滑膜细胞在骨关节炎中的研究进展

骨关节炎(osteoarthritis,OA)作为最常见的退行性关节疾病,其主要临床特点是软骨的破坏降解,进而导致关节功能丧失,严重影响患者的生活质量.越来越多的证据表明,除了软骨组织,OA的病理改变还涉及滑膜、骨以及软骨下骨在内的多个组织系统.其中,滑膜作为组织系统的重要组成部分,其病变在OA中的作用日益突出.滑膜细...     

大鼠膝骨关节炎滑膜细胞原代培养

目的:建立原代培养膝骨关节炎大鼠滑膜细胞的方法。方法:取膝骨关节炎大鼠膝关节滑膜组织,用酶消化法消化、分离细胞,用DMEM培养液进行培养并传代,观察滑膜细胞生长状况、并用细胞形态学及细胞免疫化学鉴定。结果:用酶消化法培养膝骨关节炎滑膜细胞,方法简单、成功率高、细胞形态典型。结论:单一的胶原酶消化法是一种有效的滑膜细胞原代培养的方法,可以满足一般实验需求。     

骨关节炎相关生物标志物

骨关节炎（osteoarthritis）是关节软骨进展的退化性疾病,并累及周围组织结构的病变,是致老年人伤残主要原因之一。目前,以临床表现和影像学诊断为主,缺乏早期检测和预后评估的有效方法。生物标志物的检查是具有前景的研究方向,在关节软骨结构改变之前,各种生物标志物代谢发生变化,其能帮助诊断和预测骨关节炎的发生发展及其预后。然而,生物标志物在临床诊断和治疗相关的应用仍需加以证实。通过广泛查阅近年有关骨关节炎相关分子生物诊断的相关文献,有助于了解生物标志物对于骨关节炎的早期诊断意义和临床应用前景。本文就关节软骨、骨和滑膜等不同组织类型相关的生物标志物进行综述。     

阿尔茨海默病早期诊断的体液生物标志物

阿尔茨海默病(Alzheimer’s disease,AD)是目前发病率较高的神经退行性疾病,主要临床表现为不可逆的记忆力丧失与认知功能的衰退。AD起病隐秘,不易察觉,病程长达数十年,尚无有效治疗手段,因此AD早期的诊断与干预尤为重要;但是,当前AD早期诊断缺乏灵敏、简便的检测方案。体液(特别是血液)中的生物标志物受到了越来越多的关注,可能成为AD早期诊断的有效手段。现主要综述了与AD病理进程相关的脑脊液、血液、尿液中的生物标志物,并对其应用与前景做一展望。     

miR-21在骨关节炎中作用机制的研究进展

骨关节炎(osteoarthritis,OA)是一种好发于老年人的慢性退行性关节疾病。OA的发病机制复杂,目前尚无有效的治疗手段能够完全逆转OA的病程进展。随着老龄化社会的发展,OA患者逐渐增多,给患者家庭和社会带来的经济压力越来越大。因此,深入阐明OA的发病机制,找到有效的早期分子诊断标志物或潜在的治疗靶点意义重大。miRNA(microRNA)是近几年研究较多的一种非编码RNA。大量研究显示,miR-21在OA软骨降解和关节疼痛的发生中均发挥重要作用。因此,该文通过综述miR-21与OA的相关文献,探究lncRNA、MMPs、GDF-5、FGF和TLR7等分子在miR-21调控OA的过程中可能起到的关键作用,总结miR-21对OA的具体调控机制,以期为OA早期分子诊断标志物和潜在治疗靶点的相关研究提供方向和依据。     

骨性关节炎生物学标志物研究进展

骨性关节炎(oseteoarthritis,OA)是一种随着年龄增长发病率明显升高的退行性变,常累及脊柱、髋、膝等人体负重关节,以关节缓慢发展的疼痛、肿胀,伴功能障碍为临床表现,主要有滑膜增生、软骨破坏、软骨下骨骨化及骨赘形成等一系列病理表现。OA对人类的健康和生活质量影响很大,随着老龄化社会的到来,本病的发病率日趋升高,其研究已成为医学领域中的重要课题。目前,OA的早期诊断、病变监测和有效防治仍是骨科领域亟待解决的疑难问题。随着分子生物学的发展和研究手段的提高,许多研究者都在试图寻找用于临床评价OA的生物学标志物。本文将就OA研究中所使用的主要标志物进行综述,为深入研究OA提供方便。     

瘦素对骨关节炎影响的研究进展

瘦素(Leptin)是脂肪因子的一种,可以在骨关节炎(OA)患者的血浆、滑液、软骨中被检测到。OA是一种最常见的关节疾病,其可以发生于全身的多个关节,以骨质和滑膜组织新陈代谢的改变、关节软骨的破坏和由此引起的关节软骨的减少为特征。瘦素是一种由肥胖(0b)基因编码的一个小的非糖基化肽激素。最开始,瘦素仅仅被认为是一种脂肪细胞源性的小分子(16KD),在下丘脑中枢水平作为一个饱感因子介导食物摄入量减少,并增加能量的消耗。现在已经发现,瘦素在机体内可发挥多种生理作用,并与OA病情有关。本文通过对瘦素与OA、软骨、肥胖、生物标志物、脂联素等之间的联系做一综述,以了解瘦素与OA之间的联系,为OA的治疗方面的进一步研究提供帮助。     

2 型糖尿病患者肝损伤标志物与下肢动脉病变的相关性分析

目的：探讨2 型糖尿病患者肝损伤标志物水平与其下肢动脉病变的相关性,为2 型糖尿病并发症的防治提供参考依据。方 法：选取我院收治的2 型糖尿病患者946 例,根据下肢动脉内膜中层厚度分为以下3 组,即无动脉硬化组(276 例)、单纯性动脉硬 化组(598 例)和动脉硬化伴管腔狭窄或闭塞组(72 例)。分析和比较三组之间肝损伤标志物谷丙转氨酶(ALT)、谷草转氨酶(AST)、γ-谷氨酰转肽酶(GGT)水平的差异,及其与2型糖尿病患者下肢动脉硬化程度的相关性。结果：随着动脉硬化程度的加重,2 型糖尿病患者的ALT 水平逐渐升高,三组之间两两比较差异均有统计学意义(P<0.05)。动脉硬化伴管腔狭窄或闭塞组AST 水平显著高于非动脉硬化组和单纯性下肢动脉硬化组,而动脉硬化组和非动脉硬化组之间AST 水平比较无显著差异(P>0.05)。三组之间γ-谷氨酰转肽酶水平比较无显著性差异(P>0.05)。Spearman 等级相关分析显示ALT 与糖尿病下肢动脉硬化的相关系数为0.30484。结论：2型糖尿病患者ALT 水平与其下肢动脉硬化程度显著相关。     

胰腺癌相关肿瘤标志物研究进展

胰腺癌起病隐匿,进展快,预后差,发病率约等于死亡率。胰腺癌死亡率高的原因有发病机制不明,缺乏有效的早期诊断和预后的肿瘤标志物,进展期相关治疗效果不理想。近年来在血清标志物、基因标志物、表观遗传学标志物等分子生物技术及生物信息学方面的发展为胰腺癌的诊断,尤其是早期诊断、评估预后和监测早期复发提供了新的途径。本文就近期胰腺癌相关肿瘤标志物的研究进展作一综述。     

2型糖尿病患者肝损伤标志物与下肢动脉病变的相关性分析

目的：探讨2型糖尿病患者肝损伤标志物水平与其下肢动脉病变的相关性,为2型糖尿病并发症的防治提供参考依据。方法：选取我院收治的2型糖尿病患者946例,根据下肢动脉内膜中层厚度分为以下3组,即无动脉硬化组（276例）、单纯性动脉硬化组（598例）和动脉硬化伴管腔狭窄或闭塞组（72例）。分析和比较三组之间肝损伤标志物谷丙转氨酶（ALT）、谷草转氨酶（AST）、γ-谷氨酰转肽酶（GGT）水平的差异,及其与2型糖尿病患者下肢动脉硬化程度的相关性。结果：随着动脉硬化程度的加重,2型糖尿病患者的ALT水平逐渐升高,三组之间两两比较差异均有统计学意义（P〈0．05）。动脉硬化伴管腔狭窄或闭塞组AST水平显著高于非动脉硬化组和单纯性下肢动脉硬化组,而动脉硬化组和非动脉硬化组之间AsT水平比较无显著差异（P〉0．05）。三组之间γ-谷氨酰转肽酶水平比较无显著性差异（P〉0．05）。Spearman等级相关分析显示ALT与糖尿病下肢动脉硬化的相关系数为0．30484。结论：2型糖尿病患者ALT水平与其下肢动脉硬化程度显著相关。     

膝骨关节炎患者关节液和滑膜中IL-6、MMP-13、VEGF的表达及与病情进展的关系

目的:研究膝骨关节炎(KOA)患者关节液和滑膜中白细胞介素-6(IL-6)、基质金属蛋白酶-13(MMP-13)、血管内皮生长因子(VEGF)的表达及与病情进展的关系。方法:选择自2017年1月到2017年6月在我院就诊的KOA患者30例进行研究,其中维吾尔族和汉族各15例,分别记为维吾尔族组和汉族组。另选同期在我院接受骨折修复和截肢等手术治疗的10例无骨关节炎的患者作为对照组。对比各组IL-6、MMP-13及VEGF水平以及关节软骨中水通道蛋白3(AQP3)阳性表达率,对比维吾尔族组和汉族组软骨不同区域内AQP3阳性表达,分析KOA患者关节液和滑膜中IL-6、MMP-13、VEGF及关节软骨中AQP3的阳性表达与病情进展的相关性。结果:维吾尔族组和汉族组关节液和滑膜中IL-6、MMP-13及VEGF水平均分别高于对照组,差异均有统计学意义(均P0.05)。维吾尔族组和汉族组关节软骨中AQP3阳性表达率均分别明显高于对照组,且维吾尔族组明显高于汉族组,差异均有统计学意义(均P0.05)。维吾尔族组和汉族组浅层软骨磨损严重区的AQP3阳性表达率明显高于软骨深层区和软骨下骨区,差异均有统计学意义(均P0.05)。Spearman相关性分析显示,KOA患者关节液和滑膜中IL-6、MMP-13、VEGF及关节软骨中AQP3的阳性表达与病情进展均呈正相关(均P0.05)。结论:KOA患者关节液和滑膜中IL-6、MMP-13、VEGF水平及关节软骨中AQP3阳性表达均异常升高,以上指标参与了病情的进展,且AQP3阳性表达高低还与民族有关,临床上可考虑将这些指标作为监测KOA患者病情的靶点。     

Elevated Nerve Growth Factor Levels in the Synovial Fluid of Patients with Inflammatory Joint Disease

A novel pH shock extraction procedure was used to measure nerve growth factor (NGF) levels in both normal and inflamed synovial fluids using a sensitive and specific two-site enzyme linked immunosorbant assay. To date no data is available on NGF levels in normal synovial fluids. Synovial fluids were taken from 5 normal volunteers, 12 patients with rheumatoid arthritis and 10 patients with other inflammatory arthropathies. The mean ± SEM NGF concentration in normal synovial fluids was 95 ± 33.2 pg/ml (range 39.1–143.1 pg/ml), whereas the mean NGF concentration in the synovial fluids taken from patients with rheumatoid arthritis was 532.5 ± 123.8 pg/ml (range 152–1686 pg/ml). The mean NGF concentration in patients with other inflammatory arthropathies was also raised (430.6 ± 90 pg/ml; range 89–1071 pg/ml). The NGF concentrations were significantly higher in the synovial fluids from both inflamed groups (ANOVA p < 0.05) compared to normals. Raised levels of NGF in synovial fluid may contribute directly to joint inflammation via activation of inflammatory cells.     

Permanent Stained Preparations of Synovial Fluid for Detection of Calcium Compounds Using Alizarin Red S

Permanent preparations of air dried synovial fluids were prepared by staining calcium compounds with alizarin red S stain; each slide was coverslipped with Permount. Variables studied were: (a) concentration of the solution of alizarin red S, (b) pH of staining solution, (c) time of incubation in staining solution and aqueous and ethanolic content of staining solution. The staining effect of each solution was tested on calcium pyrophosphate dihydrate, calcium oxalate, apatite and monosodium urate (MSU). Of all the solutions, best results were obtained with 0.25% alizarin red S in 50% ethanol at pH 7.0 for 30 min. With this solution, the calcium-containing compounds were well stained. MSU did not stain and still preserved negative birefringence on polarizaton. Fixation of smears with ethanol served a double purpose: It fixed the slides without dissolving or removing MSU or the calcium compounds, yet it did dissolve five corticosteroids commonly used for intra-articular injection which may interfere with interpretation of compensated polarized light microscopy of synovial fluids.     

Association of chemerin levels in synovial fluid with the severity of knee osteoarthritis

Context: Chemerin has been implicated to be correlated with inflammation.

Objective: This study aims to determine the association of chemerin levels in serum and synovial fluid (SF) with the disease severity of patients with knee Osteoarthritis (OA).

Methods: 124 patients with knee OA and 76 healthy controls were enrolled in this study.

Results: Chemerin levels in serum were significant higher with regard to paired SF. Chemerin levles in SF of knee OA patients were correlated with disease severity evaluated by KL grading criteria.

Conclusion: Chemerin levels may be involved in the pathophysiology of the development and progression of OA.     

体液循环microRNA的研究进展

循环microRNA（miRNA）是参与细胞间信息交流的一类非编码小RNA分子,作为许多疾病的生物标记物近来受到广泛关注。简要总结了循环miRNA的来源和存在形式,同时概述了体液miRNA的制备和检测方法,最后对循环miRNA的应用前景进行了简单探讨,以期为循环miRNA在理论和实践中的深入研究和应用提供参考。     

Markers of cartilage and bone matrix metabolism differ between the sera of macaque and squirrel monkeys

BACKGROUND: Nonhuman primates develop the characteristic lesions of osteoarthritis, making them attractive biomedical models for the study of environmental factors, such as diet, which may influence the progress of the condition. METHODS AND MATERIALS: We used ELISA assays of potential markers of osteoarthritis which were developed for use in humans to see if we could determined the presence of immunoreactivity in two nonhuman primate genera - Macaca (macaque monkeys) and Saimiri (squirrel monkeys). RESULTS: Inter-generic differences were significant for most markers. Three markers (bone alkaline phosphatase, hyaluronin and YKL-40) were outside the human range and two markers (laminin and C2C) did not yield useful results because they were off-scale high. CONCLUSION: Our results indicate that most of the ELISA assays designed for use with human serum can be used in nonhuman primates. The highly significant differences we observed between the sera of Macaca and Saimiri, suggest that further examination is warranted.     

Ion channel expression and function in normal and osteoarthritic human synovial fluid progenitor cells

Osteoarthritis (OA) is a chronic disease affecting the cartilage of over 15% of Canadians. Synovial fluid mesenchymal progenitor cells (sfMPCs) are present in joints and are thought to contribute to healing. OA sfMPCs have a greater proliferative ability but decreased chondrogenic potential. However, little is known about the factors influencing/regulating the differences between normal and OA sfMPCs. Recently, our lab has shown that sfMPC chondrogenic differentiation in vitro is favorably biased toward a similar osmotic environment as they experience in vivo. The current study now examines the expression and functionality of a variety of ion channels in sfMPCs derived from normal individuals and early OA patients. Results indicated that there is differential ion channel regulation at the functional level and expression level in early OA sfMPCs. All ion channels were upregulated in early OA compared to normal sfMPCs with the exception of KCNMA1 at the mRNA level. At the protein level, TRPV4 was over expressed in early OA sfMPCs, while KCNJ12 and KCNMA1 were unchanged between normal and early OA sfMPCs. At the functional level, the inward rectifying potassium channel was under expressed in early OA sfMPCs, however the membrane potential was unchanged between normal and early OA sfMPCs. In the synovial environment itself, a number of differences in ion concentration between normal and early OA synovial fluid were observed. These findings suggest that normal and OA progenitor cells demonstrate functional differences in how they interact with the synovial ion environment.     

Pulsed Electromagnetic Field Versus Whole Body Vibration on Cartilage and Subchondral Trabecular Bone in Mice With Knee Osteoarthritis

Pulsed electromagnetic field (PEMF) and whole body vibration (WBV) interventions are expected to be important strategies for management of osteoarthritis (OA). The aim of the study was to investigate the comparative effectiveness of PEMF versus WBV on cartilage and subchondral trabecular bone in mice with knee OA (KOA) induced by surgical destabilization of the medial meniscus (DMM). Forty 12-week-old male C57/BL mice were randomly divided into four groups (n = 10): Control, OA, PEMF, and WBV. OA was induced (OA, PEMF, and WBV groups) by surgical DMM of right knee joint. Mice in PEMF group received 1 h/day PEMF exposure with 75 Hz, 1.6 mT for 4 weeks, and the WBV group was exposed to WBV for 20 min/day with 5 Hz, 4 mm, 0.3 g peak acceleration for 4 weeks. Micro-computed tomography (micro-CT), histology, and immunohistochemistry analyses were performed to evaluate the changes in cartilage and microstructure of trabecular bone. The bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N) increased, and bone surface/bone volume (BS/BV) decreased by micro-CT analysis in PEMF and WBV groups. The Osteoarthritis Research Society International (OARSI) scores in PEMF and WBV groups were significantly lower than in the OA group. Immunohistochemical results showed that PEMF and WBV promoted expressions of Aggrecan, and inhibited expressions of IL-1β, ADAMTS4, and MMP13. Superior results are seen in PEMF group compared with WBV group. Both PEMF and WBV were effective, could delay cartilage degeneration and preserve subchondral trabecular bone microarchitecture, and PEMF was found to be superior to WBV. Bioelectromagnetics. 2020;41:298–307 © 2020 Bioelectromagnetics Society