| Anabolic and Anti-apoptotic Effects of Pamidonate on
Human Intervertebral Disc Cells in Vitro
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| 인간 추간판 세포에서 파미드로네이트의
기질 생성 및 세포 사멸 효과
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| 전지애1,김 향1,권언혜1,이광일1,박시영,김학선,이환모,신규호,윤성환,문성환 |
| 연세대학교 의학대학 정형외과학교실, 연세대학교 BK21 의과학사업단1 |
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| Abstract |
Background
As potent inhibitors of bone resorption, bisphosphonates (BPs) are widely used for the treatment of bone disorders that are due to increased osteoclast activity i.e. postmenopausal osteoporosis, Paget's disease, tumoral bone disease. BPs are also broad-spectrum matrix metalloproteinase (MMP) inhibitors which involves cation chelation. Apoptosis appears to provide an important mechanism that contributes to the death of disc cells and thus disc degeneration. In articular chondrocytes, BPs appear to prevent dexamethasone induced growth retardation and apoptosis. Accordingly, the objective of this study is to investigate the effect of pamidronate, N-containing BP, on human intervertebral disc cells in terms of cellular proliferation, matrix synthesis, mRNA expression of matrix components, apoptosis, and mRNA expression of MMP.
Methods
Human disc tissues were obtained from three patients with degenerative disc disease. Intervertebral disc cells were isolated by sequential enzymatic method. Apoptosis was induced by using serum starvation method with 1% FBS. Each culture was allocated to 1) control cultures with 10% FBS; 2) experimental cultures in 1% FBS without pamidronate; 3) experimental cultures with 1% FBS with various dose of pamidronate (10-12, 10-9, 10-6 M). [3H]-Thymidine for DNA synthesis and [35S]Sulfate incorporation for proteoglycan synthesis were performed. The cells were stained Propidium Idodide Buffer to detect apoptotic cells and then analyzed by flow cytometry. Reverse transcription polymerase chain reaction for mRNA of beta-actin, collagen type I, collagen type II, aggrecan, MMP-1, MMP-3, and MMP-13 was performed.
Results
Disc cell cultures with various concentrations of pamidronate (10-12, 10-9, 10-6 M) showed no significant increase in DNA synthesis compared to control culture. However disc cell cultures with pamidronate showed significant increase in proteoglycan synthesis, 40% increase with 10-9 M pamidronate (p<0.05), compared |
| Key Words:
Intervertebral disc cells, Apoptosis, Bisphosphonate, Proteoglycan |
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