肾型软骨是一种富含胶原蛋白的海洋海绵,被认为是生产哺乳动物来源胶原蛋白替代品的可持续和可行的选择。然而,缺乏关于从不同海绵部位分离的胶原蛋白特性的知识,即外部区域,或者皮质,(ectosome)和内部区域(choanosome),以及它如何影响生物材料的发展。在这项研究中,对肾形梭菌胶原的空间分布进行了简要的组织学分析,并对从外植体和小囊体分离的胶原进行了全面的比较分析。分离的胶原蛋白表征基于分离产量,傅里叶变换红外光谱(FTIR),圆二色性(CD),SDS-PAGE,斑点印迹,和氨基酸组成,以及它们的细胞相容性设想了未来生物医学应用的发展。两个海绵部分的分离收率约为20%,以及FTIR,CD,和SDS-PAGE图谱,这表明两种分离的胶原蛋白均具有较高的纯度,并保留了其三螺旋和纤维状构象。外植体胶原蛋白具有较高的OHpro含量,并具有I型和IV型胶原蛋白,而choanosome主要由IV型胶原蛋白组成。使用L929成纤维细胞细胞系进行的体外细胞毒性测定显示,在2mg/mL时,choanosome胶原具有显着的细胞毒性作用,而外体胶原蛋白增强细胞代谢和增殖,从而表明后者更适合于生物材料的发展。这项研究代表了肾形梭菌身体部位的独特比较研究,作为进一步建立这种海洋海绵作为生物医学应用的未来发展有前途的替代胶原蛋白来源的支持。
Chondrosia reniformis is a collagen-rich marine sponge that is considered a sustainable and viable option for producing an alternative to mammalian-origin collagens. However, there is a lack of knowledge regarding the properties of collagen isolated from different sponge parts, namely the outer region, or cortex, (
ectosome) and the inner region (choanosome), and how it affects the development of biomaterials. In this study, a brief histological analysis focusing on C. reniformis collagen spatial distribution and a comprehensive comparative analysis between collagen isolated from
ectosome and choanosome are presented. The isolated collagen characterization was based on isolation yield, Fourier-transformed infrared spectroscopy (FTIR), circular dichroism (CD), SDS-PAGE, dot blot, and amino acid composition, as well as their cytocompatibility envisaging the development of future biomedical applications. An isolation yield of approximately 20% was similar for both sponge parts, as well as the FTIR, CD, and SDS-PAGE profiles, which demonstrated that both isolated collagens presented a high purity degree and preserved their triple helix and fibrillar conformation.
Ectosome collagen had a higher OHpro content and possessed collagen type I and IV, while the choanosome was predominately constituted by collagen type IV. In vitro cytotoxicity assays using the L929 fibroblast cell line displayed a significant cytotoxic effect of choanosome collagen at 2 mg/mL, while
ectosome collagen enhanced cell metabolism and proliferation, thus indicating the latter as being more suitable for the development of biomaterials. This research represents a unique comparative study of C. reniformis body parts, serving as a support for further establishing this marine sponge as a promising alternative collagen source for the future development of biomedical applications.