| BackgroundOzone (O3) with strong oxidation and instability, in the air at normal temperature and pressure easily reduce to oxygen (has half-life about 45 minutes). Ozone can soluble in water, in the air at normal temperature and pressure,it's solubility in water is 13 times higher than oxygen,and 25 times higher than air.However,the speed of decomposition in water is much faster than that in the air.For example,if the concen-tration of ozone in water is 6.25×10-5 mol/L(3mg/l), the half-decay period is 5～30min,but it decompose slowly in purified water, the half-decay period is 20min in distilled water or tap water.When in the redistilled water,only 10%of ozone decompose after 85 min,if the temperature of the water close to 0℃,ozone will be more stability.Medical ozone (O2-O3) is the mixture of ozone and pure oxygen,which can quickly kill bacterium under oxidation-reduction reaction,it's consider as one kind of broad and utility bactericide duo to its stronge oxygenation ability. Along with recognizant and lucubrate oxygen-ozone to characteristic of biopharmacy, oxygen-ozone gradually expand to midecal area. In Europe, medical ozone are widely used for the treatment of diabetic foot wide, trauma and the healing of skin ulcers, stroke, ulcerative colitis, arthritis, prolapse of intervertebral disc, et al, its clinical efficacy has been fully affirmed.Ozone water, solubled in water by ozone gas,is more stronger in sterilizing, decomposition and promotion, organics and heavy metalcan be break dowm or transform into harmless material.Considering the character of ozone water, it is widely used in debrideing infected wound,mouthwash, vagina washing by hakeem domestic and overseas,and gain wonderful effect.On the other side,the facilities of prepare ozone water are small, the process is simple and quick,it couble bo conservation in the frige for 3-4days,all of which make ozone water much superior than traditional antiseptics.Clinical results show that ozone therapy has significant advantages, but why it has not really promoted, and even be refused by many doctors and patients? The most fundamental reason is that the basic research of ozone therapy is lagging far behind in clinical practice, many roles and mechanisms of the therapy still unknown, what limit its expansion of the field of clinical treatment. In particular, the lack of basic research on safety of the ozone therapy blocks its development. So the safety of medical ozone therapy studies appears to be every important.At present, the related research focuses on the ozone in the blood cells, in particular the impact of erythrocytes. Ozone has a strong sense of oxidation, contact a certain concentration and duration of ozone will cause the body damage. Most research on the ozone focus on the affection on blood erythrocytes.Although plasma has series of buffer system, such as antioxidant enzymes and glutathione, to prevent accumulation of the superoxide anion and hydrogen peroxide, which may produce alterations of tissue structure secondary to cell membrane degradation, but there is still part of the ozone in the plasma generates reactive oxygen species (ROS). In vitro experiments, the ROS can trigger a number of biochemical pathways. ROS degradation in 0.5 to 1.0 minutes by antioxidant system in the plasma. As soon as contact and dissolved in the plasma, ozone react with anti and polyunsaturated fatty acids, and cascade generated a series of complexes which had been definited, such as hydrogen peroxide and lipid peroxidation products. Studies by Bocci, etc, suggested that when concentration of medical ozone was less than 80μg/ml, buffer systems and anti-oxidation system in the blood and red blood cell, can resistant the ozone oxidization and avoid cells and tissue damage, thus ozone can play a role as drug. When ozone concentrations more than 8μg/ml, it causes tissue damage easily, with the concentration of ozone increasing, there is more obvious damage.However, the past reseashs are mainly concentrate on the anti-inflammatory, analgesia and immunoloregulation,the fewer studies on ozone water are localize on clinic.For this reason,it's necessary to find out,compare to ozone water,does different density of ozone water has different effert? What is the transmutation of organism contribute to ozone water,and what is the mechanicsm? Clinical physicians use ozone for treatments common concern these questions and the latter affect patients on the acceptability of the ozone treatment in a certain extent, and block the development of ozone therapy. But we have not yet seen relevant reports.Purpose1.To study the healing effect of different density of ozone water on the infected wound;2.To explore the changes in endogenous growth factors and their relation with wound healing by ozone water.Method1. First part:The role of ozone water on antiinflammatory and reparation of infected wound. 50 vista rats were randomly divided into five groups:chlorhexidine treatment group,5mg/L,10mg/L,20mg/L ozone water treatment group and contrast group(without any special treatment). The back of rats were artificially formed the infected wound. After 48 hours of infection, the chlorhexidine or ozone water was given, once a day,for 9 times. Before and after the 3rd,6th,9th day of treatment, the alteration of infected wound was observed. The WBC of tail venous blood was detected and the incised skin and musle tissue was lstopathologically analysed by light microscope.2. Second part:The changes in endogenous growth factors and their relation with wound healing by ozone waterBased on the first part of the experiment, before and after the 3rd,6th,9th day of treatment, harvested the muscle and skin tissue samples of the wound.The amounts of endogenous epidermal growth factor(EGF) and tumor necrosis factor-α(TNF-α) in infected wound were determined and their relation with the wound healing by ozone water were studied.3. Statistics analysis:The experimental data was analyzed by the software of SPSS 13.0, and an associated probability of 5%(P<0.05) was considered significant.Results1. Artificial infected woud findings:Compare with post-treatment, ozone water have the better effect to promote the healing of infected wound, especially the result of 10mg/L and 20mg/L are better.2. Total white blood cell changes:After the 3rd,6th,9th day of treatment, no significant difference in the total number of white blood cell betweem 10mg/L and 20mg/L ozone water treatment group (P> 0.05).But the first two have significant difference with 5mg/L ozone water treatment group after the 3rd,6th day of treatment.After the 9th day of treatment, the total number of white blood cells was no significant difference amon three kinds of concentrations of ozone water treatment group.3. Light microscopy findings:After the 3th day of treatment, Ozone water treatment group reduce inflammation in pathological, newborn epidermal cells can be seen, especially in the result of 10mg/L and 20mg/L groud; Inflammatory changes in the contrast group was increased, muscle fiber necrosis can be seen. After the 3th day of treatment,reduce inflammation in pathological of chlorhexidine treatment groud was not obvious, the inflammatory pathology of ozone water treatment group were further reduced. Epithelial repair integritily in the 10mg/L and 20mg/L groud; Skin, muscle necrosis increased in the contrast group. After the 9th day of treatment, inflammation in pathological of chlorhexidine treatment groud was reduced, Some epidermal cells appeard to connect; Ozone water treatment group have only mild inflammation and no effusion, 10mg/L and 20mg/L groud mostly return to normal skin, The growth of granulation tissue and muscle fibers were significant, The contrast group appeared to chronic inflammation.4.Content changes of EGF:In all stages of treatment, the treatment group compared with the control group and the chlorhexidine group compared with the ozone water treatment group,there were significantly difference (P<0.05). After the 3rd,6th and the 9th day of treatment,ther was no significant difference betweem 10mg/L and 20mg/L groud (P> 0.05).But the amounts of EGF in the first two were much greater than those in 5mg/L groud after the 3rd,6th day of treatment. After the 9th day of treatment, the Content changes of EGF were no significant difference amon three kinds of concentrations of ozone water treatment group.5. Content changes of TNF-α:In all stages of treatment, the treatment group compared with the control group and the chlorhexidine group compared with the ozone water treatment group,there were significantly difference (P<0.05). After the 3rd,6th and the 9th day of treatment,ther was no significant difference betweem 10mg/L and 20mg/L groud (P> 0.05).But the amounts of EGF in the first two were significant different from 5mg/L groud,5mg/L groud was higher.Conclusion1. Medium and high concentration of ozone water groud have the better effect to promote the healing of infected wound than the low one,and the first have similar effect,which indicated that 10-20mg/L ozone water is Enough to control and even eradication of wound infection.2. There is a positive correlation betweem EGF and wound repairment in rats,ozone water treatment can increase the content of EGF,and have a relationship with its concentration,10mg/L and 20mg/L groud have the best effect,the content of EGF gradually increased during the treatment process.3. Opposite to the EGF,the content of TNF-αin ozone water treatment groud was lower than that in the chlorhexidine treatment group and contrast group,10mg/L and 20mg/L groud were the lowest. Ozone water treatment can reduce the content of TNF-α, thereby inhibiting the activity of the NF-κB system,finaly reach the purpose of antiinflammatory and reparation. |
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