脱细胞真皮支架复合骨髓基质干细胞构建前交叉韧带的实验研究

Research of acellular dermal matrix scaffold combined with bone marrow stem cells to construct tissue engineering anterior cruciate ligament

[目的]研究骨髓基质干细胞(BMSCs)复合脱细胞真皮(ADM)构建组织工程膝前交叉韧带(ACL),以及特定浓度转化生长因子TGF-β1和碱性成纤维细胞因子bFGF对骨髓基质干细胞生长、粘附及分化的影响。[方法]用ADM制备4股编织柱形及4股束形支架材料,将兔BMSCs接种于ADM支架上,分别用普通培养液(对照组)及含有特定浓度的TGF-β1和bFGF因子培养液(实验组)培养,通过绘制细胞增殖MTT曲线,碱性磷酸酶ALP测定,荧光显微镜及激光共聚焦扫描显微镜观察,分析ADM支架力学性能,细胞在支架材料中的生长增殖情况,以及其在ADM上的粘附情况。[结果]ADM的一般力学性能符合ACL要求,实验组BMSCs在ADM上生长数量及粘附效果明显优于对照组(P<0.05),实验组相对于对照组细胞能够分泌更多的ALP(P<0.05)。[结论]BMSCs复合ADM支架可能成为组织工程ACL材料替代移植材料,本浓度的TGF-β1及bFGF因子培养液对骨髓基质干细胞在ADM上的生长、粘附及分化有明显的刺激增强效果。

[ Objective ] To study the bone marrow stromal cells （BMSCs） combined with acellular dermal matrix （ ADM ） for tissue engineered anterior cruciate ligament（ ACL）, as well as the effects of transforming growth factor TGF - β1 and basic fibro- blast growth factor bFGF on bone marrow stromal stem cell growth, adhesion and differentiation. [ Method ] ADM was used to prepare 4 strands braided column and 4 shares of beam shape scaffolds, rabbits BMSCs inoculated in ADM support, respectively, with ordinary medium（ control group）and containing a given concentration of TGF- β1 and bFGF factor culture fluid （experi- mental group）culture. The cell proliferation MTY curve, alkali phosphatase ALP activity,fluorescence microscopy and scanning electron microscopy（SEM） ,mechanical properties analysis of ADM scaffolds, ceils in the scaffolds in growth, proliferation, and adhesion were recorded and analyzed. [ Result] ADM general mechanical properties of ADM met the requirements of ACL. The growth quantity and adhesion effect of BMSCs on ADN in the experimental group were better than in the control group（P 〈0. 05 ）. Compared with control group, the cells in experimental groups were able to secrete more ALP（ P 〈 0. 05 ）. [ Conclusion ] BMSCs composite ADM stents can become tissue engineering ACL material,TGF - β1 and bFGF have distinct stimulus effect on cultured bone marrow stromal cells in ADM growth, adhesion and differentiation.