Enhanced Chondrogenic Potential and Osteoarthritis Treatment Using Cyaonoside A-Induced MSC Delivered via a Hyaluronic Acid-Based Hydrogel System

Osteoarthritis (OA) is a prevalent degenerative joint disease that significantly impacts the quality of life in the elderly. Traditional Chinese medicine, particularly Medicinal Cyathula Root and its active component Cyaonoside A (CyA), has been utilized to treat OA by promoting chondrocyte proliferation, inhibiting inflammatory factors, and maintaining joint homeostasis. Concurrently, mesenchymal stem cells (MSC) derived from placental umbilical cord, bone marrow, and adipose tissue have gained attention for their potential in OA treatment due to their chondrogenic differentiation capabilities. This study explored the therapeutic synergy of CyA and MSC for enhanced cartilage regeneration. Optimal chondrogenic differentiation was achieved by treating MSC with 0.5 mg/mL CyA for 3 days, significantly increasing the expression of key cartilage-specific genes ACAN, COL2A, and SOX9. Comparative gene expression and pathway analyses revealed that CyA-induced MSC (C-MSC) modulate critical signaling pathways, including TGF-β, PI3K-Akt, and Wnt, demonstrating their potential in cartilage repair. Furthermore, C-MSC-derived exosomes exhibited superior anti-inflammatory and anti-apoptotic effects compared to MSC-derived exosomes in IL-1β-treated human chondrocytes, enhancing chondrogenic gene expression and reducing cartilage degradation. To enable targeted delivery, a novel injectable hydrogel system (HAMA@C-MSC) was developed using methylacrylated hyaluronic acid (HAMA). This hydrogel facilitated uniform cell distribution, maintained structural integrity, and demonstrated excellent biocompatibility and biosafety, with no cytotoxic or hemolytic effects. In vivo studies using a rat destabilization of the medial meniscus OA model confirmed that HAMA@C-MSC significantly improved cartilage structure, enhanced chondrocyte regeneration, and restored collagen integrity, outperforming other treatment groups as validated through imaging, histology, and molecular analyses. These findings highlight HAMA@C-MSC as a promising therapeutic strategy for OA, leveraging the synergistic effects of C-MSC and advanced hydrogel technology to achieve enhanced cartilage regeneration and joint protection.