The Influence Of Bio-active Zirconia Coating On The Adhesion,Proliferation And Mineralization Of MC3T3-E1

Objective:This study was to investigate the influence of arginine-glycine-aspartic acid(RGD)and magnesium(Mg+)decorated zirconia coating on the adhesion,proliferation and mineralization of MC3T3-E1.Methods:In this study,scanning electron microscope(SEM)was employed to observe the surface morphology of RGD and Mg+ decorated zirconia,and the surface roughness and wettability were measured.MC3T3-E1 cells were seeded on the samples,the actin bundles and focal adhesion were ovserved by immunofluorescence.In addition,the cell nucleus were dyed after the incubation of 24.48.72 hours on samples to perform cell counting,evaluating the proliferation of cells.RNA was extracted from cells after the 5 days' incubation to detect the expression of integrin.Total protein was extracted after the osteogenic induction of 7.14?21 days and the expression of ALP and OCN was measured by Western blot.The mineralized nodules were observed by SEM to visualize the mineralization of MC3T3-E1.Results:The nano-scaled surface character could be found on Mg+ decorated zirconia samples,its surface roughness(1.76±0.18?m)was higher than that of regular zirconia sample(1.73±0.15?m)(P>0.05).The contact angle of RGD connected zirconia sample was 26.4±1.64° and the number in regular zirconia was 65.3±2.32°(p<0.05).Cells on RGD and Mg+ decorated zirconia samples possessd better spreading property and more focal adhesion could be seen.The higher expression of ITGA5 and ITGAv(p<0.05)was found on RGD connected zirconia by RT-PCR.Mg+ decorated zirconia possessed higher expression of ITGA1 and ITGA2(p<0.05)and Western blot results proved that the introduction of Mg+ heighten the expression of OCN of osteoblasts in osteogenic incubation.In addition,more mineralized nodules could be seen on Mg+ and RGD decorated zirconia by SEM and big mineralized nodules could be seen which was consisted with a few of little mineralized nodules.Conclusions:Compared with regular zirconia samples,RGD and Mg+ decorated zirconia possesses better wettability and the introduction of Mg+ showed nano-scaled surface character.RGD could improved the adhesion,spreading and proliferation of MC3T3-E1 and Mg+ is beneficial for the mineralization of osteoblasts,synergistic effect was found between RGD and Mg+.