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 28d,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.%Objective The purpose of this study was to establish an indirect co-culture system of rat apical tooth 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 ceils'mesenchymal derivation. Co-culture system of APTG-CM and PDLCs for 28 days, osteocalcin(OCN), collagen type Ⅰ(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 APTC-CM.
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