The Effect Of Antimicrobial Peptide Biocoating On Osteoblast Activity And Antibacterial Activity Of Porphyromonas Gingivalis

Objective: This study by micro arc oxidation and silane coupling with different concentrations of antibacterial peptide KSL on titanium surface by osteoblast compatibility testing and Porphyromonas gingivalis antimicrobial testing and evaluation.The biocompatibility and antimicrobial properties and a preparation with good biocompatibility and has multiple functions of antibacterial composite coating.Methods: The ultrasonic micro-arc oxidation-alkali treatment-silane film was as control group(group A)and the experimental group contains the antimicrobial peptides KSL0.25 mg/ml(group B)、 KSL0.50 mg/ml(group C)、 KSL0.75 mg /ml(group D).Appearance characteristics of specimens were observed by the Scanning electron microscopy(SEM).CCK 8 tested cell adhesion and proliferation in different time periods.ALP tested alkaline phosphatase activity in different time periods.The number of proportion of living and dead bacteria was observed on the specimens by laser scanning confocal microscope.Results: The testing on four groups in the CCK 8 and ALP indicate that the sequence of proliferation,adhesion and alkaline phosphatase activity is group D >group C>group B>group A,and the comparative difference of four groups possessed the statistics significance.Through the CCK-8 adhesion test,the density of osteoblast adhered to the surface of antibacterial peptide KSL was higher than that of the non additive group,which indicated that the surface layer of the membrane treated with antimicrobial peptide KSL was more favorable for cell adhesion.Through the detection of the proliferation of CCK-8 cell proliferation was observed,is larger than that of pure titanium micro arc oxidation and alkali treatment group-silane surface of cell proliferation rate of antibacterial peptide KSL after combined treatment,suggesting that the antibacterial peptide KSL composite coating group was obviously superior to the biocompatibility of pure titanium micro arc oxidation and alkali treatment-silane group.Determination of alkaline phosphatase activity of alkaline phosphatase in bone cells,with prolonged incubation time and antibacterial peptide KSL concentration increased,alkaline phosphatase activity increased gradually,that the cecropin KSL composite coating can promote the osteoblast function expression of the above results showed that the antibacterial peptide KSL composite coating,adhesion of osteoblasts the proliferation and alkaline phosphatase activity have a certain role.This experiment uses Porphyromonas gingivalis with different surface coating processing,through the test of inhibiting bacteria growth was observed in the antibacterial peptide KSL composite coating specimen of Porphyromonas gingivalis number number of viable and dead bacteria.Laser confocal microscopy showed that the effective bactericidal concentration range of antibacterial peptide KSL and the higher the concentration of the antibacterial effect of antibacterial peptide KSL was significantly better,the biofilm formation of the effect of 1 weeks,can effectively kill bacteria Porphyromonas gingivalis in biofilm.Among them,the number of dead bacteria was the highest,and the bactericidal effect was the most significant in the ultrasonic micro arc oxidation alkali treatment group KSL0.75mg/mlD.The results showed that the antibacterial peptide KSL composite coating group was more conducive to the growth of osteoblasts on the surface of the material,the difference between the four groups was statistically significant.Conclusion: The micro arc oxidation coating on pure titanium by alkali treatment of ultrasonic-silane coupling with different concentrations of antibacterial peptide KSL,more conducive to the early osteoblast adhesion and spreading,promote osteoblast proliferation on the surface of the material,has good biological activity.Among them,loading KSL 0.75mg/ml is the best.With the increase of the concentration of antibacterial peptide KSL,the antibacterial ability of the film was increased.In the range of effective bactericidal concentration,KSL 0.75mg/ml was the final modified concentration,which could effectively kill Porphyromonas gingivalis.Antibacterial peptide KSL has good bactericidal ability,no cell toxicity.