Disinfectant Mechanism And The Oxidative Effect On Skin Of Electrolyzed Oxidizing Water

Objective1.To investigate the disinfectant mechanism of electrolyzed oxidizing water on Pseudomonas aeruginosa ATCC15442 in comparison with 2% glutaraldehyde and normal saline(NS,as the control group).2.To obsever the oxidative effect of electrolyzed oxidizing water(EOW)on skin tissue of C57BL/6 mice in comparison with 3% hydrogen peroxide(H2O2)and normal saline(NS,as the control group).Method1.Firstly,the bactericidal rate was determined to compare the bactericidal effect between EOW and 2% glutaraldehyde.The Pseudomonas aeruginosa ATCC15442 was treated by each disinfectant at preset time.Then the intracellular protein leakage,DNA damage and calcium ion channels of bacteria were evaulated respectively to determine the disinfectant mechanism of EOW.2.Totally 30 female mice aged 6-8 weeks were selected in this study,which were randomly allocated into 3 group design: NS group as the negitive control group(n=10),EOW group as the experiment group(n=10),H2O2 group as the positive control group(n=10).All the mice in each group were treated with the corresponding disinfectant for 7 consecutive days at the same time.To observe and measure the skin tissue epidermal thickness by the Hematoxylin and Eosin(HE)staining;the enzyme activities of total superoxide dismutase(SOD)and the content of Malondialdehyde(MDA)in the skin tissue were tested by SOD and MDA kits;the IL-6,TNF-α expression levels were evaluated by Real Time-q PCR.Results1.The bactericidal rate on Pseudomonas aeruginosa ATCC15442 was as high as 99.00% after 30 s duration of EOW and 2% glutaraldehyde,as 100% after 60 s.The bactericidal efficacy showed no difference between these two disinfectants(p>0.05).2.After 60 s contact with EOW,2% glutaraldehyde and NS,the protein leakage detected by BCA were(96.00±7.42)、(216.97±10.35)、(94.15±7.49)μg/m L,respectively.The protein leakage showed no difference after been treated by EOW and NS(p>0.05),while the 2% glutaraldehyde group displayed significant difference compared with others(p<0.001).3.EOW and 2% glutaraldehyde led to the fluorescence intensity decreased obviously compared with the NS group(p<0.001).4.HE staining displayed the epidermal thickness had no differences between EOW and NS group(p>0.05),3% H2O2 group were thickened than both of EOW and NS group(p<0.01).5.The enzyme activities of SOD and the content of MDA all showed no differences between EOW and NS group(p>0.05),but they were significantly different with 3% H2O2 group,which presented enzyme activities of SOD reduceing(p<0.005),MDA content increasing(p<0.001).6.The expression of IL-6 and TNF-α were dramatically downregulated both in EOW and 3% H2O2 groups in comparison with NS group(p<0.001),meanwhile,no differences were observed between these two groups(p>0.05).Conclusion1.The disinfection effect on Pseudomonas aeruginosa ATCC15442 of EOW is the same as 2% glutaraldehyde.2.These results indicate that EOW probably affects the calcium ion channel of cell membrane and changes membrane permeability.3.EOW cannot cause the protein leakage,moreover,DNA of these bacterial may not the target site of EOW.4.The oxidative effect results indicate that EOW presents less oxidative damage on skin tissue as much as NS does,however,it’s not safe enough to use 3% H2O2 continuously due to the oxidative damage.EOW is considered milder and less stimulus than 3% H2O2.5.The application of disinfectant EOW and 3% H2O2 on dorsa skin taha induces the changes of bacterial flora may contribute to the supression of T-cell mediated immune response,resulting in down-regulating the expression of cytokines such as IL-6 and THF-α.