Pilot Study Of The Effect Of Micromolecule Drug On The Proliferation And Osteogenesis Differentiation Of Human Periodontal Ligment Stem Cells And Gingival Mesenchymal Stem Cells

Experiment1:Effects of SB431542 on the proliferation and osteogenesis differentiation of human periodontal ligment stem cellsObjective:To investigate the effects of SB431542 on the proliferation and osteogenesis differentiation of human periodontal ligament mesenchymal stem cells(hPDLSCs)in vitro.Methods:1.hPDLSCs were cultured in vitro.2.The second generation hPDLSCs examined by flow cytometry for the detection of stem cell markers CD73,CD90,CD34,CD45.hPDLSCs were randomized into 3 groups:Test group were cultured by osteogenic induction medium with 1μMSB431542,whereas the control group received no SB431542.MSC conventional medium was added in the blank group.3.The proliferation of hPDLSCs were observed by CCK8 kit,subsequently plotting cell growth curve.4.hPDLSCs osteogenic differentiation was examined through matrix mineralization by alizarin red staining.5.mRNA of hPDLSCs expression of ALP,OCN,OPN,COLLEGE 1 was detected by RT-PCR.Results:hPDLSCs was positive for the expression of CD73,CD109 and negative of CD34 and CD45.Compared to control group,OD value is higher in test group(P>0.05)which means the proliferation of test group is faster.The expression of osteogenic revalent gene were significantly higher(P<0.05),and test group showed more and larger mineralized nodules.Conclusion:1μMSB431542 could enhance the osteogenic differentiation of hPDLSCs in vitro.Nevertheless,there was no effect on the proliferation of hPDLSCs.Conclusions:This research provides a new thought to the application of hPDLSCs in bone regeneration clinical translational medicine.Experiment2:Effects of SB431542 on the proliferation and osteogenesis differentiation of human gingival mesenchymal stem cells.Objective:To detect the effects of SB431542 on the proliferation and osteogenesis differentiation of human gingival mesenchymal stem cells(hGMSCs)in vitro.Methods:1.hGMSCs from young implant patients were cultured in vitro.2.The second generation hGMSCs examined by flow cytometry for the detection of stem cell markers CD73,CD90,CD34,CD45.hGMSCs were randomized into 3 groups:Test group were cultured by osteogenic induction medium with 1μMSB431542,whereas the control group received no SB431542.MSC conventional medium was added in the blank group.3.The proliferation of hGMSCs were observed by CCK8 kit,subsequently plotting cell growth curve.4.mRNA of hGMSCs expression of ALP,OCN,OPN was detected by RT-PCR.5.hGMSCs osteogenic differentiation was examined through matrix mineralization by alizarin red staining.Results:hGMSCs was positive for the expression of CD73,CD 109 and negative of CD34 and CD45.Compared to control group,OD value is higher in test group(P>0.05)which means the proliferation of test group is faster.The expression of osteogenic revalent gene(OCN、OPN、ALP、COLLEGE1)were significantly higher(P<0.05),and test group showed more and larger mineralized nodules.Conclusion:In vitro,1μMSB431542 could enhance the osteogenic differentiation of hGMSCs.Nevertheless,there was no effect on the proliferation of hGMSCs.Discussion:This research provides a new thought to the application of hGMSCs in bone regeneration clinical translational medicine.