Department of Marine Bio-Pharmacology, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Department of Fish Quality Assurance and Management, Fish Quality Monitoring and Certification Centre, Fisheries College and Research Institute, Tamil Nadu Dr.J.Jayalalitha Fisheries University, Tuticorin 628008, India.
Cells. 2019 May 11;8(5):446. doi: 10.3390/cells8050446.
Collagen is the most abundant extracellular fibrous protein that has been widely used for biomedical applications due to its excellent biochemical and biocompatibility features. It is believed that the smaller molecular weight collagen, i.e., collagen peptide (CP), has more potent activity than native collagen. However, the preparation of CP from fish bone collagen is a complex and time-consuming process. Additionally, the osteogenic effect of CP depends on its molecular weight and amino acid composition. Considering the above concept, the present work was undertaken to extract the CP directly from Mahi mahi fish () bones and test its osteogenic potential using bone marrow mesenchymal stem (BMMS) cells. The hydrolyzed collagen contained triple alpha chains (110 kDa) and a peptide (~1 kDa) and the peptide was successfully separated from hydrolyzed collagen using molecular weight cut-off membrane. CP treatment was up-regulated BMMS cells proliferation and differentiation. Interestingly, CP accrued the mineral deposition in differentiated BMMS cells. Protein and mRNA expression revealed that the osteogenic biomarkers such as collagen, alkaline phosphatase, and osteocalcin levels were significantly increased by CP treatment in differentiated BMMS cells and also further elucidated the hypothesis that CP was upregulated osteogenesis through activating Runx2 via p38MAPK signaling pathway. The above results concluded that the CP from Mahi mahi bones with excellent osteogenic properties could be the suitable biomaterial for bone therapeutic application.
胶原蛋白是最丰富的细胞外纤维状蛋白质，由于其优异的生化和生物相容性特征，已被广泛应用于生物医学领域。人们认为，分子量较小的胶原蛋白，即胶原蛋白肽（CP），比天然胶原蛋白具有更强的活性。然而，从鱼骨胶原蛋白中制备 CP 是一个复杂且耗时的过程。此外，CP 的成骨作用取决于其分子量和氨基酸组成。考虑到上述概念，本工作旨在直接从 Mahi mahi 鱼（）鱼骨中提取 CP，并使用骨髓间充质干细胞（BMMS）细胞测试其成骨潜力。水解胶原蛋白含有三股 α 链（110 kDa）和一条肽（~1 kDa），肽可使用分子量截止膜从水解胶原蛋白中成功分离。CP 处理可上调 BMMS 细胞的增殖和分化。有趣的是，CP 可促进分化的 BMMS 细胞中的矿物质沉积。蛋白质和 mRNA 表达表明，CP 处理可显著增加分化的 BMMS 细胞中胶原、碱性磷酸酶和骨钙素等成骨生物标志物的水平，这进一步证明 CP 通过激活 p38MAPK 信号通路来上调成骨作用。上述结果表明，具有优异成骨性能的 Mahi mahi 鱼骨 CP 可能是骨治疗应用的合适生物材料。
