Biotechnology Group, Chemical Engineering Department, Tarbiat Modares University, Tehran, Iran.
Biomimetic Materials and Tissue Engineering Laboratory, Department of Chemical Engineering, University of South Carolina, Columbia, SC, USA.
Int J Biol Macromol. 2020 Jan 1;142:790-802. doi: 10.1016/j.ijbiomac.2019.10.020. Epub 2019 Oct 14.
Given the great demand for biopolymer and protein-based products from renewable resources, synthesis of a keratin-based hydrogel is presented herein. In this work, a novel hydrogel of poly(γ-glutamic acid) (γ-PGA) and keratin was synthesized through facile EDC·HCl/HOBt chemistry. Since keratin main chain is rich in amino side groups, carboxyl groups in γ-PGA were crosslinked with multi terminated amine groups in keratin. In the following, the hydrogel characteristics, including swelling ratio (2010% at molar ratio of HOBt/EDC = 0.105), in vitro degradation and mass loss (about 20% at day 21 for the aforementioned sample), chemical decomposition and the rheological properties were investigated. The chemical activator agents, enhanced the elastic modulus of swollen hydrogel from around 1000 to 4000 Pa by increasing the crosslinking degree. Despite good biocompatibility for cell growth, some kind of self-assembled keratin hydrogels are not suitable for microscopic observation while the γ-PGA-Keratin hydrogel in our study is transparent. The γ-PGA-Keratin hydrogels possess significant features of rapid hydrogel formation in seconds, maximum swelling ratio of about 2500% maximum elastic modulus (stiffness) of about 4.5 kPa (for the swollen sample) with controllable matrix pore size. For further application, the biocompatibility of the γ-PGA-Keratin hydrogel was assessed by live/dead assay. Recent studies have demonstrated the effect of hydrogel porosity, water absorbing and stiffness on cell spreading, proliferation and differentiation of mesenchymal stem cells. Bone marrow mesenchymal stem cells could be differentiated into various cell fates depending on the elastic modulus of materials they are cultured on. We carried out a statistical study (to skip the cell work labor) to predetermine the proper working span in which we can gain a hydrogel to cover all features needed to be applied for some application like cartilage repair.
鉴于对可再生资源的生物聚合物和蛋白质基产品的巨大需求,本文提出了一种基于角蛋白的水凝胶的合成方法。在这项工作中,通过简便的 EDC·HCl/HOBt 化学方法合成了一种新型的聚(γ-谷氨酸)(γ-PGA)和角蛋白水凝胶。由于角蛋白的主链富含氨基酸侧基,γ-PGA 中的羧基与角蛋白中的多末端伯胺基交联。接下来,研究了水凝胶的特性,包括溶胀比(摩尔比 HOBt/EDC = 0.105 时为 2010%)、体外降解和质量损失(上述样品在第 21 天约为 20%)、化学分解和流变特性。化学激活剂增强了溶胀水凝胶的弹性模量,从约 1000 Pa 增加到 4000 Pa,从而增加了交联度。尽管对细胞生长具有良好的生物相容性,但某些自组装角蛋白水凝胶不适合微观观察,而我们研究中的 γ-PGA-角蛋白水凝胶是透明的。γ-PGA-角蛋白水凝胶具有快速水凝胶形成的显著特征,在几秒钟内形成,最大溶胀比约为 2500%,最大弹性模量(刚度)约为 4.5 kPa(对于溶胀样品),并且具有可控的基质孔径。为了进一步应用,通过活/死测定评估了 γ-PGA-角蛋白水凝胶的生物相容性。最近的研究表明,水凝胶的孔隙率、吸水性和刚度对间充质干细胞的细胞扩散、增殖和分化有影响。骨髓间充质干细胞可以根据其在材料上培养的弹性模量分化为各种细胞命运。我们进行了一项统计研究(跳过细胞工作劳动),以预先确定适当的工作范围,在该范围内,我们可以获得一种水凝胶,以涵盖应用于软骨修复等应用所需的所有特征。