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Effects of the Supernatant from Ankylosing Spondylitis Synovial Cell Primary Culture on the Osteogenic Differentiation of Ligament Fibroblasts

Author: LiangZhiMin
Tutor: XuWeiDong；WangZiMin；WangZhiWei
School: Second Military Medical University
Course: Surgery
Keywords: Ankylosing spondylitis Synovial cells Ligament fibroblasts Osteogenic differentiation of Alkaline phosphatase Osteocalcin
CLC: R593.23
Type: Master's thesis
Year: 2010
Downloads: 97
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Abstract

Background of ankylosing spondylitis (Ankylosing Spondylitis, AS) is a common chronic inflammatory rheumatic diseases. Chronic inflammation of the axial skeleton and new bone formation is its iconic pathological features. In recent years, with the improvement and enhancement of imaging techniques and treatment from fundamentally changed the diagnosis and treatment of the disease. However, the lack of effective drugs in inhibiting structural damage to choose from. At present, the traditional non-steroidal anti-inflammatory drugs is still one of the main drug treatment of ankylosing spondylitis. Tumor necrosis factor antagonists, achieved unprecedented results in the improvement of clinical symptoms and signs, but in recent years, clinical follow-up study did not find that they are able to stop the progress of the new bone formation. The starting and ending points and articular cartilage damage at the main part of the new bone formation in AS. The pathology study found that there are a lot of abnormal proliferation of fibroblasts in these parts. Therefore, we speculate that the the AS new bone formation process, there may be fibroblasts transformed cells into bone cells. In addition, the in AS synovial tissue can be detected to promote osteogenic differentiation capacity factor expression, such as transforming growth factor-beta and bone morphogenetic protein. Therefore, we further speculated to the AS synovial cells may be able to some promote osteogenic differentiation of factors, thus contributing to the transformation of fibroblasts into osteoblasts. The purpose of ankylosing spondylitis hip ligament source of fibroblasts in vitro osteogenic potential, as well as synovial cell culture supernatants its osteogenic differentiation. The experimental derived from ankylosing spondylitis, aseptic necrosis of the femoral head (Aseptic necrosis of head of femur of ANFH) in patients with hip ligaments and synovial tissue samples were required. Respectively ligament fibroblasts and synovial cells in primary culture, while collecting the first generation of synovial cell culture supernatant. Four different conditions were applied for induction medium on the third generation of the ligament fibroblasts, namely: the OIM: bone induction medium (containing 2 × 10-8mmol / L dexamethasone, 0.1mg/ml vitamin C, 20mmol/Lβ- glycerophosphate and 10% FBS DMEM/F12 medium) containing 10% FBS DMEM/F12 medium the; OIMBMP 2: bone-inducing medium containing 0.3ug/ml BMP-2 medium DMED/F12; OIMANFH: bone induction medium ANFH synovial cell culture supernatant; PIMAS: bone induction medium the AS synovial cell culture supernatant. Were 0, 5, 10, 15, and 20 days after the induction, the use of quantitative and qualitative changes in alkaline phosphatase activity. Osteocalcin expression levels in the cell culture supernatant was detected by ELISA. Von Kossa staining was used to detect the formation of calcium nodules. The results of the application of four different medium ligament fibroblasts after induction, the expression levels of alkaline phosphatase activity and osteocalcin as the induction time extended gradually increased, wherein the alkaline phosphatase in the first 10 days significantly increased, 15 reached the peak, osteocalcin in the first 15 days was significantly higher and reached a peak. Induction in the first 20 days in each group of cells can be detected in the formation of calcium nodules. Comparison of four different conditions of medium ligament derived fibroblast-like cells in vitro induced osteogenic potential, OIMBMP-2 >> OIMAS >> OIMANFH> OIM, performance in alkaline phosphatase activity and osteocalcin expression level of the level of between the four treatment groups relative to more statistically significant, P lt; 0.05. AS ligament into of fibroblasts and ANFH ligament fibroblasts alkaline phosphatase activity and osteocalcin expression level of the former than the latter, the two groups were statistically significant, P lt; 0.05. Conclusion ankylosing spondylitis hip ligament sources fibroblasts transformed into osteoblasts in vitro, and this transformation is stronger than the control group. Ankylosing spondylitis synovial cell culture supernatant osteogenic differentiation ligament fibroblasts has a certain role, suggesting that ankylosing spondylitis synovial cells can secrete promote osteogenic differentiation factor. The research confirmed ankylosing spondylitis hip joint ligament derived fibroblast-like cells in vitro to differentiate into bone cells transformed, also found that the AS synovial cell culture supernatant the ligament fibroblasts osteogenic differentiation has a certain role in promoting. The study, we recognize that synovial to play a destructive role in the process of arthritis injury, is also in the process of new bone formation in ankylosing spondylitis play a certain role in promoting, for further study of ankylosing spondylitis new bone formation mechanism laid the foundation.

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CLC: > Medicine, health > Internal Medicine > Systemic disease > Autoimmune diseases > Autoimmune diseases, connective tissue disease > Ankylosing spondylitis
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