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通过将骨形态发生蛋白(BMP)-2 与具有生物活性的新型纳米复合材料相结合来诱导成骨。

Osteoinduction by combining bone morphogenetic protein (BMP)-2 with a bioactive novel nanocomposite.

机构信息

University of Cambridge, Department of Materials Science, Pembroke Street, Cambridge CB2 3QZ, UK.

出版信息

Bone Joint Res. 2012 Jul 1;1(7):145-51. doi: 10.1302/2046-3758.17.2000082. Print 2012 Jul.

Abstract

OBJECTIVES

There is increasing application of bone morphogenetic proteins (BMPs) owing to their role in promoting fracture healing and bone fusion. However, an optimal delivery system has yet to be identified. The aims of this study were to synthesise bioactive BMP-2, combine it with a novel α-tricalcium phosphate/poly(D,L-lactide-co-glycolide) (α-TCP/PLGA) nanocomposite and study its release from the composite.

METHODS

BMP-2 was synthesised using an Escherichia coli expression system and purified. In vitro bioactivity was confirmed using C2C12 cells and an alkaline phosphatase assay. The modified solution-evaporation method was used to fabricate α-TCP/PLGA nanocomposite and this was characterised using X-ray diffraction and scanning electron microscopy. Functionalisation of α-TCP/PLGA nanocomposite by adsorption of BMP-2 was performed and release of BMP-2 was characterised using an enzyme-linked immunosorbent assay (ELISA).

RESULTS

Alkaline phosphatase activity of C2C12 cells was increased by the presence of all BMP-2/nanocomposite discs compared with the presence of a blank disc (p = 0.0022), and increased with increasing incubation concentrations of BMP-2, showing successful adsorption and bioactivity of BMP-2. A burst release profile was observed for BMP-2 from the nanocomposite.

CONCLUSIONS

Functionalisation of α-TCP/PLGA with BMP-2 produced osteoinduction and was dose-dependent. This material therefore has potential application as an osteoinductive agent in regenerative medicine.

摘要

目的

骨形态发生蛋白(BMPs)在促进骨折愈合和骨融合方面具有重要作用,其应用日益广泛。然而,仍需要确定最佳的递送系统。本研究旨在合成具有生物活性的 BMP-2,将其与新型α-磷酸三钙/聚(D,L-乳酸-co-乙醇酸)(α-TCP/PLGA)纳米复合材料结合,并研究其从复合材料中的释放。

方法

采用大肠杆菌表达系统合成 BMP-2 并进行纯化。使用 C2C12 细胞和碱性磷酸酶测定法来确认其体外生物活性。采用改良的溶液蒸发法制备α-TCP/PLGA 纳米复合材料,并通过 X 射线衍射和扫描电子显微镜进行表征。通过吸附作用将 BMP-2 功能化到α-TCP/PLGA 纳米复合材料上,并通过酶联免疫吸附测定(ELISA)来表征 BMP-2 的释放。

结果

与空白盘相比,所有 BMP-2/纳米复合材料盘的存在均增加了 C2C12 细胞的碱性磷酸酶活性(p = 0.0022),并且随着 BMP-2 孵育浓度的增加而增加,表明 BMP-2 成功吸附并具有生物活性。从纳米复合材料中观察到 BMP-2 的快速释放。

结论

将 BMP-2 功能化到α-TCP/PLGA 上可产生成骨诱导作用,并呈剂量依赖性。因此,该材料具有作为再生医学中骨诱导剂的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be8f/3629446/718275756cfa/2000082-galleyfig1a.jpg

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