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可定制的明胶-丝素水凝胶及其激活/交联作用,以增强成纤维细胞的增殖。

Tailorable hydrogel of gelatin with silk fibroin and its activation/crosslinking for enhanced proliferation of fibroblast cells.

机构信息

School of Medical Science & Technology, IIT Kharagpur, Kharagpur 721302, West Bengal, India.

Department of Chemical Engineering, IIT Kharagpur, Kharagpur 721302, West Bengal, India.

出版信息

Int J Biol Macromol. 2020 Dec 1;164:4073-4083. doi: 10.1016/j.ijbiomac.2020.09.016. Epub 2020 Sep 6.

Abstract

Gelatin based hydrogel (Gel) possess remarkable cytocompatibility profile rendering it appropriate for tissue engineering applications. Herein, the questionable mechanical property of Gel was tuned by tailoring with different loading concentrations of silk fibroin (SF). The as tailored matrix was reconnoitred for its physico-mechanical, chemical and biological properties in order to investigate the effect of SF loading. Ethanol treatment lead to enhance β-sheet formation of silk and subsequently, carbodiimide coupling was deployed to covalently crosslink the matrix. Substantial increase in cohesive energy with amplifying concentration of SF in the Gel matrix. As evidenced by Raman spectroscopy, right shift of the Amide I peak and stretching of COO confirms activation of fibroin moiety along with crosslinking of gelatin, respectively. Moreover, with addition of SF, surface properties were tuned to attain maximum cell adhesion and proliferation. Further, MTT assay corroborated the same with definite increase in mitochondrial activities of L929 fibroblast cells for SF containing matrix as compared to its bare counterfeit while enhanced proliferation was confirmed by Rhodamine-DAPI staining. The alteration in mechanical and textural tribology of SF tailored Gel matrix certainly portrayed improved stability along with excellent cytocompatibility thus making it a plausible alternative in regenerative medicine.

摘要

基于明胶的水凝胶(Gel)具有出色的细胞相容性,非常适合组织工程应用。在此,通过调整丝素蛋白(SF)的不同加载浓度来调整 Gel 的可疑机械性能。对定制的基质进行物理力学、化学和生物学特性的研究,以研究 SF 负载的影响。乙醇处理会促进丝的β-折叠形成,随后使用碳二亚胺偶联将基质进行共价交联。随着 Gel 基质中 SF 浓度的增加,内聚能显著增加。如拉曼光谱所示,酰胺 I 峰的右移和 COO 的拉伸证实了丝素部分的活化以及明胶的交联。此外,随着 SF 的加入,表面性质得到了调整,以获得最大的细胞粘附和增殖。此外,MTT 分析证实,与裸对照相比,含有 SF 的基质中 L929 成纤维细胞的线粒体活性明显增加,而罗丹明-DAPI 染色则证实了其增殖增强。SF 定制的 Gel 基质的机械和纹理摩擦学的改变确实提高了其稳定性和优异的细胞相容性,使其成为再生医学中一种可行的替代方法。

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