Dipartimento di Ingegneria, Università degli Studi di Palermo, Viale delle Scienze, Edificio 6, 90128 Palermo, Italy.
Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Via U. La Malfa, 153, 90146 Palermo, Italy.
Int J Biol Macromol. 2022 Jun 30;211:639-652. doi: 10.1016/j.ijbiomac.2022.05.068. Epub 2022 May 13.
Polymeric hydrogels are increasingly considered as scaffolds for tissue engineering due to their extraordinary resemblance with the extracellular matrix (ECM) of many tissues. As cell adhesion is a key factor in regulating important cell functions, hydrogel scaffolds are often functionalized or loaded with a variety of bioactive molecules that can promote adhesion. Interesting biomimetic approaches exploit the properties of mussel-inspired recombinant adhesive proteins. In this work, we prepared hydrogel scaffolds with a 50%w mixture of k-carrageenan (kC) and polyvinyl alcohol (PVA), by a two-step physical gelation process, and we coated them with Perna viridis foot protein-5β (Pvfp5β). The mechanical and morphological properties of hydrogels were investigated both after conditioning with typical cell culture media and also after coating with the Pvfp5β. The protein resulted strongly adsorbed onto the surface of the hydrogel and also able to penetrate in its interiors to a certain depth, mainly interacting with the kC component of the scaffold as resulted from the confocal analysis. Mouse embryonic fibroblasts NIH-3T3 were seeded on top of the hydrogels and cultured up to two weeks. The role of Pvfp5β in promoting cell adhesion, spreading and colonization of the scaffold was demonstrated.
水凝胶由于其与许多组织的细胞外基质（ECM）非常相似，因此越来越多地被认为是组织工程的支架。由于细胞黏附是调节重要细胞功能的关键因素，水凝胶支架通常经过功能化或负载各种生物活性分子，以促进黏附。有趣的仿生方法利用贻贝启发的重组黏附蛋白的特性。在这项工作中，我们通过两步物理凝胶化过程，制备了 50%w 的角叉菜胶（kC）和聚乙烯醇（PVA）混合物的水凝胶支架，并在其上涂覆了贻贝足蛋白-5β（Pvfp5β）。在使用典型的细胞培养基进行调理后，以及在涂覆 Pvfp5β 后，我们研究了水凝胶的机械和形态特性。该蛋白强烈吸附在水凝胶的表面上，并且能够渗透到一定深度的内部，主要与支架的 kC 成分相互作用，如共聚焦分析所示。将小鼠胚胎成纤维细胞 NIH-3T3 接种在水凝胶顶部，并培养长达两周。证明了 Pvfp5β 在促进细胞黏附、铺展和支架定殖方面的作用。
