【摘要】 目的:体外评价壳聚糖季铵盐/甘油磷酸(CS-HTCC/GP)温敏水凝胶的细胞相容性。方法:体外培养人牙周膜成纤维细胞(HPDLCs),采用四甲基偶氮唑盐(MTT)法测定不同浓度的CS-HTCC/GP温敏凝胶浸提液对1、3、5d细胞增殖和碱性磷酸酶(ALP)活性的影响。采用扫描电镜和透射电镜观察对细胞超微结构的影响。采用SSPS13.0软件包对数据进行统计学分析。结果:免疫组化显示,培养的细胞波形丝蛋白染色阳性,角蛋白染色阴性,证明体外原代培养细胞及传代细胞均为中胚层组织来源,且无上皮来源细胞混杂。不同浓度的CS-HTCC/GP温敏水凝胶浸提液均有促进HPDLCs增殖的作用,在第3天和第5天,差异有高度显著性(P<0.001)。各组均可提高ALP活性。第2组和第3组在第5天与第1天ALP表达的提高有高度显著差异(P<0.001)。扫描电镜和透射电镜观察水凝胶浸提液对HPDLCs的超微结构无影响。结论:CS-HTCC/GP温敏水凝胶具有良好的细胞相容性,具有用于牙周局部药物释放系统载体及组织工程支架的潜力。更多还原

【Abstract】 PURPOSE: The aim of this investigation was to evaluate the cytocompatibility of an in situ chitosan-quaternized chitosan / α,β-glycerophosphate(CS-HTCC/GP) thermosensitive hydrogel in vitro.METHODS: The primary cells were isolated from human periodontal ligament and cultured.The role of different concentrations of CS-HTCC/GP extract to HPDLCs was evaluated by MTT assay and alkaline phosphatase(ALP) activity.Also,the ultra-architecture of HPDLCs was determined by scanning electron microscopy(SEM) and transmission electron microscopy(TEM) respectively.SPSS13.0 software package was used for statistical analysis.RESULTS: By immunocytochemical method,the cells were stained positively to antibodies against vimentin,and negatively to antibodies against cytokeratin,which indicated that they were external embryo mesenchymal cell without epithelial cell mixure.CS-HTCC/GP thermosensitive hydrogel promoted proliferation of HPDLCs,especially at 3d and 5d,the results was significantly different(P<0.001).ALP activity was significantly greater in group 2 and 3 than in group 4 after 5d(P<0.001).Also,no negative influence to ultrastructure of HPDLCs was found through SEM and TEM.CONCLUSIONS: The results indicate that CS-HTCC/GP thermosensitive hydrogel exhibits excellent cytocompatibility and has potential to be used as an in situ injectable local periodontal drug delivery vehicle and a tissue-engineering scaffold for periodontal disease therapy.更多还原