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通过生化方法评估蓝鲨皮胶原蛋白对分化骨细胞增殖的影响及其在骨组织工程中的应用。

Evaluation of Differentiated Bone Cells Proliferation by Blue Shark Skin Collagen via Biochemical for Bone Tissue Engineering.

机构信息

Department of Marine Bio-Pharmacology, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.

Division of Marine Bioscience, Korea Maritime and Ocean University, Busan 606791, Korea.

出版信息

Mar Drugs. 2018 Sep 25;16(10):350. doi: 10.3390/md16100350.

Abstract

Collagen from a marine resource is believed to have more potential activity in bone tissue engineering and their bioactivity depends on biochemical and structural properties. Considering the above concept, pepsin soluble collagen (PSC) and acid soluble collagen (ASC) from blue shark () skin were extracted and its biochemical and osteogenic properties were investigated. The hydroxyproline content was higher in PSC than ASC and the purified collagens contained three distinct bands α₁, α and β dimer. The purity of collagen was confirmed by the RP-HPLC profile and the thermogravimetric data showed a two-step thermal degradation pattern. ASC had a sharp decline in viscosity at 20⁻30 °C. Scanning electron microscope (SEM) images revealed the fibrillar network structure of collagens. Proliferation rates of the differentiated mouse bone marrow-mesenchymal stem (dMBMS) and differentiated osteoblastic (dMC3T3E1) cells were increased in collagen treated groups rather than the controls and the effect was dose-dependent, which was further supported by higher osteogenic protein and mRNA expression in collagen treated bone cells. Among two collagens, PSC had significantly increased dMBMS cell proliferation and this was materialized through increasing RUNX2 and collagen-I expression in bone cells. Accordingly, the collagens from blue shark skin with excellent biochemical and osteogenic properties could be a suitable biomaterial for therapeutic application.

摘要

从海洋资源中提取的胶原蛋白被认为在骨组织工程中具有更大的潜在活性,其生物活性取决于生化和结构特性。基于上述概念,从蓝鲨皮中提取了胃蛋白酶可溶性胶原蛋白(PSC)和酸溶性胶原蛋白(ASC),并对其生化和成骨特性进行了研究。PSC 的羟脯氨酸含量高于 ASC,且纯化的胶原蛋白含有三条明显的 α₁、α 和 β 二聚体带。反相高效液相色谱(RP-HPLC)图谱证实了胶原蛋白的纯度,热重数据显示其具有两步热降解模式。ASC 在 20-30°C 时粘度急剧下降。扫描电子显微镜(SEM)图像显示了胶原蛋白的纤维状网络结构。与对照组相比,胶原蛋白处理组的分化小鼠骨髓间充质干细胞(dMBMS)和分化成骨细胞(dMC3T3E1)的增殖率增加,且呈剂量依赖性,胶原蛋白处理的成骨细胞中骨形成蛋白和 mRNA 表达增加进一步支持了这一结果。在这两种胶原蛋白中,PSC 显著增加了 dMBMS 细胞的增殖,这是通过增加成骨细胞中 RUNX2 和胶原蛋白-I 的表达来实现的。因此,具有优异生化和成骨特性的蓝鲨皮胶原蛋白可能是一种适合治疗应用的生物材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2885/6212988/455d6fdfe170/marinedrugs-16-00350-g001.jpg

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