Kubinová Šárka, Horák Daniel, Vaněček Václav, Plichta Zdeněk, Proks Vladimír, Syková Eva
Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic.
J Biomed Mater Res A. 2014 Jul;102(7):2315-23. doi: 10.1002/jbm.a.34910. Epub 2013 Aug 30.
Superporous poly(2-hydroxyethyl methacrylate) is successfully used as a scaffold material for tissue engineering; however, it lacks functional groups that support cell adhesion. The objective of this study was to investigate the cell-adhesive properties of biomimetic ligands, such as laminin-derived Ac-CGGASIKVAVS-OH (SIKVAV) peptide and fibronectin subunits (Fn), as well as small molecules exemplified by 2-mercaptoethanol (ME) and cysteine (Cys), immobilized on a copolymer of 2-hydroxyethyl methacrylate (HEMA) with 2-aminoethyl methacrylate (AEMA) by a maleimide-thiol coupling reaction. The maleimide group was introduced to the P(HEMA-AEMA) hydrogels by the reaction of their amino groups with N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS). Mesenchymal stem cells (MSCs) were used to investigate the cell adhesive properties of the modified hydrogels. A significantly larger area of cell growth as well as a higher cell density were found on Fn- and SIKVAV-modified hydrogels when compared to the ME- and Cys-modified supports or neat P(HEMA-AEMA). Moreover, Fn-modification strongly stimulated cell proliferation. The ability of MSCs to differentiate into adipocytes and osteoblasts was maintained on both Fn- and SIKVAV-modifications, but it was reduced on ME-modified hydrogels and neat P(HEMA-AEMA). The results show that the immobilization of SIKVAV and Fn-subunits onto superporous P(HEMA-AEMA) hydrogels via a GMBS coupling reaction improves cell adhesive properties. The high proliferative activity observed on Fn-modified hydrogels suggests that the immobilized Fn-subunits maintain their bioactivity and thus represent a promising tool for application in tissue engineering.
超大孔聚甲基丙烯酸2-羟乙酯已成功用作组织工程的支架材料;然而,它缺乏支持细胞黏附的官能团。本研究的目的是研究仿生配体的细胞黏附特性,这些仿生配体包括层粘连蛋白衍生的Ac-CGGASIKVAVS-OH(SIKVAV)肽和纤连蛋白亚基(Fn),以及以2-巯基乙醇(ME)和半胱氨酸(Cys)为代表的小分子,它们通过马来酰亚胺-硫醇偶联反应固定在甲基丙烯酸2-羟乙酯(HEMA)与甲基丙烯酸2-氨基乙酯(AEMA)的共聚物上。通过其氨基与N-γ-马来酰亚胺丁酰氧基琥珀酰亚胺酯(GMBS)反应,将马来酰亚胺基团引入到P(HEMA-AEMA)水凝胶中。使用间充质干细胞(MSC)来研究改性水凝胶的细胞黏附特性。与ME和Cys改性的载体或纯P(HEMA-AEMA)相比,在Fn和SIKVAV改性的水凝胶上发现细胞生长面积显著更大且细胞密度更高。此外,Fn改性强烈刺激细胞增殖。在Fn和SIKVAV改性上,MSC分化为脂肪细胞和成骨细胞的能力得以维持,但在ME改性的水凝胶和纯P(HEMA-AEMA)上则降低。结果表明,通过GMBS偶联反应将SIKVAV和Fn亚基固定在超大孔P(HEMA-AEMA)水凝胶上可改善细胞黏附特性。在Fn改性水凝胶上观察到的高增殖活性表明,固定化的Fn亚基保持了其生物活性,因此是组织工程应用中有前景的工具。