Biphasic Scaffolds from Marine Collagens for Regeneration of Osteochondral Defects.

Anne Bernhardt,Birgit Paul,Michael Gelinsky

doi:10.3390/md16030091

Anne Bernhardt, Birgit Paul + Show 1 more

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https://doi.org/10.3390/md16030091

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Journal: Marine Drugs	Publication Date: Mar 13, 2018
Citations: 40	License type: CC BY 4.0

Affiliation: University Hospital Carl Gustav Carus, TU Dresden

Abstract

Background: 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). Methods: 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. Results: 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. Conclusions: Biphasic, but monolithic scaffolds from exclusively marine collagens are suitable for the development of osteochondral constructs.

Highlights

Collagen is one of the most frequently applied biomaterials for biomedical research as well as clinical applications [1,2]
Marine collagens which can be obtained from both invertebrates and vertebrates [3,4] show promising features and have the potential to overrule mammalian collagens in biomedical applications for several reasons
Scanning electron microscopy (SEM) images show the smooth surface of the non-mineralized jellyfish collagen pore walls, as well as the rough morphology of the mineralized salmon collagen, which originates in the presence of hydroxyapatite nanocrystals, decorating the surface (Figure 2)

Summary

Introduction

Collagen is one of the most frequently applied biomaterials for biomedical research as well as clinical applications [1,2]. Scaffolds for tissue engineering applications are increasingly developed from collagens of marine origin, such as from fish collagen, collagen of marine sponges, jellyfish collagen and collagen from marine gastropods [5]. 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). Different experiments were performed to analyze the influence of cell density and TGF-β on osteogenic differentiation of the cells in the scaffolds. 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;

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