Gene-enhanced tissue engineering: Applications for bone healing using cultured periosteal cells transduced retrovirally with the BMP-7 gene
Gene-enhanced tissue engineering: Applications for bone healing using cultured periosteal cells transduced retrovirally with the BMP-7 gene
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DOI：
10.1097/00000637-199905000-00005
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发表时间：
1999-05-01
影响因子：
1.5
通讯作者：
Grant, RT
中科院分区：
文献类型：
--
作者：
Breitbart, AS;Grande, DA;Grant, RT
骨膜具有细胞群，包括骨祖细胞和软骨祖细胞，其可以在细胞培养物中生长并在适当条件下形成骨和软骨。作者先前已经表明，培养的骨膜细胞可用于骨的组织工程，并且他们在兔颅骨缺损模型中证明了大量的骨形成。在目前的研究中，组织工程的原理与基因治疗的原理相结合，以产生培养的骨膜细胞与骨形态发生蛋白7（BMP-7）基因转导逆转录病毒用于治疗骨缺损。使用逆转录聚合酶链反应从细胞系产生人BMP-7互补脱氧核糖核酸，并克隆到逆转录病毒载体质粒中。然后使用逆转录病毒载体颗粒接种新西兰白色兔骨膜细胞。转导的骨膜细胞表现出大量生产的BMP-7信使核糖核酸的北方印迹分析和BMP-7蛋白的酶联免疫吸附测定。然后将这些细胞接种到聚乙醇酸（PGA）基质中，并用于修复关键尺寸的兔颅骨缺损。在12周时，与阴性对照转导细胞/PGA、未转导细胞/PGA、单独PGA或未修复的缺损修复的缺损部位相比，BMP-7转导骨膜细胞/PGA修复的缺损部位骨修复的放射学和组织学证据显著增加。因此，本研究成功地证明了一种利用基因修饰细胞进行骨修复的组织工程方法。
Periosteum has cell populations, including osteoprogenitor and chondroprogenitor cells, that can be grown in cell culture and form both bone and cartilage under appropriate conditions. The authors have shown previously that cultured periosteal cells can be used in the tissue engineering of bone, and they demonstrated substantial bone formation in a rabbit cranial defect model. In the current study, principles of tissue engineering were combined with principles of gene therapy to produce cultured periosteal cells transduced retrovirally with the bone morphogenetic protein 7 (BMP-7) gene to be used in the treatment of bone defects. Human BMP-7 complementary deoxyribonucleic acid was generated from a cell line using reverse transcription polymerase chain reaction and cloned into a retroviral vector plasmid. Retroviral vector particles were then used to transduce New Zealand White rabbit periosteal cells. Transduced periosteal cells demonstrated substantial production of both BMP-7 messenger ribonucleic acid by Northern blot analysis and BMP-7 protein by enzyme-linked immunosorbent assay. These cells were then seeded into polyglycolic acid (PGA) matrices and used to repair critical-size rabbit cranial defects. At 12 weeks, defect sites repaired with BMP-7-transduced periosteal cells/PGA had significantly increased radiographic and histological evidence of bone repair compared with those defect sites repaired with negative control-transduced cells/PGA, nontransduced cells/PGA, PGA alone, or unrepaired defects. Thus, this study demonstrates successfully a tissue engineering approach to bone repair using genetically modified cells.