| Objective:In this paper we investigate the effects of astragalus polysaccharide(APS)on biological behavior of human gingival fibroblasts(HGF).Its effects of fibroblast proliferation,migration and type I collagen secretion were discussed and compared with Epidermal Growth Factor(EGF).A new research method combining traditional Chinese and western medicine is proposed for how to promote gingival tissue increment and accelerate wound healing.Method:1 ? HGFs were primary cultured,subcultured,frozen and thawed.The gingival tissue were circular separated from dental implant surgery patients.2 ? Cells were identified by immunocytochemical analysis and immunofluorescence analysis.3?Drawing cell-growth curve and observing cell-growth law.4?The effects of different concentrations of APS on proliferation of HGFs were determined by CCK-8 assay in 12 h,24h,48 h respectively.(The concentrations of APS were divided into A group 0?g /ml,B group12.5?g /ml,C group 25?g /ml,D group 50?g /ml,E group 100?g /ml,F group200?g /ml,G group 400?g /ml.)5?The effects of different concentrations of APS on the total content of protein synthesized of HGFs were determined by Coomasssie brilliant blue assay in 72 h.6?The effects of different concentrations of EGF on proliferation of HGFs were determined by CCK-8 assay in 12 h,24h,48 h respectively.7 ?The effects of APS and EGF on the migration of HGFs were determined by wound healing model in 12 h,24h,48 h respectively.8?ELISA assay detected the protein expression of Collagen?after using the APS and EGF to treat the HGFs for 24 h,48h or 72 h.9 ? The experimental results statistics were used by SPSS17.0software,one way analysis of variance.Results:1 ? HGFs were primary cultured by tissue explant culture technique.The cells showed long spindle,angular or star appearance.The cell nucleus was in the center of the cell and showed round or oval appearance.The cells grew well.2?The cells tested positive to anti-vimentin antibody and negative to anti-keratin antibody.The results showed that the cells were derived from mesenchyme rather than epithelium.3?Drawing cell-growth curve and it was similar to “s” shape.The cells grew slowly in the first day to second day,logarithmically in the third day to fifth day and flat in the sixth day to seventh day.The result was consistent with general cell growth.4?The CCK 8 results show that the experimental groups and controlgroup all can promote proliferation.The B group compared with control group was not significant(P > 0.05).The C group and D group promoting cell proliferation were better than that of control group in 48 h and 72 h(P < 0.05).The E group,F group and G group promoting cell proliferation were better than that of control group in 24 h,48 h and 72 h(P <0.05).The F group showed the most significant effect on promoting cell proliferation in 72h(P < 0.001).The proliferation ability of G group was less compared with F group.5?Using the APS to treat the HGFs for 72 h then it shows that the difference of the total content of protein synthesized between B group and control group was not significant(P>0.05).The total content of protein synthesized of the C group to G group were higher than that of control group(P<0.05).The expression of F group was most.6? Compared with the control group,the EGF concentration was 40ng/ml and 80 ng/ml,which significantly promoted cell proliferation in48 h and 72h(P<0.05),and 80 ng/ml EGF had the most significant effect on cell proliferation in 48h(P<0.001).7? Scratch test showed that in 24 h the wound healing rate of there groups had no obvious difference(P > 0.05).In 48 h wound healing rate of APS group and EGF group were higher than that of ?-MEM group(P <0.01).The majority of cell were disruptive and the results can't calculate.8 ? Elisa assay result showed that the type ?collagen expression of APS group was higher than that of control group in 24 h,48h and 72 h.(P<0.05).The type? collagen expression of EGF group was higher than that of control group in 48 h and 72h(P<0.01).Conclusion:APS could promote HGFs proliferation,the total content of protein synthesized increase and type?collagen expression.Also it promotes the cell migration and accelerates wound healing. |
PDF Full Text Request