| Objective: The purpose of this study is adding copper element to the conventional medical cobalt-based alloy material and useing selective laser melting technology to produce the new copper-containing cobalt-based alloy.Then explore its biology compatibility and biofunctionalization.Materials and methods: Use two new copper-containing cobalt-based alloys(Co Cr W-mix,Co Cr W-alloy)produced by 3D printing technology as experimental group,conventional medical products without copper elements produced by 3D printing technology cobaltbased alloy as control group.Co-cultured PDLF,MC3T3-E1,and HUVEC with the experimental materials.Then using RTCA technology,cck8 method,phalloidin cell staining experiments to detect the effect of materials on cell proliferation,apoptosis,cell adhesion.HUVEC cells using in transwell migration assay,small tube experiments and PCR to detect the migration ability of cells on the material.Mouse embryonic osteoblasts(MC3T3-E1)were used in ALP activity experiment,the alizarin red staining experiment and PCR technology to detects their ability of promote bone formation.Results: 1.The results of CCK-8 test showed that the new 3D printing copper-cobalt-based alloy materials has good biocompatibility with PDLF,MC3T3-E1,and HUVEC.Their cell proliferation rate was greater than 75%,belonging to level 1 Cytotoxicity.2.The results of RTCA experiments showed that after 120 hours,all 3 cells co-cultured with experimental materials were well cultured,and the cell growth curve did not block or decrease.In the same situation,the growth curves of the experimental group and the control group were basically the same.3.The results of apoptosis experiment showed that there was no significant difference between the apoptosis rate of new copper-containing cobalt-based alloy and the apoptosis rate of conventional medical cobalt-based alloys.4.After Staining PDLF,HUVEC and MC3T3-E1 cells with phalloidin for 4 hours and 24 hours.We can observe the cells of all groups at each time point under the microscope were similar to each other and no significant differences were observed.5.The results of Transwell migration experiment showed that the number of cell migration in the two experimental groups was higher than that in the control group,but there was no significant difference between the experimental group and the control group(P>0.05).The results of small tube experiments showed that the number of tube and length of tube formed in the experimental group were significantly higher than those in the control group(P<0.05).7.The results of PCR experiments showed that compared with the control group,the expression of cell-related tubule genes in the experimental group was significantly higher than that in the control group(P<0.05).8.The results of ALP activity test showed that there was no significant difference in ALP activity between the Co Cr W-alloy group and the Co Cr W group,but the ALP activity of Co Cr W-mix group was significantly higher than that of the mixed powder group.The alizarin red staining experiment showed that the formation of calcium nodules in the Co Cr W-alloy group was slightly higher than that in the Co Cr W group and the Co Cr W-mix group,but there was no significant difference(P>0.05).10.The results of PCR experiments showed that the expression of bone-related genes in the Co Cr W-alloy group was significantly higher than that in Co Cr W-mix group(P<0.05),but there was no significant difference in the expression of bone-related genes between the Co Cr W-alloy group and the Co Cr W group.Conclusion: 1.Compared with the control group,the experimental group did not have any obvious cytotoxicity to PDLF,HUVEC and MC3T3-E1.All test indicators meet the standards for the use of biomedical materials.2.Co Cr W-alloy have significant provascularization effects compared to Co Cr W material.3.The effect of Co Cr W-alloy on osteogenesis is not obvious,but Co Cr W-alloy osteogenesis is better than Co Cr W-mix. |
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