Fluorescence histochemical studies of adrenergic nerve fibres and serotonin containing mast cells of the knee joint synovial membrane in normal and experimental arthritic rats

Summary Distribution of noradrenergic nerve fibres and serotonin containing mast cells in the normal and experimental arthritic synovial membrane of the rat knee joint has been demonstrated using the fluorescence histochemical method of Falck and associates [3]. Noradrenergic nerve fibres, usually associated with the arteries and arterioles, as well as mast cells were mainly found to be in the subsynovial layer close to the ligamentous layer. They are increased in the synovial membrane of rats with adjuvant arthritis.

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Résumé Grâce à la technique de fluorescence histochimique de Falck et coll. [3], les auteurs ont précisé la distribution des fibres nerveuses noradrénergiques et des mastocytes renfermant de la sérotonine dans la membrane synoviale du genou du rat normal, ainsi que dans celle de l'animal atteint d'une arthrite expérimentale. Les fibres nerveuses noradrénergiques, en relation ou non avec les artères et les artérioles, ainsi que les mastocytes ont été retrouvés essentiellement dans la couche sous-synoviale, à proximité du plan ligamentaire. Ils augmentent dans la membrane synoviale des rats atteints d'arthrite.

     

CD44H and the isoforms CD44v5 and CD44v6 in the synovial fluid of the osteoarthritic human knee joint

OBJECTIVE: To determine whether the concentrations of CD44H and v5 and v6 in the synovial fluid are correlated with the presence of accompanying synovitis in the osteoarthritic joint and with the grade of osteoarthritis. DESIGN: Using antero-posterior and lateral X-rays of the knee joint and patellar view of 46 patients were graded with the Kellgren & Lawrence scale. Synovial fluid from these patients with different grades of primary osteoarthritis of the knee joint with and without synovial inflammation (synovitis) was collected during surgical procedures. A horseradish peroxidase conjugated anti-CD44H-, anti-sCD44v5- or anti-sCD44v6-antibody was added and labeled with tetramethylbenzidine. The absorbance was measured at wavelengths of 450/620 nm. Regression analysis was performed and the statistical significance was assessed, using the Student t-test for unequal variance. RESULTS: CD44H and v5 and v6 were detected in the synovial fluid of all 46 patients. Osteoarthritic patients with synovial inflammation showed significantly higher levels of CD44H and v6, but not v5, than osteoarthritic patients without synovial inflammation. With progression of osteoarthritis from Kellgren grade II to III, CD44v5 decreased significantly. All other isoform concentrations remained statistically unchanged. CONCLUSIONS: CD44H and the isoforms v5 and v6 were present in the synovial fluid of osteoarthritic patients. Their concentrations do not reflect the osteoarthritic stage in the Kellgren grading scale. CD44H and CD44v6, but not CD44v5, are significantly up-regulated in osteoarthritic synovial inflammation.     

Significance of chondroitin sulfate isomers in the synovial fluid of osteoarthritis patients

To test whether chondroitin sulfate (CS) isomers in synovial fluid are related to the radiographic severity of osteoarthritis (OA) and the age and sex, we investigated the concentrations of chondroitin 6-sulfate (C6S) and chondroitin 4-sulfate (C4S) in patients with OA of the knee joint and age-matched healthy controls. Synovial fluid was obtained from 133 patients with OA of the knee and 27 volunteers (controls). Twenty-seven patients were radiographically classified as grade 1, 59 as grade 2, 34 as grade 3, and 13 as grade 4 according to a modified version of the Kellgren and Lawrence scale. Compared with findings in the controls, values for the concentrations of C6S and C4S, and the C6 : C4S ratio, were significantly higher in the grade 1 patients, whereas in grade 3 and 4 patients, the C6S concentration and the C6 : C4S ratio were lower. There was an obvious sex difference, and all values for the women were significantly lower than those for the men in both the control and the patient groups. In the control group, multiple regression analysis showed a moderate inverse correlation between age and all three of the above biochemical variables, and sex was moderately associated with the C6S concentration and the C6S : C4S ratio. In the patient group, age and sex showed a poor correlation with any of the three variables. However, the radiographic severity of OA showed a relatively strong inverse correlation with the C6S concentration and the C6S : C4S ratio, and a weak inverse correlation with the C4S concentration. Received: June 11, 2001 / Accepted: October 22, 2001     

膝关节液中生物学标记物Ⅱ型胶原羧基端端肽的检测及临床意义

目的探讨膝关节液中Ⅱ型胶原羧基端端肽（CTX-Ⅱ）含量与关节软骨损伤和滑膜炎两种病变程度的关系。方法采用ELISA法对膝关节骨性关节炎37例患者（骨性关节炎组，即OA组）、膝关节其他疾病34例患者（非OA组）关节液中CTX-Ⅱ含量进行检测，并在关节镜下利用Outerbridge软骨损伤评分法和Ayral滑膜炎评分法对软骨损伤程度和滑膜炎程度进行评价。结果OA组关节液中CTX-Ⅱ含量高于非OA组（t=-7．558，P〈0．05）；Outerbridge软骨损伤累计评分≥10分组关节液CTX-Ⅱ的含量高于Outerbridge软骨损伤累计评分〈10分组（t=-7．235，P〈0．05）；Ayral滑膜炎评分≥60分组关节液CTX-Ⅱ的含量高于Ayral滑膜炎评分〈60分组（t=-5．113，P〈0．05）；71例膝关节疾病患者CTX-Ⅱ含量除主要与Outerbridge软骨损伤累计评分呈正相关外（r=0.743，P〈0．05），还与Ayral滑膜炎评分呈正相关（r=0．706，P〈0．05）；OA组中Out—erbridge软骨损伤累计评分与Ayral滑膜炎评分呈正相关（r=0．418，P〈0．05）；OA组中CTX-Ⅱ含量与Outerbridge软骨损伤累计评分（r=0．533，P〈0．05）和Ayral滑膜炎评分均呈正相关（r=0．522，P〈0．05）。结论膝关节液中CTX-Ⅱ的含量除主要反映关节软骨损伤累计程度外，在一定程度上还反映关节滑膜炎程度；膝关节液CTX-Ⅱ含量升高提示软骨损伤程度较重。     

Monocyte chemoattractant protein 1 expression in synovial fluid of patients with total hip arthroplasty

Monocyte chemoattractant protein 1 (MCP-1) is a member of the chemokines family that stimulates monocyte chemotaxis and whose expression is induced during osseous inflammation. Total hip arthroplasty (THA) now uses ceramic-on-ceramic (cer-cer) or metal-on-metal (met-met) coupling that is considered the bearing of new generation, with improved performance compared with the previous polyethylene bearings. While the ceramic on ceramic is universally considered highly biocompatible, met-met coupling can induce hypersensitivity correlated to immunological response. This study aimed at quantifying the MCP-1 levels in synovial fluid of THA patients with these two articular couplings; synovial fluid of osteoarthritic patients who underwent primary THA was used as a control. The presence of wear particles originating from the prostheses was also assessed in the synovial fluid by scanning electron microscopy and energy dispersive X-ray spectroscopy. The MCP-1 levels in the synovial fluid of 17 patients with met-met coupling were significantly higher than in 13 patients with cer-cer coupling and in seven controls; median MCP-1 level was 919 pg/mL in the cer-cer group, 4274 pg/mL in the met-met group, and 326 pg/mL in the control group. No correlation was found between MCP-1 levels and the presence of wear particles in the synovial fluid, time from surgery, or periprosthetic osteolysis. We conclude that the presence of the prosthesis is responsible for an increase in the level of MCP-1; moreover, the significantly higher level of this chemokine in met-met coupling can be related to inflammatory and possibly hypersensitivity processes induced by metal ions.     

Two-dimensional gel electrophoresis of synovial fluid: method for detecting candidate protein markers for osteoarthritis

Synovial fluid (SF) is a dynamic reservoir for proteins originating from serum, synovial tissue, and cartilage. The composition of the SF proteome may reflect the pathophysiological conditions affecting the circulatory system and cartilage. Our long-term goal is to identify reliable protein markers for osteoarthritis (OA) in SF. We first evaluated the pattern of SF proteins on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) as a function of protein loading, pH range for isoelectric focusing, and concentration of acrylamide in SDS-PAGE. Removal of albumin and -globulins from the samples did not improve the detection of protein spots on 2D-PAGE. The repeatability of protein spot intensity was tested by triplicate 2D-PAGE of a given sample; these experiments showed low intrasample variability (correlation coefficients 0.89–0.95). Differences between multiple samples were tested by comparing the 2D-PAGE of four samples. These experiments showed slightly greater variation between samples (correlation coefficients 0.85–0.93) and a number of differentially expressed proteins. The intensity of 18 protein spots differed more than fivefold, and the intensity of nine protein spots differed more than 100-fold. These results show that 2D-PAGE can be used under standard conditions to screen SF samples and identify a small subset of proteins in SF that are potential markers associated with OA.     

人精浆中的胰岛素样生长因子及其结合蛋白的测定 (在正常和输精管结扎男性间的比较研究)

目的　为探讨精浆中几种主要的胰岛素样生长因子 (IGFs)及其IGF 结合蛋白 (IGFBPs)的来源、存在特征和含量。方法　用免疫放射分析法 (IRMA) ,WesternLigandBlotting(WLB)和WesternImmunoblottig(WIB)法分析 18例正常和 15例输精管结扎男性精浆中的IGFs及其IGFBPs。结果　IRMA测定表明正常男性精浆中游离的IGF Ⅰ浓度 (3.3± 0 .6 μg/L)显著高于输精管结扎男性 (1.4± 0 .3μg/L) (P <0 .0 5 ) ;在正常和结扎男性精浆中游离的IGF Ⅱ浓度 (分别为 12 .2± 0 .2 μg/L和 12 .8± 0 .8μg/L)间和IGFBP 3浓度 (分别为 5 6 .5± 1.7μg/L和5 7.6± 1.4μg/L)间均无显著意义的差别 (P >0 .0 5 )。用WLB和WIB分析法证实了精浆中亦存在IGFBP 2、IGFBP 4和低分子量的IGFBP 3水解片段。结论　据推测精浆中的IGF Ⅰ可能主要来源于睾丸组织 ,而精浆中IGF Ⅱ和IGFBP 3不是主要从睾丸来源的 ;精浆中存在大量低分子量的IGFBP 3,提示在精浆中有对IGFBP 3特异的水解活性物质。     

Serum insulin-like growth factor (IGF)-1 and IGF-binding protein-3 do not correlate with Gleason score or quantity of prostate cancer in biopsy samples

OBJECTIVE: To examine the relationship of serum insulin-like growth factor (IGF)-1 and IGF binding protein-3 (IGFBP-3) with histological cancer characteristics in men undergoing transrectal ultrasonography (TRUS)-guided biopsy. PATIENTS AND METHODS: Patients (652), with either an elevated serum prostate-specific antigen level or an abnormal digital rectal examination, were initially evaluated by TRUS and sextant prostatic needle biopsy. Blood was drawn before biopsy, serum extracted and stored frozen until IGF-1 and IGFBP-3 were measured. In all, 241 patients had prostate cancer (37%) and were included in this study. The number of positive biopsies, the volume of tumour in each positive biopsy and the Gleason score were recorded. RESULTS: Of the 241 patients, 37 had five or six positive biopsies (from six), 128 had two to four and 76 had one. Serum IGF-1 did not correlate with the number of positive biopsies, with means of 176.7, 178.3 and 164.4 ng/mL, respectively (P = 0.3), while the mean IGFBP-3 was 2695, 2795 and 2572 ng/mL, respectively (P = 0.09). The additive percentiles of tumour volume in positive biopsies were assessed for each patient but serum IGF-1 and IGFBP-3 did not correlate (P = 0.7 and 0.9, respectively). In all, 92 patients had a Gleason score of < 7, 80 a score of 7 and 69 a score of > 7; the mean (sd) IGF-1 levels for the three groups were 181 (39), 174.6 (35) and 176 (26) ng/mL, and the mean IGFBP-3 2798 (240), 2735 (284) and 2647 (221) ng/mL, respectively, none of the differences being statistically significant. CONCLUSIONS: Serum IGF-1 and IGFBP-3 do not correlate with quantity of cancer or Gleason score in biopsy samples from patients with prostate cancer.     

Growth factors, insulin-like growth factor-1 and growth hormone, in synovial fluid and serum of patients with rheumatic disorders

OBJECTIVE: Synovial fluid (SF) plays an important role in joint function. We evaluated the growth factors, insulin-like growth factor-1 (IGF-1) and growth hormone (GH) in SF and serum from patients with osteoarthritis (OA), rheumatoid arthritis (RA), gout, pseudogout and diffuse idiopathic skeletal hyperostosis (DISH). DESIGN: Standard radioimmunoassay techniques were used to measure concurrent levels of IGF-1 and GH. SF samples and serum samples were obtained concomitantly from 27 patients with OA, 22 patients with RA, nine men with gout, 14 patients with pseudogout and eight men with DISH. RESULTS: In the case of IGF-1, a comparison of serum and SF levels shows that SF levels of IGF-1 are lower than serum levels in all groups. Men and women gave similar values. In contrast, in the case of GH, all groups, except males with RA, had higher GH values in SF when compared with serum values. Individual patients with other forms of arthritis demonstrated similar relationships. CONCLUSION: The finding that IGF-1 is present in levels about one-half as great in SF as compared with serum suggests that IGF-1 may be produced in lesser amounts or is utilized by the patient in customary joint function. The finding that GH is present in SF at values twice as high, or more, of serum levels in inflammatory arthritides suggests that GH may play a role in the pathophysiology of arthritic disorders.     

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.     

A low-fat diet and/or strenuous exercise alters the IGF axis in vivo and reduces prostate tumor cell growth in vitro

BACKGROUND: Prostate cancer is the most common solid-tumor cancer in US males but is rare in Asian males. When Asian men adopt the US lifestyle, clinical prostate cancer increases greatly. Epidemiological data from men in the US indicate that regular activity may reduce the risk for prostate cancer. METHODS: Serum was obtained from three groups of similar-aged men, Control, Diet and Exercise, and Exercise alone were used to stimulate LNCaP cells in culture. Growth and apoptosis of tumor cells were measured. Serum samples were also used to measure insulin, IGF-1, IGFBP-1. RESULTS: The Diet and Exercise and the Exercise alone groups had lower serum insulin and IGF-1 but higher IGFBP-1 compared to Controls. LNCaP cell growth was reduced in both groups compared to Control and there was a major increase in apoptosis of tumor cells. CONCLUSIONS: A low-fat diet and/or intensive exercise results in change in serum hormones and growth factors in vivo that can reduce growth and induce apoptosis of LNCaP prostate tumor cells in vitro.