Dynamic loading enhances chondrogenesis of human chondrocytes within a biodegradable resilient hydrogel

Hyaline cartilage in the knee joint is a soft tissue that is both stiff and elastic, which raises unique challenges in developing scaffolds for the repair of cartilage injury. In this study, we mixed poly-d,l-lactic acid/polyethylene glycol/poly-d,l-lactic acid (PEG-PDLLA-DA) with polycaprolactone-poly(ethylene glycol)-polycaprolactone (PEG-PCL-DA) with the aim to create a cartilage-like hydrogel. Results indicated that the hydrogel made from PEG-PDLLA-DA/PEG-PCL-DA (50/50) was biodegradable and resilient, able to bear compressive loads with strains up to 50%. Human chondrocytes maintained high viability after being seeded in the hydrogel and underwent robust chondrogenesis upon stimulation. The application of dynamic compressive loading further promoted the generation of cartilage matrix and increased the compressive moduli of engineered cartilage tissues. Then engineered cartilage tissues, with or without being stimulated by dynamic loading, were implanted subcutaneously in mice, and results showed that the cartilage matrices and chondrocyte phenotypes were well preserved. Lastly, we conducted the mechanistic study to understand how dynamic loading influenced chondrogenesis. Specifically, the levels p-Erk and p38 kinases were found to remarkably increase on day 1 upon dynamic compressive loading, decrease on day 3, and then slightly elevate on day 7. In comparison, the expression of YAP and RhoA peaked on day 3 after mechanical loading. Levels of PIEZO1 and TRPV4 protein increased with the extension of dynamic loading culture time. Taken together, this newly developed resilient hydrogel represents a robust scaffold for cartilage regeneration. Moreover, based on the time their levels reach the peak, three groups of proteins are identified in mediating chondrocyte response to dynamic loading, which has not been previously reported.