Effects Of IL-10 And Mechanical Stimulation On Expression Of OPG And RANKL In Human Periodontal Ligament Cells Under High Concentration Of Glucose In Vitro

Objective: To observe the effects of IL-10 on the expression of OPG and RANKL in human periodontal ligament cells(HPDLCs)under mechanical stimulation and high glucose concentration,and to explore the effect of IL-10 on alveolar bone remodeling in diabetic patients under orthodontic force.Methods:1.HPDLCs was cultured by tissue culture method and passaged,and identified the cultured cells by immunohistochemical method.2.The expression of OPG and RANKL in cell culture supernatant of each group was detected by ELISA.The cells were divided into pressure group and tension group according to the mechanical stimulation methods.The pressure group were divided into 4 groups: Group A:HPDLCs+low glucose DMEM+pressure stimulation;Group B:HPDLCs+low glucose DMEM+pressure stimulation+IL-10;Group C:HPDLCs+high glucose DMEM+pressure stimulation;Group D:HPDLCs+high glucose+pressure stimulation +IL-10.The tension group were divided into 4 groups: Group E:HPDLCs+low glucose DMEM+tension stimulation;Group F:HPDLCs+low glucose DMEM+ tension stimulation+IL-10;Group G:HPDLCs+high glucose DMEM+tension stimulation;Group H:HPDLCs+high glucose DMEM+tension stimulation+IL-10.HPDLCs in each group was were subjected to pressure or tension stimulation for 0,12,24,48,and 72 h,respectively.Meanwhile,in Group B,D,F snd H,IL-10 was used to intervene.Results:1.The cultured cells were spindle or polygon,and growth around the tissue.After passage,cells growth rapidlly and were distributed in fascicular,swirl or radial.After immunohistochemical staining,the cells were positive for Vim and negative for CK,which was consistent with the cell characteristics of mesenchymal,and the cultured cells could be determined as HPDLCs.2.Pressure group:After HPDLCs stimulated by pressure,OPG and RANKL concentration changed in the same trend.OPG firstly increased and then decreased,RANKL gradually increased with time,and OPG/RANKL ratio gradually decreased with time.Comparison between group A and B,group A and C,group C and D respectively,there were significant differences in OPG,RANKL and OPG/RANKL ratio(P < 0.05).Tension group: After HPDLCs stimulated by tension,OPG and RANKL concentration changed in the same trend.OPG gradually increased with time,RANKL first slightly increased and then decreased,OPG/RANKL ratio gradually increased with time.Comparison between group E and F,group E and G,group G and H respectively,group E and F have no significant differences in RANKL concentration(P >0.05).OPG,RANKL and OPG/RANKL ratio among other groups were significant differences(P < 0.05).Conclusion:1.HPDLCs were successfully cultured by tissue culture method.2.Under the condition of mechanical stimulation of HPDLCs by pressure or tension,high glucose environment can decrease the expression of OPG,and increase the expression of RANKL and decrease OPG/RANKL ratio in HPDLCs,bone resorption was greater than bone formation.Local application of IL-10 in periodontal tissue may improve alveolar bone resorption induced by periodontal hyperglycemic environment during diabetic orthodontics.