North Carolina Oral Health Institute, The University of North Carolina at Chapel Hill, USA.
J Dent Res. 2014 Apr;93(4):406-11. doi: 10.1177/0022034514521237. Epub 2014 Jan 30.
The aim of this study was to determine the effects of glutathione-S-transferase-fused recombinant biglycan (GST-BGN) on craniofacial bone regeneration. We recently demonstrated a positive effect of tissue-derived BGN on bone morphogenetic protein 2 (BMP-2) function, which is exerted likely via the BGN core protein. Here, we investigated the effects of GST-BGN lacking any posttranslational modifications on BMP-2 function in vitro and in vivo. In the C2C12 cell culture system, BMP-2-induced Smad 1/5/8 phosphorylation and alkaline phosphatase activity were both enhanced by the addition of GST-BGN. For the in vivo effect, we employed a Sprague-Dawley rat mandible defect model utilizing 1 µg (optimal) or 0.1 µg (suboptimal) of BMP-2 combined with 0, 2, 4, or 8 µg of GST-BGN. At 2 weeks post-surgery, newly formed bone was evaluated by microcomputed tomography and histologic analyses. The results revealed that the greatest amounts of bone within the defect were formed in the groups of suboptimal BMP-2 combined with 4 or 8 µg of GST-BGN. Also, bone was well organized versus that formed by the optimal dose of BMP. These results indicate that recombinant BGN is an efficient substrate to promote low-dose BMP-induced osteogenesis.
本研究旨在确定谷胱甘肽-S-转移酶融合重组 biglycan(GST-BGN)对颅面骨再生的影响。我们最近证明了组织来源的 BGN 对骨形态发生蛋白 2(BMP-2)功能具有积极影响,这种作用可能是通过 BGN 核心蛋白发挥的。在这里,我们研究了 GST-BGN 在体外和体内对 BMP-2 功能的影响,这种 GST-BGN 缺乏任何翻译后修饰。在 C2C12 细胞培养系统中,添加 GST-BGN 增强了 BMP-2 诱导的 Smad 1/5/8 磷酸化和碱性磷酸酶活性。为了评估体内作用,我们采用了 Sprague-Dawley 大鼠下颌骨缺损模型,使用 1 µg(最佳)或 0.1 µg(亚最佳)的 BMP-2 与 0、2、4 或 8 µg 的 GST-BGN 结合。在手术后 2 周,通过 microCT 和组织学分析评估新形成的骨。结果表明,在亚最佳 BMP-2 与 4 或 8 µg GST-BGN 结合的组中,缺陷内形成的骨量最大。此外,与最佳剂量的 BMP 形成的骨相比,骨的组织更为有序。这些结果表明,重组 BGN 是一种有效的促进低剂量 BMP 诱导成骨的底物。