构建一种新型牙周膜牙囊复合细胞膜片的研究  

Study of Building a type of composite cell sheets of human dental follicle cells (hDFCs)and human periodontal ligament stem cells(hPDLSCs)

作　　者：王丽颖 金作林 宋扬[2] 刘佳 金岩[3] 刘文佳[3] 吴凡 

机构地区：[1]第四军医大学口腔医院正畸科 [2]解放军第323医院口腔科 [3]第四军医大学组织工程中心 [4]解放军68332部队

出　　处：《临床口腔医学杂志》2013年第4期195-199,共5页Journal of Clinical Stomatology

基　　金：国家自然科学基金(30973358)

摘　　要：目的:拟构建一种牙周膜牙囊复合细胞膜片,并比较其与单一膜片的性能差异,以期寻找一种更加优越的牙周缺损修复材料。方法:将第三代牙周膜干细胞及牙囊干细胞直接混合共同培养,将牙周膜牙囊混合细胞、牙周膜干细胞及牙囊干细胞分别制备成细胞膜片,应用HE染色,扫描电镜比较三种膜片形态学差异,应用茜素红染色,RT-PCR比较三种膜片成骨能力差异。结果:扫描电镜结果显示,牙囊牙周膜混合细胞膜片分泌更多的细胞外基质。HE染色结果也显示牙囊牙周膜混合细胞膜片细胞层数更多,基质分泌量更大。同时,牙囊牙周膜混合细胞膜片ALP、Runx2、OCN表达显著高于单一细胞膜片。成骨诱导21天后,茜素红染色结果显示,与单一细胞膜片相比,牙囊牙周膜混合细胞膜片染色更深。结论:牙周膜牙囊复合细胞膜片较单一细胞膜片,细胞层数更多,基质分泌量更大,骨向分化能力更强,为牙周再生提供了新的思路。Objective：To build the composite cell sheets of DFCs and PDLSCs to improve the competence of regener- ation of the complex periodontal structure. Method：The two types of stem cells hPDLSCs and hDFCs were isolated and mixed for co-cuhure. Then we used the mixed cells of hPDLSCs and hDFCs,hPDLSC and hDFCs to make cell sheets and compared the osteogenic ability of these cell sheets. HE dyes and scanning electron microscopy were used to observe the morphology and structure of these three cell sheets. Result： Compared with hPDLSCs cell sheet.hDFCs cell sheet, the RT- PCR showed that the mixed cells of composite cell sheets of hDFCs and hPDLSCs express more ALP,Runx2 and OCN. Alizarin red staining results also showed that, after 21 days of the osteogenic induction,the staining of the mixed cells of composite cell sheets of hDFCs and hPDLSCs are deeper than single cell sheet. HE dyes and scanning electron microscopy indicated abundant extracellular matrix （ECM） existed in the composite cell sheets of hDFCs and hPDLSCs, which are more than them in hPDLSCs cell sheet and hDFCs cell sheet. HE dyes also indicated there are more layers of cells in the compos- ite cell sheets of hDFCs and hPDLSCs than in hPDLSCs cell sheet and hDFCs cell sheet. Conclusion： The osteogenic ability of composite cell sheets of hDFCs and hPDLSCs is stronger than hDFCs cell sheet and hPDLSCs cell sheet. Besides, there are more layers of cells and extracellular matrix （ECM） xisted in the composite cell sheets of hDFCs and hPDLSCs, so composite cell sheets of hDFCs and hPDLSCs are good for tissue regenerate the complex periodontal structure.

关 键 词：细胞膜片 牙囊细胞 牙周膜干细胞 牙周再生 

分 类 号：R813.1[医药卫生—放射医学]