黄芪-壳聚糖/聚乳酸多孔支架对犬骨髓基质细胞生物学行为的影响

Effects of astragalus polysaccharides-chitosan / polylactic acid composite material on biological behavior of canine bone marrow stromal cells cultured in vitro

目的:观察黄芪壳聚糖/聚乳酸、壳聚糖/聚乳酸多孔支架复合材料对犬骨髓基质细胞(BMSCs)的黏附、生长及分化的影响,寻找较合适的牙周骨组织工程复合物支架材料。方法:将诱导分化的犬BMSCs分别与黄芪壳聚糖/聚乳酸、壳聚糖/聚乳酸支架体外培养。第5天取材,测定2种材料的吸附率,用扫描电镜观察超微结构,以免疫组织化学检测I型胶原,放射免疫法检测骨钙素的分泌情况。结果:BMSCs与2组材料均能形成良好贴附,黄芪壳聚糖/聚乳酸复合材料上贴附的细胞吸附率、Ⅰ型胶原及骨钙素的分泌量高于壳聚糖/聚乳酸组。结论:黄芪壳聚糖/聚乳酸能促进BMSCs生长、分化及基质分泌,是一应用前景良好的组织工程骨构建复合材料。

Objective To observe the biological behavior of canine bone marrow stromal cells (BMSCs) cultured in vitro with the astragalus polysaccharides-chitosan/polylactic acid (AP-C/PLA) and with the chitosan/polylactic acid (C/PLA) and to find a suitable compound material for periodontal tissue engineering. Methods BMSCs (induced 14 days by 50mg/L vitamine C, 10 -8 mol/L dexamethasone, 10mmol/L β-sodium glycerylphosphate) were cultured on AP-C/PLA or C/PLA for 5 days respectively. The BMSCs attachment and the morphology were observed with scanning electronic microscope and the combining rates were counted.Type I collagen synthesis was examined with immunohistochemistry staining and the content of osteocalin was determined with radio-immunological method.Results Combining rates, type I collagen synthesis, and the content of osteocalin of BMSCs on AP-C/PLA were significantly higher than those on C/PLA. Conclusion AP-C/PLA may promote the BMSC proliferation, differentiation and extracellular matrix synthesis, and it can be used as a good scaffold material for bone tissue engineering.