Nagai Nobuhiro, Mori Kazuo, Satoh Yasuharu, Takahashi Noritaka, Yunoki Shunji, Tajima Kenji, Munekata Masanobu
Creative Research Initiative "Sousei" (CRIS), Hokkaido University, N21-W10, Kita-ku, Sapporo, Hokkaido 001-0021, Japan.
J Biomed Mater Res A. 2007 Aug;82(2):395-402. doi: 10.1002/jbm.a.31110.
Marine-derived collagen is expected to be a much safer alternative to calf collagen, which in medical applications carries the risk of bovine spongiform encephalopathy. In this study, acid-soluble collagen was extracted from salmon skin and crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide during fibril formation to produce a crosslinked salmon collagen (SC) gel. The growth rates and the differentiated functions of human periodontal ligament fibroblasts (HPdLFs) cultured on the SC gel were investigated. Growth was faster on the SC gel than on porcine collagen (PC) gel. In addition, the HPdLFs cultured on the SC gel exhibited higher alkaline phosphatase (ALP) activity than those cultured on the PC gel. Quantitative RT-PCR revealed higher mRNA expression of type I collagen, ALP, and osteocalcin in the HPdLFs cultured on the SC gel. HPdLFs had a flat shape on the SC gel and a spindle shape on the PC gel, as revealed by observation with scanning electron microscopy and immunostaining with cytoskeletal protein and vinculin. The results showed that HPdLFs could grow and show highly differentiated activity on the SC gel as well as on the PC gel.
海洋来源的胶原蛋白有望成为小牛胶原蛋白更安全的替代品，小牛胶原蛋白在医学应用中存在牛海绵状脑病的风险。在本研究中，从鲑鱼皮中提取酸溶性胶原蛋白，并在原纤维形成过程中与1-乙基-3-(3-二甲基氨基丙基)-碳二亚胺交联，以制备交联鲑鱼胶原蛋白（SC）凝胶。研究了在SC凝胶上培养的人牙周膜成纤维细胞（HPdLFs）的生长速率和分化功能。在SC凝胶上的生长速度比在猪胶原蛋白（PC）凝胶上快。此外，在SC凝胶上培养的HPdLFs比在PC凝胶上培养的HPdLFs表现出更高的碱性磷酸酶（ALP）活性。定量逆转录聚合酶链反应显示，在SC凝胶上培养的HPdLFs中I型胶原蛋白、ALP和骨钙素的mRNA表达更高。通过扫描电子显微镜观察以及用细胞骨架蛋白和纽蛋白进行免疫染色发现，HPdLFs在SC凝胶上呈扁平状，在PC凝胶上呈纺锤状。结果表明，HPdLFs在SC凝胶以及PC凝胶上均能生长并表现出高度分化的活性。
