Steenvoorden M M C, Bank R A, Ronday H K, Toes R E M, Huizinga T W J, DeGroot J
Bio Sciences, TNO Quality of Life, Leiden, The Netherland.
Clin Exp Rheumatol. 2007 Mar-Apr;25(2):239-45.
In vitro models for joint diseases often focus on a single cell type, such as chondrocytes in osteoarthritis (OA) or fibroblast-like synoviocytes (synoviocytes) in rheumatoid arthritis (RA). However, these joint diseases affect the whole joint and interaction between chondrocytes and synoviocytes may play an important role in disease pathology. The current study was designed to study the use of the alginate recovered chondrocyte method as a model for cartilage degradation and to study interaction between chondrocytes and synoviocytes.
Bovine chondrocytes were cultured in alginate beads for 1 week, subsequently chondrons were retrieved and seeded into transwells. Every two days cartilage-slices were analysed for proteoglycan content (colorimetric, Blyscan GAG kit), collagen content (HPLC) and collagen HP and LP crosslinking (HPLC). For degradation experiments, monocultures of cartilage-slices labelled with (35)S and cocultures with synoviocytes were stimulated with IL-1beta or TNF-alpha. After 7 days, (35)S release was measured taken as a measure of cartilage degradation.
After biochemical analysis, three week old cartilage-like slices were chosen to perform cartilage-degradation experiments. Synoviocytes were able to induce cartilage degradation only in the presence of living chondrocytes. In addition, the cytokines interleukin 1 (IL-1beta) and tumor necrosis factor (TNF-alpha) were only able to induce cartilage degradation by chondrocytes, not by synoviocytes.
These data indicate that the alginate recovered chondrocyte method provides a novel model for cartilage degradation in which the interaction between synoviocytes and chondrocytes can be studied.
关节疾病的体外模型通常聚焦于单一细胞类型,如骨关节炎(OA)中的软骨细胞或类风湿关节炎(RA)中的成纤维样滑膜细胞(滑膜细胞)。然而,这些关节疾病会影响整个关节,软骨细胞与滑膜细胞之间的相互作用可能在疾病病理过程中发挥重要作用。本研究旨在探讨使用藻酸盐回收软骨细胞法作为软骨降解模型,并研究软骨细胞与滑膜细胞之间的相互作用。
将牛软骨细胞在藻酸盐珠中培养1周,随后取出软骨粒并接种到Transwell小室中。每两天分析软骨切片的蛋白聚糖含量(比色法,Blyscan GAG试剂盒)、胶原蛋白含量(高效液相色谱法)以及胶原蛋白HP和LP交联(高效液相色谱法)。对于降解实验,用白细胞介素-1β(IL-1β)或肿瘤坏死因子-α(TNF-α)刺激标记有(35)S的软骨切片单培养物以及与滑膜细胞的共培养物。7天后,测量(35)S释放量作为软骨降解的指标。
经过生化分析后,选择三周龄的软骨样切片进行软骨降解实验。滑膜细胞仅在存在活软骨细胞的情况下才能诱导软骨降解。此外,细胞因子白细胞介素1(IL-1β)和肿瘤坏死因子(TNF-α)仅能诱导软骨细胞而非滑膜细胞导致软骨降解。
这些数据表明,藻酸盐回收软骨细胞法为软骨降解提供了一种新模型,可用于研究滑膜细胞与软骨细胞之间的相互作用。