Controlled domain gels with a biomimetic gradient environment for osteochondral tissue regeneration

In order to repair an osteochondral defect, it is critical to advance a bi-lineage constructive scaffold with gradient transition. In this study, we developed a simple and straightforward approach for fabricating a multi-domain gel scaffold through the assembly/disassembly of low-molecular-weight gels (LMWGs) inside a stable PEGDA network by photopolymerization. The versatility of this technology enabled to vary biological, topological, and mechanical properties through material selection and to generate a chondrogenic-osteogenic gradient transition. The multi-domain gel exhibited an increasing stiffness gradient along the longitudinal direction from the cartilage layer at approximately 20 kPa to the bone layer at approximately 300 kPa as well as spatial variation at the gradient interface. Moreover, the transitional layer with a condensed structure and intermediate stiffness prevented delamination of the contrasting layers and maintained microenvironmental differences in the upper and lower layers. The in vitro results indicated that each domain had an individual capacity to spatially control the differentiation of MSCs toward osteoblastic lineage and chondrocytic lineage. This was mainly because the mechanical and topographical cues from the respective domains played an important role in modulating the Rho-ROCK signaling pathway, whereas the blockage of ROCK signals significantly impaired domain-modulated osteogenesis and enhanced chondrogenesis. Additionally, the quantity and quality of osteochondral repair were evaluated at 4 and 8 weeks through histological analysis and micro-computed tomography (micro-CT). The results indicated that the multi-domain gels distinctly improved the regeneration of subchondral bone and cartilage tissues. Overall, the outcomes of this study can motivate future bioinspired gradient and heterogeneity strategies for osteochondral tissue regeneration. STATEMENT OF SIGNIFICANCE: The regeneration of osteochondral defects remains a major challenge due to the complexity of osteochondral structure and the limited self-repair capacity of cartilage. The gradient design to mimic the transition between the calcified cartilage and the subchondral bone plate as well as the zones of cartilage is an effective strategy. In this study, controlled multi-domain gels were fabricated through the assembly/disassembly of low-molecular-weight gels inside a stable PEGDA network by photopolymerization. The prepared multi-domain gels showed a chondrogenic-osteogenic gradient transition, which decreased the possibility of delamination and stimulated osteochondral tissue regeneration in vivo. Overall, our study promotes new strategies of bioinspired gradients and heterogeneities for more challenging applications.