Synthesis and characterization of an engineered dual crosslinked hydrogel system based on hyaluronic acid, chondroitin sulfate, and carboxymethyl chitosan with platelet‐rich plasma

Abstract Hydrogels play a key role in cartilage tissue engineering (CTE) and cell transplantation. Idealy, the hydrogels designed for CTE should facilitate the formation of cartilage ECM, cilinically. However, challenges remain to engineer hydrogels with controllable responses, fast‐self recovery, and high stability. In the present study, a dual crosslinked hydrogel system based on the electrostatic and ionic bonds by mixing polymers composed of hyaluronic acid (HA), chondroitin sulfate (CS), and carboxymethyl chitosan (CMC) combined with platelet‐rich plasma (PRP) was successfully prepared. To increase the stability of hydrogel, 1‐Ethyl‐3‐(3‐dimethylamino propyl‐carbodiimide hydrochloride (EDC)/N‐hydroxy‐succinimide (NHS) was used as the crosslinking agent. The results revealed more stability of dual crosslinking hydrogel (HC54) due to proper stoichiometric ratio between polymers components and the crosslinking agent. According to the rheology results, a non‐Newtonian and viscoelastic behavior was recognized for all hydrogels and the highest mechanical property (modulus) of HC54 hydrogel was confirmed. Based on the SEM micrographs and weight loss analysis, the lowest degradation rate was observed for the HC54 hydrogel after immersion in phosphate buffered saline solution. PRP release from the hydrogel was analyzed with the Bradford assay method and a cumulative release of 50 percent during 15 days was found. Finally, the MTT assay conducted on dual crosslinked HA/CS/CMC hydrogel with and without PRP both demonstrated cytocompatibility of hydrogel while the presence of PRP enhanced the cell viability.