| Animal ExperimentPercutaneous intradiscal ozone(O3)-injection:an experimental study in caninesBackground Ozone is known as one of the most intense oxidants. For the last decade,ozone has been used in some western-European countries for treatment of lumbar discal herniation,and good results were reported. But no experimental and pathological literatures on it were found. Purpose To evaluate the influence of ozone on normal nucleus pulposus and the safety of intradiscal ozone-injection for treatment of herniated lumbar disc by the study of animal experiment and pathology. Methods Nine canines were divided into 2 groups. Five canines in group A were subjected medical ozone injection into selected lumbar discs ( 3 ml ) and the paraspinal space ( 7 ml ) with 20 G Chiba needle under fluoroscopy. The medical ozone concentration was 30ug/ml and 50 ug/ml respectively. Two discs were selected for each concentration. Total 20 discs were injected. Three of the canines were given one-time ozone-injection and sacrified for pathology one week,one month and two months respectively after the procedure,and the other two canines were given two-time ozone-injection with one week interval and sacrified one month and two months respectively. Four canines (16 discs in total ) in group B were experienced the same procedure,but the medical ozone with concentration of 6 ug/ml and medical purified oxygen were injectedrespectively. The specimens including nucleus pulposus,end-plate,spinal cord,nerve root and greater psoas muscle were observed macroscopically and microscopically. Results No serious behavior abnormalities were observed in all animals. In group A,the apparent atrophy of nucleus pulposus could be observed macroscopically one month and two months after ozone-injection due to significant reduction of water and extensive proliferation of collagenous fiber in the matrix. Majority of the cells appeared necrosis and dissolution,and the survival cells were sparsely seen. The influence on the atrophy of nucleus pulposus demonstrated no significant difference between the selected two concentrations of medical ozone,but was more apparent with two-time ozone-injection compared to that with one-time ozone-injection. Sixteen items of end-plates increased slightly or moderately in thickness and a few of fibers in greater psoas muscle suffered slight atrophy in 5 items. In group B,slight atrophy of nucleus pulposus could be visualized in the experimental discs with ozone-injection. Observed under light and electron microscope,the distance between cells was increase. Pyknotic nucleuses were observed in a small part of cells,but no necrotic cells were discovered. Mild hyperplasia of collagenous fiber was presented in the matrix. The nucleus pulposus with medical oxygen injection demonstrated no significant changes. All samples from end-plate and greater psoas muscle and nerve and spinal cord showed no apparent abnormalities macroscopically and microscopically. Conclusions This study suggested that percutaneous intradiscal ozone-injection was a safe method with the above three concentrations. The ozone with concentrations of 30 ug/ml and 50 ug/ml could cause significant atrophy of nucleus pulposus,but the ozone with concentration of 6 ug/ml was much more less effective. No apparent change of nucleus pulposus was revealed for the medical purified oxygen injection. This study provided the evidence of the feasibility and value of the procedure of percutaneous intradiscal ozone-injection in clinics.
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