背景 前期研究通过静电纺丝技术获得的聚乳酸/羟基磷灰石膜有利于细胞的贴附和生长.目的 分析电纺聚乳酸/羟基磷灰石膜和人羊膜基质细胞构建骨组织工程细胞/支架复合体的可行性.方法 利用MTT 法检测聚乳酸和聚乳酸/羟基磷灰石膜浸提液对人羊膜基质细胞增殖的影响;将第3 代人羊膜基质细胞培养于含聚乳酸和聚乳酸/羟基磷灰石膜的成骨诱导培养液中,进行组织学检查及免疫荧光细胞化学染色检测.结果 与结论 聚乳酸和聚乳酸/羟基磷灰石膜浸提液对人羊膜基质细胞均无明显细胞毒性.与两种膜材料复合培养后,人羊膜基质细胞细胞增殖明显,可观察到钙化结节的形成,钙化结节处细胞Ⅰ型胶原和碱性磷酸酶表达阳性,且聚乳酸/羟基磷灰石膜组细胞钙化结节数量及成熟程度优于聚乳酸组.说明电纺聚乳酸/羟基磷灰石膜与人羊膜基质细胞可以共同构建成细胞/支架复合体,具有应用于骨组织工程的潜力.%BACKGROUND: Previous study has shown that electrospun poly (L-lactic acid)/hydroxyapatite (PLLA/HA) membranes aresuitable for cellular adhesion and proliferation.OBJECTIVE: To investigate the feasibility of the human amniotic mesenchymal cells (hAMCs) and PLLA/HA membranesconstructing the complex of seed cells/scaffold in bone tissue engineering.METHODS: Effect of leaching liquor from the PLLA and PLLA/HA membranes on the proliferation of hAMCs was detected by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay method. The third passage of hAMCs were cultured in theosteogenic induce media containing PLLA and PLLA/HA membranes, and then were detected by histological andimmunofluorescence cytochemical staining.RESULTS AND CONCLUSION: The leaching liquor of the PLLA and PLLA/HA had no cytotoxicity on the hAMCs. After seededonto two kinds of the membranes, hAMCs proliferated actively and calcified nodules were observed. Cells in calcified noduleswere shown positive expression of collagen type I and alkaline phosphatase, and the number and maturity of the calcifiednodules in the PLLA/HA group were better than those of the PLLA group. It is showed that the hAMCs and PLLA/HA canconstruct cells/scaffold complex and show a great potential of application in bone tissue engineering.
展开▼
