Effect Of Strontium-containing Cross-linked Sodium Alginate Gel On Adhesion And Proliferation Of Pre-osteoblasts

Objective: To synthesize a new type of alginate gel containing ruthenium chloride by using ruthenium chloride as a cross-linking agent,to detect the physicochemical properties of glutamic acid-containing gel,and to observe and detect the MC3T3-E1 cells in mice.Adhesion and proliferation in the gel,it is proposed to use ruthenium chloride as a cross-linking agent to improve the bio-performance of the gel,and to provide a theoretical basis for the alginate injectable tissue engineering scaffold.METHODS: The cytotoxicity of alginate solution was analyzed by MTT cell proliferation and cytotoxicity test kit.The ratio of calcium ion and strontium ion was A(4:0)B(3:1)C(2:2)D(1: 3)E(0:4)prepared 5 groups of electrolytes with a final concentration of 4 g / L(Group A is the control group),a final concentration of 2% wt sodium alginate solution,p H adjusted to 7.4,will Five groups of composite precursor solutions were autoclaved and mixed with an equal amount of sodium alginate.Five sets of gel discs with a diameter of 1 cm were prepared by specific methods.The swelling ratio of each group of gel in double distilled water and phosphate buffer solution(PBS)was measured,the compressive modulus of the gel was measured,and the microstructure of each group of gels was observed by ultra-high resolution scanning electron microscopy(SEM).The leachate was collected on days 1,2,and 3,and the concentration of barium ions in the solution was measured by inductively coupled plasma spectrometry(ICP).The mouse bone-like MC3T3-E1 cell line was taken as seed cells and inoculated on the surface of each group.After 1 day of inoculation,ultra-high-resolution scanning electron microscopy was used to observe the cell adhesion morphology,and the same period of day 1,2,and 3 was inoculated.The cells were stained with fluorescein diacetate/canonidine(FDA and PI),and the proliferation activity of the cells on the gel surface was observed.The cell count was counted and the cell survival rate was calculated.The data was statistically analyzed using the Graph Pad Prism 6.0 software package.RESULTS: The results of cytotoxicity test showed that the sodium alginate powder used in this experiment had good biocompatibility and could be used as a follow-up experimental study.The swelling rate of alginate gel in double distilled water and phosphate buffer solution(PBS)increased with the increase of strontium ion concentration,which was statistically significant compared with the control group(P<0.05).Scanning electron microscopy showed with the increase of strontium ion concentration,the pores of the gel gradually increased.The gel pores of group C were uniform and uniform.The groups D and E showed that the gel fiber network was loose and the pores were uneven.The compression modulus results showed that with the enthalpy.The ion concentration increased,and the gel strength of group C was the highest.With the increase of strontium ion concentration,the gel strength showed a downward trend,but it was higher than the strength of the control gel.The ICP results showed that the cesium ion concentration of each group gradually increased.Decrease,the amount of strontium ions released into the solution decreases and is proportional to the total strontium ions initially introduced into the gel;it can be seen that the A,B,and C groups have a larger number of extensions,and the adhesion of the C group is better.Most of the microscopes and FDA/PI staining showed that the proliferation of seed cells on the gel surface of group C was significantly improved.On the third day of cell seeding,the survival,death and survival rates of the cells in each group showed that the survival rate of group C was the highest.Group E The survival rate was the lowest and the difference was statistically significant(P<0.05).Conclusion: Compared with the control group,as the concentration of strontium ions increases,the mechanical strength of the gel increases,and the swelling rate increases.Among them,group C has a more significant effect on cell adhesion and proliferation.Therefore,the composite gel formed by the incorporation of equal proportion of strontium ions provides a theoretical basis for the novel injectable bone tissue engineering scaffold.