关节腔内注射不同蛋白酶建立兔膝骨关节炎模型的对比研究

目的 探讨两种不同蛋白酶诱导的骨关节炎(OA)模型的异同.方法 12只新西兰兔被随机均分为两组,分别经皮右膝关节腔内注入1.6%木瓜蛋白酶0.3 ml (4.8 mg)或Ⅱ型胶原蛋白酶溶液0.5 ml(2 mg)各两次建立模型,末次注射后第6周行右膝放射学及病理学检查.结果 两组模型大体及显微镜病理检查均显示内侧胫骨平台关节软骨呈骨关节炎样退变,其中胶原蛋白酶组的软骨退变程度较木瓜蛋白酶组严重,Mankin评分分别为(10.2±1.9)分及(4.3±1.0)分(t=-4.699, P<0.01),胶原蛋白酶组的关节边缘还可见骨赘形成,而木瓜蛋白酶组无可见骨赘.结论 两种蛋白酶关节腔内注射均可成功建立起骨关节炎动物模型,低剂量的胶原蛋白酶可较稍高剂量的木瓜蛋白酶诱导出更为严重的软骨退变.     

综合治疗及护理膝骨关节炎疗效观察

目的:观察关节腔冲洗、低浓度臭氧联合玻璃酸钠关节腔注射治疗膝骨关节炎临床疗效及不良反应.方法:215例病人随机分两组,治疗组(A组):112例,每个膝关节腔冲洗后注射30μg/l的低浓度臭氧10ml,2-3次,玻璃酸钠2.5ml,共5次;对照组(B组):103人,关节腔注射曲安奈德20mg,玻璃酸钠2.5ml,共 5次.结果:治疗组:总有效率88.39%.对照组:总有效率86.41%.结论:关节腔冲洗、低浓度臭氧联合玻璃酸钠膝关节腔注射治疗膝骨关节炎疗效肯定,且不良反应发生率低.     

臭氧液对兔膝骨性关节炎滑膜炎症的影响

目的评价臭氧液对兔膝骨性关节炎滑膜炎症的影响。方法新西兰兔32只,采用随机数字表法分为四组,每组8只。A组不予任何处理;B组仅造模;C组造模成功后,关节腔注射20μg/ml臭氧液2ml,每周1次,连续3周;D组不造模,仅行关节腔注射臭氧液,方法同C组。观察膝关节滑膜形态,检测滑膜组织中IL-6和TNF-α含量。结果 A组滑膜组织无充血水肿,滑膜细胞无增生现象,滑膜组织结构正常;B组滑膜组织正常结构破坏,细胞重度增生,炎性细胞浸润明显,血管增生充血,滑膜厚度较A组明显增加;C组滑膜组织炎症细胞减少,充血水肿减轻,滑膜厚度较B组明显降低;D组滑膜无炎症表现。与A组比较,D组IL-6和TNF-α含量无明显增高,B、C组IL-6和TNF-α含量明显增高(P0.05),但C组明显低于B组(P0.05)。结论关节腔注射20μg/ml臭氧液2ml可以减轻滑膜炎症,降低滑膜中IL-6和TNF-α含量,且对正常关节滑膜组织无明显损害。     

关节腔注射云南白药对SD大鼠骨关节炎CGRP和ED1表达的影响

[目的]通过观察关节腔注射云南白药对SD大鼠骨关节炎滑膜降钙素基因相关肽(calcitonin gene related peptide,CGRP)、滑膜组织单克隆抗体(ED1)表达的影响及关节形态的改变,探索关节腔注射云南白药对SD大鼠骨关节炎的抗疼痛及抗炎症的作用机理。[方法]35只健康SD大鼠,15只作为对照组(正常组),其余20只全部手术剪断双侧前交叉韧带,于手术后第3 d起连续每天关节腔内分别于左侧注射生理盐水0.1 ml,右侧注射云南白药液0.1 ml。于注射后3 d,1、2、4、8周分5次分别处死3只对照组大鼠和注射药物SD大鼠4只。取左右全膝关节。对比同体左右膝关节滑膜CGRP和ED1的变化及左右膝形态的改变,并与对照组比较。[结果]术后云南白药治疗组(右膝)降钙素基因相关肽(CGRP)较术后生理盐水治疗组(左膝)明显减少,8周时对照最为明显,手术组左右膝滑膜CGRP对比有显著性差异(P<0.05);术后1、2、4周时滑膜均可见左右膝滑膜组织ED1阳性细胞升高,8周时,术后云南白药治疗组(右膝)ED1阳性细胞较术后生理盐水治疗组(左膝)明显减少,左右膝ED1阳性细胞有显著性差异(P<0.01)。全膝软骨形态肉眼观,术后生理盐水治疗组(左膝)软骨破坏较术后云南白药治疗组(右膝)严重。[结论]关节腔注射云南白药,白药通过抑制CGRP阳性神经细胞的增生,抑制滑膜组织中ED1阳性炎症细胞数量,对骨关节炎疼痛有明显的治疗作用,并对骨关节炎炎症有一定作用。     

臭氧对兔膝关节滑膜中IL-1、TNF-α的影响

[目的]观察不同浓度臭氧对兔膝关节滑膜中致炎因子IL-1、TNF-α的影响,探讨臭氧治疗骨关节炎的机制。[方法]将64只新西兰大白兔随机分为8组,分别为臭氧10～50μg/ml组及正常组、纯氧组和模型组,正常组不予任何处理,模型组仅造模,余6组分别在兔膝关节腔中注射相应浓度的臭氧或纯氧2 ml,1次/周,共3次。观察各组关节滑膜形态,并检测滑膜中IL-1及TNF-α的含量。[结果]臭氧各组膝关节的滑膜数量明显减少,病理切片显示滑膜组织明显萎缩、细胞凋亡,有少量炎性细胞浸润;IL-1及TNF-α两种因子的分布一致,均为臭氧50μg/ml组>纯氧组和模型组>臭氧40μg/ml组>臭氧10μg/ml、30μg/ml组>臭氧20μg/ml组>正常组。[结论]臭氧可影响关节滑膜细胞中IL-1及TNF-α的水平,较低浓度的臭氧可能可以延缓骨关节炎病程的进展,而较高浓度的臭氧能加重关节的损伤。     

青藤碱降低兔膝骨关节炎模型瘦素含量及其受体表达

目的观察青藤碱关节腔注射对兔膝骨关节炎模型血清及关节液瘦素水平以及关节软骨瘦素受体(Ob-Rb)表达的影响。方法 38只新西兰大白兔随机分为空白组、模型组、透明质酸钠组和青藤碱组。空白组不予任何处理,模型组、透明质酸钠组和青藤碱组通过木瓜蛋白酶关节腔注射法诱导膝骨关节炎模型后,分别用生理盐水、透明质酸钠和青藤碱干预5周,双抗体夹心酶联免疫吸附(ELISA)法检测血清及关节液瘦素含量,实时荧光定量PCR(q PCR)法检测Ob-Rb的m RNA表达,Western blot法检测Ob-Rb的蛋白表达。结果模型组和透明质酸钠组血清、关节液瘦素水平较空白组均显著升高(P0.05),青藤碱组血清、关节液瘦素水平明显低于模型组(P0.01)和透明质酸钠组(P0.05);模型组Ob-Rb的m RNA和蛋白表达较空白组均显著升高(P0.05,P0.01),青藤碱组Ob-Rb的m RNA和蛋白表达明显低于模型组(P0.05,P0.01)。结论青藤碱膝关节腔注射可降低兔膝骨关节炎模型血清及关节液瘦素水平,下调关节软骨Ob-Rb的表达。     

青藤碱降低兔膝骨关节炎模型痩素含量及其受体表达

目的 观察青藤碱关节腔注射对兔膝骨关节炎模型血清及关节液瘦素水平以及关节软骨瘦素受体（Ob-Rb)表达的影 响。方法 38只新西兰大白兔随机分为空白组、模型组、透明质酸钠组和青藤碱组。空白组不予任何处理,模型组、透明质酸 钠组和青藤碱组通过木瓜蛋白酶关节腔注射法诱导膝骨关节炎模型后,分别用生理盐水、透明质酸钠和青藤碱干预5周,双抗 体夹心酶联免疫吸附（ELISA)法检测血清及关节液瘦素含量,实时荧光定量PCR(qPCR)法检测Ob-Rb的mRNA表达, Western blot法检测Ob-Rb的蛋白表达。结果 模型组和透明质酸钠组血清、关节液瘦素水平较空白组均显著升髙（P < 0. 05),青藤碱组血清、关节液瘦素水平明显低于模型组（P <0. 01)和透明质酸钠组（P <0. 05);模型组Ob-Rb的mRNA和蛋 白表达较空白组均显著升髙（P <0.05,P <0.01),青藤碱组Ob-Rb的mRNA和蛋白表达明显低于模型组（P <0.05,P < 0. 01)。结论 青藤碱膝关节腔注射可降低兔膝骨关节炎模型血清及关节液瘦素水平,下调关节软骨Ob-Rb的表达。     

当归阿魏酸钠局部用药治疗骨关节炎的实验研究

[目的]探讨当归、阿魏酸钠关节腔注射对大鼠骨关节炎(OA)的疗效.[方法]40只大鼠木瓜蛋白酶法建立双膝OA模型,造模后分为模型对照组,25%及5%当归治疗组,0.5%及0.1%阿魏酸钠治疗组,另取8只大鼠作正常对照;各组关节腔注射相应浓度药物或生理盐水0.1 ml,3日一次;6周后检测血浆、关节液SOD、MDA,关节软骨取材光镜观察,检测软骨MMP-1、TIMP-1、Bcl-2、Bax的表达.[结果]25%当归和0.5%阿魏酸钠可增加血浆、关节液SOD活性(均P＜0.05)并降低其MDA水平(均P＜0.01);降低关节软骨Mankin's评分(均P＜0.01),降低软骨细胞MMP-1、Bax的表达(P＜0.05或P＜0.01),同时使TIMP-1、Bcl-2的表达增加(P＜0.05或P＜0.01);5%当归无显著效果,0.1%阿魏酸钠对部分检测指标有统计意义.[结论]当归、阿魏酸钠关节腔注射治疗大鼠OA均有疗效,其作用具有剂量依赖性,阿魏酸是当归注射液治疗OA的有效单体成分.     

医用臭氧对膝骨关节炎患者疼痛程度、关节功能及血清NO、SOD、MDA的影响

目的:探讨医用臭氧关节腔注射对膝骨关节炎患者疼痛程度、关节功能及血清一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平的影响.方法:将80例膝骨关节炎患者随机分为治疗组和对照组,每组40例.治疗组采用医用臭氧关节腔内注射治疗,对照组采用玻璃酸钠关节腔内注射治疗.2组均每周治疗1次,5次为1个疗程.比较2组...     

兔膝骨性关节炎模型肠道菌群的16S rRNA检测分析

[目的]探讨肠道菌群在兔膝骨关节炎(knee osteoarthritis,KOA)与正常对照之间的差异.[方法]20只大耳白兔,随机分成2组,12只关节内注射2％木瓜蛋白酶溶液制备KOA模型,另外8只关节内注射等量生理盐水为正常对照.采集动物粪便样本,利用16SrRNA基因靶向测序技术进行检测,分析各自粪便中肠道菌群...     

关节腔内注射臭氧对骨关节炎软骨作用的实验研究

目的探讨医用臭氧直接在关节腔内注射对骨关节炎（OA）软骨的作用。方法32只新西兰兔随机均分为4组：正常对照组（N）、模型对照组（M）、低剂量臭氧组（L）和高剂量臭氧组（H）,每组8只。M、L、H三组分别于第1、4实验日经右膝关节腔内注射1．6％papain溶液0．3ml制备OA动物模型。成功建模1周后,L、H组右膝关节腔内注射浓度分别为20μg/ml和40μg/ml的医用臭氧2ml,每周两次,共4周。末次注射后1周处死全部实验兔,截取胫骨内侧平台做病理学观察并进行Mankin评分。结果M、L、H三组胫骨内侧平台软骨均出现骨关节软骨损害改变,L、H组损害比M组严重,而H组又比L组严重;Mankin评分从正常组－模型组－低剂量臭氧组－高剂量臭氧组逐步递增,组间差异有统计学意义（P〈0．01）。结论关节腔内反复注射臭氧后,对膝关节OA软骨产生了进一步损害的作用,损伤程度随臭氧浓度的增加而加重。     

Effects of SKI 306X,a new herbal agent,on proteoglycan degradation in cartilage explant culture and collagenase-induced rabbit osteoarthritis model

OBJECTIVE: Protective effects of SKI 306X, a natural herbal product extracted from three herbs Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris, on articular cartilage was examined and compared with other osteoarthritis (OA) drugs using in vitro and in vivo models. METHODS: In vitro culture of rabbit articular cartilage explants was used as a model to measure the effects of drugs on the matrix degradation. The recombinant human interleukin-1alpha (rhIL-1alpha, 5 ng/ml) was added to induce proteoglycan (PG) degradation and the degree of PG degradation was assessed by measuring the amount of glycosaminoglycan (GAG) released into the culture medium. In in vivo experiment, collagenase was intraarticularly injected twice into the right knee joint of rabbits to induce OA-like change, and test agents were orally administered once a day for 28 days. The degrees of OA-like changes were evaluated through a histological examination. RESULTS: In vitro study revealed SKI 306X inhibited the degradation of PG in a concentration-dependent manner. Trichosanthes kirilowii, which is one of the major components of SKI 306X, also significantly inhibited the GAG release in cartilage explant culture at 0.3 and 0.1 mg/ml. Dexamethasone and NSAIDs, such as diclofenac and rofecoxib, had no significant effects on the suppression of PG degradation. In in vivo studies, OA-like degeneration of the articular cartilage and synovial tissue was induced by injecting collagenase into the right knee joint of mature rabbits. At a dose of 200 mg/kg, SKI 306X reduced the OA-like histological changes, whereas diclofenac had no effect at 10 mg/kg. CONCLUSION: These results indicate that SKI 306X inhibited PG degradation in cartilage explant culture, and its prophylactic administration significantly protected the knee joint of rabbit from OA-like change in collagenase-induced experimental OA model. This strongly suggests that SKI 306X can be a good OA agent with some cartilage protection activity.     

MR T2 mapping成像评估止痛健骨方治疗膝骨性关节炎关节软骨损伤疗效的实验研究

目的探讨MR T2mapping成像评价止痛健骨方治疗骨性关节炎软骨损伤疗效的价值。方法采用木瓜蛋白酶关节腔注射法制作新西兰大白兔膝骨关节炎模型,根据治疗方法不同将48只新西兰大白兔随机分为正常对照组、模型对照组、盐酸氨基葡萄糖治疗组、止痛健骨方治疗组。各组治疗4周后行膝关节MR T2mapping成像并对关节软骨基质金属蛋白酶1(MMP-1)含量进行免疫组化分析。比较各组间关节软骨T2值及关节软骨MMP-1含量的差异,并分析不同干预方法、关节软骨MMP-1含量与关节软骨T2值的相关性。结果模型对照组关节软骨T2值高于其他3组(P均0.01),其他3组间关节软骨T2值差异无统计学意义。盐酸氨基葡萄糖治疗模型组、止痛健骨方治疗模型组间关节软骨MMP-1含量差异无统计学意义,但2组关节软骨的MMP-1含量低于模型对照组(P均0.05)、高于正常对照组(P均0.05)。关节软骨MMP-1含量与T2值间为非线性关系,随着前者的升高,后者先缓慢上升再快速上升(P0.05)。结论 MR T2mapping成像有助于评价止痛健骨方治疗模型兔骨关节炎软骨损伤的疗效,关节软骨MMP-1含量与T2值间可能为非线性关系。     

治疗型关节炎护膝对实验性骨关节炎软骨细胞凋亡基因Bcl-2及p53的影响

目的：通过检测OA护膝对日本大耳白兔膝骨性关节炎软骨细胞凋亡基因Bcl-2、p53mRNA表达的影响,探讨OA护膝防治兔膝骨性关节炎软骨细胞凋亡的分子生物学机制。方法：健康6月龄日本大耳白兔54只,雌雄各半,空腹体重2～2．2kg,采用改良Huhh法复制膝骨性关节炎模型,随机分为6组,即正常组、模型组、对照组（微波组）、实验1组（电组）、实验2组（热组）、实验3组（护膝组）．正常组10只,常规饲养;模型组9只,造模后常规饲养;对照组9只,微波仪治疗30min,每日1次;实验1组9只,电（疏密波）治疗30min,每日1次;实验2组8只,热（热软膜）治疗30min,每日1次;实验3组9只,电热（OA护膝）治疗30min,每日1次,连续治疗16周时处死。采用荧光定量RT—PCR法检测各组膝关节软骨细胞Bcl-2、p53mRNA的表达水平。结果：16周时,各组所有抽提的兔关节软骨组织总RNA的OD260／OD280值均在1．80～2．00范围内,表明RNA纯度高：模型组、对照组、实验1组、实验2纽、实验3组关节软骨细胞p53的mRNA相对呈高表达,而关节软骨细胞Bcl-2的mRNA相对低表达,与正常组差异有统计学意义（P〈0．01）：关节软骨细胞Bcl-2、p53的mRNA相对表达水平,对照组、实验1组、实验2组、实验3组与模型组差异有统计学意义（P〈0．01）;对照组、实验1组、实验2组与实验3组差异有统计学意义（P〈0．01）。结论：OA护膝能提高关节软骨细胞Bcl-2 mRNA表达,减弱软骨细胞p53mRNA表达,从而抑制软骨细胞凋亡,延缓膝关节软骨的退变：     

Intra-articular injections and articular cartilage metabolism

Summary We studied the effects of repeated intra-articular injections of sterile 140 mM NaCl solution on articular cartilage in adult rabbits. After 20 injections into the knee joints over a period of 4 weeks, chondrocyte glucosaminoglycan synthesis was evenly reduced in all cartilage layers, accompanied by a significant proteoglycan depletion of the matrix which was most marked in the superficial half of the cartilage. These and other changes only partially reversed during a further 4-week period after the injections had been stopped. Our data underline the need for a clear-cut indication for intra-articular injections. The microtrauma caused by injection, in conjunction with the introduction of a carrier solution into the joint, may, at least when repeated at short intervals, lead to measurable damage to the articular cartilage.Recipient of grant no. Ne 308/1-1 from the Deutsche Forschungsgemeinschaft, Bonn, Germany     

关节内注射富含血小板血浆在兔膝骨关节炎动物模型中的作用

目的评估关节内注射富含血小板血浆(PRP)对胶原酶诱导的兔膝骨关节炎(OA)动物模型的影响。方法 20只新西兰大白兔在超声引导下行右侧膝关节内胶原酶注射,建立兔OA动物模型。实验的第4周随机分为PRP组和生理盐水组,每组10只,每周给药1次,共4次。实验第9周取关节软骨观察软骨总体形态及软骨微形态的变化。结果 PRP组软骨总体形态评分为(1.7±0.3)分,明显低于生理盐水组的(3.5±0.6),分,差异有统计学意义(P0.05),提示PRP组软骨总体形态损伤较小;PRP组软骨微形态评分为(3.2±1.0)分,明显低于生理盐水组的(7.4±1.2)分,差异有统计学意义(P0.05),提示PRP组软骨微形态损伤较小。结论关节内注射PRP有保护软骨作用。     

Intra-articular injections and articular cartilage metabolism. An experimental study in rabbits.

We studied the effects of repeated intra-articular injections of sterile 140 mM NaCl solution on articular cartilage in adult rabbits. After 20 injections into the knee joints over a period of 4 weeks, chondrocyte glucosaminoglycan synthesis was evenly reduced in all cartilage layers, accompanied by a significant proteoglycan depletion of the matrix which was most marked in the superficial half of the cartilage. These and other changes only partially reversed during a further 4-week period after the injections had been stopped. Our data underline the need for a clear-cut indication for intra-articular injections. The microtrauma caused by injection, in conjunction with the introduction of a carrier solution into the joint, may, at least when repeated at short intervals, lead to measurable damage to the articular cartilage.     

Cyclodextrin polysulphate protects articular cartilage in experimental lapine knee osteoarthritis

OBJECTIVE: To evaluate the in vivo chondroprotective effect of cyclodextrin polysulphate (CDPS) in a rabbit model of experimental osteoarthritis (OA). DESIGN: Experimental OA was induced in rabbits by anterior cruciate ligament transection (ACLT). Forty-eight hours post-surgery, the rabbits were randomised into three treatment groups (n=15 in each group) and a sham-operated control group. The rabbits were either injected subcutaneously with saline, 0.25mg/kg CDPS or 1mg/kg CDPS once a week for a period of 12 weeks, and their weight was monitored as a parameter for their general status. The animals were then sacrificed for macroscopic and histological assessment of the knee joints. RESULTS: At the lowest dose, CDPS treatment was unable to induce a significant improvement of cartilage degradation vs the saline control in the experimentally induced knee OA. However, subcutaneous injections of 1mg/kg CDPS induced a marked inhibition (P<0.05) of osteophyte formation. Additionally, a significant reduction of cartilage degradation revealed an overall chondroprotective effect of CDPS at a concentration of 1mg/kg. No significant effects on weight gain were noted. CONCLUSIONS: Systemic administration of CDPS is able to protect cartilage in vivo and can therefore be considered as a chondroprotective agent with structure modifying capacities.     

Fibroblast growth factor-18 stimulates chondrogenesis and cartilage repair in a rat model of injury-induced osteoarthritis

OBJECTIVE: Osteoarthritis (OA) is the most common form of arthritis and a primary cause of disability, however, there are no treatments that can slow disease progression or repair damaged joint cartilage. Fibroblast growth factor-18 (FGF18) has been reported to have significant anabolic effects on cartilage. We therefore examined its effects on repair of cartilage damage in a rat meniscal tear model of OA. DESIGN: Surgical damage to the meniscus in rats leads to joint instability and significant damage to the articular cartilage at 3 weeks post-surgery. At this time, animals received bi-weekly intra-articular injections of FGF18 for 3 weeks, and the knee joints were then harvested for histologic examination. RESULTS: FGF18-induced dose-dependent increases in cartilage thickness of the tibial plateau, due to new cartilage formation at the articular surface and the joint periphery. The generation of new cartilage resulted in significant reductions in cartilage degeneration scores. The highest dose of FGF18 also induced an increase in chondrophyte size and increased remodeling of the subchondral bone. CONCLUSIONS: The results of this study demonstrate that FGF18 can stimulate repair of damaged cartilage in a setting of rapidly progressive OA in rats.     

风湿骨痛药酒药槌外治法防治关节软骨退变的实验研究

目的 探讨风湿骨痛药酒药槌外治法防治关节软骨退变的作用机理。方法 选用日本大耳白兔18只，随机将12只通过结扎右股静脉造成膝关节骨性关节炎模型。未造模兔6只为正常组，12只造模兔随机分为造模组和治疗组。造模8周后，治疗组施以风湿骨痛药酒槌外治法进行治疗。造模11周后，观察软骨病理组织形态学、滑膜中NO浓度、软骨中PA及PAI活性和关节液中HA含量变化。结果 治疗组滑膜中NO浓度、软骨中PA及PAI活性和关节液中HA含量与正常组相比有显著性差异。造模组所测各项指标与治疗组相比有显著性差异。治疗组滑膜中NO浓度及软骨中PA含量明显低于造模组，而软骨中PAI含量及关节液中HA含量明显高于造模组。治疗组关节软骨退变不明显，类似正常组，而造模组关节软骨退行性变明显。结论 风湿骨痛药酒药槌外治法能通过改善骨内及周围组织的微循环，抑制滑膜组织中NO的过度产生，降低软骨中PA的活性，提高PAI的活性，消除导致关节软骨退变的内在因素，提高关节液中HA的含量，从而达到保护关节软骨、抗关节软骨退变的目的。