Regeneration of calvarial defects with Escherichia coli-derived rhBMP-2 adsorbed in PLGA membrane
Regeneration of calvarial defects with Escherichia coli-derived rhBMP-2 adsorbed in PLGA membrane
复制标题
PLGA 膜吸附大肠杆菌来源的 rhBMP-2 修复颅骨缺损
DOI：
10.1159/000356947
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发表时间：
2013
影响因子：
2.7
通讯作者：
Kuboki T
中科院分区：
文献类型：
--
作者：
Ono M;Sonoyama W;Nema K;Hara ES;Oida Y;Pham HT;Yamamoto K;Hirota K;Sugama K;Sebald W;Kuboki T
目的大肠杆菌衍生的重组人骨形态发生蛋白-2 （E-BMP-2）已被证明与哺乳动物细胞衍生的BMP-2一样有效。然而，由于E-BMP-2的合成过程存在差异，主要与蛋白质非糖基化有关，因此还需要进行一些体外和体内实验来验证其有效性。本研究的目的是研究可生物降解聚乳酸-羟基乙酸酯（PLGA）膜是否适合作为E-BMP-2的载体，用于大鼠颅骨临界尺寸缺损的骨再生。材料与方法首先，通过分析骨钙素mRNA水平证实E-BMP-2在体外培养小鼠骨髓基质细胞中的成骨作用，并通过茜素红染色检测钙沉积。在体内实验前，采用ELISA法测定E-BMP-2在PLGA膜上的释放谱。E-BMP-2（0、1、5、10 μg/μl）应用于异位和正位骨形成，x线、显微ct和组织学分析。结果释放谱测试表明，PLGA膜可保留初始应用的E-BMP-2的94%。异位骨形成实验显示，E-BMP-2/PLGA膜的结合强烈诱导骨形成。5和10 μg/ l E-BMP-2吸附的PLGA膜具有较强的骨诱导能力，可完全修复临界尺寸的缺陷，而不给膜组和0 μg/ l E-BMP-2的组未见骨形成。结论PLGA膜是E-BMP-2缓释的合适载体，E-BMP-2/PLGA膜复合可有效促进骨缺损模型的再生。
ObjectiveEscherichia coli-derived recombinant human bone morphogenetic protein-2 (E-BMP-2) has been shown to be as effective as mammalian cell-derived BMP-2. However, several in vitro and in vivo experiments are still necessary to validate the effectiveness of E-BMP-2 due to the difference in synthesis process, mainly related to protein nonglycosylation. The objective of this study was to investigate whether biodegradable polylactide-co-glycolide (PLGA) membrane is a suitable carrier for E-BMP-2 delivery for bone regeneration of critical-sized defects in rat calvaria.Materials and MethodsFirst, the osteoinductive effect of E-BMP-2 was confirmed in vitro in mouse bone marrow stromal cells by analysis of osteocalcin mRNA levels, and calcium deposition was detected by alizarin red staining. Before in vivo experiments, the release profile of E-BMP-2 from PLGA membranes was determined by ELISA. E-BMP-2 (0, 1, 5 and 10 μg/μl) was applied for ectopic and orthotopic bone formation and was analyzed by X-ray, micro-CT and histology.ResultsRelease-profile testing showed that PLGA membrane could retain 94% of the initially applied E-BMP-2. Ectopic bone formation assay revealed that combination of E-BMP-2/PLGA membrane strongly induced bone formation. Stronger osteoinductivity with complete repair of critical-sized defects was observed only with PLGA membranes adsorbed with 5 and 10 μg/μl of E-BMP-2, whereas no bone formation was observed in the groups that received no membrane or 0-μg/μl dose of E-BMP-2.ConclusionPLGA membrane was shown to be a suitable carrier for sustained release of E-BMP-2, and the E-BMP-2/PLGA membrane combination was demonstrated to be efficient in bone regeneration in a model of critical-sized defects.