| Background and ObjectiveAt present, there is no ideal treatment for periodontitis and peri-implantitis in clinical, meanwhile, plasma technology has been attracting significant attentions, and our prior researcher demonstrated that the atmospheric pressure non-equilibrium plasma jet (APNPs) had huge potential for curing oral desease. The aim of this study was to evaluate the effectiveness of APNPs in destroying Periodontal advantage pathogenic bacteria and to assess the biological safety of APNPs by animal and cell experiment.Material and methodsAPNPs was driven by a kilohertz pulsed DC power (the settings of the equipment were 10 kHz, 1600 ns and 8 kV, the working gas was helium or helium and oxygen mixtures( volume ration is 2:0.02) ).1. Periodontal advantage pathogenic bacteria PG (ATCC 33277) in the petridish was dealt with by APNPs (the working gas was helium or helium and oxygen mixtures( volume ration was 2:0.02)), observed the size of antibacterial circle and the surface morphology chang of PG by using transmission electron microscopy. PG biofilms in coverslip was dealt with by APNPs (the working gas was helium and oxygen mixtures( volume ration was 2:0.02)), observed and photographed the biofilms which were processed with Dead/live fluorescence staining by laser scanning confocal microscope.2. the rabbit's buccal and abdomen tongue mucosas were treated with APNPs (the working gas was helium and oxygen mixtures( volume ration was2:0.02)), the rabbits were killed on the first and fifth day respectively, clinical observation and histopathologic scores were used to assess mucous membrane irritation.3. The murine osteoblastic cell line MC3T3-E1 was treated with APNPs (the working gas was helium and oxygen mixtures( volume ration was 2:0.02)), to simulation of cell normal or exposed cell in periodontal surgery physiological condition, the treated cell was covered with a ting layer of DMEM, cell viability was determined by MTT assay, by flow cytometry and AO/EB staining analysis.Result1. in sterilization experiment, we could observe obvious antibacterial circle, the PG cell was broken and death by transmission electron microscopy. The biofilms that was treated by APNPs for 5 min became red and yellow by laser scanning confocal microscope, that was to say, the bacteria had been death.2. in mucous membrane irritation experiment, the clinical observations showed that both the experimental and control groups (on the first day and fifth day after processed by APNPs, separately)were no hyperemia, edema, ulcers, erosions and other oral mucositis, and the results of histopathological evaluation indicated that the mucous membrane irritation index of the first-day group was negative 0.83 and the fifth-day group was negative 0.67, thus both groups were no intense mucous membrane irritation responses.3. in cell experiment, the cell that was covered with a ting layer of DMEM was treated by APNPs for 0.5min, 1.0min,or 1.5min, the relative growth rate respective were 109.5%, 103.2% and 105.7% (P>0.05), there was no significant difference between each two groups. In the group that cell was covered with a ting layer of DMEM and in blank group,the percentage of living cell were 96.56±2.60% and 95.56±1.12% (P>0.05), there was no significant difference between the two groups. The cell of two group were green after AO/EB staining, that was to say, the cell was living.ConclusionAPNPs can effectively kill Periodontal advantage pathogenic bacteria PG, and there was no mucous membrane irritation responses to normal mucosa. In simulation of normal physiological condition, it had no effect on bioactivity of the cell. In the case of the current experiment results, the APNP can cure periodontitis and peri-implantitis in theory, of course, the APNPs was authentically applied in clinic, further basic and clinical researchs were necessary elucidate this subject. |
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