The Effects On Proliferation And Differentiation To Osteoblast Of SD Rat Bone Mesenchymal Stem Cells In Vitro Of Quercetin

Objective:In my study,I try to establish cultivation system in vitro of bone marrow stromal cells from SD rats,observe their growth and characteristics and investigate the effects ofquercetin on their proliferation and osteoblast-differentiation ability.And to investigate the expression of BMP-2 mRNA in induced procedure with quercetin.In order to provide a probable action mechanism of quercetin prevent and cure OP in stem cell aspect.Then we can further have knowledge of the Chinese traditional medicine and put it into a brand-new application in bone tissue engineering.Methods:1.MSCs was isolated and purified by differential time adherent method and to obtain MSCs maximally;2.MSCs was identified by morphologic characteristics,surface antigens detected by flow cytometer and differentiation into osteoblasts,adipocytes and chondrocytes;3.MTT method was used to confirm the most effective concentration of quercetin and cytokines on the proliferation of MSCs;Groups were divided:The blank group,classics group(DEX+β-GP+VitC) and quercetin group(induced by the most effective Concentrations );The ALP activity and the number of mineralized nodesls were measured to indicate the osteoblast function of different groups.4.Semi-quantitative analysis was used to detect the expression of BMP-2 during osteodifferentiation through RT-PCR method.Results:1.Primary cultured MSCs adhered to plastic surface within 24 hours and reached confluence within 11-15days.It looks like fibroblast.MSCs have great abilityto prolifera.2.OD values of MTT shows quercetin groups of 5μmol/L and 10μmol/L were higher than the blank groups.3.Results of ALP staining for ALP-positive cell counts showed quercetin of 10μmol/L addition had a definit induction to osteoblast differentiation compared with the blank withsignificant difference(P＜0.05).4.ALP activity values in MSCs 15 days of quercetin groups were higher than that in blank groups.And there were significant Differences between classics group and quercetin group(P＜0105).The average number of mineralized nodes in MS Cs 21 days of quercetin groups were higher than that in blank group.But there were no significant differences between classics group and quercetin group(P＞0.05).5.The expression of the BMP-2 mRNA with quercetin was stronger than bank group.There were significant differences between quercetin group with classics group and the blank group(P＜0.05).And there were significant differences between quercetin group and classics group(P＜0.05).Conclusion:MSCs can be isolated from rat bone marrow by the method of Differential attachment.The obtained MSCs are pure and have great ability to proliferate;In this study,we demonstrated that the Quercetin could stimulate the proliferation and osteogenic differentiation of bone marrow mysenchymal stem cells.In the osteoblast-differentiation progress,Quercetin in the concentration of 10μmol/L can enhance the expression of BMP-2 And mRNA.Perhaps the effect of Quercetin on BMSCs is one of cytology mechanisms of promoting the concrescence of bone fracture and preventing and treating osteoporosis.