Dexamethasone induces human spinal ligament derived cells toward osteogenic differentiation

Ossification of spinal ligament is characterized by heterotopic bone formation in the spinal ligaments that are normally composed of fibrous tissues. The pathogenesis of ossification of spinal ligament has been suggested to be associated with osteogenic differentiation of the spinal ligament cells. In order to address this hypothesis, cells derived from human spinal ligament were investigated for their osteogenic potential by the treatment of dexamethasone in vitro. Yellow ligaments were obtained from patients with spinal disorders except ossification of spinal ligament during surgery, and the adhering tissues were removed completely. Most of the ligament cells treated with vehicle exhibited a fibroblast-like spindle shape, while the dexamethasone-treated cells acquired a polygonal morphology. Growth of the ligament cells was suppressed by dexamethasone at a high concentration. Some of the vehicle treated-cells were alkaline phosphatase-positive, and dexamethasone increased the alkaline phosphatase-positive cells and alkaline phosphatase activity in the cells. Northern blot analysis demonstrated that mRNAs expression of pro-alpha1(I) collagen and alkaline phosphatase were promoted by dexamethasone. Analysis by reverse transcription-polymerase chain reaction showed that expression of osteocalcin mRNA was detected in the dexamethasone-treated cells but not in the vehicle-treated cells, and dexamethasone-induced osteocalcin mRNA expression was promoted by 1,25-dihydroxyvitamin D(3). Finally, mineralization of extracellular matrix in the cells was induced by the presence of dexamethasone and 1,25-dihydroxyvitamin D(3). These results suggest for the first time that dexamethasone has a possible involvement in the osteoblastic differentiation of human spinal ligament cells.