膝关节骨性关节炎血液和滑液MMP-3 水平与其病变程度的相关性

目的探讨膝关节骨关节炎（osteoarthritis’OA)患者血液和滑液MMP-3水平与其病变严重程度的关系。方法随机选取33例膝关节0A患者[男20例,女13例;年龄55?87岁,平均（65. 8 ±6. 8)岁]和15名健康对照者[男9例,女6例;年龄53?83岁,平均（62. 3 ± 7. 2)岁]作为研究对象。采用Kellgren-Lawrence标准进行放射学分级,酶联免疫吸附法（ELISA)测定关节血液和滑液MMP-3 水平。结果0A患者与对照者相比,关节血液MMP-3水平（178. 61 ±83. 82) ： (73. 26 ±21.51)ng/ml 和滑液MMP-3水平（5612.31 ± 3013.93)： (782. 63 ±426. 22)ng/ml均明显升高。关节血液和滑液 MMP-3水平均与0A病变严重程度（KL分级）呈正相关卜=0. 627,P<0. 001;r=0. 55,P =0.001)。关节血液MMP-3水平与滑液MMP-3水平呈正相关（r =0. 509,P =0. 002)。结论0A患者血液和滑 液MMP-3水平都与病变严重程度相关。因此,MMP-3在0A的病理过程中起关键作用,并且可以作 为0A病变严重程度的生物标记。     

女性骨关节炎患者关节液visfatin水平与CTX-Ⅱ的相关分析

目的研究女性骨关节炎患者关节液visfatin水平与CTX-Ⅱ的相关分析。方法利用ELISA方法测定30例OA患者及12例对照组血浆和关节液visfatin及CTX-Ⅱ水平。结果 OA患者关节液visfatin浓度明显高于对照组（8.95±2.51ng/ml vs 4.48±2.49ng/ml,P〈0.001）,两组血浆visfatin浓度无统计学意义（5.37±1.45ng/ml vs 4.93±1.4ng/ml,P=0.466）。关节液visfatin水平与Ⅱ型胶原降解产物CTX-Ⅱ呈正相关（r=0.497,P〈0.01）,与hsCRP无相关（r=-0.246,P=0.19）。K-LⅣ级OA患者关节液visfatin浓度明显高于K-LⅢ级visfatin浓度（10.57±2.49ng/ml vs 7.54±1.5ng/ml,P〈0.01）。关节液visfatin水平明显与OA严重程度正相关（r=0.421,P=0.021）。结论 Visfatin可能通过关节腔局部作用参与OA关节软骨降解通路。     

Human mesenchymal stem cells in synovial fluid increase in the knee with degenerated cartilage and osteoarthritis

We investigated whether mesenchymal stem cells (MSCs) in synovial fluid (SF) increased in the knee with degenerated cartilage and osteoarthritis. SF was obtained from the knee joints of 22 patients with anterior cruciate ligament (ACL) injury during ACL reconstruction, and cartilage degeneration was evaluated arthroscopically. SF was also obtained from the knee joints of 6 healthy volunteers, 20 patients with mild osteoarthritis, and 26 patients with severe osteoarthritis, in which the grading was evaluated radiographically. The cell component in the SF was cultured for analyses. Synovium (SYN) and bone marrow (BM) were also harvested during total knee arthroplasties. The MSC number in SF was correlated with the cartilage degeneration score evaluated by arthroscopy. The MSC number in the SF was hardly noticed in normal volunteers, but it increased in accordance with the grading of osteoarthritis. Though no significant differences were observed regarding surface epitopes, or differentiation potentials, the morphology and gene profiles in SF MSCs were more similar to those in SYN MSCs than in BM MSCs. We listed 20 genes which were expressed higher in both SYN MSCs and SF MSCs than in BM MSCs, and 3 genes were confirmed by quantitative RT-PCR. MSCs in SF increased along with degenerated cartilage and osteoarthritis.     

Exercise and injury increase chondroitin sulfate chain length and decrease hyaluronan chain length in synovial fluid

OBJECTIVES: (1) To investigate the effects of exercise and osteochondral (OC) injury on synovial fluid (SF) chondroitin sulfate (CS) and hyaluronan (HA) concentration and chain length, (2) to compare SF and cartilage CS data from joints with OC fragmentation, and (3) to compare SF CS and HA profiles with those seen in serum from the same horses. METHODS: Serum and SF were obtained from (1) normal horses after 8 weeks rest, (2) the same horses after 9 months treadmill training, and (3) horses with OC injury from racing. Articular cartilage was also collected from group 3 horses. Concentrations and chain lengths of CS and HA were determined by gel chromatography and fluorophore-assisted carbohydrate electrophoresis. RESULTS: SF CS peak chain length in the OC injury group increased significantly (18.7kDa) when compared to rested horses (11.6kDa), with exercise producing an intermediate chain length (15.6kDa). Cartilage and serum from the OC injury group had the abnormally long CS chains seen in SF from these horses. Total SF HA was significantly lower in the OC injury group compared to the rested group. Both the OC injury group and the exercised group had significant decreases in SF HA chain length compared to the rested group. CONCLUSIONS: Chain length of SF CS was increased by exercise and OC injury. Exercise resulted in a modest increase, whereas OC injury caused a marked increase. In contrast to CS, SF HA chain length was decreased by OC injury, and to a lesser extent by exercise. Chain length analysis of SF CS and HA may provide a useful tool for evaluation of joint health.     

Differential levels of synovial fluid aggrecan aggregate components in experimental osteoarthritis and joint disuse

The levels of proteoglycan aggregate components (link protein, keratan sulfate epitope, and total sulfated glycosaminoglycan) were determined in the synovial fluid lavages of dogs with experimental osteoarthritis or disuse atrophy. A model of experimental osteoarthritis was created by transection of the anterior cruciate ligament of the right knee; studies were carried out 6 and 12 weeks after surgery. Joint disuse was studied at 4 and 8 weeks after initiation of the disuse. Recovery after disuse also was studied in joints that had 3 weeks of remobilization after 4 or 8 weeks of disuse. Synovial fluid lavages from the right knee joints of untreated animals were used as controls. The concentrations of keratan sulfate epitope, sulfated glycosaminoglycan, and link protein in the synovial fluid lavages at 6 and 12 weeks after transection of the anterior cruciate were elevated compared with the control values. Similar analysis of the fluid after disuse showed that the levels of keratan sulfate epitope and sulfated glycosaminoglycan were increased compared with the control levels and the levels after transection. However, the concentration of link protein in the fluid after disuse was not significantly different from the control level. The levels of keratan sulfate epitope and sulfated glycosaminoglycan in the synovial fluid lavages after disuse with recovery were high, but the levels of link protein remained low. The results indicate that the catabolism of proteoglycan aggregates in articular cartilage during early osteoarthritis and disuse is different. The determination of keratan sulfate epitope in synovial fluid lavages appears to provide a relatively general indication of proteoglycan catabolism, whereas increased levels of link protein may be more indicative of cartilage degeneration.     

基质金属蛋白酶家族在骨关节炎软骨组织中表达的研究

[目的]观察骨关节炎关节软骨中MMP-7、MMP-9、MMP-13和TIMP-1的表达,探讨其与软骨退变的关系及可能的作用机制。[方法]选取20例因骨关节炎行关节置换的软骨组织,常规HE染色观察其组织学形态,ABC免疫组化法观察关节软骨MMP-7、MMP-9、MMP-13和TIMP-1的表达,2例因意外受伤截肢患者的正常膝关节软骨标本作为对照。统计采用Mann-Whitney U非参数检验及相关分析。[结果]骨关节炎关节软骨出现裂隙、纤维化,软骨细胞增多、排列紊乱,并出现大量簇聚软骨细胞和肥大软骨细胞。MMP-7和MMP-13在正常软骨全层均呈低表达,但在退变软骨中的表达则明显增多,光密度值行U检验,两组差异有显著性(P<0.01)。在正常与OA软骨的浅层,MMP-9和TIMP-1的表达无显著性差异(P>0.05);但在深层软骨中,OA软骨MMP-9和TIMP-1的表达较正常软骨明显增多,两组差异有显著性(P<0.01)。[结论]MMP-7,13在OA软骨全层表达均多于正常软骨;MMP-9,13仅在OA软骨深层出现过多表达。MMPs与TIMPs的失衡是导致关节软骨发生组织学退变的原因之一。     

血清和关节液中钙结合蛋白（S100A12)的水平与膝关节骨关节炎严重程度的相关性分析

目的探讨患者关节液和血清中钙结合蛋白(S100A12)浓度和膝关节骨性关节炎严重程度相关性,以评估它在骨性关节炎(osteoarthritis,OA)发病中的作用。方法 40例骨性关节炎(OA组)与40例需关节镜手术的半月板、韧带损伤的患者及行髓内钉固定的股骨及胫骨骨折患者(对照组);对两组患者静脉采血及取关节液,采用ELISA法测S100A12含量;采用Kellgren-Lawrence(K-L)标准对膝关节X线片进行评估分级。结果 OA组的血清中S100A12水平(35.10±26.43)ng/ml显著高于对照组含量(22.13±15.67)ng/ml,有统计学差异(P=0.026);OA组关节液S100A12含量(14.09±8.08)ng/ml显著高于对照组关节液中的含量(9.30±6.85)ng/ml(P=0.02)。OA组血清中的S100A12水平显著高于关节滑液中的水平(P0.01)。关节液中的S100A12含量和血液中的S100A12水平显著正相关(r=0.42,P=0.02),但相关关系并不密切。OA组关节滑液中S100A12与K-L分级成正相关(r=0.65,P0.01)。血清中的S100A12与K-L分级关联不显著(r=0.29,P=0.24)。结论 S100A12在OA患者的血液及关节液中都有表达;关节液的S100A12表达与患者骨关节炎严重程度成正相关;检测关节液中的S100A12有助于骨性关节炎的早期诊断,提示病变严重程度,为临床治疗提供参考。     

Relationship of plasma and synovial fluid BMP-7 with disease severity in knee osteoarthritis patients: a pilot study

The objective of this study was to investigate bone morphogenetic protein-7 (BMP-7) levels in both plasma and synovial fluid of patients with primary knee osteoarthritis (OA) and to determine their relationship to disease severity. Thirty-two patients with knee OA and 15 healthy subjects were enrolled in the study. Anteroposterior knee radiographs were taken to determine the disease severity of the affected knee. The radiographic grading of OA in the knee was performed using the Kellgren-Lawrence criteria. BMP-7 levels in the plasma and synovial fluid were measured using enzyme-linked immunosorbent assay. The mean plasma BMP-7 concentration of the knee OA patients was significantly higher compared with that of healthy controls (12.1 ± 1.6 vs 3.5 ± 0.9 pg/ml, P = 0.001). Although BMP-7 levels in plasma were higher with respect to paired synovial fluid samples, the difference was not statistically significant (12.1 ± 1.6 vs 10.5 ± 2.2 pg/ml, P = 0.3). Subsequent analysis showed that plasma BMP-7 levels significantly correlated with disease severity (r = 0.77, P < 0.001). Furthermore, the synovial fluid levels of BMP-7 also correlated with disease severity (r = 0.60, P < 0.001). In addition, plasma BMP-7 levels showed a positive correlation with synovial fluid BMP-7 levels (r = 0.71, P < 0.001). Overexpression of BMP-7 in plasma and synovial fluid is related to progressive joint damage in knee OA. These findings suggest that BMP-7 might serve as a biochemical parameter for determining disease severity in primary knee OA and could play a potential role in cartilage protection and repair of OA.     

关节液中脂联素水平与膝关节骨关节炎严重程度的相关性研究

背景：众所周知,肥胖是引起骨关节炎发生、发展的危险因素之一。肥胖人群容易发生关节炎的原因是超载作用和代谢因素。脂肪组织是一个活跃的器官,它的分泌脂肪细胞因子包括脂联素进入全身循环,参与了骨关节炎的发病。 目的：测量骨关节炎患者血清和关节滑液中脂联素水平,分析其与膝关节骨关节炎严重程度的相关性。 方法：64例膝关节骨关节炎患者（关节炎组）和19例因半月板或韧带损伤患者（对照组,行关节镜检查后排除软骨损伤）被纳入本研究。采用Kellgren-Lawrence（KL）标准对关节炎患者的膝关节前后位X线片进行评估分级。使用酶联免疫吸附试验检测脂联素在对照组和关节炎组患者血清和关节滑液中的表达水平。使用WOMAC量表（Western On-tario and McMaster Universities Arthritis Index）评估患者膝关节功能。 结果：骨关节炎组患者血清中的脂联素水平[（6177.8±991.0）ng/ml]高于对照组[（5793.4±673.3）ng/ml],但差异无统计学意义（P=0.12）。但在骨关节炎组患者,血清中的脂联素浓度[（6177.8±991.0）ng/ml]显著高于关节滑液中的脂联素[（824.3±304.7）ng/ml,P〈0.001）。关节炎患者关节液中脂联素浓度与评估疾病的严重程度呈负相关（r=-0.51,P〈0.001）,但血清脂联素浓度与疾病的严重程度关联不显著（r=-0.17,P=0.18）。血清脂联素水平、滑膜液脂联素水平与MOMAC疼痛评分均不显著相关（r=-0.01和r=-0.13,P＞0.05）。 结论：骨关节炎患者滑膜液中脂联素水平与膝关节炎严重程度呈负相关。检测关节滑液中的脂联素可能有助于早期发现关节炎。     

Rapid increase in serum levels of matrix metalloproteinase-9 (MMP-9) postoperatively is associated with a decrease in the amount of intracellular MMP-9

We have previously demonstrated a significant decrease in the serum concentration of intact insulin-like growth factor-binding protein (IGFBP-3) after laparotomy. IGFBP-3, a major IGF binding protein, inhibits the growth of tumor cells via several mechanisms. Our goal was to determine, in a murine model, whether matrix metalloproteinase-9 (MMP-9), a known protease of IGFBP-3, is responsible for the postoperative decrease in serum IGFBP-3 levels. Six IGFBP-3 transgenic mice on a CD-1 background were used in this study. These mice over-express human IGFBP-3. Sham laparotomy, in the form of a midline abdominal incision, was the test procedure. General anesthesia was established using ketamine and xylazine immediately before a 30-minute sham laparotomy and before preoperative blood sampling, done via retro-orbital venipuncture, 48 hours before surgery. The animals were sacrificed and blood was drawn 24 hours postoperatively. Plasma MMP-9 activity was measured using zymography at each time point (48 hours before and 24 hours after operation). MMP-9 activity was also measured in mononuclear cell lysates at both time points. Zymography analysis demonstrated significantly higher plasma levels of MMP-9 postoperatively compared with preoperative levels (81 RU vs 40 RU; P < .05). In contrast, mononuclear cell levels of MMP-9 were significantly higher preoperatively compared with postoperative levels (37.5 RU vs. 0.75 RU, P < .05). Plasma levels of MMP-9, a known protease of IGFBP-3, are significantly elevated postoperatively. In addition, mononuclear cells that store MMP-9 are depleted of it postoperatively. This suggests that rapid MMP-9 release by mononuclear cells leads to an increase in serum levels of this protease postoperatively. Further studies will elucidate mechanisms of MMP-9-related IGFBP-3 depletion.     

膝骨关节炎患者关节滑液中骨桥蛋白和软骨寡聚基质蛋白水平与疾病严重程度相关性

目的:探讨膝骨关节炎（knee osteoarthritis,KOA）患者关节滑液中骨桥蛋白（osteopontin,OPN）和软骨寡聚基质蛋白（cartilage oligomeric matrix protein,COMP）水平与疾病严重程度的相关性。方法：选取2018年2月至2020年5月收治的59例KOA患者作为KOA组,其中男25例,女34例;年龄60~75（65.57±1.56）岁;体质量指数（body mass index,BMI）21.4~30.7（26.12±1.54） kg/m²。采用Kellgren-Lawrence（K-L）分级对X线结果进行评估,其中Ⅱ级（K-L2组）14例,Ⅲ级（K-L3组）27例,Ⅳ级（K-L4组）18例。另选取18例因韧带或半月板疾病进行关节镜检查且无软骨损伤患者作为对照组,男7例,女11例;年龄61~78（64.88±1.60）岁;BMI 22.8~29.9（25.89±1.49） kg/m²。治疗前采集研究对象关节滑液样本,采用酶联免疫吸附试验检测关节滑液OPN、COMP水平,比较KOA组与对照组关节滑液OPN、COMP水平。比较不同K-L分级KOA患者性别、年龄、BMI等临床资料,采用酶联免疫试验检测其关节滑液中白细胞介素-1β（interleukin -1β,IL-1β）,OPN,COMP,基质金属蛋白酶3（matrix metalloproteinase -3,MMP-3）水平,比较不同K-L分级KOA患者临床资料和生化指标,采用Logistic回归分析影响KOA患者K-L分级的因素,采用ROC曲线下面积（area under the curve,AUC）预测KOA疾病严重程度。结果：59例KOA患者获得随访,时间8~27（15.75±3.27）个月。KOA组关节滑液OPN、COMP水平高于对照组（P<0.001）。K-L2组、K-L3组、K-L4组IL-1β、OPN、COMP、MMP-3水平比较差异有统计学意义（P<0.001）;与K-L2组比较,K-L3、K-L4关节滑液关节滑液IL-1β、OPN、COMP、MMP-3 水平均升高（P<0.05）;与K-L3组比较,K-L4关节滑IL-1β、OPN、COMP、MMP-3水平均升高（P<0.05）。多因素Logistic回归分析结果显示：关节滑液OPN、COMP、MMP-3水平是影响KOA患者K-L分级的独立危险因素（OR=6.653, 4.229,1.579,P<0.001）。关节滑液OPN预测K-L4级KOA的AUC为0.720[95%CI（0.588-0.851）],灵敏度为94.4%,特异度为65.9%;关节滑液COMP预测K-L4级KOA的AUC为0.731[95%CI（0.592-0.870）],灵敏度为88.9%,特异度为63.4%;关节滑液OPN联合COMP预测K-L4级KOA的AUC为0.839[95%CI（0.724-0.953）],灵敏度为94.4%,特异度为51.2%;OPN联合COMP预测K-L4级KOA的AUC大于单独OPN、COMP的AUC（Z=4.037,3.540,P<0.05）。结论：KOA患者关节滑液OPN、COMP水平升高,并随着K-L分级增加而升高。关节滑液OPN、COMP是影响KOA患者K-L分级的独立危险因素,二者预测K-L4级KOA 的AUC、灵敏度、特异度高,可用于评估KOA疾病进展。     

关节液YKL-40水平与骨关节炎的相关性研究

[目的]探讨膝骨关节炎患者关节液中YKL-40水平与膝关节病变程度的相关性,为关节炎的早期诊断提供新的思路和临床依据。[方法]114例骨关节炎患者,WOMAC评分系统评估患者临床症状程度,X线K-L分级方法评估影像改变程度。抽取患膝关节滑液,采用双抗夹心酶联免疫吸附法检测关节滑液中YKL-40的水平。采用秩相关分析的多元相关分析YKL-40水平与骨性关节炎程度的相关性。[结果]不同K-L分期组滑液YKL-40水平差异有统计学意义,Ⅳ级患者高于Ⅲ级患者,Ⅲ高于Ⅱ级患者(P<0.05),秩相关分析显示,滑液SYKL-40水平与患者X线K-L分级呈正相关(r=0.542,P<0.001)。此外,秩相关分析显示,滑液YKL-40水平与WOMAC疼痛(r=0.517,P=0.001)、肢体功能(r=0.380,P=0.025)以及总分(r=0.407,P=0.010)呈正相关,但与WOMAC关节僵硬评分无明显相关性(r=0.093,P=0.133)。多元相关分析进一步证实,滑液YKL-40水平与WOMAC疼痛、肢体功能评分评分及总分呈正相关(P<0.05)。[结论]膝骨关节炎患者关节液YKL-40水平与骨关节炎严重程度的相关性,可作为一种潜在的评估膝骨关节炎症状严重性的生物标记物。     

膝关节骨关节炎滑液和软骨骨桥蛋白水平与其病变程度的相关性

Objective To examine osteopontin (OPN) levels in both synovial fluid and articular cartilage of patients with primary knee osteoarthritis (OA) and to investigate their relationship with severity of the disease. Methods Fifty patients with knee OA and 10 healthy controls were enrolled in this study.There were 15 males and 35 females with an average age of 61.8±7.4 years in OA group. The control group included 4 males and 6 females with an average age of 63.2±6.0 years. Mankin score were taken to determine the disease severity of the affected knee. The radiographic grading of OA in the knee was performed using the Kellgren-Lawrence criteria. OPN levels in synovial fluid were measured using enzyme-linked immunosorbent assay. OPN levels in articular cartilage were assessed by immunohistochemical methods. Results Compared to healthy controls, the knee OA patients had higher OPN concentration in synovial fluid ([4519.60±1830.37] pg/ml vs. [1179.70±303.39] pg/ml) and articular cartilage([0.60±0.06] vs. [0.43 ±0.07]). In addition, synovial fluid OPN levels showed a positive correlation with articular cartilage OPN levels (r=0.411,P=0.003). Subsequent analysis showed that the OPN levels in synovial fluid significantly had been correlated with severity of disease using Kellgren-Lawrence criteria (r=0.581, P＜ 0.001). Furthermore,the levels of OPN in the articular cartilage also were correlated with disease severity using Mankin score (r=0.675, P＜ 0.001).Conclusion The data suggest that OPN in synovial fluid and articular cartilage is related to progressive joint damage and could be a predictive biomarker respect to disease severity and progression in knee OA.     

膝关节骨关节炎滑液和软骨骨桥蛋白水平与其病变程度的相关性

Objective To examine osteopontin (OPN) levels in both synovial fluid and articular cartilage of patients with primary knee osteoarthritis (OA) and to investigate their relationship with severity of the disease. Methods Fifty patients with knee OA and 10 healthy controls were enrolled in this study.There were 15 males and 35 females with an average age of 61.8±7.4 years in OA group. The control group included 4 males and 6 females with an average age of 63.2±6.0 years. Mankin score were taken to determine the disease severity of the affected knee. The radiographic grading of OA in the knee was performed using the Kellgren-Lawrence criteria. OPN levels in synovial fluid were measured using enzyme-linked immunosorbent assay. OPN levels in articular cartilage were assessed by immunohistochemical methods. Results Compared to healthy controls, the knee OA patients had higher OPN concentration in synovial fluid ([4519.60±1830.37] pg/ml vs. [1179.70±303.39] pg/ml) and articular cartilage([0.60±0.06] vs. [0.43 ±0.07]). In addition, synovial fluid OPN levels showed a positive correlation with articular cartilage OPN levels (r=0.411,P=0.003). Subsequent analysis showed that the OPN levels in synovial fluid significantly had been correlated with severity of disease using Kellgren-Lawrence criteria (r=0.581, P＜ 0.001). Furthermore,the levels of OPN in the articular cartilage also were correlated with disease severity using Mankin score (r=0.675, P＜ 0.001).Conclusion The data suggest that OPN in synovial fluid and articular cartilage is related to progressive joint damage and could be a predictive biomarker respect to disease severity and progression in knee OA.     

膝关节骨关节炎滑液和软骨骨桥蛋白水平与其病变程度的相关性

目的 探讨膝关节骨关节炎(osteoarthritis,OA)患者滑液和关节软骨骨桥蛋白(osteopontin,OPN)水平及其与病变严重程度的关系.方法 随机选取50例膝关节OA患者[男15例,女35例;年龄48～81岁,平均(61.8±7.4)岁]和10名健康对照者[男4例,女6例;年龄59～68岁,平均年龄(63.2±6.0)岁]作为研究对象.采用Mankin评分评价疾病严重程度,Kellgren-Lawrence标准进行放射学分级,酶联免疫吸附法测定关节滑液OPN水平,免疫组化方法测定关节软骨OPN光密度值.结果 OA患者与对照者相比,关节滑液OPN水平[(4519.60±1830.37)pg/ml:(1179.70±303.39)pg/ml)和关节软骨OPN光密度值[(0.60±-0.06):(0.43±0.07)]均明显升高.关节滑液OPN水平与关节软骨OPN表达呈正相关(r=0.411,P=0.003).关节滑液OPN水平与OA病变严重程度(KL分级)呈正相关(r=0.581,P＜0.001).关节软骨OPN表达与OA病变严重程度(Mankin评分)呈正相关(r=0.675,P＜0.001).结论 关节滑液和关节软骨OPN水平与病变严重程度相关.