Farcasanu Marc V, Ruiz Thais de Las Heras, Brito Francesca M Johnson de Sousa, Soul Jamie, Coxhead Jonathan, German Matthew J, Young David A, Ferreira-Duarte Ana M, Piróg Katarzyna A
Biosciences Institute, Newcastle University, Newcastle upon Tyne, UK.
Computational Biology Facility, University of Liverpool, Liverpool, UK.
Biotechnol Bioeng. 2025 Sep;122(9):2574-2591. doi: 10.1002/bit.29026. Epub 2025 May 25.
Hyaline cartilage is a dense avascular tissue with low regenerative potential, present at the ends of the diarthrodial joints and in the cartilage growth plate. Skeletal diseases often result from extracellular changes in this tissue; however, studies of these are hindered by the tissue complexity, the difficulty in obtaining human material, and the cost of generating animal models. Recent developments in tissue engineering are opening possibilities to develop mechanoresponsive zonally stratified models of cartilage in vitro. In this study, we optimized a 3D model of cartilage using chondroprogenitor cells cultured for 21 days in 2% agarose hydrogel constructs with daily dynamic compression. Our hydrogel constructs developed pericellular matrices with nanostiffness comparable with native murine tissue and showed increased production of extracellular matrix components and expression of chondrogenic and differentiation markers. Daily dynamic compression resulted in progressive increase in mechanoresponsive gene expression and promoted a juvenile cartilage phenotype, decreasing expression of dedifferentiation and cartilage degradation markers. Our study highlights the potential of hydrogel-enhanced chondrogenesis and proposes an adaptable and scalable in vitro model to study mechanoresponses, intracellular signals, and pericellular matrix involvement in cartilage development and disease.
透明软骨是一种致密的无血管组织，再生潜力低，存在于滑膜关节的末端和软骨生长板中。骨骼疾病通常源于该组织的细胞外变化；然而，由于组织复杂性、获取人类材料的困难以及生成动物模型的成本，对这些疾病的研究受到阻碍。组织工程的最新进展为在体外开发机械响应性分层软骨模型开辟了可能性。在本研究中，我们使用软骨祖细胞在含2%琼脂糖水凝胶构建体中培养21天，并每日进行动态压缩，优化了一种软骨三维模型。我们的水凝胶构建体形成了具有与天然小鼠组织相当的纳米硬度的细胞周基质，并显示出细胞外基质成分的产量增加以及软骨生成和分化标志物的表达。每日动态压缩导致机械响应基因表达逐渐增加，并促进了幼年软骨表型，降低了去分化和软骨降解标志物的表达。我们的研究突出了水凝胶增强软骨生成的潜力，并提出了一种可适应且可扩展的体外模型，用于研究机械响应、细胞内信号以及细胞周基质在软骨发育和疾病中的作用。
