Preliminary Study On The Effect Of Hydrogen-Rich Water On Peri-Implant Inflammation

Research background:In recent years,more and more reports have shown that as a novel antioxidant,hydrogen has protective effects on transient cerebral ischemia,neonatal cerebral hypoxia,cold ischemia-reperfusion injury,and inflammation-related deterioration of allogenic heart and aortic receptors,etc.Molecular hydrogen is an emerging therapeutic medical gas,often used in the treatment of degenerative diseases;including Parkinson’s disease,diabetes,osteoporosis,etc..It has been reported that the injection of hydrogen-rich normal saline reduces left ventricular hypertrophy and vascular dysfunction in hypertensive rats.At present,there are mainly hydrogen inhalation method,injection of hydrogen rich brine method and drinking hydrogen rich water method as common treatment methods.Compared with hydrogen,hydrogen-rich brine and water are safe,economical,and readily available.In dentistry,animal studies have shown that HW can reduce gum oxidative stress caused by aging and periodontal disease.A recent study also showed that drinking HW activates the expression of antioxidant defense genes and helps promote the healing of oral mucosal wounds in rats.Gingival oxidative stress is involved in the process of alveolar bone resorption.Obesity suppressed by HW consumption can reduce alveolar bone resorption by limiting oxidative stress in the gingival.Research objectives:The optimum concentration time range of hydrogen-rich water was determined by titration experiment.The peri-implant inflammation model of beagle mandibular bone was established to determine the inhibitory effect of hydrogen-rich water on the peri-implant inflammation of soft and hard tissues.The changes of bacterial content in the presence of hydrogen-rich water were explored by RNA sequencing in the samples.Materials and methods:(1)Around the implant infection model:the three little beagle bilateral mandibular first,second,third,fourth premolar immediately after tooth extraction plant,two three months after operation,the healing base stations,in the same period in the fine silk ligation,after every two weeks with periodontal probe for internal bleeding,excessive concentration of the implant ditch and depth measurement and X-ray observation of verify modeling success(2)Determination of the concentration of hydrogen ions in the saturated hydrogen-rich water at different times:the machine prepares the saturated hydrogen-rich water,and drops the hydrogen-rich water titrator into it after placing the water for different periods of time,and calculates its concentration,so as to obtain the most appropriate experiment time:Hydrogen-rich water and deionized water were washed around the implants twice a day.Gingival index(GI),modified bleeding index(MSBI)and average probing depth(PD)of the implants were measured every two weeks,and gingival crevicular fluid was extracted(3)The expression gradient of IL-1,IL-6 and MMPs was determined by ELISA kit.After the beagles were killed,the expressions of IL-1,IL-6 and MMPs in the two groups were detected by PCR in the peri-implant bone tissue.He staining was used to verify the distribution trend in the two groups(4)The gingival crevicular fluid around the implant was taken in the sixth week for PCR amplification,16S RNA sequencing,and genetic analysis to detect the differences in bacteria between the two groups.Results:(1)After 6 weeks,clinical observation showed bleeding and overflow of soft tissue around the implant,and the probing was more than 5 mm.X-ray film showed angular absorption of alveolar bone.(2)Three times of hydrogen-rich water titration experiments showed that the optimal concentration time range was within 5 min after the preparation of hydrogen-rich water;Periimplant clinical indexes(GI,MSBI,PD)were measured at the 0,2,4 and 6 weeks of the experiment,and the results showed that all clinical indexes in the experimental group decreased,but there was no statistical significance compared with the control group.(3)Quantitative analysis of inflammatory cytokines(IL-1,IL-6,MMPs)using ELISA kit showed that there was significant difference between the experimental group and the control group(P≤0.01),especially in the first two weeks of the experimental group inflammatory cytokines decreased significantly;The results of HE staining showed that the inflammatory cells in the experimental group had a decreasing trend compared with the control group.The expression levels of IL-1,IL-6 and MMPs in the control group were significantly higher ’than those in the experimental group by PCR,and the measured results were statistically significant.(4)16S RNA sequencing in gingival crevicular fluid showed that the content of porphyromonas;fusobacteria and other pathogenic bacteria in the experimental group was lower than that in the control group(P<0.01).Conclusions:(1)Hydrogen rich water machine,using solid polymer proton exchange pure water electrolysis technology to prepare hydrogen rich water should be ready on the spot;(2)Experimental peri-implantitis induced by filament ligation plus high sugar soft food feeding can successfully establish peri-implantitis model;(3)Hydrogen-rich water has an inhibitory effect on peri-implant inflammation...