| Background:Chronic osteomyelitis is a significant problem of patients and orthopedic surgeon. It is very difficult in treatment of this kind of disorder because of its character. One of the basic principles of the treatment of chronic osteomyelitis is adequate antibiotic treatment on the basis of completely clear of lesions. Despite the use of a thorough debridement method, there are still a number of bacterial residues in local osteomyelitis region. The treatment is easy to fail because of the amount of reproduction of bacterium. Because of bad blood supply in the local region, it is difficult for the body immune function to kill the bacterium. Local drug delivery system can be directly on the lesion site, and didn't carry the drug to the lesions by blood and can be sustained in a high local drug concentration. With the unreasonable application of antibiotics in recent years, the types of pathogens are changing in chronic osteomyelitis, which result in bacterial mutation and bacterial resistance. The therapeutic effect of antibiotics is decreasing. So one of the purposes of this project is to find an effective fungicide to treat chronic osteomyelitis. Ozone water can effectively treat wound infection and accelerate wound healing, and it is difficult to produce bacterial resistance. VSD is smooth for suction drainage. Ozone water infusion and vacuum aspiration is a new and effective method for treatment of osteomyelitis.Objective:To investigate bactericidal effect of different concentrations of ozone water on Staphylococcus aureus and Pseudomonas aeruginosa.The mechanism of anti-inflammatory effects of ozone water was studied. Rat models of chronic osteomyelitis at proximal tibia were established. The effect of ozone water was studied on the base of rabbit model of chronic osteomyelitis. Ozone water provide not only a new method of sterilization but also anti-inflammation to treat the chronic osteomyelitis. It can provide experimental evidence for further clinical applications.Method: The experiment was divided into four parts.1,The antimicrobial activity of ozone water on Staphylococcus aureus and Pseudomonas aeruginosa.Staphylococcus aureus, Pseudomonas aeruginosa bacteria were prepared in sterile saline suspension, respectively. Adjust the amount of bacteria by McNamara bacterial turbidimetric method. Different concentrations and time points of ozone water were used in bacteria suspended solution, and the bacteria killing rate were calculated.2,Experimental study of ozone water on Anti-inflammation.On the basis of models of xylene-induced by acute inflammatory edema of mice ear, the mice were devided randomly into 3 groups: ozone water group, voltaren group and normal saline group. The observations of model animals included thickness and weight difference of mice ear, detection of serum TNF-a, histopathology manifestation, which can illuminate the results of anti-inflammation.3,The establishment and evaluation of experimental chronic osteomyelitis induced by staphylococcus aureus in ratsFive percent sodium morrhuate and serial dilutions of the bacteria of Staphylococcus aureus suspended in saline were inoculated into the proximal metaphysis of the tibia.The relationship between the inoculation dose of the bacteria of Staphylococcus aureus and the severity of experimental Staphylococcus aureus osteomyelitis in rats was studied to provide a suitable animal model of chronic osteomyelitis.4,Experimental study of treatment of chronic osteomyelitis with ozone water infusion and vacuum aspiration.In order to observe the therapeutic effects of ozone water on osteomyelitis of rat models, modeled chronic osteomyelitis SD rats were devided randomly into 3 groups: ozone water group, antiseptic solution group and normal saline group. All were lavaged for 2 weeks. The observations of model animals included blood leukocyte count, bone X rays,detection of TNF-a and histopathology manifestation, which can illuminate the results of sterilization and anti-inflammation.Results:1,The antimicrobial activity of ozone water on Staphylococcus aureus and Pseudomonas aeruginosa.The ozone water has the similar bactericidal effect on the 2 bacteria .The bacteria in the suspension was completely killed with the treatment of 2mg/L ozone water for 30s or 1mg/L ozone water for 60s(Pseudomonas aeruginosa) and 90s (Staphylococcus aureus).2,Experimental study of ozone water on Anti-inflammation Statistical analysis indicate that there was a significant difference between group A (ozone water group),group B(voltaren group) and group C(normal saline group), no significant difference between group A and group B of mice ear weight difference after 2 hours. Concentration of TNF-a of mice blood serum indicates that group A and group B significantly less than group C(P<0.01) , no significant difference between group A and group B (P>0.05). Histological observation group C showed that there were significant epidermal cells and intracell- ular edema infiltration of mononuclear cells, dermal connective tissue edema. Histological observation group A and B find the reduction of edema and inflammatory cell infiltration3,The establishment and evaluation of experimental chronic osteomyelitis induced by staphylococcus aureus in ratsFour weeks after modeling of A group(0.1ml Normal Sodium was inoculated),B group(0.1ml 1×106 cfu/ml staphylococcus aureus ),C group (0.1ml 1×108 cfu/ml staphylococcus aureus),D group (0.1ml 1×106 cfu/ml staphylococcus aureus), radiography show that there were different degrees of soft tissue swelling, bone mineral density reduced, bone trabecular rare, cortical thinning, osteolysis, sequestra formation in experimental groups;histomorphology indicated that there were varying degree of neutrophil infiltration, necrosis of marrow cells and interstitial hemorrhage。It can establish successfully the rat model of acute staphylococcal osteomyelitis with the staphylococcus aureus of 1.0×l08CFU inoculating directly into tibial bone of rat.4,Experimental study of treatment of chronic osteomyelitis with ozone water infusion and vacuum aspirationStatistical analysis indicate that there was a significant difference between group A(ozone water group),group B(antiseptic solution group) and group C(normal saline group), no significant difference between group A and group B of Norden osteomyelitis scores after 2 weeks. Bacterial count per gram indicate that group A and group B significantly less than group C(P<0.01), group B less than group A,,and there was a significant difference(P<0.05). Concentration of TNF-a of local bone indicates that group A and group B significantly less than group C(P<0.01), group B less than group A, and there was a significantly difference(P<0.05). Histological observation showed that the inflammation response of group A and group B was significantly lighter than group C, group A lighter than group B. ConclusionOzone water can effectively kill Staphylococcus aureus and Pseudomonas aeruginosa. Ozone water can also inhibit the inflammatory response process and promote the extinction of inflammation. In the treatment of osteomyelitis with ozone water infusion and vacuum aspiration, ozone water can skill the bacterium, prohibit local inflammation response, promote tissue reparation and the bone pathological changes of chronic osteomyelitis. VSD is easy and smooth for suction drainage, which can avoid environmental pollution caused by ozone leak. Ozone water can gradually replace the application of antibiotics and provides a new method for the treatment of chronic osteomyelitis.
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