| Objectives The effects of graphene oxide(GO)and graphene(G)on the proliferation,differentiation and osteogenic gene expression of mouse embryonic osteoblast precursor cells MC3T3-E1 were studied.The biocompatibility and osteogenic ability of GO and G were compared,which provided a theoretical basis for the clinical application of graphene oxide.Methods 1 The GO film and G film were produced by the modified Hummers method and vacuum filtration method,and characterized and analyzed by scanning electron microscopy,Raman and X-ray photoelectron spectroscopy;2 MC3T3-E1 cells were seeded on the membrane respectively,and the differences of cell morphology and growth on different membranes were compared.In GO group,cells were seeded on GO membrane,In G group,cells were seeded on G membrane,After 24 hours,the morphology of cells on different membranes was observed by scanning electron microscope;DAPI staining was performed on 3,5 and 7 days,and the proliferation and adhesion of cells were observed by inverted fluorescence microscope;At 1,3,5,and 7 days,CCK-8 method was used to detect cell proliferation activity;at 7 days,alkaline phosphatase(ALP)detection kit was used to detect the ALP activity of the three groups of cells.3 Use Alizarin Red staining method(GO group is normal medium + GO dispersion,G group is normal medium + G dispersion,control group is normal medium)to observe the formation of cell mineralization nodules.4 The m RNA expression levels of BMP-2,Runx2 and ALP were detected by real-time fluorescent polymerase chain reaction(q RT-PCR).5 Use SPSS23.0to statistically analyze the data.Results 1 The prepared GO film and graphene film are characterized by scanning electron microscopy,Raman analysis,and X-ray photoelectron spectroscopy,which are consistent with the characteristics of GO and G.2 Using scanning electron microscope to observe the morphology of the cells on the scaffold material,the results show that the cell morphology is good and can be well attached to the surface of the material;DAPI staining results show that the number of cells is increasing at 3,5,and 7 days,GO group > G group>control group,there is a statistical difference between the pairwise comparisons(P< 0.05);CCK-8 results show that there is no significant change on the 1st day,and the absorbance values of each group on the 3rd,5th,and 7th day Change occurs,GO group >G group>control group,and the difference between the control group and the GO group is obvious(P < 0.01);at 3d,the difference between the control group and G group is not obvious(P < 0.05);at 5d,There was no significant difference between the GO group and the GO group(P < 0.05);on the 7th day,there was a statistical difference between the pairwise comparisons(P < 0.01).The ALP activity test showed that the expression level of ALP in the GO group and the G group was stronger than that of the control group,and the GO group > G group.3 The results of the Alizarin Red staining experiment showed that the size and staining degree of calcified nodules in the GO group and the G group were stronger than those in the control group,and the GO group > G group.4 The m RNA expression results of osteogenic related genes BMP-2,Runx2,ALP showed that the gene expression level of GO group was higher than that of G group and the control group,and the difference between the groups was significant(P < 0.01);ALP gene expression level G group > control Group,the difference was not significant(P < 0.05).Conclusions 1 The characterization of GO and G film meets the requirements and has good biocompatibility,which is conducive to the growth and attachment of MC3T3-E1 cells.2 Both GO and G can promote the proliferation and differentiation of mouse embryonic osteoblast precursor cells MC3T3-E1,and graphene oxide has a better promoting effect.Figure 11;Table 11;Reference 138... |
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