含固有RGD的丝素蛋白-聚(ε-己内酯)纳米纤维基质用于骨组织工程的潜力

Potential of inherent RGD containing silk fibroin-poly (Є-caprolactone) nanofibrous matrix for bone tissue engineering.

作者信息

Bhattacharjee Promita, Kundu Banani, Naskar Deboki, Kim Hae-Won, Bhattacharya Debasis, Maiti T K, Kundu S C

机构信息

Materials Science Centre, Indian Institute of Technology Kharagpur, West Bengal, 721302, India.

Department of Biotechnology, Indian Institute of Technology Kharagpur, West Bengal, 721302, India.

出版信息

Cell Tissue Res. 2016 Feb;363(2):525-40. doi: 10.1007/s00441-015-2232-6. Epub 2015 Jul 15.

Abstract

The current study deals with the fabrication and characterization of blended nanofibrous scaffolds of tropical tasar silk fibroin of Antheraea mylitta and poly (Є-caprolactone) to act as an ideal scaffold for bone regeneration. The use of poly (Є-caprolactone) in osteogenesis is well-recognized. At the same time, the osteoconductive nature of the non-mulberry tasar fibroin is also established due to its internal integrin binding peptide RGD (Arg-Gly-Asp) sequences, which enhance cellular interaction and proliferation. Considering that the materials have the required and favorable properties, the blends are formed using an equal volume ratio of fibroin (2 and 4 wt%) and poly (Є-caprolactone) solution (10 wt%) to fabricate nanofibers. The nanofibers possess an average diameter of 152 ± 18 nm (2 % fibroin/PCL) and 175 ± 15 nm (4% fibroin/PCL). The results of Fourier transform infrared spectroscopy substantiates the preservation of the secondary structure of the fibroin in the blends indicating the structural stability of the neo-matrix. With an increase in the fibroin percentage, the hydrophobicity and thermal stability of the matrices as measured from melting temperature Tm (using DSC) decrease, while the mechanical strength is improved. The blended nanofibrous scaffolds are biodegradable, and support the viability and proliferation of human osteoblast-like cells as observed through scanning electron and confocal microscopes. Alkaline phosphatase assay indicates the cell proliferation and the generation of the neo-bone matrix. Taken together, these findings illustrate that the silk-poly (Є-caprolactone) blended nanofibrous scaffolds have an excellent prospect as scaffolding material in bone tissue engineering.

摘要

本研究涉及对印度柞蚕(Antheraea mylitta)的热带柞蚕丝素蛋白与聚(ε-己内酯)的混合纳米纤维支架进行制备与表征,使其成为骨再生的理想支架。聚(ε-己内酯)在骨生成中的应用已得到广泛认可。同时,非桑蚕丝素蛋白的骨传导性质也已得到证实,这归因于其内部的整合素结合肽RGD(精氨酸-甘氨酸-天冬氨酸)序列,该序列可增强细胞相互作用和增殖。鉴于这些材料具有所需的良好性能,使用丝素蛋白(2 wt%和4 wt%)与聚(ε-己内酯)溶液(10 wt%)的等体积比形成混合物来制备纳米纤维。纳米纤维的平均直径为152±18 nm(2%丝素蛋白/聚己内酯)和175±15 nm(4%丝素蛋白/聚己内酯)。傅里叶变换红外光谱结果证实了混合物中丝素蛋白二级结构的保留,表明新基质的结构稳定性。随着丝素蛋白百分比的增加,通过熔点温度Tm(使用差示扫描量热法)测量的基质的疏水性和热稳定性降低,而机械强度提高。混合纳米纤维支架具有生物可降解性,并且通过扫描电子显微镜和共聚焦显微镜观察发现,其支持人成骨样细胞的活力和增殖。碱性磷酸酶测定表明细胞增殖以及新骨基质的生成。综上所述,这些发现表明丝素蛋白-聚(ε-己内酯)混合纳米纤维支架作为骨组织工程中的支架材料具有极好的前景。

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