Hertwig's上皮根鞘与牙囊共培养细胞膜片的构建及初步研究  

Study of building a co-culture cell sheets of Hertwig's epithelial root sheath cells and dental follicle cells

作　　者：葛雅能 张静怡 田卫东[3] GE Yaneng;ZHANG Jingyi;TIAN Weidong(Department of Stomatology, Nanjing Drum Tower Hospital, The Affiliated Stomatological Hospital of Nanjing University, Nanjing 210029, China;Chengdu Shilian Health Biotechnology Co. Ltd,Chengdu 610041, China;State Key Laboratory of Oral Diseases, National Engineering Laboratory for Oral Regenerative Medicine, Department of Traumatic and Plastic Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China)

机构地区：[1]南京大学医学院附属鼓楼医院口腔科,江苏南京210029 [2]成都世联康健生物科技有限公司,四川成都610041 [3]口腔疾病研究国家重点实验室,四川大学华西口腔医院口腔再生医学技术国家地方联合工程实验室,四川大学华西口腔医院创伤与整形外科,四川成都610041

出　　处：《口腔生物医学》2022年第2期92-95,101,共5页Oral Biomedicine

基　　金：四川省科技计划重点研发项目(2019YFS0515)。

摘　　要：目的:模拟牙周膜/牙骨质发育微环境,构建一种新型的赫特维希上皮根鞘(HERS)细胞-牙囊细胞(DFC)直接混合共培养细胞膜片,为上皮参与牙周膜/牙骨质再生的调控提供一种新策略。方法:将处于同一牙胚、同一牙根发育期的HERS细胞与DFC,直接混合共培养后制备成细胞膜片,观察其形态学的特点,免疫组化染色检测共培养细胞膜片与DFC膜片之间成牙周膜/牙骨质能力的差异。结果:成功构建出牙源性上皮-间充质共培养细胞膜片,共培养细胞膜片层数较多,能分泌较多的细胞外基质。免疫组化染色结果显示,共培养细胞膜片骨钙蛋白(OCN)、Ⅰ型胶原蛋白(COLI)表达阳性,而DFC膜片的OCN、COLI表达则为阴性,共培养细胞膜片碱性磷酸酶(ALP)、骨涎蛋白(BSP)表达较DFC膜片更强。结论:共培养细胞膜片成牙周膜/牙骨质能力更强,HERS细胞在牙周膜/牙骨质发育过程中的调控作用不可忽视。Objective: To build a direct co-culture cell sheets of Hertwig’s epithelial root sheath(HERS) cells and dental follicle cells(DFC) by simulating the microenvironment, so as to provide a new strategy for epithelium to participate in the regulation of periodontal ligament/cementum regeneration.Methods: The HERS cells and DFC in the same development stage and the same tooth germ were directly co-cultured to prepare the cell sheets. The morphological and histological characteristics were observed. Then we compared the differences of periodontal ligament/cementum formed abilities between them and dental follicle cell sheets.Results: The composite cell sheets were successfully constructed, which had much cell layers and could secrete abundant extracellular matrix. Compared with DFC sheets, the expressions of mineralization related proteins alkaline phosphatase(ALP) and bone sialoprotein(BSP) were higher.At the same time, the expressions of osteocalcin(OCN) and periodontal ligament associated proteins collagen Ⅰ(COLI) were positive,while the expressions of OCN and COLI in DFC sheets were negative.Conclusions: The co-culture cell sheets have stronger abilities to form periodontal ligament/cementum. The HERS cells play a crucial regulatory role in the development of periodontal ligament/cementum.

关 键 词：赫特维希上皮根鞘细胞 牙囊细胞 细胞膜片 牙再生 

分 类 号：R780.2[医药卫生—口腔医学]