Diogo Gabriela S, Carneiro Filipa, Freitas-Ribeiro Sara, Sotelo Carmen G, Pérez-Martín Ricardo I, Pirraco Rogério P, Reis Rui L, Silva Tiago H
3B's Research Group, I3Bs - Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017 Barco, Guimarães, Portugal; ICVS/3B's - PT Government Associate Laboratory, Braga/Guimarães, Portugal.
Instituto de Investigaciones Marinas (CSIC), Eduardo Cabello 6, 36208 Vigo, Spain.
Mater Sci Eng C Mater Biol Appl. 2021 Jan;120:111587. doi: 10.1016/j.msec.2020.111587. Epub 2020 Sep 30.
Representing a strategy of marine by-products valorization, based on isolation of biocompounds and assessment of biomedical applicability, the potential of blue shark (Prionace glauca (PG)) skin collagen to induce chondrogenic differentiation of human adipose stem cells (hASC) was investigated, with and without exogenous stimulation. For that, a cryogelation method was applied to produce highly interconnected porous 3-dimensional (3D) constructs made of collagen and collagen:hyaluronic acid (20:1). In vitro studies reveal that hASC adhere abundantly to the constructs which then suggests the early chondrogenic differentiation of those cells. These findings are supported by the mRNA expression encoding chondrogenic-related markers like Coll II and Sox-9 that are markedly upregulated at an early stage for both conditions, with and without exogenous stimulation. The introduction of hyaluronic acid (Hya) seems to play a crucial role at later time points, as shown by the evident immunodetection of aggrecan (ACAN), even without exogenous stimulation. It is hypothesized that the PG collagen itself can support chondrogenic differentiation at early time points, but exogenous stimulation is required to ensure phenotype maintenance. The present work highlights the relevance of using blue shark collagen biopolymer as a building block to produce highly effective temporary matrices for cartilage applications.
基于生物化合物的分离和生物医学适用性评估，本研究代表了一种海洋副产品增值策略，研究了蓝鲨（Prionace glauca，PG）皮肤胶原蛋白在有无外源性刺激的情况下诱导人脂肪干细胞（hASC）软骨分化的潜力。为此，采用冷冻凝胶化方法制备了由胶原蛋白和胶原蛋白：透明质酸（20:1）制成的高度互连的多孔三维（3D）构建体。体外研究表明，hASC大量粘附于构建体，这表明这些细胞早期发生软骨分化。这些发现得到了编码软骨生成相关标志物（如Coll II和Sox-9）的mRNA表达的支持，在有无外源性刺激的两种情况下，这些标志物在早期均显著上调。透明质酸（Hya）的引入似乎在后期起着关键作用，即使没有外源性刺激，聚集蛋白聚糖（ACAN）的明显免疫检测也表明了这一点。据推测，PG胶原蛋白本身可以在早期支持软骨分化，但需要外源性刺激来确保表型维持。本研究强调了使用蓝鲨胶原蛋白生物聚合物作为构建块来生产用于软骨应用的高效临时基质的相关性。
