首页> 外文期刊>Acta biomaterialia >Non-mulberry silk gland fibroin protein 3-D scaffold for enhanced differentiation of human mesenchymal stem cells into osteocytes.
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Non-mulberry silk gland fibroin protein 3-D scaffold for enhanced differentiation of human mesenchymal stem cells into osteocytes.

机译:非桑蚕丝腺纤维蛋白蛋白3-D支架,用于增强人间充质干细胞向骨细胞的分化。

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摘要

This study investigates the potential of three-dimensional (3-D) scaffolds of wild non-mulberry tropical tasar silk gland fibroin protein as substratum for osteogenic differentiation of human mesenchymal stem cells (hMSCs). The novelty of the study lies in the fabrication of scaffolds from non-bioengineered silk fibroin directly extracted from the glands of non-mulberry tropical tasar silkworms using sodium dodecyl sulfate dissolution protocol and its osteogenic application using single- and double-seeding methods. The scaffolds were mechanically robust and showed homogenous pore distribution within the scaffold. hMSCs were seeded on the scaffolds and were cultured for up to 28days under static conditions in osteogenic media. Osteogenic differentiation of hMSCs seeded on fibroin scaffolds resulted in extensive mineralization with the formation of large calcium nodules, higher alkaline phosphatase activity and intense von Kossa staining. Real-time studies revealed higher transcript levels for osteopontin (OS) and bone sialoprotein (IBSP) under double-seeded conditions as compared to single-seeded scaffolds. Histological analysis showed the development of osteoblastic cells and large calcified nodules. The development and spreading of nuclei and actin filaments on fibroin matrices were revealed through confocal studies. The results suggest the suitability of non-mulberry silk-fibroin protein 3-D scaffolds as natural biomaterial for potential in vitro bone-tissue engineering applications.
机译:这项研究调查了野生的非桑树热带塔萨尔丝绸腺素纤维蛋白的三维(3-D)支架作为基质为人类间充质干细胞(hMSCs)成骨分化的潜力。该研究的新颖性在于使用十二烷基硫酸钠溶解方案直接从非桑树热带塔莎蚕腺体中提取的非生物工程化丝素蛋白制造支架,并通过单播和双播方法将其用于成骨。支架在机械上坚固并且在支架内显示均匀的孔分布。将hMSCs接种在支架上,并在成骨培养基中的静态条件下培养长达28天。植入纤维蛋白支架的hMSC的成骨分化导致广泛的矿化作用,形成大的钙结节,更高的碱性磷酸酶活性和强烈的von Kossa染色。实时研究显示,与单种子支架相比,双种子条件下骨桥蛋白(OS)和骨唾液蛋白(IBSP)的转录水平更高。组织学分析显示成骨细胞的发展和大钙化结节。通过共聚焦研究揭示了核素和肌动蛋白丝在纤维蛋白基质上的发育和扩散。结果表明非桑蚕丝素蛋白3-D支架作为天然生物材料的潜在体外骨组织工程应用的适用性。

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