Effects of Purified Bioactive Peptides on Bone Regeneration

胶原蛋白是在组织工程、化妆品和功能性食品等领域应用最广泛的生物材料之一。最近，鱼胶原蛋白肽（FCP）已被用作膳食补充剂，但其对细胞功能的影响知之甚少。本研究采用成骨细胞MC 3 T3-E1培养体系，研究了FCP对胶原合成和质量的影响。当细胞用0. 2%（w/ v）FCP，胶原修饰酶的基因表达水平，i. e.赖氨酰羟化酶（LH 1 -3）、赖氨酰氧化酶（LOX和LOXL 1 -4）和含有25个结构域的糖基转移酶1（GLT 25 D1），除了LOX和LOXL 1，在培养48小时时都显著上调。此外，当分析培养14天时沉积的胶原时，与未处理组相比，FCP处理组显示出显著更高的胶原含量、更大程度的赖氨酸羟基化（Hyl）、更高水平的Hyl-醛衍生的交联和加速的交联成熟。此外，还发现FCP处理加速了培养物中基质的矿化。这些结果表明，FCP在成骨细胞培养体系中对胶原合成、胶原质量和矿化产生积极影响，表明FCP在骨组织工程中的潜在用途。

Collagen is one of the most widely used biomaterials in the fields of tissue engineering, cosmetics and functional foods. Recently, fish collagen peptides(FCP) have been used as a dietary supplement, but its effects on cellular functions are poorly understood. In this study, the effects of FCP on the collagen synthesis and quality were investigated using an osteoblastic MC3T3-E1 cell culture system. When cells were treated with 0. 2%(w/ v) FCP, gene expression levels of collagen modifying enzymes, i. e. lysyl hydroxylases(LH1-3), lysyl oxidases(LOX and LOXL1-4) and glycosyltransferase 25 domain containing 1(GLT25D1) were all, except LOX and LOXL1, significantly upregulated at 48 hrs of culture. In addition, when collagen deposited at 14 days of culture was analyzed, the FCP treated group showed significantly higher collagen content, a greater extent of lysine hydroxylation(Hyl), higher levels of Hyl-aldehyde derived cross-links and accelerated cross-link maturation, in comparison to the untreated group. Furthermore, it was found that FCP treatment accelerated the matrix mineralization in the cultures. These results indicate that FCP exerts positive effects on collagen synthesis, collagen quality and mineralization in an osteoblastic cell culture system, suggesting the potential utility of FCP for bone tissue engineering.