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采用 PLA 和八目鳗鱼线角蛋白复合材料制备纳米纤维支架;其对细胞黏附、生长和成骨细胞分化的影响。

Fabrication of nanofibrous scaffold using a PLA and hagfish thread keratin composite; its effect on cell adherence, growth, and osteoblast differentiation.

机构信息

Wonkwang Bone Regeneration Research Institute, Wonkwang University, Iksan 570-749, Korea.

出版信息

Biomed Mater. 2013 Aug;8(4):045006. doi: 10.1088/1748-6041/8/4/045006. Epub 2013 Jun 4.

Abstract

Electrospinning is a useful method for the production of nanofibrous scaffolds in the field of tissue engineering. Keratin has been used as a biomaterial for electrospinning and can be used in a variety of biomedical applications because it is a natural protein, giving it the ability to improve cell affinity of scaffolds. In this study, keratin was extracted from hagfish slime thread (H-keratin) and blended with polylactic acid (PLA) polymer solution to construct a nanofibrous scaffold. Wool keratin (W-keratin) was used as a control for the comparison of morphological, physical, and biological properties. The results of Fourier transform infrared spectroscopy showed the presence of both W-keratin and H-keratin in the electrospun PLA/keratin. Observations with a scanning electron microscope revealed that PLA, PLA/W-keratin, and PLA/H-keratin had similar average diameters (~800 nm). Cell attachment experiments showed that MG-63 cells adhered more rapidly and spread better onto PLA/H-keratin than onto the pure PLA or PLA/W-keratin. Cell proliferation assay, DNA content, live/dead, and alkaline phosphatase activity assays showed that PLA/H-keratin scaffolds could accelerate the viability, proliferation, and osteogenesis of MG-63 cells relative to pure PLA or PLA/W-keratin nanofibrous scaffolds. These findings suggest that H-keratin can improve cellular attraction and has great potential to be used as a biomaterial in bone tissue engineering.

摘要

静电纺丝是组织工程领域生产纳米纤维支架的一种有用方法。角蛋白已被用作静电纺丝的生物材料,并且由于它是一种天然蛋白质,因此具有提高支架细胞亲和力的能力,可用于多种生物医学应用。在这项研究中,从盲鳗粘液丝(H-角蛋白)中提取角蛋白,并与聚乳酸(PLA)聚合物溶液混合,构建纳米纤维支架。羊毛角蛋白(W-角蛋白)被用作比较形态,物理和生物特性的对照。傅里叶变换红外光谱的结果表明,电纺 PLA/角蛋白中存在 W-角蛋白和 H-角蛋白。扫描电子显微镜的观察结果表明,PLA、PLA/W-角蛋白和 PLA/H-角蛋白的平均直径相似(约 800nm)。细胞附着实验表明,MG-63 细胞在 PLA/H-角蛋白上的附着速度更快,铺展效果更好,而在纯 PLA 或 PLA/W-角蛋白上则较差。细胞增殖测定、DNA 含量、死活和碱性磷酸酶活性测定表明,与纯 PLA 或 PLA/W-角蛋白纳米纤维支架相比,PLA/H-角蛋白支架可以加速 MG-63 细胞的活力、增殖和成骨作用。这些发现表明 H-角蛋白可以提高细胞吸引力,并且在骨组织工程中用作生物材料具有很大的潜力。

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