Curcumin-loaded Engineered Exosomes For The Intervention Study On Rheumatoid Arthritis

Objective Uncontrolled inflammatory response is a key pathological characteristic in various inflammatory diseases,which include rheumatoid arthritis.Consequently,inhibition of inflammatory response is considered to be one of the most crucial and effective strategies for the treatment of rheumatoid arthritis.The purpose of this study is to constructed a targeted drug delivery system with prolonged circulation by modifying polyethylene glycol(PEG)and hyaluronic acid on the surface of exosomes carrying the anti-inflammatory drug curcumin,and investigate its therapeutic effects on rheumatoid arthritis.Methods Mesenchymal stem cells were obtained from the femur and tibia of mice,and mesenchymal stem cells-derived exosomes were obtained by ultracentrifugation.After externally modifying targeted component hyaluronic acid and the long-circulation component PEG,as well as the internally loading anti-inflammatory drug curcumin,a targeted drug delivery system(Cur@EXs-HP)with prolonged circulation was constructed.The prolonged circulation ability was analyzed by flow cytometry and immunofluorescence.The targeted ability was analyzed by in vivo imaging system and tissue section.At the cellular level,its anti-inflammatory ability was evaluated by immunofluorescence,enzyme-linked immunosorbent assay(ELISA)and flow cytometry.After intravenous administration of Cur@EXs-HP to rheumatoid arthritis mice model,the in vivo therapeutic effects were evaluated by arthritis score,the swelling of paws,micro-CT and ELISA.This study also evaluated the safety of Cur@EXs-HP in mice.Results The experiment of prolonged circulation showed that the exosomes modifying by PEG could effectively prolong the blood circulation time.The experiment of targeting distribution showed that the modification of hyaluronic acid could enhance the accumulation of drug delivery carrier in the inflammatory site.At the cellular level,after treating with Cur@EXs-HP,the level of inflammasome and pro-inflammatory cytokines were effectively downregulated,while the level of anti-inflammatory cytokine and the anti-inflammatory phenotype(M2 macrophage)were upregulated.In the evaluation of therapeutic efficacy in vivo,the arthritis score was significantly reduced in Cur@EXs-HP group.In addition,paws swelling and bone erosion were significantly relieved,and the levels of pro-inflammatory cytokines were also significantly reduced.In the safety evaluation,mice receiving treatment with Cur@EXs-HP showed no abnormality in major organs(heart,liver,spleen,lung,and kidney),and the levels of red blood cell(RBC),platelet(PLT),and aminotransferase(AST)within the normal ranges.Conclusions Experiments in vivo and in vitro show that Cur@EXs-HP could effectively target to the inflammatory lesion site,and equipped with prolonged circulation ability.By observing the therapeutic effects of Cur@EXs-HP in rheumatoid arthritis mice model,we found that Cur@EXs-HP could exert significant anti-inflammatory effects and alleviate the progress of diseases.In addition,Cur@EXs-HP exhibited the satisfied safety.Cur@EXs-HP displayed a new strategy for anti-inflammatory therapy,and provide a new perspective for the treatment of rheumatoid arthritis.