Centre for Translational Bone, Joint and Soft Tissue Research, University Hospital Carl Gustav Carus and Faculty of Medicine of Technische Universität Dresden, Fetscherstraße 74, 01307 Dresden, Germany.
Mar Drugs. 2018 Mar 13;16(3):91. doi: 10.3390/md16030091.
Collagens of marine origin are applied increasingly as alternatives to mammalian collagens in tissue engineering. The aim of the present study was to develop a biphasic scaffold from exclusively marine collagens supporting both osteogenic and chondrogenic differentiation and to find a suitable setup for in vitro chondrogenic and osteogenic differentiation of human mesenchymal stroma cells (hMSC).
Biphasic scaffolds from biomimetically mineralized salmon collagen and fibrillized jellyfish collagen were fabricated by joint freeze-drying and crosslinking. Different experiments were performed to analyze the influence of cell density and TGF-β on osteogenic differentiation of the cells in the scaffolds. Gene expression analysis and analysis of cartilage extracellular matrix components were performed and activity of alkaline phosphatase was determined. Furthermore, histological sections of differentiated cells in the biphasic scaffolds were analyzed.
Stable biphasic scaffolds from two different marine collagens were prepared. An in vitro setup for osteochondral differentiation was developed involving (1) different seeding densities in the phases; (2) additional application of alginate hydrogel in the chondral part; (3) pre-differentiation and sequential seeding of the scaffolds and (4) osteochondral medium. Spatially separated osteogenic and chondrogenic differentiation of hMSC was achieved in this setup, while osteochondral medium in combination with the biphasic scaffolds alone was not sufficient to reach this ambition.
Biphasic, but monolithic scaffolds from exclusively marine collagens are suitable for the development of osteochondral constructs.
海洋来源的胶原蛋白越来越多地被应用于组织工程中，作为哺乳动物胶原蛋白的替代品。本研究的目的是开发一种由纯海洋胶原蛋白制成的双相支架，既能支持成骨分化，又能支持软骨分化，并找到一种合适的方法来体外诱导人间充质基质细胞（hMSC）的成软骨和成骨分化。
通过联合冷冻干燥和交联，制备仿生矿化三文鱼胶原蛋白和纤维水母胶原蛋白的双相支架。进行了不同的实验来分析细胞密度和 TGF-β 对支架中细胞成骨分化的影响。进行了基因表达分析和软骨细胞外基质成分的分析，并测定碱性磷酸酶的活性。此外，还分析了分化细胞在双相支架中的组织学切片。
成功制备了两种不同海洋胶原蛋白的稳定双相支架。开发了一种用于骨软骨分化的体外方法，包括：（1）在不同相中接种不同的细胞密度；（2）在软骨部分额外应用藻酸盐水凝胶；（3）支架的预分化和顺序接种；（4）骨软骨培养基。在这种设置中实现了 hMSC 的空间分离的成骨和成软骨分化，而仅使用骨软骨培养基和双相支架不足以实现这一目标。
仅由海洋胶原蛋白制成的双相、但整体的支架适合开发骨软骨构建体。
