Synthesis Of Chondroitin Sulfate Magnesium And Its Anti-osteoarthritis Effects

Objectives:1.To synthesis chondroitin sulfate magnesium and investigate its physical and chemical properties;2.To explore the effect of chondroitin sulfate magnesium on OA cartilage;3.To preliminarily explore the repair effect of chondroitin sulfate magnesium on the OA animal model.Methods:1.The sodium ions was adsorbed by ion exchange resin and the magnesium ions then combined with the chondroitin sulfate;2.The SEM/EDS was taken to detect the surface morphology of chondroitin sulfate magnesium and the distribution of magnesium ions on the surface of chondroitin sulfate magnesium.The ICP-MS was used for detecting the content of magnesium and the FTIR was used to analyze the functional groups that magnesium ions might bind to;3.Cell proliferation assay was taken to detect the effect of chondroitin sulfate magnesium on the proliferation of OA chondrocytes.The anti-apoptosis effect of chondroitin sulfate magnesium on OA chondrocytes was analyzed by Flow cytometry assay.RT-qPCR was taken to detected the effects of chondroitin sulfate magnesium on OA chondrocytes at the transcriptional level;4.C57 mice undergone DMM surgery to construct OA animal model.After 4 weeks or 6 weeks injecting of chondroitin sulfate magnesium into the joint,the mice were sacrificed by neck dissection.Additionally,the knee was taken for Micro-CT examination and histological staining.Results:1.We successfully synthesis the chondroitin sulfate magnesium,in which magnesium ions,bonded to chondroitin sulfate by sulfonate group and carboxyl group,successfully replaced sodium ions and distributed evenly on the surface of chondroitin sulfate.The content of magnesium ions in the product increases with the amount of magnesium chloride;2.MgCS can promote the proliferation and inhibit the apoptosis rate of chondrocytes in osteoarthritis(OA).RT-qPCR analysis showed that MgCS significantly increased the expression of COLII in OA chondrocytes and decreased the expression of Il-1?and iNOS.In addition,it can down-regulate the expression of p53 and up-regulate the expression of Bcl-2;3.Animal experiments showed that chondroitin sulfate magnesium could increase the cartilage thickness of OA mouse model and inhibit the destruction of subchondral bone in the early stage of OA.Conclusion:Compared with chondroitin sulfate,MgCS has a certain anti-osteoarthritis effect including increasing of the thickness of OA cartilage and reducing the bone loss on the subchondral bone in the early stage of OA.Hence it is expected to be a new choice for the osteoarthritis treatment in the future.