雪旺细胞与PDLLA/CS/CHS自组装复合材料生物相容性研究

Biocompatibility of Schwann Cells with PDLLA/CS/CHS Self-assembled Composite Materials

目的利用自行培养的雪旺细胞（SCs）与具有良好可生物降解性的高分子聚乳酸／硫酸软骨素／壳聚糖（PDL—LA／CS／CHS）复合材料进行生物相容性研究，评价该材料应用于周围神经修复的可能性。方法利用双差速贴壁法进行SCs的培养与纯化，并观察其生长曲线，苏木精-伊红染色和免疫组织化学染色观察细胞表征；将纯化后的SCs接种在PDLI，A／CS／CHS复合材料上进行材料生物相容性研究，用MTT法和环境扫描电镜检测和观察细胞生长情况。结果SCs生长平台期为1d，对数期为5～7d，倍增时间为5d，其纯度超过90％。MTT实验显示，细胞接种0、2、4d，PDLLA组、CHS组和PDLLA／CS／CHS组的吸光度值均低于SCs对照组，至7d和10d，PDLLA、CHS组吸光度值亦低于SCs对照组，PDLLA／CS／CHS组高于SCs对照组，但差异无统计学意义（P〉O．05），PDLLA／CS／CHS组与PDLLA组相比差异有统计学意义（P〈O．05）。结论PDLLA／CS／CHS复合材料是具有较理想的生物相容性的生物材料，有良好的材料－细胞界面，较利于SCs黏附及生长、增殖，可应用于周围神经修复的研究。

Objective To investigate the biocompatibility of self-cultured Schwann cells with polylactic acid/chondroitin sulfate/chitosan（PDLLA/CS/CHS）composite material with good biodegradability,and evaluate the possibility of the PDLLA/CS/CHS material applied to the repair of peripheral nerve.Methods By using pairs of differential adhesion,Schwann cells were cultured and purified.The growth curve of Schwann cells was drawn,and their phenotypes were examined by using HE staining and immunohistochemical staining.The purified Schwann cells were seeded in the PDLLA/CS/CHS composite materials and the biocompatibility was studied by using MTT method and environmental scanning electron microscopy.Results The growth platform period of Schwann cells was 1 day,the logarithmic period was 5-7 days,and the doubling time was 5 days with the purity of more than 90%.MTT assay revealed that at 0,2nd,4th day after inoculation,The absorbance（A）values in PDLLA group,CHS group,and PDLLA/CS/CHS group were lower than in control group.At the 7th and 10th day,the A values in PDLLA and CHS groups were also lower than in control group,and those in PDLLA/CS/CHS group were higher than in control group with no significant difference（P〈0.05）.But there was significant difference in the A values between PDLLA/CS/CHS group and PDLLA group（P〈0.05）.Conclusion PDLLA/CS/CHS composite is biological materials with ideal biocompatibility and good material-cell interface,and facilitates the adhesion,growth and proliferation of Schwann cells.