【摘要】 目的探讨牙周膜细胞(PDLCs)膜片的构建方法,并观察膜片聚合体的组织学特征。方法通过组织块培养法获得PDLCs,经体外鉴定扩增后,用添加50μg/mL的L-抗坏血酸-2-磷酸酯的DMEM培养基连续培养2周,构建PDLCs膜片,将构建的PDLCs膜片在该培养条件下自行聚合获得膜片聚合体,HE染色进行形态学观察。结果 PDLCs被成功分离、培养、鉴定。在加入L-抗坏血酸-2-磷酸酯的DMEM培养基中连续培养2周后获得白色膜状PDLCs膜片。倒置显微镜下观察显示,细胞呈复层生长,且呈典型的纺锤状。PDLCs膜片在该培养条件孵育7d,皱缩形成约米粒大小的球样。经HE染色显示,细胞密度较大,且细胞与细胞之间存在着大量细胞外基质。结论 L-抗坏血酸-2-磷酸酯可以通过增加细胞外基质而促进PDLCs膜片的形成,为利用PDLCs膜片修复牙周组织及牙周缺损提供了技术保证。更多还原

【Abstract】 Objective To study the effect of L-ascorbic acid 2-phosphate in the construction of PDLCs sheet and characterize the biological property of the polymer of sheet.Methods PDLCs were obtained by human periodontal ligament tissues,PDLCs were subcultured to construct PDLCs sheet culturing in the DMEM containing 50 ug/ml L-ascorbic acid 2-phosphate for two weeks,examined by inverted microscope.and obtain the cell sheet.Then let the cell sheet shrink to obtain the polymer,HE staining.Results After 2 weeks in vitro culture,a white membranaceous PDLC sheet formed.Under inverted microscope,it was shown that PDLSCs grew in multilayer and retained their fibroblastic spindle shape.After shrinking for seven days,the cell sheet changed into a ball.Histological examination indicated high density of cells and abundant extracellular matrix(ECM) existed among the cells.Conclusion L-ascorbic acid 2-phosphate can promote the construction of PDLCs sheet by increasing the ECM,which provided technological foundation for future studies in regeneration and repair of periodontal tissue destruction.更多还原