| BackgroundCelastrol.derived from traditional chinese medicine Tripterygium root barkis.is one of the natural products with a variety of biologically activities possessing a strong antioxidant,anti-rheumatic,and anti-angiogenesis effect of cancer.However,its water solubility is poor,and it has toxic side effects, which limits its clinical application.Considerable efforts need to be made to solve the poor solubility and toxic effects of Celastrol.PLGA,poly (lactic-co-glycolic acid),is a function of biodegradable organic polymer,which has good biocompatibility, good film-forming properties into a capsule,be non-toxic,and is widely used in pharmaceutical,medical engineering materials and modern industries.FDA has approved in the United States that PLGA was included into the United States Pharmacopoeia,which was formally used as pharmaceutical excipients.Chitosan,with good biocompatibility,low toxicity,is widely used in the field of medicine.However,its solubility is poor.Carboxymethyl chitosan fcmCHI) is carboxymethyl chitosan derivative with water-soluble improved,no antigenicity,good in adhesion,which is widely used in the field of Medicine. AimTo overcome the disadvantages of Celastrol and expand its clinical application, we developed cmCHI/PLGA/Celastrol nano system.Celastrol-loaded carboxymethyl chitosan PLGA nanoparticles were successfully synthesized with a solvent evaporation method.Then the obtained nanoparticles were characterized by scanning electron microscope (SEM), differential scanning calorimetry (DSC),X-ray powder diffractometry (XRD) and fourier transform infrared (FTIR) techniques.Finally,their characteristics of pharmacy were analyzed.Finally,the in vivo and in vitro efficacy experiments were carried out.Methods1.Preparation and characterization:cmCHI/PLGA/Celastrol nanoparticles were prepared by using the emulsion solvent evaporation method.We used DSC,XRD,FTIR to characterize the physicochemical properties of the prepared nanoparticles.Then,the drug loading efficiency,encapsulation efficiency and in vitro release behavior of nanoparticles were analyzed.2.In vivo effects of nanoparticles:The arthritis models of mice induced by ovalbumin were prepared, and then we use the drug to them.At last.we evaluated the joint swelling score and investigated the animal joints via X optical digital imaging system of Faxitron MX-20.3.In vitro effects of nanoparticles:We selected Caco-2, SW982 and Raw264.7 cell lines, and then carried out the cellular uptake,cytotoxicity.flow cytometry apoptosis of nanoparticles,and ELISA inflammatory cytokine test.Results1 The SEM images showed the obtained NPs had regular spherical shape with mean diameter of 80 nm.The DSC,XRD,FTIR results revealed that the novel nanoparticles were successfully synthesized.The encapsulation efficiency and drug loading content is 83.33% and 7.78%, respectively. In vitro release study demonstrated a sustained release of the encapsulated drug from the NPs.2.The results suggested that the in vivo anti-inflammatory effect of the prepared nanoparticles is stronger than that of celastrol. The results of Cellular uptake test showed that nanoparticle materials can promote the accumulation of tripterine in cells in a time-dependent manner. Cytotoxicity test indicated that nanoparticles can reduce the toxicity of tripterine.Flow cytometry suggested nanoparticle materials can promote apoptosis of synovial cellsin when the concentration is higher.ELISA detection of inflammatory factors showed that Celastrol can significantly reduce the expression of TNF-α and IL-6 induced by LPS in SW982 cells and inhibit the expression of TNF-α,G-CSF, Rantes, GM-CSF induced by LPS in Raw 264.7 cells.ConclusionThe synthesized nanoparticles can significantly improve the solubility of tripterine and decrease its toxicity, and the in vivo,in vitro anti-inflammatory effects have been raised in a certain degree.So the prepared cmCHI/PLGA nanoparticles would be a reliable and effective drug delivery systemon for some insoluble drugs.
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