Effects Of Injectable BG/SA Hydrogel On Proliferation And Osteogenic Differentiation Of MC3T3-E1

Objectives Periodontitis can destroy the integrity of the soft and hard tissues around the teeth,which is the main factor of adult tooth loss,and the initiating factor of periodontitis is thought to be dental plaque.The purpose of this study was to investigate the morphology of bioactive glass/sodium alginate(BG/SA)hydrogel,which was obtained by loading BG onto SA hydrogel,and its effect on the proliferation and osteogenic differentiation of MC3T3-E1 cells.It was a preliminary discussion on the feasibility of injecting BG/SA hydrogel in periodontist treatment.Methods 1 Preparation and biocompatibility of SA hydrogel for injection: 5%(w/v%)SA hydrogel was prepared and freeze-dried in vacuum.The morphology of SA hydrogel was observed by scanning electron microscope.MC3T3-E1 cells were cultured in vitro to prepare the hydrogel extract,which was diluted in the ratio of 1/2,1/4,1/8 and 1/16.The experiment was divided into six groups,the original extract group was named SA group,gradient dilution group(1/2,1/4,1/8,1/16)and negative control group(N Group).CCK-8kit,fluorescence microscope,ALP kit and alizarin red S staining solution were used to explore the effects of different concentrations of extracts on MC3T3-E1 cell proliferation and promoting bone formation.2 Preparation and biocompatibility of BG/SA hydrogels for injection.Based on the 5%(w/v%)SA hydrogels prepared in Experiment 1,BG of different quality was mixed by physical mixing method.The mass ratio of BG/SA(w/w%)was 5%,10%,15%,20%,25%,recorded as 5%,10%,15%,20% and 25% group respectively,the negative control group was SA group without BG.The microstructure of freeze-dried hydrogel was observed by electron microscope scanning.MC3T3-E1 cells are cultured in vitro and the hydrogel extracts from each group are prepared to test the cytocompatibility of BG/SA hydrogel extracts with different content and the osteogenic Ability of MC3T3-E1 cells.The detection method is the same as experiment 1.All the data were analyzed by statistical software SPSS 26.0,the test standard was set as α=0.05,and the difference was statistically significant when P<0.05.Results 1 The SA hydrogel was successfully fabricated.Scanning microscope showed that the sample was porous,the material skeleton was interlinked with each other and had a certain surface area.Although the OD value measured by CCK-8 and the absorbance value of high concentration exudate were lower than those of contrast and low concentration exudate,but the proliferation of each gradient extract was not significantly different from that of the negative control group,and there was no significant difference among the extract groups(P>0.05).Fluorescence microscope showed that the cells grew well on the lifting plate in 24 hours,and there was no significant difference in cell distribution between groups.ALP activity,SA group were slightly higher than other groups,but there was no significant difference among the groups(P>0.05).According to alizarin S staining,there was no difference in the formation of calcium nodules between the original extract group and the control group,with few nodules and light color.2 The preparation of injectable BG/SA hydrogels has successfully been achieved,and the surface sample of the BG group with a high content is smaller in size and smaller in pores than in the group with lower content.As a result of an in vitro cell experiment,BG was loaded into the SA hydrogel,and cell proliferation was promoted,and the low content BG group showed the ability to promote better multiplication compared with the group in high content,and the 10%BG group was the best promoting the cell proliferation.The ALP activity was significantly higher than that of the negative control group which loaded the BG,and the ALP activity rose with the increase in BG content,and the 25%BG Group promoted the bone capacity.Each BG/SA hydrogel group in red stained S staining showed a broad distribution of red deep colored calcium nodules.Conclusions 1 The prepared SA hydrogel is injectable,has no toxic side effects to MC3T3-E1 cells and good biocompatibility,but does not possess the ability to promote osteogenic differentiation of cells.2 The prepared injectable BG/SA hydrogel showed good cytocompatibility,with low content BG showing better cell proliferation ability and high content BG showing better cell osteogenesis ability,but cell proliferation inhibition would be expected if the BG was too high.Figure 13;Table 7;Reference 155...