Abstract:
Collagen filaments derived from the two marine demosponges Ircinia oros and Sarcotragus foetidus were for the first time isolated, biochemically characterised and tested for their potential use in regenerative medicine. SDS-PAGE of isolated filaments revealed a main collagen subunit band of 130 kDa in both of the samples under study. DSC analysis on 2D membranes produced with collagenous sponge filaments showed higher thermal stability than commercial mammalian-derived collagen membranes. Dynamic mechanical and thermal analysis attested that the membranes obtained from filaments of S. foetidus were more resistant and stable at the rising temperature, compared to the ones derived from filaments of I. oros. Moreover, the former has higher stability in saline and in collagenase solutions and evident antioxidant activity. Conversely, their water binding capacity results were lower than that of membranes obtained from I. oros. Adhesion and proliferation tests using L929 fibroblasts and HaCaT keratinocytes resulted in a remarkable biocompatibility of both developed membrane models, and gene expression analysis showed an evident up-regulation of ECM-related genes. Finally, membranes from I. oros significantly increased type I collagen gene expression and its release in the culture medium. The findings here reported strongly suggest the biotechnological potential of these collagenous structures of poriferan origin as scaffolds for wound healing.
摘要:
首次分离出来自两种海洋demospongesIrciniaoros和Sarcotragusfoetidus的胶原蛋白丝,生物化学表征和测试它们在再生医学中的潜在用途。分离的细丝的SDS-PAGE显示,在所研究的两个样品中,主要的胶原蛋白亚基带都为130kDa。用胶原海绵丝生产的2D膜的DSC分析显示出比商业哺乳动物衍生的胶原膜更高的热稳定性。动态机械和热分析证明,从S.foetidus长丝获得的膜在升高的温度下更有抵抗力和稳定性。与源自I.oros细丝的那些相比。此外,前者在生理盐水和胶原酶溶液中具有较高的稳定性和明显的抗氧化活性。相反,它们的水结合能力结果低于从I.oros获得的膜。使用L929成纤维细胞和HaCaT角质形成细胞的粘附和增殖测试导致两种开发的膜模型的显着生物相容性。和基因表达分析表明ECM相关基因明显上调。最后,来自I.oros的膜显着增加了I型胶原蛋白基因的表达及其在培养基中的释放。此处报道的发现有力地表明了这些孔虫起源的胶原结构作为伤口愈合支架的生物技术潜力。