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通过谷氨酸修饰的 BMP-2 肽的组合来改善体外磷酸钙支架的成骨作用。

Improvement of calcium phosphate scaffold osteogenesis in vitro via combination of glutamate-modified BMP-2 peptides.

机构信息

Department of Orthopaedics, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002, Jiangsu, PR China; School of Medicine, Nanjing University, Nanjing 210002, Jiangsu, PR China.

Department of Orthopaedics, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002, Jiangsu, PR China.

出版信息

Mater Sci Eng C Mater Biol Appl. 2019 Mar;96:412-418. doi: 10.1016/j.msec.2018.11.048. Epub 2018 Nov 27.

Abstract

Alpha-tricalcium phosphate (α-TCP) based porous scaffolds have superior osteoconduction and osteoinduction in bone tissue engineering, furthermore, these 3D porous scaffolds can be used as efficient drug delivery carriers. In the concept of tissue engineering, the "drugs" could be defined as drug molecules or biomacromolecules, even cells. These "drugs" have endowed the scaffolds which were laden improved abilities compared with the blank scaffolds. In this study, we anchored osteogenic bone morphogenetic protein-2 (BMP-2) derived peptides to α-TCP 3D porous scaffolds by linking the E7 domain to the target peptides, constructed the modified active peptides (E7BMP-2 peptides) delivery system, which finally achieved the modified peptides sustaining release and enhanced rat bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation in vitro. The α-TCP 3D porous scaffolds had micropores and interconnected micropores which expanded surface area of the scaffolds. The release test testified the constructed the delivery system had realized long-term release in which the peptides dosage could be detected by the BCA protein assay kit after 10 days compared with BMP-2 proteins which absorbed on the same α-TCP 3D porous scaffolds. The constructed E7BMP-2 peptides delivery system supported rat BMSCs osteogenic differentiation in the form of improving the genes expression levels of Runx2, ALP and OCN. Based on electrostatic interactions, E7 domain fastened combination between the active BMP-2 derived peptides and the α-TCP 3D porous scaffolds, the sustaining E7BMP-2 peptides release promoted the BMSCs osteogenesis as BMP-2 proteins did, which endowed α-TCP 3D porous scaffolds enhanced osteoinductive abilities in vitro.

摘要

α-磷酸三钙(α-TCP)基多孔支架在骨组织工程中具有优异的骨传导性和骨诱导性,此外,这些 3D 多孔支架可用作有效的药物输送载体。在组织工程的概念中,“药物”可以定义为药物分子或生物大分子,甚至是细胞。这些“药物”使负载的支架具有比空白支架更好的性能。在这项研究中,我们通过将 E7 结构域连接到靶肽,将成骨骨形态发生蛋白-2(BMP-2)衍生肽锚定到α-TCP 3D 多孔支架上,构建了改良的活性肽(E7BMP-2 肽)输送系统,最终实现了改良肽的持续释放,并增强了大鼠骨髓间充质干细胞(BMSCs)的体外成骨分化。α-TCP 3D 多孔支架具有微孔和相互连通的微孔,扩大了支架的表面积。释放试验证明,构建的输送系统实现了长期释放,在 10 天后可以通过 BCA 蛋白试剂盒检测到肽的剂量,而不是与相同的α-TCP 3D 多孔支架结合的 BMP-2 蛋白。构建的 E7BMP-2 肽输送系统以提高 Runx2、ALP 和 OCN 基因表达水平的形式支持大鼠 BMSCs 成骨分化。基于静电相互作用,E7 结构域使活性 BMP-2 衍生肽与α-TCP 3D 多孔支架之间的结合更加牢固,持续的 E7BMP-2 肽释放促进了 BMSCs 的成骨作用,与 BMP-2 蛋白一样,赋予了α-TCP 3D 多孔支架在体外增强的成骨能力。

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