摘要
目的:探讨不同密度的人牙髓细胞(HDPCs)在可注射聚乳酸/羟基乙酸(PLGA)微球支架上的粘附生长情况.方法:取第4代HDPCs,以3种不同密度(2×105/mL、1×105/mL、5×104/mL)接种于PLGA微球(A、B、C组),于不同时间点倒置显微镜下观察细胞粘附生长情况;检测各组细胞双链脱氧核糖核酸(dsDNA)含量及碱性磷酸酶(ALP)活性.体外培养8周后,扫描电镜观察HDPCs和PLGA微球材料的形貌结构.结果:倒置显微镜下可见细胞粘附于微球材料,随着时间延长,细胞增殖情况良好,与材料融合生长.培养1周,3组细胞dsDNA含量及ALP活性均无显著差异;培养2周,A、B两组细胞dsDNA含量显著高于C组,B组ALP活性水平最高.培养8周, 扫描电镜可见细胞与微球完全融合,表面有基质样物质沉积,局部有凹陷形成.结论:1×105/mL HDPCs接种于PLGA微球,细胞dsDNA含量及ALP活性水平较高.PLGA微球作为一种可注射支架材料,有利于HDPCs粘附生长,可望用于构建组织工程化牙髓.
Abstract
AIM:To observe the adhesion and proliferation of human dental pulp cells (HDPCs) at different density on an injectable polylactic-co-glycolic acid (PLGA) microsphere scaffold.METHODS:HDPCs of fourth passage were seeded onto PLGA microsphere scaffolds at 2×105, 1×105 and 5×104cells/mL respectively (Group A, B and C).Cell adhesion and proliferation were observed under inverted microscope.Double stranded deoxyribonucleic acid (dsDNA) content and alkaline phosphatase (ALP) activity were examined.After 8 weeks of culture, the ultrastructure was observed under SEM.RESULTS:Adhesion of HDPCs on PLGA microspheres was observed, the cells proliferated well and fused together.After 1 week, there was no significant differences in the cells dsDNA content and ALP activity among 3 groups.After 2 weeks, Group A and B showed significantly higher dsDNA content than Group C.Group B showed the highest ALP specific activity.After 8 weeks of culture, SEM showed that cells and scaffolds were completely fused together.There were matrix deposition and hollow sinking on the scaffolds.CONLUSTION:1×105cells/mL of HDPCs seeded onto PLGA microsphere scaffolds may produce better results of dsDNA content and ALP activity.
基金项目
天津市口腔医院博硕士重点攻关项目(2015BSZD01)
首都医科大学附属北京口腔医院重点实验室开放课题(KFKT2017008)
NETL
NSTL
维普
万方数据