背景:目前膝骨关节炎的研究重点多在关节软骨、软骨下骨及滑膜,国内外对动物模型半月板超微结构的研究较少.目的:在观察半月板白区软骨细胞超微结构及其类型基础上,观察早期骨关节炎动物模型半月板白区细胞超微结构变化,探讨半月板微观结构变化与生理功能改变的关系.方法:SD大鼠分空白对照组、模型组.模型组通过膝关节腔注射木瓜蛋白酶制作早期骨关节炎动物模型;造模5周后取半月板,电镜制样、定位,观察白区细胞超微结构.实验方案经福建中医药大学动物实验伦理委员会批准.结果与结论:①空白对照组大鼠半月板白区浅表层细胞多呈梭形,里层细胞多呈类三角形,含有丰富的粗面内质网、线粒体,并见高尔基体等细胞器;细胞周围胶原原纤维主要为有序密集的Ⅱ型胶原原纤维;半月板白区细胞为透明软骨细胞.模型组半月板组织表面粗糙;细胞肿胀,粗面内质网等细胞器减少且肿胀变性,糖原蓄积;细胞核多有变形,异染色质凝集;细胞周围胶原原纤维变得杂乱而稀疏;②结果提示,早期骨关节炎半月板软骨细胞与基质轻度退变,将使细胞合成分泌基质成分的能力降低,半月板生理功能将受到影响.

背景:前期研究结果显示,透骨消痛胶囊能降低软骨细胞炎症因子表达,减少软骨细胞凋亡,促进软骨细胞增生,延缓软骨、软骨下骨退变,从而缓解骨关节炎疾病进程,减轻炎症反应,但该方对滑膜病变的作用机制尚不明确.目的:从早期骨关节炎滑膜炎症水肿与相关水通道蛋白调节的角度,进一步探讨透骨消痛胶囊抗炎消肿的作用机制.方法:24只SD大鼠随机分成空白组、模型组、中药组3组,后两组应用木瓜蛋白酶法制备早期骨关节炎模型1周后,分别以透骨消痛胶囊(中药组)或生理盐水(空白组和模型组)灌胃4周.小动物MRI观察膝关节形态结构,放射免疫法测定血清和关节液白细胞介素1β和肿瘤坏死因子α水平,石蜡切片观察滑膜组织显微结构,透射电镜观察滑膜细胞超微结构,免疫组化/荧光染色定量、定位水通道蛋白1和水通道蛋白3的表达.实验方案经福建中医药大学动物实验伦理委员会批准(批准号为[2019]福中医伦理审字第024号).结果与结论:①与模型组相比,中药组滑膜水肿减轻、滑液分泌减少;血清和关节液白细胞介素1β和肿瘤坏死因子α水平明显下降;滑膜细胞增生减少,组织纤维化和血管增生明显减轻;滑膜细胞粗面内质网、高尔基体、囊泡和溶酶体等减少;水通道蛋白1和水通道蛋白3表达显著下调,散在分布于细胞膜和细胞质,无聚集现象;②结果说明,透骨消痛胶囊可能通过减少滑膜水通道蛋白1和水通道蛋白3含量,减少滑液分泌,改善关节水肿,这可能是该药物抗炎消肿,延缓骨关节炎进展的作用机制之一.

The aim of the study was to observe the effects of Tougu Xiaotong capsule (TGXTC) on the microstructure and ultrastructure of meniscus in rats with early knee osteoarthritis (KOA). A total of 27 Sprague Dawley rats were randomly divided into three groups: The normal group (non-papain-induced KOA; received saline only), the model group (papain-induced KOA; received saline only) and the TGXTC group [papain-induced KOA; received TGXTC (0.31g·kg-1·d-1)]. After 4 weeks treatment, the animals were anesthetized and the sagittal plane of the intact knees (n=6 per group) was obtained and prepared in paraffin section. Following hematoxylin and eosin staining, the degeneration of cartilage structure was evaluated via Mankin score, the microstructure of meniscus was observed and the area of calcification in meniscus was analyzed. Following toluidine blue staining, the content of proteoglycan in meniscus was analyzed. Three samples in each group were obtained and the ultrathin sections of meniscus were observed through a transmission electron microscope. The results showed that compared with the normal group, in the model group the joint space became narrow and the cartilage layer was slightly damaged and the Mankin score was 4.17±0.76, suggesting that the early KOA model was successfully established. After TGXTC treatment, the joint space stenosis and cartilage damage were improved as the Mankin score significantly decreased. Compared with the normal group, in the model group the surface of meniscal cartilage was much more uneven, the area of calcification was significantly increased and the content of proteoglycan of cartilage matrix was significantly decreased. However, following TGXTC treatment, the surface of the meniscal cartilage was much more smooth and flat, and the damage of tissue structure and the calcified area were significantly reduced, and the proteoglycan of cartilage matrix content was significantly increased. Compared with the normal group, the number of cellular processes and organelles, including the rough endoplasmic reticulum, mitochondria and Golgi apparatus of meniscal cartilage were reduced and swollen in the model group. In addition, the nuclei were deformed and heterochromatin agglutinated. The extracellular collagen fibrils became slender, disordered and sparse. Compared with the model group, the TGXTC group had more cell processes and organelles, alleviated swelling and heterochromatin agglutinating. Additionally, the collagen fibrils around the cells were thicker, larger and arranged in an orderly manner. In conclusion, TGXTC exerted its therapeutic effects on the development of KOA via reducing the destruction of the cartilage structure of the meniscus and improving the composition and function of the meniscus cartilage matrix.

The aim of the study was to observe the effects of Tougu Xiaotong capsule (TGXTC) on the microstructure and ultrastructure of meniscus in rats with early knee osteoarthritis (KOA). A total of 27 Sprague Dawley rats were randomly divided into three groups: The normal group (non-papain-induced KOA; received saline only), the model group (papain-induced KOA; received saline only) and the TGXTC group [papain-induced KOA; received TGXTC (0.31g·kg-1·d-1)]. After 4 weeks treatment, the animals were anesthetized and the sagittal plane of the intact knees (n=6 per group) was obtained and prepared in paraffin section. Following hematoxylin and eosin staining, the degeneration of cartilage structure was evaluated via Mankin score, the microstructure of meniscus was observed and the area of calcification in meniscus was analyzed. Following toluidine blue staining, the content of proteoglycan in meniscus was analyzed. Three samples in each group were obtained and the ultrathin sections of meniscus were observed through a transmission electron microscope. The results showed that compared with the normal group, in the model group the joint space became narrow and the cartilage layer was slightly damaged and the Mankin score was 4.17±0.76, suggesting that the early KOA model was successfully established. After TGXTC treatment, the joint space stenosis and cartilage damage were improved as the Mankin score significantly decreased. Compared with the normal group, in the model group the surface of meniscal cartilage was much more uneven, the area of calcification was significantly increased and the content of proteoglycan of cartilage matrix was significantly decreased. However, following TGXTC treatment, the surface of the meniscal cartilage was much more smooth and flat, and the damage of tissue structure and the calcified area were significantly reduced, and the proteoglycan of cartilage matrix content was significantly increased. Compared with the normal group, the number of cellular processes and organelles, including the rough endoplasmic reticulum, mitochondria and Golgi apparatus of meniscal cartilage were reduced and swollen in the model group. In addition, the nuclei were deformed and heterochromatin agglutinated. The extracellular collagen fibrils became slender, disordered and sparse. Compared with the model group, the TGXTC group had more cell processes and organelles, alleviated swelling and heterochromatin agglutinating. Additionally, the collagen fibrils around the cells were thicker, larger and arranged in an orderly manner. In conclusion, TGXTC exerted its therapeutic effects on the development of KOA via reducing the destruction of the cartilage structure of the meniscus and improving the composition and function of the meniscus cartilage matrix.

BACKGROUND:Inflammatory cytokines enhanced the progress of the pathogenesis of osteoarthritis, however the mechanisms remain unclear. The objective is to determine aquaporins (AQPs) in the pathogenesis of osteoarthritis.METHODS AND FINDINGS:Primary rat articular chondrocytes were treated with IL-1β to mimic the early stage of osteoarthritis in vitro. Early osteoarthritis animal model was established by intra-articular injection of 4% papain. Micro- or ultra-structure histopathologic changes, cell viability, apoptosis cells and cell membrane permeability, locations and expressions of AQP1 and AQP3 and matrix were detected in the cartilage or in the chondrocytes of knee. IL-1β could reduce the chondrocytes viability, increase the apoptosis cells, and also impair the cell membrane and organelles. IL-1β significantly induced the up-regulation of AQP1 and AQP3 in the chondrocytes. In the chondrocytes, AQPs were mainly clustered in both membrane and perinuclear region of cytoplasm, while higher AQPs were detected in the superficial and middle layers of the cartilage. With the up-regulation of AQPs, the cartilage matrix was considerably decreased in both the chondrocytes and in the osteoarthritis cartilage. In the early osteoarthritis rat model, serum and synovial fluid confirmed that higher IL-1β could increase the expressions of AQPs, and decrease the cartilage matrix in both the chondrocytes and the cartilage.CONCLUSIONS:Inflammatory cytokine IL-1β via up-regulation of AQPs caused the abnormal metabolism of water transport and loss of the cartilage matrix in the chondrocytes, and ultimately exacerbated the pathogenesis of early osteoarthritis. Therefore, AQPs may be a candidate therapeutic target for prevention and treatment of osteoarthritis.

Objective:To investigate the effects of Tougu Xiaotong capsule (TXC) on chondrocytes ultrastructure and cartilage matrix in rats with early stage osteoarthritis.Methods:Twenty four male rats were randomized into non-osteoarthritis group, osteoarthritis model group and TXC treated osteoarthritis model group, with eight rats in each group. The rat model of osteoarthritis was established by papain injection into knee joint cavity, and the rats received TXC aqueous solution intragastric for four weeks, then rats were sacrificed to extract the femoral condyle cartilage for sample preparation. The chondrocytes microstructure and ultrastructure in different regions were observed by electromicroscopy. The contents of proteoglycan and collagen II in cartilage matrix were analyzed by histochemical approaches.Results:Chondrocytes ultrastructure showed that the chondrocyte proliferation and clustering, and substantial reduction or swelling organelles were observed in osteoarthritis model group but not in non-osteoarthritis group. The clustering of chondrocytes was reduced, and the organelles of golgi complexes, rough endoplasmic reticulum and mitochondria were increased in TXC treated osteoarthritis model group compared to osteoarthritis model group. Cartilage matrix quantitative analysis showed that the contents of proteoglycan and collagen II in osteoarthritis model group were significantly decreased compared to non-osteoarthritis group (P<0.01); the contents of proteoglycan and collagen II were significantly increased in TXC treated osteoarthritis model group compared to osteoarthritis model group (P<0.01).Conclusion:TXC could ameliorate chondrocytes degeneration, endoplasmic reticulum and mitochondria damage, and increase the synthesis of cartilage matrix to delay the cartilage degeneration of osteoarthritis rats at early stage.

Imaging diagnostic methods of osteoarthritis (OA) mainly include X-ray films,CT,MRI and ultrasound,etc.,which have their own characteristics in terms of the specificity,sensitivity and the utilization.Compared with the above traditional imaging examination means,high-resolution small animal imaging technology such as Micro-CT and nuclear magnetic imaging developed in recent years is showing obvious advantages in image resolution,microscopic structure display,as well as quantitative analysis,and playing important role in the study of OA animal models.In this paper,the establishment of OA animal model and the characteristics of various imaging techniques were introduced,the roles of bony structure imaging and cartilage imaging of Micro-CT in OA animal model were reviewed,the application values of small animal nuclear magnetic imaging in OA animal model were discussed from aspects of cartilage imaging sequences selection,cartilage thickness measurement,cartilage composition analysis and other tissue lesions analysis.At the same time,future prospects of small animal imaging technology in OA animal model were expected.

Object:To investigate the effection of Tougu Xiaotong Granules in early and mid osteoarthritis of rabbits and the application value of imaging technology in OA experiment. Methods:48 New Zealand white rabbits were randomly divided into normal group,model group,Tougu Xiaotong group,and bolus group. All models were based on the improved Hulth. DR,CT,MRI screening technology was taken in 4W,8W,12W,16W after operation,observing the changes of joint space narrowing,subchon-dral bone,osteophytes,arthroedema,cartilage,et al. Results:Model group had more arthroedema each time,early experiments (4 and 8 weeks) could observe subchondral sclerosis,late experiments (12 and 16 weeks) slightly joint space narrowing could be seen. Tougu Xiaotong group of mid-dose and high-dose had lesser arthroedema than model group each time,with cartilage sur-face coarse and subchondral sclerosis and lightest high-dose group lesions in late experiments. The performance of bolus group was between high-and low-dose Tougu Xiaotong group. Conclusion:Tougu Xiaotong Granules can reduce inflammation of OA,de-lay lesion of subchondral bone and recover the cartilage.