Objectives: This study aims to investigate the effects and mechanisms of chondroitin sulfate (CS), dermatan sulfate (DS), and heparin (HEP) on chondrogenesis of murine chondrogenic cell line (ATDC5) cells and the maintenance of murine articular cartilage in vitro.
Methods: ATDC5 and articular cartilage tissue explant were cultured in the medium containing different sulfated glycosaminoglycans. Cell proliferation, differentiation, cartilage formation, and mechanism were observed using cell proliferation assay, Alcian blue staining, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blot, respectively.
Results: Results showed that HEP and DS primarily activated the bone morphogenetic protein (BMP) signal pathway, while CS primarily activated the protein kinase B (AKT) signal pathway, further promoted ATDC5 cell proliferation and matrix production, and increased Sox9, Col2a1, and Aggrecan expression.
Conclusions: This study investigated the differences and mechanisms of different sulfated glycosaminoglycans in chondrogenesis and cartilage homeostasis maintenance. HEP promotes cartilage formation and maintains the normal state of cartilage tissue in vitro, while CS plays a more effective role in the regeneration of damaged cartilage tissue.
目的: 本研究旨在探讨硫酸软骨素（CS）、硫酸皮肤素（DS）与肝素（HEP）对软骨形成细胞成软骨分化和小鼠关节软骨状态维持的作用及其机制。方法: 小鼠软骨发生细胞系（ATDC5）和小鼠关节软骨组织块在含不同硫酸化程度糖胺聚糖的培养基中培养后，使用细胞增殖试验、阿利新蓝染色、实时荧光定量聚合酶链反应（RT-qPCR）和蛋白质印迹（Western blot）分析来观察细胞增殖、成软骨分化、软骨形成、软骨组织维持的作用，并进一步探讨其潜在机制。结果: HEP和DS主要通过激活骨形态发生蛋白（BMP）信号通路，CS主要通过激活蛋白激酶B（AKT）信号通路促进软骨形成细胞的增殖能力、提高基质蛋白多糖的生成、增加Sox9、Ⅱ型胶原蛋白（Col2a1）和聚集蛋白聚糖（Aggrecan）的表达水平。结论: 本研究探讨了不同硫酸化程度的糖胺聚糖对细胞成软骨与软骨稳态的维持作用差异及其机制，HEP有助于促进软骨形成和维持软骨组织正常状态，CS在损伤软骨组织再生方面效果更好。.
Keywords: cartilage repair; chondrogenesis; degree of sulfation; sulfated glycosaminoglycan.