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新型表面改性聚甲基丙烯酸2-羟乙酯水凝胶在组织工程中的应用:用纤连蛋白亚基与Ac-CGGASIKVAVS-OH、半胱氨酸和2-巯基乙醇改性处理表面

The use of new surface-modified poly(2-hydroxyethyl methacrylate) hydrogels in tissue engineering: treatment of the surface with fibronectin subunits versus Ac-CGGASIKVAVS-OH, cysteine, and 2-mercaptoethanol modification.

作者信息

Kubinová Šárka, Horák Daniel, Vaněček Václav, Plichta Zdeněk, Proks Vladimír, Syková Eva

机构信息

Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic.

出版信息

J Biomed Mater Res A. 2014 Jul;102(7):2315-23. doi: 10.1002/jbm.a.34910. Epub 2013 Aug 30.

Abstract

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.

摘要

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。

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