口腔医学研究

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (3): 239-243.DOI: 10.13701/j.cnki.kqyxyj.2016.03.007

• 基础研究论著 • 上一篇    下一篇

bFGF/BMP-2转染大鼠骨髓干细胞体内成牙的研究

张悦, 胡杨, 沈玉凤, 单健良, 何雨桐, 何惠宇   

  1. 新疆医科大学第一附属医院口腔修复科 新疆 乌鲁木齐 830054
  • 收稿日期:2015-08-24 出版日期:2016-03-28 发布日期:2016-03-29
  • 通讯作者: 何惠宇,电话:0991-4365663
  • 作者简介:张悦 (1988~ ),女,侗族,广西横县人,硕士在读,主要从事口腔修复学临床及基础的研究工作。
  • 基金资助:
    国家自然科学基金项目(编号:81060088)
    新疆维吾尔自治区科技厅科技支疆项目(编号:201291l73)
    新疆维吾尔自治区科技厅自然科学家基金(编号:2014211C037)

Study on Rat Bone Marrow Stem Cells Co-transfected with bFGF/BMP2 to Differentiate into Teeth in Vivo.

ZHANG Yue, HU Yang, SHEN Yu-feng, SHAN Jian-liang, HE Yu-tong,HE Hui-yu*   

  1. Department of Prosthodontics,First Hospital Affiliated to Xinjiang Medical University, Urumqi 830054, China
  • Received:2015-08-24 Online:2016-03-28 Published:2016-03-29

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摘要： 目的: 牙胚细胞和成纤维生长因子(Fibroblast growth factor bFGF)/骨形成蛋白2(bone morphogenetic protein 2 BMP-2)联合转染的骨髓间充质干细胞 (bone marrow stem cells,BMSCs)混合培养,复合明胶海绵支架构建牙组织工程植入大鼠体内,探究其成牙能力。方法: 取健康SD大鼠128只,随机分为4组:细胞团块组;明胶海绵组;细胞团块+明胶海绵组;空白对照组,不做任何干预,均在全麻下植入大鼠肾被膜下。术后按4个时间点(5、10、14、28 d各8只)分期取材,进行大体组织观察,Masson染色和免疫组化染色。结果: 大体组织和Masson染色:术后5、10、14、28 d形态学观察显示细胞团块+明胶海绵组形成牙样组织。免疫组化:析因分析显示细胞团块+明胶海绵组在5、10、14 d DMP-1、BMP-4、DSP表达量最高且均高于其余各组,随着时间推移表达量逐渐有减小趋势,差异有统计学意义(P＜0.05)。结论: 牙胚细胞和基因转染BMSCs混合培养,复合明胶海绵构建牙组织工程的体内实验结果形成牙样组织,为组织工程牙的成功构建奠定了一定的基础。

关键词: 牙胚细胞, 骨髓间充质干细胞, 牙本质基质蛋白-1, 骨形成蛋白4, 牙本质涎蛋白

Abstract: Objective: To investigate the ability of heterogeneous tooth tissue engineering (HTTE) which was established by germ cells co-cultured with bone marrow stem cells (BMSCs) co-transfected with fibroblast growth factor bFGF/BMP-2 to differentiate into teeth through integrally implanted in rats. Methods: 128 healthy SD rats were randomly divided into four groups: the cell pellet group, gelatin sponge group, cell clumps + gelatin sponge group, and control group. The HTTE was implanted in the renal subcapsular of rats under general anesthesia. Samples were collected after 5, 10, 14, and 28 days and each group included 8 samples. Gross tissue observation, Masson staining, and immunohistochemical staining were performed. Results: Histological gross observation and Masson staining indicated that there was dental tissue formed in the cell clumps + gelatin sponge group. Factorial analysis showed that, at each time point, the expression of cell clumps + gelatin sponge group reached the peak and was significantly higher than those of other groups. Then, the expression level declined. The difference was statistical significance (P<0.05). Conclusion: Mixing culture of germ cells and gene co-transfected BMSCs seeded on the gelatin sponge could form tooth-like organization, which might lay a certain foundation to construct the tissue engineering teeth.

Key words: Tooth germ cell , Bone marrow mesenchymal stem cells , Dentin matrix protein-1 , Bone morphogenetic protein 4, Dentin sialoprotein

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