The Study Of The Effects Of Ozone Inactivating Hepatitis B Virus In Dental Unit Water Lines

Objective:To study different concentrations of ozone solution which active on the Hepatitis b virus (HBV) in dental unit waterlines (DUWLs) and find out the minimum ozone water concentration which can inactivate HBV in DUWLs. This experiment plans to compound different concentrations of ozone solutions and then put the different concentrations of ozone solution into the2.2.15cell of HepG2cell line. After cultureing together with each other, collect the cell supernate and do the polymerase chain reaction (PCR) amplification.Next by the fluorescence quantitive polymerase chain reaction (FQ-PCR), detect the concentration of HBV-DNA. Finally use Statistics software to analysis these ozone water treams" effects of inactivation. Observe and compare the influence of normal oral epithelial cells by actived with different concentrations of ozone solutions. All of these, the objective is that sutdy the effect and method of ozone steriling DUWLs and provide theoretical basis for the application of ozone in DUWLs on the control of infection.Method:1-, Compound different concentrations of ozone solutions and concentration attenuation experiment;2、Culture the HepG2.2.15cell line:routine culture the HepG2.2.15cell line;3、The different concentrations of ozone solutions affect on the expression of the HepG2.2.15cell line;4、The cytomorphology and cytotoxicity of normal oral epithelial cells acted with ozonated water;5、statistics analysis:adopt one-way ANOVA, and compare tow groups with each other with Student-Newman-Keuls (SNK-q).Results:1、Different sealed conditions and standing times about the changes of ozonanted water concentration:after standing30min, there are Statistically Significant meaning on concentration with every groups of ozonated water (F=9.77,P<0.01). In sealed condition. for5℃and water temperature water for10℃when comparing ozone concentration was not statistically significant (q=1.36, P>0.05); in sealed condition, for water temperature5℃and water temperature20℃about when comparing ozone concentration has statistical significance (q=4.88, P<0.01). In5℃water temperature, there was not statistical significance between sealing containter and not sealing containter (q=0.59,P>0.05); in20℃water temperature, there was statistical significance between sealing containter and not sealing containter (q=3.37,P<0.01)2、 Ozonated water to the liquid supernatant of HepG2.2.15cells HBV-DNA PCR fluorescence results show that:Ozone solution can significantly reduce HBV-DNA content in the liquid supernatant of HepG2.2.15cells. Different concentrations of ozone solution has different influences about HBV-DNA content the greater the solution concentration of ozone, the more obvious expressive inhibition on HepG2.2.15cells.3、HVB-DNA test results in the liquid supernatant of HepG2.2.15cells:ozone solution group than in the control group did not add ozone ultra-pure water hbv-dna quantity is decreased obviously. In1min of ozone reacting, compared with other groups, the group of1.5mg/L ozone concentration all have statistically significant differences (P<0.01). Respectively at5、10、15、30min of ozone reacting, each group can reduce HVB-DNA content, there are statistically significant meaning between blank controller and other ozonated groups (each P<0.01); after15min of ozone reacting,there are not statistically significant differences between the each groups (each P>0.05).4、Ozone solution to the role of cell Hela cells morphological observation:ues the inverted microscope to observe the Hela cell line that:the cells adhere wall, be stellate,be diamond-shaped, be eumorphism, the body of cells are transparent, be a typical stick wall irregular form. After added in low concentration (0.5mg/L,0.6mg/L) ozonated water, there is no significant effect of cells form; after added in moderate concentration (1.5mg/L) ozonated water, there are emerged some granules in cell sap and small part of cells’body become diminish, which point out the state of cells by certain degree influence. The form of cells are obviously damaged and emerge lots of cell debris when the FC solution was joined into it. All of that show a large number cells are dead and obviously damaged.5、Ozone solution to the influence of cell proliferation:the MTT results show that. the cell relative growth rate of the negative control group and low concentration ozone solution (0.5mg/L,0.6mg/L) is between100～103%and the toxicity rate is0level; the cell relative growth rate of moderate concentration ozonated water (1.5mg/L) is between75～99%and the toxicity rate is1level. The cell relative growth rate of the positive control group is between1～24%and the toxicity rate is4level. The security of3groups ozonated water is qualified.Conclusion:1、water temperature and whether the container is sealing or not can affect the ozone concentration in water.The hotter the water temperature is, the more obviously changes the solution has. when the container is sealing, the ozonated water is stable.2、Ozone solution can be effective inactivated HBV-DNA and the higher the concentration is, the better the inactivation of HBV-DNA is.3、Low concentration of ozone solution on oral normal tissue non-toxic function, and can be used in DUWLs disinfection.