衍生肌腱支架材料的细胞相容性研究

目的 探讨衍生肌腱支架材料（tendon derivation biomaterials，TDBM）与肌腱细胞的细胞相容性及肌腱细胞在此三维支架上培养的生物学行为，为肌腱组织工程新型支架材料的应用提供依据。方法将肌腱细胞与TDBM体外复合培养，设置单纯肌腱细胞培养为对照组，进行形态学观察，并检测细胞增殖、细胞周期、细胞DNA倍体水平及肌腱细胞凋亡率。通过^3H-脯氨酸（^3H-Proline）掺入试验了解材料对肌腱细胞胶原合成的影响。结果 肌腱细胞在TDBM上呈梭形生长，TDBM组细胞第2天进入对数增长期，倍增时间为3d，而单纯肌腱细胞培养对照组倍增时间为3．75d。TDBM组与单纯肌腱细胞培养对照组比较，^3H-Proline掺入值差异无统计学意义（P〉0．05），说明细胞功能未受影响。与支架材料复合培养的肌腱细胞DNA指数为0．96，增殖指数较对照组高10．1％，提示肌腱细胞在三维胶原支架上生长速度快，增殖能力强。结论TDBM具有良好的细胞相容性，可作为肌腱细胞的有效载体应用于组织工程化肌腱的构建。

Experimental studies on tendon derivation biomaterials combined with cultured tenocytes in vitro

Objective To study the biocompatibility of tendon derivation biomaterials (TDBM) and biological behavior of cultured tenocytes combined with this three-dimension scaffold in tissue engineering. Methods Tenocytes were separated from Roman chicken's flextor digitorum profoundus, and cultured combining with TDBM in vitro, tenocytes cultured in plates as control group. The morphological characters were observed at 8 h, 24 h, 72 h, 7 d and 2 months, cell proliferation was tested at 1, 2, 3, 4, 5, 6, 7 and 8 d, mitotic cycle, DNA index and apoptosis were detected. Collagen synthesis was tracked by incorporated radioactive 3H labeled proline into culture. Results The tenocytes attached to TDBM at 8 h and grew along fibers in spindle-like manner at 24 h. Extracellular matrix was founded among materials fibers at 3 d. Cells in TDBM group reached logarithmic increasing period at the second day. Its' doubling time was 3 d, while cells in control group was 3.75 d. There was no difference of 3H-Proline value between the two groups(P >0.05), hinted that the tenocyte's function was not disturbed. The DNA index of cells of TDBM group was 0.96, 10.1% higher than the control group, indicating that the tenocytes grow and proliferate faster when being combine cultured on TDBM. Non malproliferation of tenocytes were founded, and lots of collagen formed among TDBM fibers within 2 months in vitro culture. Conclusion TDBM show good biocompatibility combined with tenocytes because of more similarity of surface and component with tendon. It could be promising extracellular matrix scaffold for cell transplantation in tendon tissue engineering.