Platelet-rich plasma-hyaluronic acid/chondrotin sulfate/carboxymethyl chitosan hydrogel for cartilage regeneration

In this study, the chondrogenic potential of hyaluronic acid/chondrotin sulfate/carboxymethyl chitosan hydrogels with adipose-derived mesenchymal stem cells (ADMSCs) was evaluated. Here, hyaluronic acid, chondrotin sulfate, and carboxymethyl chitosan were used as the substrate for cartilage tissue engineering in which the hydrogel is formed due to electrostatic and hydrogen bonds through mixing the polymers. Because of the instability of this hydrogel in the biological environment, 1-ethyl-3-(3-dimethylaminopropyl-carbodiimide hydrochloride/N-hydroxy-succinimide was used as a crosslinker to increase the hydrogel stability. The hydrogels showed reasonable stability due to the combined effect of self-crosslinking and chemical crosslinking. The cells were treated with the prepared hydrogel samples for 14 and 21 days in nondifferentiation medium for evaluation of the cellular behavior of ADMSCs. Gene expression evaluation was performed, and expression of specific genes involved in differentiation was shown in the crosslinked hydrogel with platelet-rich plasma (PRP) (H-EN-P) had increased the gene expression levels. Quantification of immunofluorescence intensity indicated the high level of expression of SOX9 in H-EN-P hydrogel. Based on the results, we confirmed that the presence of PRP and the similarity of the hydrogel constituents to the cartilage extracellular matrix could have positive effects on the differentiation of the cells, which is favorable for cartilage tissue engineering approaches.