Characterization and Potential of a Bilayered Hydrogel of Gellan Gum and Demineralized Bone Particles for Osteochondral Tissue Engineering

Osteochondral (OC) tissue engineering (TE) is a promising strategy to regenerate acute or degenerative chondral and OC lesions. However, advancing a proper model for OC TE is still under way. Herein, a bilayer hydrogel (BH) based on gellan gum (GG) hydrogel and demineralized bone particles (DBPs) is suggested as a new model. The BH composite can be fabricated easily with a cell-friendly biomaterial and cross-linker. The BH composite was characterized by a morphological method and physicochemical aspect. The mechanical and rheological characters were further confirmed to verify its applicability in OC TE. The thermodynamic property of the composite was determined to analyze thermal stability and interaction among matrices. The bioactivity of the material was studied by treating simulated body fluid (SBF) solution for 28 days to examine the formation of crystalline structure in the BH construct. In vitro studies were carried out to study the viability and biochemical characters of the developed biomaterial. An in vivo study was performed to analyze the biocompatibility of the material and regeneration of the injured OC region implanted with BH composites. The data displayed stable physicochemical properties and mechanical characters when the DBPs were incorporated with a proper amount. The bioactivity of the DBP-loaded hydrogels displayed a high amount of apatite formation. The cytotoxicity of the fabricated material was low, which allows application in vitro and in vivo. The biochemical studies displayed a high level of alkaline phosphatase (ALP) activity and gene expression, which shows promising application of DBP-loaded GG in the bone layer of the BH model. The long-term in vivo study displayed excellent biocompatibility and great potential in the OC defected region. Overall, these results suggest the significance of combined and innovative approaches to improve the therapeutic strategies for OC regeneration, and the BH model suggested in this study can be a promising biomaterial model for OC TE.