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骨形态发生蛋白-2(BMP-2)和血管内皮生长因子(VEGF)转染人骨膜细胞可增强成骨细胞分化和骨形成。

Bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) transfection to human periosteal cells enhances osteoblast differentiation and bone formation.

作者信息

Samee Mayurach, Kasugai Shohei, Kondo Hisatomo, Ohya Keiichi, Shimokawa Hitoyata, Kuroda Shinji

机构信息

Section of Oral Implantology and Regenerative Dental Medicine, Department of Masticatory Function Rehabilitation, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, Japan.

出版信息

J Pharmacol Sci. 2008 Sep;108(1):18-31. doi: 10.1254/jphs.08036fp. Epub 2008 Sep 6.

Abstract

Periosteum has been demonstrated to contain mesenchymal progenitor cells differentiating to osteoblasts, and both bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) may play important roles in cell-based approaches to bone regeneration. The purpose of this study was to evaluate the feasibility and efficacy of BMP-2 and/or VEGF on periosteal cell differentiation to osteoblasts in vitro and ectopic bone formation in vivo. Human periosteum-derived cells were transfected with BMP-2, VEGF, BMP-2 + VEGF, or vehicle as a control by non-viral gene transfer and then cultured and implanted to nude mice intramuscularly. Real-time polymerase chain reaction analysis of the culture revealed that transgenes for BMP-2 and BMP-2 + VEGF induced more mRNA expression of alkaline phosphatase, collagen type I, and osteocalcin than VEGF and vehicle treatments; additionally, alizarin red S staining, alkaline phosphatase staining, and alkaline phosphatase activity were significantly higher in the BMP-2 + VEGF transgene than in the other versions. After implantation, ectopic bone was observed at 4 weeks and greatly increased at 8 weeks in all groups. In particular, the combination of BMP-2 and VEGF formed significantly more bone at 4 weeks, and VEGF transfection resulted in more blood vessels relative to the conditions without VEGF. Thus, VEGF might enhance BMP2-induced bone formation through modulation of angiogenesis.

摘要

骨膜已被证明含有可分化为成骨细胞的间充质祖细胞,骨形态发生蛋白-2(BMP-2)和血管内皮生长因子(VEGF)在基于细胞的骨再生方法中可能都发挥着重要作用。本研究的目的是评估BMP-2和/或VEGF在体外诱导骨膜细胞分化为成骨细胞以及在体内诱导异位骨形成方面的可行性和有效性。通过非病毒基因转移,将人骨膜来源的细胞用BMP-2、VEGF、BMP-2 + VEGF或作为对照的载体进行转染,然后进行培养并肌肉注射到裸鼠体内。对培养物进行实时聚合酶链反应分析发现,与VEGF和载体处理相比,BMP-2和BMP-2 + VEGF的转基因诱导碱性磷酸酶、I型胶原蛋白和骨钙素的mRNA表达更多;此外,BMP-2 + VEGF转基因组的茜素红S染色、碱性磷酸酶染色和碱性磷酸酶活性显著高于其他组。植入后,所有组在4周时均观察到异位骨,8周时异位骨显著增加。特别是,BMP-2和VEGF的组合在4周时形成的骨明显更多,相对于无VEGF的情况,VEGF转染导致更多的血管生成。因此,VEGF可能通过调节血管生成来增强BMP2诱导的骨形成。

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