| BackgroundOsteoarthritis(OA) is a chronic degenerative disease with very high morbidity among middle-aged and old people, which could lead injuries to knees most frequently with high teratogenesis rate, and makes patients suffered very much. The mechanism of OA is very complicated, and the cartilago articularis lesion is the most prominent with the effects of both mechanical factors and biological factors. As one of the important manifestation of osteoarthritis, arthromeningitis could lead to early stage of osteoarthritis symptoms and then affect the changes of cartilago articularis. There were varieties of therapies for Knee Osteoarthritis(KOA) in clinic at present, including taking anti-inflammatory analgesic or opioid drugs to relieve the pain, injecting sodium hyaluronate or medical ozone in joint cavity to improve the symptoms of OA, operating on the advanced OA patients with severe pain, still some other treatment methods, such as traditional Chinese medicine(TCM) therapy, are frequently applied. It has reached confirmed curative effects in clinic to do intra-articular injection with ozone water to cure osteoarthritis, but there hasn’t been any report about experiment research of it. This experiment intervened the knee osteoarthritis of rabbits by injecting with ozoned water to intra-articular. By comparativing the experimental animals joint mobility, observing the pathological changes of articular cartilage and synovial membrane, detecting the inflammatory factor levels in synovial of all the four groups rabbits, we evaluate the effectiveness of ozoned water for knee osteoarthritis in rabbits, and discusse the mechanism of the ozoned water for osteoarthritis. ObjectiveTo observe the effects of ozone water to cartilage and synovium as well as IL-6,TNF-α and VEGF in synovium, and to discuss the mechanisms of ozoned water curing osteoarthritis of knees. Methods1.the establishment and intervention of animal model Thirty-two rabbits male and female in half were randomly and evenly divided into four groups by random number method, named blank control group(A group), model control group(group B), ozoned water control group(C group), ozoned water treatment group(group D),each had 8 rabbits. Rabbits in A group had not been handled before they were killed; all the rabbits in B group were only made osteoarthritis models; rabbits in C group only received intra-articular injection with ozoned water of 20 μg/ml(2 ml) once a week for three weeks; After the osteoarthritis models were made successfully, rabbits in D group received intra-articular injection with ozoned water of 20 μg/ml(2 ml) once a week for three weeks also.2. HE staining of cartilage and synovial specimens Cartilage and synovial specimens were made into pathological sections, then pathological histology of joint cartilage and synovium was observed by optical microscope, including the integrity of the synovial tissue, whether there is tissue proliferation or degeneration, and whether there is inflammatory cell infiltration or cell necrosis in synovial.3. ELISA The expression levels of IL-6, TNF-α and VEGF in the joint synovium were compared among the four groups.4. Statistics processing The relevant experimental data was handled by SPSS 17 statistical software, taking a = 0.05 as the level test. The difference was considered to be statistically significant when P ≤ 0.05. Results1. Joint mobility Compared with A group, the rabbits joint mobility of B and D groups is smaller(P < 0.05); which is similar that the joint mobility at the two different time points when rabbit were killed and osteoarthritis models were made in the B group(P > 0.05); the rabbit joint mobility in D group is bigger than in B group at the animals were killed time point(P < 0.05); the joint mobility was significantly improved when the animals were killed in D group(P < 0.05).2. Knee joint synovial thickness Compared with A group, the synovial thickness in B and D group is chicker(P < 0.05);the difference of synovial thickness between C and A groups is non-statistical significance; synovial thickness in D group is obviously thinner than in B group(P < 0.05).3. HE dyeing observations of samples of cartilage and synovium In group A, there were complete surfaces of cartilage, four layers of clear structures, cells in alignment, complete wires. In group B, there were rough and irregular surfaces of cartilage, obvious cracks which could be seen, thinner cartilage cell layers,smaller number of cells and turbulent arrangements. In group C there were four clear layers of cartilage structures, still regular arrangements of cartilage cells, no increase of cartilage fiber cells. In group D there were less irregular defect on cartilage surface than in group B, less numbers and diffuse distribution of cartilage cells, structure arrangement in order. In group A there were no hyperemia and edema of synovium tissue, no hyperplasia of synovium cells, and normal structure of synovium tissue. In group B there were broken structure of synovium tissue,serious hyperplasia of cells, obvious inflammatory cells infiltration, vascular hyperplasia and hyperemia, remarkable thicker synovium than that in group A. In group D the inflammatory cells in synovium tissue reduced, the hyperemia and edema relieved and the thickness of synovium obviously reduced compared with that in group B. It didn’t show inflamatory symptom of synovium in group C.4. results of ELISA Compared with group A, the expressions of IL-6, TNF-α and VEGF in synovium didn’t increase obviously in group C(P > 0.05) and they increased in the rest two groups(P < 0.05). The expressions of IL-6, TNF-α and VEGF in synovium decreased in group D compared with group B(P < 0.05). Conclusions1. Injecting 2 ml ozoned water of 20 μg/ml into articular cavity can significantly inhibit the pathological damage and reduce the expression of IL-6, TNF-α and VEGF in synovium tissue of rabbit osteoarthritis, improve the joint mobility;2. Injecting ozoned water of 20 μg/ml into articular cavity has no damage to normal joint tissues, nor effects the expression of IL-6, TNF-α and VEGF in synovium tissue of normal rabbit, which prove the treatment is safe. |
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