The Effect Of Myricetin On The Proliferation Ability,Osteogenic Differentiation And Secretion Of IL-1 Beta,IL-6 Of Stem Cells From Human Exfoliated Deciduous Teeth

Objective The stem cells from human exfoliated deciduous teeth(SHED)was isolated and cultured by tissue explant method,to investigate the effect of myricetin on proliferation and osteogenic ability of SHED under non inflammatory conditions.And the effects of myricetin on the proliferation and secretion of cytokines in SHED under inflammatory conditions induced by porphyromonas endodontalis lipopolysaccharides.Methods Tissue block method was used to culture SHED.Inverted microscope was used to observe the morphological characteristics of SHED.Flow cytometry was used to detect the expression of SHED surface marker CD29 and CD105.And the expression of CD34 and CD45,the expression of immunofluorescence detection of cytokeratin and vimentin was also detected.MTS assay was used to detect the effects of different concentrations of myricetin on the proliferation ability of SHED,such as 1 ?mol/L,5 ?mol/L,10 ?mol/L,15 ?mol/L,20 ?mol/L,50 ?mol/L and 100 ?mol/L.ALP assay was used to detect the effects of different concentration of myricetin on deciduous dental pulp stem cells in early osteogenic ability,such as 1 ?mol/L,5 ?mol/L,10 ?mol/L,15 ?mol/L and 20 ?mol/L.The none myricetin group as control group.For experimental groups,we set up as following: 1),200mg/L LPS + DMEM;2),200mg/L LPS + 20?mol/L myricetin + DMEM;3),200mg/L LPS + 100mg/L metronidazole + DMEM.Only DMEM was used as scramble.OD values of SHED were measured in 24 h,48 h and 72 h after giving different interventions.Also,ELISA was introduced to detect the protein level of IL-1? and IL-6 in culture supernatant after culture for 48 h.Results The inverted microscope showed that primary cultured SHED isolated from tissues in 9-12 days.Most of the cells were long fusiform or triangular.The cells extending from both ends of the cell were elongated,some of them forked.The cytoplasm was homogeneous.The nucleus was centered,round or oval.Other cells were polygonal and smaller.Immunofluorescence staining showed that the fourth generation of human periodontal ligament cells showed strong positive expression of vimentin and negative staining of keratin.The growth curve of the fourth generation of periodontal ligament cells showed an inverted S shape.After 30 days of mineralization induced,a large number of mineralized nodule formed,calcium salt deposited,and alizarin red staining showed red.Flow cytometry showed that the expression of CD29 and CD105 was positive,and the expression of CD34 and CD45 was negative.Effect of myricetin on proliferation and osteogenic ability of SHED.In 24 h,48 h and 72 h,10 ? mol/L,15 ? mol/L and 20 ?mol/L myricetin promoted the proliferation of SHED.In 3 d,5 d,7 d,myricetin promoted the ALP activity of SHED at 10?mol/L,15?mol/L and 20?mol/L,and was positively correlated.Effects of myricetin on proliferation and secretion of cytokines in SHED under porphyromonas endodontalis lipopolysaccharides.20 ?mol/L myricetin reduced the inhibitory effect of porphyromonas endodontalis lipopolysaccharide on the proliferation of SHED.20 ?mol/L myricetin inhibited the secretion of IL-1? and IL-6 by SHED under porphyromonas endodontalis lipopolysaccharides.Conclusion Appropriate concentration of myricetin promoted the proliferation and osteogenic differentiation ability of SHED.20?mol/L myricetin reduced the inhibitory effect of porphyromonas endodontalis polysaccharides on the proliferation of SHED.20?mol/L myricetin inhibited the secretion of IL-1? and IL-6 by SHED under porphyromonas endodontalis lipopolysaccharides.