Cys （GRGDSPC） was immobilized on the surface of PLGA, TGF-β1 was transfected into BMSCs, and simple PLGA and simple BMSCs were used as controls. The 2 kinds of BMSCs were respectively seeded onto the 2 kinds of PLGA. After incubation for 1, 2 and 3 d, the cell densities were cal-culated and used as indexes of cell proliferation. After incubation for 4 and 12 h, the numbers of adhesive cells were counted to evaluate the efficiency of cell adhesion. The FITC-conjugated phalloidin was used to stain the cytoskeleton of cells incubated for 24 h. After the BMSCs were cultured on the scaffolds for 7, 14 and 21 d with osteogenic supplements, their ALP activities were measured. Results Transfected BMSCs had larger cell density than simple BMSCs, and expressed higher ALP activity in any spots. Modified PLGA had higher efficiency and better quality in cell adhesion than simple PLGA, and also expressed higher ALP activity in 14 d. Conclusion TGF-β gene and RGD peptide may enhance cell adhesion, increase cell proliferation and promote differentiation of BMSCs into osteoblasts.

关 键 词：组织工程 骨 修饰 TGFΒ 骨髓细胞 

分 类 号：R318.1[医药卫生—生物医学工程]