关节内注射医用臭氧对大鼠膝退行性关节炎的影响

Effect of intra-articular ozone injection on degenerative cartilage of the rat's knee

[目的]观察医用臭氧膝关节腔内注射对大鼠退行性关节炎的影响,探讨不同浓度医用臭氧对退行性关节炎的防治效应及可能机理。[方法]将SD大鼠40只随机分为A(正常对照组)、B(模型组)、C(空气组)、D(35μg/ml医用臭氧组)、E(70μg/ml医用臭氧组)5组,后4组复制出膝退行性关节炎(OA)模型,D、E组依据浓度不同,隔日1次给予膝关节腔内注射医用臭氧,每次1ml,间隔一周再注射,共2周。4周后处死动物,HE染色观察光镜下关节软骨的一般形态、Masson染色进行软骨Mankin’s评分,关节灌洗液超氧化物岐化酶(SOD)、丙二醛(MDA)作为检测指标。[结果]B组光镜下关节软骨退变明显,Masson染色可见蓝绿色丢失较重,SOD下降,MDA升高;D组显微镜下关节软骨形态观察有明显改善,Mankin’s评分较A组无显著性差异,与B、C组比较有显著性差异(P<0.05),SOD升高,MDA下降;E组显微镜下及Mankin’s评分较B组无明显改善,SOD下降,MDA升高。[结论]浓度为35μg/ml医用臭氧组能够减轻退行性关节炎软骨退变,增强机体清除自由基的能力,从而有效防治退行性关节炎,而浓度为70μg/ml医用臭氧组引起组织细胞脂质过氧化反应,导致关节软骨组织结构破坏。

[ Objective] To observe the effect of medical ozone on cartilage in the model of osteoarthritis （OA） to explore its therapeutic mechanism. [ Method] Forty healthy adult SD rats were randomly divided into five groups ： A （group normal）, B （groupmodel）, C （ group air）, D （group O2 -O3 35 μg/ml） and E （group 02 -O3 70 μg/ml） （n=8 in each group） . The OA model was duplicated by performed anterior eruciate ligament transection （ACLT） and partial meniscectomy of 1/3 inside. Air was injected into joints of group C, according to concentration different, which O2 - O3 35 μg/ml and O2 - O3 70 μg/ml were respectively injected into joints to D and E group every other day , every time 1 - 1.5 ml , and after a week injected again. After the model was successfully made, the condylar cartilage of femur was excised after 4 weeks for the observation by transmissive microscope of HE staining. Mankin＇ s index was recorded. At the end of the experiment, douche fluid of the diseased joints were obtained for the measurements of superoxide dismutase （SOD） and malondialdehyde （MDA） . [ Result] The OA model in B group founded obviously degenerative changes under microscopy, resulting in mostly blue and green color destaining of Masson staining at superficial cartilage. The activity of SOD in the douche fluid of the diseased joints degraded and the quantity of MDA in the serum in the OA model group increased. The Mankin＇s index in the group D was significantly higher than that in the group B, C （ P 〈 0. 05 ）, which had no significant differences between group A, and obvious improvement were found in group D under microscopy, SOD levels increased and MDA levels degraded. There was no significant differences to group B in the Mankin＇s index. SOD levels degraded and MDA levels increased. [ Conclusion] Thirty -five μg/ml O3 can prevent degeneration of articular cartilage, improve the ability to clear free radical. There is reasonable effect for the treatment of OA, but 70 μg/ml O3 resultes peroxidatic reaction tissue and cell, which resultes to destroy of articular cartilage.