Effects Of Morinda Officinalis Oligosaccharides On The Osteogenic Differentiation Of Bone Marrow Mesenchymal Stem Cells

ObjectiveTo study the effects of Morinda officinalis oligosaccharides on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells and to discuss its mechanism of action.MethodsBone marrow-derived mesenchymal stem cells were isolated and purified for culture in vitro and serial passage using a whole bone marrow adherent culture technique.The morphological changes of the bone marrow-derived mesenchymal stem cells were continuously observed under the invered microscope and the membrane antigens were deteced with flow cytometry.CCK-8 method was used to detect the effect of different concentration of Morinda officinalis oligosaccharides(0.5-5000ug/ml)in the intervention Groups and control group on bone marrow mesenchymal stem cells at 48 hours.Alkaline Phosphatase Assay Kit was used to detect the effect of morinda officinalis oligosaccharides on the activity of alkaline phosphatase in the intervention groups and control group after 7 days of osteogenic induction.Staining of alizarin red was used to detect the effect of morinda officinalis oligosaccharides on the number of mineral node in intervention groups and control group after 7 days of osteogenic induction.The gene expression of ALP,Col-?,Runx2,OCN in intervention groups and control group were detected by RT-PCR after 7 days of osteogenic induction.ResultsThe whole bone marrow adherent culture technique can be effectively used to isolate and purify SD rat bone marrow-derived mesenchymal stem cells.The cultured bone marrow-derived mesenchymal stem cells are stable in growth with active proliferation.The FCM results indicated that the obtained cells expressed CD44?CD73?CD90?CD29,but were negative for CD34?CD45.The cultured bone marrow-derived mesenchymal stem cells share the general biological characteristics of bone marrow-derived mesenchymal stem cells.The OD values showed that there were no inhibitory or cytotoxic effects of morinda officinalis oligosaccharides on BMSCs at doses(0.5-500 ug/ml).But the dose 5000ug/ml showed cytotoxic effects on BMSCs.0.5-500ug/ml morinda officinalis oligosaccharides inhibited the expression of ALP in the BMSCs on day 7,compared with the untreated control.Little plaques of calcified extracellular matrix could be detected in 5 and 50 ug/ml morinda officinalis oligosaccharides on day 7,compared with the untreated control.The gene expression of ALP,Col-?,Runx2,OCN were inhibited by morinda officinalis oligosaccharides at the concentration of 5 and 50 ug/ml,compared with the untreated control.ConclusionMorinda officinalis oligosaccharides might inhibite the osteogenic differentiation of bone marrow mesenchymal stem cells.