Chondroitin sulfate modified chitosan nanoparticles as an efficient and targeted gene delivery vehicle to chondrocytes.

Conventional treatments for osteoarthritis (OA), including drug delivery and tissue engineering approaches, could not offer a high yield of cartilage repair due to the compact and exclusive structure of cartilage. Targeted and high-efficiency delivery of gene sequences is necessary to rebalance the lost homeostatic properties of the cartilage in OA. Herein, we synthesized chitosan (CH)-chondroitin sulfate (CS) nanoparticles (NPs) as a platform for delivering gene sequences. These new nanoparticles benefit from two natural polymers that minimize the toxicity, and the presence of CS can be in favor of targeted delivery. The CAG-GFP plasmid was used as a gene sequence model, and the nanoparticles could successfully encapsulate approximately all of them in their structure. Loaded nanoparticles were characterized in terms of morphology, size, zeta potential, the efficiency of encapsulation and, DNA release pattern. Cell viability and uptake of new nanoparticles were compared to the chitosan nanoparticles and Lipofectamine. After substituting TPP with CS, NPs exhibited a significant decrease in size. In addition, there was little difference in zeta potential between nanoparticles. Furthermore, a tremendous increase in plasmid uptake and cell viability was observed by CH-CS NPs compared to CH-TPP NPs and Lipofectamine. In the final stage, the knockdown level of MMP13 was evaluated with real-time RT-PCR for confirming the potential uptake of CH-CS NPs. The results revealed cellular uptake of siRNA loaded NPs and effective knockdown of MMP13 in chondrocytes. In conclusion, CH-CS nanoparticles can be considered as a candidate for gene therapy purposes in cartilage diseases.