骨形成蛋白2基因转染的人牙周膜成纤维细胞的骨诱导作用
Bone Induction by Human Periodontal Ligament Fibroblasts Transfected with Bone Morphogenetic Protein 2 Gene
作者:司晓辉,刘正
Author:SIXiaohui,LIUZheng
收稿日期:2003-10-25 年卷(期)页码:2003,21(05):347-
期刊名称:华西口腔医学杂志
Journal Name:West China Journal of Stomatology
关键字:骨形成蛋白,人牙周膜成纤维细胞,基因转染,诱导,成骨,
Key words:bone morphogenetic protein,human periodontal ligament fibroblasts,gene transfection,induction, osteogenesis,
基金项目:
本课题为国家自然科学基金(编号30000191)、中国博士后科学基金(编号中博基1999第17号)及上海市博士后基金(编号沪科2000 第487号)资助项目
中文摘要
目的 建立表达骨形成蛋白2 (BMP-2)的人牙周膜成纤维细胞(HPDLFs),观察其体内成骨作用,为牙周缺损修复基因疗法的建立提供实验依据。方法 采用脂质体载体将人BMP-2噬菌粒表达载体pBK-B2转染至 HPDLFs。利用免疫组化ABC法检测转染细胞内BMP-2蛋白的表达,并于无菌条件下将转染细胞注射于裸鼠股部肌肉内,21 d后取材,苏木精—伊红染色观察肌肉组织内骨形成情况。结果 BMP-2基因转染后HPDLFs内有BMP- 2蛋白的表达。在注射BMP-2基因转染细胞的肌肉组织内可见明显的新骨形成。结论 BMP-2基因成功转入HP- DLFs中并得到有效表达,转染BMP-2基因的HPDLFs能够在体内诱导新骨形成。
英文摘要
Objective To establish human periodontal ligament fibroblasts (HPDLFs) that express bone morphogenetic protein 2 and observe their osteogenesisin vivo, so as to provide fundamental data for future study on gene therapy of periodontal defect treatment.Methods The phagemid expression vector for BMP-2 (pBK-B2) was transfected intoHPDLFs by usingLipofect AMINE. The BMP-2 expressionwas determined by immunohistochemical ABCmethod. The BMP-2 gene transfectedHPDLFswere injected into thigh muscle of nude mice under sterile condition. Specimenswere collected in 21 days after injection and underwent normal procedures forfixation, decalcification, embedding, incising and stainingwithHE.Results The results demonstrated that BMP-2 protein expressed in HPDLFs after BMP-2 gene transfection. The HPDLFs transfected with BMP-2 gene could induce new bone formation at the injection site.Conclusion The results indicated that BMP-2 gene was transfected successfully and ex- pressed efficiently in HPDLFs. The HPDLFs transfected with BMP-2 gene are capable of inducing new bone formationin vivo.
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