p38蛋白激酶抑制剂对大鼠膝骨关节炎的软骨保护作用

目的观察p38丝裂原活化蛋白激酶(p38 MAPK)抑制剂SB203580对大鼠膝骨关节炎(OA)的软骨保护作用。方法将40只大鼠随机均分为A、B、C、D组,均行单侧膝关节前交叉韧带切断。A、B组术后分别于关节腔注射0.1ml浓度为100、10μm/L的SB203580,每周1次,连续6周;C组注射等量生理盐水;D组不做任何处理。术后6周处死动物,比较各组软骨的大体变化、OA评分、软骨表面分级及Mankin炎性分级。结果A、B组软骨大体评分、软骨表面分级及Mankin炎性分级均轻于C、D组。结论关节腔注射p38 MAPK抑制剂能延缓软骨退变程度,具有软骨保护作用。     

兔膝骨关节炎负重模型的建立

目的 建立兔膝骨关节炎（osteoarthfitis，OA）负重模型，为骨关节炎实验研究奠定基础。方法 兔右侧膝关节前交叉韧带切断建成ACLT模型后，以石膏将左腿固定于腹部，右腿负重锻炼建成重模型，通过大体形态观察、关节液白介素-1（IL-1）、一氧化氮（NO）及软骨基质金属蛋白酶-1（MMP-I）含量的检测反映关节损害程度，比较负重模型与ACLT模型在不同造模时间关节损害的差别。结果 负重模型组较同期ACLT模型组关节退变更为明显；与ACLT模型组相比，负重模型能显著增加OA关节液IL-1、NO的产生和软骨MMP-1的表达（P〈0．05；P〈0．01），在造模4w升高6．5％、22．8％和26．2％，在8w升高3．0％、5．0％和3．6％。结论 兔膝骨关节炎负重模型可加快关节退行性变，缩短造模时间，是一种自然有效且造模时间较短的OA模型。     

关节腔内注射医用臭氧对兔膝骨性关节炎的影响及最佳浓度探讨

目的观察膝关节腔内注射医用臭氧对兔骨性关节炎（OA）的影响。方法将健康家兔32只随机分为空气组及臭氧低、中、高剂量组各8只,每组均采用Hulth法制备OA模型,左侧关节作为对照。造模成功后,臭氧低、中、高剂量组分别予膝关节腔内注射25、40、80μg/ml医用臭氧2 ml,1次/周,共4周;空气组注射空气2 ml。末次注射后3 d处死动物,取股骨内侧髁附近软骨组织行病理学观察及Mankin评分;取关节液行NO检测;分别在造模前、造模成功和处死前测定膝关节活动度。结果各组造模成功时右膝关节活动度较造模前降低,处死前臭氧低、中、高剂量组右膝关节活动度较造模成功时升高,低剂量组处死前即刻右膝关节活动度较中剂量和高剂量组明显升高（P〈0.05）;各组右膝关节软骨病理检查均出现关节软骨损害改变,低剂量组关节软骨退变程度轻,Mankin评分较左膝无显著性差异（P〉0.05）,中、高剂量组关节软骨退变明显,Mankin评分较左膝有显著性差异（P〈0.05）;各组右膝关节关节液中NO含量升高,低剂量组关节液中NO含量相对于中、高剂量组降低程度更大。结论三种浓度的医用臭氧均能够减轻骨性关节炎的炎症反应,而40、80μg/ml的医用臭氧会导致关节软骨组织结构破坏,臭氧浓度越高损害越重。     

维骨力联合透明质酸钠关节内注射治疗膝骨性关节炎的疗效观察

目的 探讨硫酸氨基多糖（维骨力）联合透明质酸钠关节内注射治疗膝骨性关节炎（OA）的疗效。方法 门诊选取膝关节骨关节炎患者240例，随机分为维骨力组、透明质酸钠组以及维骨力联合透明质酸钠用药组，各80例，单盲给药。应用平均WOMAC关节炎指数评分和平均OA严重程度指数来评定两组用药前后临床症状改善情况以及关节磁共振成像（MRI）进行检测关节软骨结构与功能的改变情况，并作统计学分析。结果 经12月治疗观察。联合组治疗后1、3、6月，症状改善最为明显，关节评分3组间比较差异显著（P〈0．01）；且治疗6月后，透明质酸钠组的评分明显升高；到12月，其结果与该组治疗前比较无显著差异（P〉0.05）。而维骨力组与联合组治疗6月后关节评分还在持续下降，但以联合组为甚，两组与治疗前比较均有显著性差异（P〈0.01），且12月后两组间比较有显著性差异（P〈0．05）。关节MRI均显示治疗6月后，联合组与维骨力组软骨磨损出现明显好转与减少，两组间无显著性差异（χ^2=0．087，P〉0．05），但它们与透明质酸钠组比较差异显著（χ^2=38．6，P〈0．01）；到12月后，透明质酸钠组显示软骨磨损比治疗前有所加重，而联合组的MRI的表现结果明显好于维骨力组，且两组间变化差异显著（χ^2=5．91，P〈0．05）。结论 维骨力联合透明质酸钠关节内注射对膝OA的疗效肯定，既能早期控制症状、改善关节功能，又能长期维持疗效，还能保护关节软骨磨损和促进软骨的修复为临床治疗膝老年性OA提供了一条好的治疗方案。     

甲状旁腺激素相关肽在兔骨关节炎模型软骨中的表达

目的 观察甲状旁腺激素相关肽（PTHrP)在骨关节炎（OA）软骨和正常关节软骨中的表达规律。方法 采用膝关节制动法制备OA动物模型。取石膝股骨髁关节标本，以改良的Mankin评分法对OA模型进行评分。经免疫组织化学染色检测PTHrP的表达。并用光镜和图像分析仪测定软骨中PTHRP阳性细胞百分率和PTHrP的表达强度。结果 OA组关节软骨PTHrP阳性细胞百分率和表达强度均比正常关节软骨高，两者间的差异有显著意义。结论 关节软骨中PTHrP的表达可能参与了OA的病理生理过程。     

不同分子量玻璃酸钠关节腔注射治疗轻中度膝骨关节炎疗效比较

目的比较不同分子量玻璃酸钠（SH）关节腔注射治疗轻中度膝骨关节炎（OA）的疗效。方法将120例膝OA患者随机分为两组,治疗组关节腔内注射高分子量SH,对照组注射低分子量SH,1次/周,共5次。比较两组的临床疗效,测定关节液中基质金属蛋白酶（MMP）含量。结果治疗后两组关节液中MMP-1、MMP-3和MMP-9含量均明显降低,治疗组MMP-1、MMP-3含量明显低于对照组。术后6个月治疗组临床疗效优于对照组。结论高分子量SH可明显降低关节液中MMP-1、MMP-3和MMP-9含量,关节腔内注射治疗膝OA的疗效明显优于低分子量SH。     

玻璃酸钠和复方倍他米松治疗膝骨关节炎疗效观察

目前临床上治疗骨关节炎（OA）以服用非甾体抗炎药物（NSAIDs）为主，其主要作用为抗炎、止痛，对关节软骨没有保护作用，甚至某些药物还可引起软骨的分解。透明质酸（玻璃酸）是关节滑液的主要活性成分，具有保护关节软骨，减少软骨的破坏，延缓病情的进展，改善病人生活质量的作用。我科采用玻璃酸钠（SH）和复方倍他米松进行膝关节腔注射治疗OA。     

关节腔冲洗辅助关节镜手术治疗膝关节骨关节炎疗效观察

目的观察关节腔持续冲洗辅助关节镜手术治疗膝关节骨关节炎（OA）的疗效。方法将40例膝关节OA患者随机分为两组,各20例。冲洗组在行关节镜手术治疗后给予生理盐水2 000 ml持续冲洗关节腔,6 h内冲完。非冲洗组行关节镜手术治疗后不冲洗关节腔。术前及术后1周分别抽取关节液2 ml,检测过氧化物歧化酶（SOD）活力。术前及术后1、2周根据JOA标准对患膝进行评分。结果术后1周时,冲洗组关节液中的SOD活力明显高于非冲洗组（P〈0.05）,术后2周时,冲洗组JOA评分高于非冲洗组（P〈0.05）。结论膝关节OA行关节镜手术后予持续冲洗关节腔在短期内可提高关节液中SOD活力,抑制关节内氧自由基的产生,有助于提高手术疗效。     

降钙素对兔膝骨性关节炎Ⅱ型胶原和基质蛋白酶-1的影响

目的探讨降钙素(CT)对兔膝骨性关节炎(KOA)关节软骨、Ⅱ型胶原和基质金属蛋白酶(MMP)-1的影响。方法将36只健康大耳白兔随机分为空白对照组(A组)、模型组(B组)、CT治疗组(C组),B、C两组采用改良Hulth法复制KOA模型,A组除切开皮肤和关节腔外,不做其他处理,从术后第6周开始C组每天皮下注射CT 5 U/kg,A、B组给予等量生理盐水皮下注射,连续注射20 d后处死动物。观察关节软骨病理形态、软骨基质Ⅱ型胶原及MMP-1的变化。结果与A组相比较,B组JP和Mankin评分明显升高,关节软骨损伤明显加重,Ⅱ型胶原表达减少而MMP-1表达增高(P<0.05);与B组比较,C组JP评分和Mankin评分明显降低,软骨损伤明显减轻,Ⅱ型胶原表达明显增多,而MMP-1表达则明显减少(P<0.05)。结论 CT可通过下调MMP-1表达,抑制Ⅱ型胶原降解,减少软骨基质破坏,从而减轻KOA软骨的损伤。     

透明质酸钠对兔骨关节炎软骨诱导型NO合酶基因表达的影响

目的观察透明质酸钠(Na—HA)关节腔注射对骨关节炎(OA)模型关节软骨中诱导型NO合酶(iNOS) mRNA表达水平的影响。方法16只大耳白兔行单侧前交叉韧带切断术，术后5周将动物随机分为实验组和对照组，实验组关节腔注射1％Na—HA 0.3ml，每周1次，连续5周，对照组则注射等量生理盐水。术后10周处死动物，观察两组股骨内髁关节软骨的大体形态学和组织学病理改变，采用反转录一聚合酶链反应(RT—PCR)方法检测软骨iNOS mRNA的表达。结果实验组软骨退变程度较对照组明显减轻，实验组软骨中iNOS mRNA的表达水平与对照组比较差异无统计学意义。结论Na—HA能有效地减轻早期OA关节软骨的退变程度，Na—HA对早期OA软骨中的iNOS的表达没有下调作用。     

盘状结构域受体2与软骨破坏程度的相关研究

目的 观察盘状结构域受体(DDR)2和基质金属蛋白酶(MMP)-13在膝骨关节炎(OA)大鼠不同时期软骨及滑膜中的表达,探讨DDR2与关节软骨破坏之间的关系.方法 采用改良膝关节腔内注射木瓜蛋白酶法制作OA大鼠模型,从蛋白水平检测造模后不同病理阶段关节软骨及滑膜中DDR2和MMP-13的表达规律和分布特点.结果 DDR2在各模型组关节软骨及滑膜中的表达较健康组增高(P＜0.01),并且各组中软骨表达高于相应滑膜,MMP-13表达呈现与DDR2相同的特点,二者相关系数r=0.93(P＜0.01).结论 初步证明"DDR2-MMP-13-软骨破坏"途径在OA病理过程中起重要作用.软骨和滑膜DDR2的表达升高共同促进软骨退变.     

Lipid peroxidation,glutathione, vitamin E,and antioxidant enzymes in synovial fluid from patients with osteoarthritis

Aim: To compare levels of lipid peroxidation and antioxidants in synovial fluid from primary knee osteoarthritis (OA) patients with severe cartilage damage undergoing total knee replacement with those in the synovial fluid from injured knee joint patients with intact cartilage undergoing knee arthroscopy. Methods: Thirty‐two OA patients and 10 injured knee joint patients were recruited. Lipid peroxidation (thiobarbituric acid reactive substances [TBARs]), iron and glutathione (GSH) were measured using a colorimetric method. Vitamin E was measured with high‐performance liquid chromatography (HPLC). Activities of antioxidant enzymes (glutathione peroxidase [GPx], superoxide dismutase [SOD]) were analyzed with the use of a kinetic method. Results: TBARs, iron and GSH levels in synovial fluid were not significantly different between OA patients and injured knee joint patients. Antioxidant enzymes such as GPx and SOD activities also indicated no significant difference. Only vitamin E level was significantly lower in the synovial fluid of OA patients than in that of the injured knee joint patients. Conclusions: Oxidative stress may have a role in pathogenesis of knee osteoarthritis. Vitamin E supplementation may have a role in the management of patients.     

Increased pentosidine, an advanced glycation end product, in serum and synovial fluid from patients with knee osteoarthritis and its relation with cartilage oligomeric matrix protein

BACKGROUND: Pentosidine, an advanced glycation end product, increasingly accumulates in articular cartilage with age, and contributes to the pathogenesis of osteoarthritis (OA). Increased pentosidine concentrations are associated with inflammatory disorders-for example, rheumatoid arthritis. OBJECTIVE: To compare pentosidine serum concentrations in patients with knee OA and in healthy volunteers and to determine a relationship between pentosidine and cartilage oligomeric matrix protein (COMP)-a marker of articular cartilage destruction. METHODS: Paired serum and synovial fluid samples were obtained by arthrocentesis from 38 patients with knee OA and from 38 healthy volunteers. Pentosidine concentration was measured by reverse phase high performance liquid chromatography with fluorescent detection and COMP was determined by sandwich ELISA. RESULTS: Significantly increased serum pentosidine (p<0.01) and COMP (p<0.05) levels were detected in the patients with OA compared with the control group. Serum pentosidine correlated significantly with synovial fluid pentosidine (p<0.001). Pentosidine in synovial fluid (p<0.05) and in serum (p<0.05) correlated significantly with synovial fluid COMP. Pentosidine and COMP concentrations did not correlate significantly with the radiological stage of the disease. CONCLUSION: Increased pentosidine serum concentration in patients with OA and its correlation with the cartilage destruction marker COMP in synovial fluid suggests that pentosidine may be important in OA pathology and is a new potential OA marker.     

Relationship between osteoprotegerin/osteoclastogenesis inhibitory factor concentration in synovial fluid and disease severity in individuals with osteoarthritis of the knee

We studied the relationship between osteoprotegerin (OPG)/osteoclastogenesis inhibitory factor (OCIF) concentration in synovial fluid from individuals with osteoarthritis (OA) of the knee and the severity of this condition. The study population included 111 Japanese women with knee OA (153 knees) and 23 normal controls. Osteoarthritic changes were graded according to the system of Kellgren and Lawrence. The concentration of OPG/OCIF in synovial fluid increased with severity of knee OA and was significantly higher in individuals with OA of grade IV than in those with OA of grade 0 or grade 1. It has been shown in a previous study that administration of OPG/OCIF prevents cartilage destruction in adjuvant-induced arthritis in rats. The increase in the concentration OPG/OCIF in synovial fluid of individuals with knee OA might thus reflect a compensatory response to degeneration of articular cartilage and serve to protect cartilage rather than be a cause of OA.     

Cartilage protection by nitric oxide synthase inhibitors after intraarticular injection of interleukin-1beta in rats.

OBJECTIVE: To evaluate the effect of nitric oxide synthase (NOS) inhibitors on proteoglycan synthesis following intraarticular administration of interleukin-1beta (IL-1beta) in rats. METHODS: Recombinant human IL-1beta and NOS inhibitors with different selectivity for inducible NOS (N-monomethyl-L-arginine [L-NMA], N-iminoethyl-L-ornithine [L-NIO], and S-methylisothiourea [SMT]) were simultaneously administered in rats by a single intraarticular injection in each knee. L-NMA was also infused for 72 hours using an Alzet mini osmotic pump implanted into the peritoneal cavity 24 hours before IL-1beta challenge. NO production was determined as nitrate and nitrite, either in synovial fluid or ex vivo in supernatants of synovium and patellae. Proteoglycan synthesis was measured by ex vivo incorporation of 35SO4(2-) into patellar cartilage. RESULTS: IL-1beta induced a time-dependent increase in NO production in synovial fluid. Synovium and patellae released large amounts of nitrate and nitrite under ex vivo conditions, indicating that both tissues are effective sources of NO within the joint. This production of NO was accompanied by a delayed inhibition of proteoglycan synthesis. The intraarticular administration of L-NMA and L-NIO reduced NO release in synovial fluid and resulted in a partial recovery of proteoglycan synthesis. Under our experimental conditions, SMT failed to reduce NO synthesis and to restore proteoglycan synthesis. The protection of cartilage was improved by the systemic and sustained delivery of L-NMA. However, the complete inhibition of NO production in synovial fluid was not sufficient to fully restore cartilage anabolism. CONCLUSION: Our findings show that in rats: 1) NO may be an early mediator of the effect of IL-1beta on cartilage, 2) NO inhibition may have therapeutic relevance, although it is not sufficient to fully reverse the deleterious effects of IL-1beta, 3) among NOS inhibitors tested, only amino acid derivatives are effective, 4) protection can be achieved by local administration of NOS inhibitors, and 5) systemic and sustained delivery of the NOS inhibitor with the highest efficacy after intraarticular injection provides the most benefit.     

Characterization of human synovial fluid cells of 26 patients with osteoarthritis knee for cartilage repair therapy

Aim: To investigate the possibility of chondrogenic differentiation and cartilage repair of synovial fluid cells of osteoarthritis (OA) knee. Methods: Synovial fluids from 26 patients with OA knee were aspirated from each knee joint and cultured in vitro. The morphology of cultured synovial fluid cells, cell proliferation rate, the phenotype, and chondrogenic differentiation were analyzed in in vitro. Also, human autologous synovial fluid cells were transplanted to OA cartilage, and the cells were traced in ex vivo. Results: In 19 of 26 materials, the cells proliferated satisfactorily. The cell proliferation in six materials was very slow and one material contaminated. Culture‐expanded synovial fluid cells had a fibroblastic morphology and the phenotype was negative for CD10, CD14, and CD45, and positive for CD13, CD44, and CD105. Pellet culture of synovial fluid cells showed chondrogenic differentiation. In the ex vivo study, autologous transplanted synovial fluid cells were observed in repaired or enhanced regenerative cartilage areas and showed a tendency to infiltrate the original degenerative cartilage of OA. Conclusions: This study proved the possibility of chondrogenic differentiation of synovial fluid cells of OA knee joints despite the pathologic environment within a diseased joint. Synovial fluid cells were actually heterogeneous cells but they showed chondrogenic differentiation, similar to that of bone marrow‐derived mesenchymal progenitor cells (BMMPCs). The Ex vivo study suggested that synovial fluid cells had a tendency to adhere to OA degenerative cartilage in humans.     

Prevalence of cartilage shards in synovium and their association with synovitis in patients with early and endstage osteoarthritis.

It has been suggested that incorporation of shards of fibrillated cartilage into the synovium is a cause of synovitis in osteoarthritis (OA). We examined the prevalence with which fragments of cartilage are seen in synovium, and their association with synovitis, in patients with endstage OA and early OA of the knee. Samples of synovium were obtained from 12 patients with endstage OA who were undergoing knee joint replacement and 30 with only mild/moderate radiographic changes of OA who exhibited articular cartilage changes of OA at arthroscopy. The presence of cartilage shards was sought in synovium from the medial and lateral tibiofemoral compartments and the suprapatellar pouch of each patient. Comparable volumes of the synovial lining from patients with endstage and early OA were examined, and tissue mononuclear cell infiltration was graded as an indicator of synovitis. Cartilage shards were seen in synovium from 7 of 12 patients with endstage OA, all of whom had synovitis. No topographic relationship was found between shards and mononuclear cell infiltration. In contrast, cartilage fragments were not seen in synovium from any of the 30 patients with early OA, although 9 of them had full thickness cartilage ulcers and 17 had evidence of synovitis.     

The quantitation of a native chondroitin sulfate epitope in synovial fluid lavages and articular cartilage from canine experimental osteoarthritis and disuse atrophy

Objective. Previous studies have shown the presence of a native chondroitin sulfate epitope in articular cartilage proteoglycans from canine knee joints with experimental early osteoarthritis (OA), but not in normal cartilage. The objective of this study was to quantitate the native epitope recognized by monoclonal antibody 3-B-3 in synovial fluids and articular cartilage of diseased joints. Methods. An immunoassay with monoclonal antibody 3-B-3, which recognizes a native chondroitin-6-sulfate structure, was developed and used to analyze synovial fluid lavage material and extracts of articular cartilage from canine knee joints with early experimental OA or with mild disuse atrophy, and from control animals. Results. The concentration of epitope in the OA fluids was elevated 33–35-fold, and in the OA articular cartilage extracts it was elevated > 200-fold, compared with samples from the control group. No significant difference was detected in the levels of 3-B-3 epitope in the synovial fluid lavage material or cartilage extracts from the joints of the disuse group versus the control group. Conclusion. The native 3-B-3 epitope in articular cartilage and synovial fluids may be a specific marker of ongoing anabolic events in early degenerative joint disease.     

Canine serum keratan sulfate and hyaluronate concentrations. Relationship to age and osteoarthritis

Elevated serum levels of keratan sulfate (KS) and hyaluronate (HA) in patients with osteoarthritis (OA) have been reported. We measured KS and HA in dogs to determine if there was an elevation of these serum glycosaminoglycans in a canine model of OA. A single intraarticular injection of 1 mg of chymopapain into a shoulder joint increased serum KS by tenfold, and HA by less than twofold, in 24 hours. Serum KS and HA levels were 3-5-fold higher in dogs younger than 2 months of age than in older dogs. Serum KS and HA concentrations and synovial fluid KS concentrations were unrelated to spontaneous cartilage degeneration in 1-year-old dogs. Higher KS levels in synovial fluid correlated with higher KS levels in serum (r = 0.54, P less than 0.025). The mean KS concentration in sera of older dogs (greater than 3 years old) with OA was 37% higher than that in disease-free controls, but the difference between the groups was not statistically significant. Thus, elevated levels of serum KS and HA do not appear to have clinical significance in this model of OA.