华西口腔医学杂志

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鼠根端乳头条件培养基诱导下的牙周膜细胞膜片与牙本质管和煅烧骨间的牙周膜再生

宁慧影 刘宏伟   

  1. 同济大学附属口腔医院牙周科, 上海200072
  • 收稿日期:2011-08-25 修回日期:2011-08-25 出版日期:2011-08-20 发布日期:2011-08-20
  • 通讯作者: 刘宏伟,Tel:021-66313739
  • 作者简介:宁慧影(1982—),女,黑龙江人
  • 基金资助:

    国家自然科学基金资助项目(30772418);上海市科委重点攻关课题基金资助项目(074119514)

Periodontal ligament regeneration using apical tooth germ cell-conditioned medium induced periodontal ligament cells sheet between dental tube and ceramic biologic bone

Ning Huiying, Liu Hongwei   

  1. Dept. of Periodontology, School of Stomatology, Tongji University, Shanghai 200072, China
  • Received:2011-08-25 Revised:2011-08-25 Online:2011-08-20 Published:2011-08-20
  • Contact: Liu Hongwei,Tel:021-66313739

摘要:

目的利用鼠根端乳头条件培养基(APTG-CM)与牙周膜细胞(PDLCs)建立间接共培养体系,探讨牙周组织再生的研究。方法胶原酶联合组织块法获得人PDLCs,用波形蛋白和角蛋白鉴定PDLCs的来源。APTG-CM诱导PDLCs 28 d,通过免疫细胞化学法检测PDLCs中骨钙素(OCN)、Ⅰ型胶原(COL Ⅰ)、骨涎蛋白(BSP)的表达;观察细胞外部形态。体外构建牙本质管与牙周膜细胞膜片和煅烧骨(CBB)颗粒移植体,植入裸鼠皮下8周,观察牙周组织再生情况。结果体内实验结果显示:经过APTG-CM诱导后,PDLCs在牙本质和CBB表面均有牙骨样基质形成,且在CBB表面有纤维组织的黏附和垂直嵌入。对比而言,PDLCs的生长在CBB的表面要好于在牙本质管。而对照组无纤维的垂直嵌入。结论经过APTG-CM孵育的PDLCs有向成牙骨质细胞谱系转化的功能,APTG-CM有利于牙周组织再生。

关键词: 根端乳头细胞, 根端乳头条件培养液, 牙周膜细胞, 细胞外基质, 微环境

Abstract:

Objective The purpose of this study was to establish an indirect co-culture system of rat apical toot germ-conditioned medium(APTG-CM) and periodontal ligament cells(PDLCs). Methods PDLCs were isolated and cultured through the method of enzyme-digestion. Vimentin and cytokeratin(CK) were used to demonstrate the cells’ mesenchymal derivation. Co-culture system of APTG-CM and PDLCs for 28 days, osteocalcin(OCN), collagen typeⅠ (COL Ⅰ) and bone sialoprotein(BSP) were detected in PDLCs by immunocytochemistry. Morphological changes were observed by inverted microscope. With building a transplant by dental tube, periodontal ligament cell sheet and ceramic biologic bone(CBB) in vitro, then, the combinations of dental tube and PDLCs incubated by APTG-CM were implanted subcutaneously into athymic mice for 8 weeks. Results This study demonstrated that cellular cementum-like tissue formed along the dentin surface and CBB, with fibrous tissue adjacent or inserted into CBB in vivo. PDLCs were grown better in the CBB than in dentin tubes. And the vertical fibers can’t embed in the control. Conclusion PDLCs, embedded within this APTG-CM, exhibite several phenotypic characteristics of cementoblast lineages. Thereby it contributes to the main processes of periodontal tissue regeneration with rat APTG-CM.

Key words: apical tooth germ cell, apical tooth germ-conditioned medium, periodontal ligament cell, extracellular matrix, microenvironment