Accidental and war trauma, burns and all sorts of acute and chronic cutaneous infections can be proficiently treated with ozonated water. In current study, 24 burned rabbits were divided into 4 groups (6 rabbits in each group): topical treatment with saline; topical treatment with ozone; topical treatment with ozone and ozone autohemotherapy (OAHT); and topical treatment with silvirin.
Journal of military pharmaco-medicine 7-2013 EVALUATION OF ANTIBACTERIAL EFFECT BY OZONE THERAPY ON BURN INJURY Ho Anh Son*; Nguyen Linh Toan*; Vu Quoc Binh** SUMMARY Accidental and war trauma, burns and all sorts of acute and chronic cutaneous infections can be proficiently treated with ozonated water In current study, 24 burned rabbits were divided into groups (6 rabbits in each group): topical treatment with saline; topical treatment with ozone; topical treatment with ozone and ozone autohemotherapy (OAHT); and topical treatment with silvirin The results showed that topical use of ozone in burn wounds present in animal model allow a rapid antibacterial effectiveness during the treatment periods * Key words: Bacteria; Ozone autohemotherapy; Burn; Rabbit INTRODUCTION Human skin is the barrier to prevent entering of bacteria to the body, when the skin is burned, these bacteria enter the wound and cause infection site Bacteria can enter the burn wounds from healthy skin around, waste of patients, from the care people, from linens, from health care workers, and the atmosphere, drugs, the medical instruments… Many reports have warned that burns hospital infections are a major cause of death in burn patients (accounting for 50 - 70%) The serious treated problem of drug-resistant bacteria present in the burn is also mentioned Besides, the cost of using systematic and tropical antibiotics accounts for a large proportion of funding treatment Therefore, we conducted this study using ozone as an antimicrobial substance with strong effect has been known for a long treated history, with a very low cost to assess their ability to antibacteria on the surface of burn wounds SUBJECTS AND METHOD Study subjects Healthy Newzealand rabbits (from a farm in Hungyen province), medium weight kg/each Rabbits were breed in experimental conditions, individual clean cages during the study period and free access to food, vegetable Total number of rabbits was 24, including both male and female, were randomized divided into groups of rabbits: - NaCl group: animals were burned and the wounds were cleaned by saline solution - NaCl + O3 group: animals were burned and the wounds were cleaned by ozonated saline solution - O3 + O3 group: animals were burned and the wounds were cleaned by saline solution and OAHT - Silvirin group: animals were burned and the wounds were treated by Silvirin The experiment were performed in Pathophysiology Department, Vietnam Military Medical University Method * Ozone autohemotherapy: The method was descibed somewhere [1], in brief: ml/kg body weigh ear vein blood was collected in heparined plastic container with mixture of ozone-oxygen The blood is exposed to the gas mixture for minutes and transfered back to the rabbit The process of OAHT was conducted in total of times, times per week, starting with 50 mcg/ml ozone dose and increased to 50% after every time of treatment [1, 2, 3] Journal of military pharmaco-medicine 7-2013 Figure 1: Transfusion process of ozone exposure to rabbit vein blood * Experimental burn rabbit model: Rabbits fur on back are removed, then they were mounted on the table Animals were anesthetised by thiophental, 50 mg/kg dose A cyclindal container was poured by boiling water and keep perpendicular to the back skin, followed by heavy weights kg set up container for 30 seconds The wound was done on both spine sides with grade III lesions, and covers about 10% of the total surface area of rabbit skin * Ozonated saline to rinse the wounds: Ozone dose of saline to wash the wound began 80 mcg/ml After the first washed day, the ozonated saline doses were reduced 10 mcg/ml per day When the dose is reduced to 10 mcg/ml, this concentration is maintained until the wounds were heal * Assess the level of infection in burn wounds: Microbiological tests in burns: rolling sterile swab on the surface burns injury, which is cm2 area, is limited by the plastic paper The test were performed before, after the of 3, and 14 days of ozone treatment The sterile cotton swab soaked in 0.9% NaCl sterile, push the swab into a glass tube to reduce water, gently rolling on the wounds, roll across the gaps in the plastic, so that the burn fluid stained on the entire circumference of the swab for 20 seconds The swab specimens were put into ml tubes available 0.9% NaCl sterile Shaking slightly the tube for about minute, transfer to lab microorganisms Specimens were staging on the different cultures agar, incubated at 370 C for 24 hours Count the number of colonies on the agar culture, then calculate the number of bacteria on the surface of cm burn wounds The technique was conducted on Bitex system in Paraclinical Department, National Institute of Burns * Data analysis: Data was analyzes by Stata software and One way-ANOVA, SPSS 12.0 RESULT Table 1: The rate of infection in rabbits burns day rd Group (n = 12/group) NaCl group (1) Rate of infection (%) b (-) b < 103 103 < b < 104 104 < b < 105 b > 105 0 0 100 Journal of military pharmaco-medicine 7-2013 NaCl + O3 group (2) 33.3 0 66.7 O3 + O3 group (3) 33.3 0 66.7 Silvirin group (4) 0 0 100 p > 0.05 (b: bacteria) Results showed that the rate of microbial infections in burn wounds after days of treatment was quite common in the study groups With two groups using ozonated saline to clean wounds, rate of burns infection was 33.3% Other burns in the remaining groups had over 105 bacteria/cm Chi-square statical analysis showes no significant diference of rate infection among agoups Table 2: The rate of infection in rabbits burns day th Group Rate of infection (%) b (-) b < 103 103 < b < 104 104 < b < 105 b > 105 NaCl group (1) 0 16.7 83.3 NaCl + O3 group (2) 0 16.7 83.3 O3 + O3 group (3) 16.7 0 50 33.3 Silvirin group (4) 0 50 50 p3-4 p3-1,2 < 0.05 < 0.005 p3,4 > 0.05 (n = 12/group) > 0.05 p4-1,2,3 < 0.005 p Data showed: infection rates increased significantly on day after the process of burn treatment Only burns (16.7%) in O + O3 treated group, bacteria did not grow in the culture dish In the other burns, population of bacteria was over 103/cm In range 103 - 104/cm bacterial infected burns, there were (50%) of 12 wounds in silvirin treated group, it is significanly lower than other groups (X2 = 8.0; p3-4 < 0.005) The range 104 - 105/cm bacterial infected burns, there were (50%) of 12 wounds in silvirin treated group, it is significanly lower than silvirin groups (X = 8.0; p3-4 < 0.005) A mong bacterial infected burns over 105/cm 2, there were four (33.3%) of 12 wounds in O3 + O3 treated group, it is significanly lower than NaCl and NaCl+O treated group (X2 = 6.17; p31,2 < 0.05) Table 3: The rate of infection in rabbits burns day 14 th Journal of military pharmaco-medicine 7-2013 Group rate of infection (%) b (-) b < 103 103 < b < 104 104 < b < 105 b > 105 0 16.7 83.3 NaCl + O3 group (2) 83.3 0 16.7 O3 + O3 group (3) 66.7 0 33.3 Silvirin group (4) 25 25 50 (n = 12/group) NaCl group (1) p2-1 < 0.001 p2-4 < 0.005 p3-1 < 0.005 p1-2