| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2010: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
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| Research Abstract |
Several studies have investigated the roles of biomechanical and metabolic factors in the development and progression of ossification of spinal ligament, based on the importance of genetic and biological factors. The process of ossification includes enchondral ossification, though such pathology remains poorly defined. We analyzed the cultured cells or paraffined sections derived from ossification of the posterior longitudinal ligament or ligamentum flavum using histological and immunohistochemical staining, Western blotting, and real-time RT-PCR. Cyclic tensile strain was produced by Flexercell(R) FX-3000, applied for 0, 6, 12, or 24 hours. Controlled samples were harvested from non-ossified ligamentum flavum of patients who underwent thoracic posterior surgeries. Under resting conditions(no tensile strain), the mRNA levels ofβ-catenin, Runx2, and VEGF in cultured cells from ossified ligament were significantly higher than in the control cells. Application of cyclic tensile strain to cells from ossified ligament resulted in significant increases in mRNA expression levels ofβ-catenin, Runx2, and VEGF at 24 hours. Chondrocytes around the calcification front were immunopositive forβ-catenin. Immunoreactivity of Wnt and Wnt receptors were strongly present in premature mesenchymal cells around blood vessel in the fibrocartilage area. Our results indicated that cyclic tensile strain applied to cells from ossified ligament activated their ossification through a process mediated by the Wnt/β-catenin signaling pathway.
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