Cyclooxygenase-2 deficiency causes delayed ossification of lumbar vertebral endplates

: Based on previous findings that cyclooxygenase-2 (COX-2) is a critical molecule in chondrocyte differentiation and skeletal repair, we hypothesized that COX-2 deficiency or inhibition affects the ossification of vertebral endplates (VEP) and degeneration of intervertebral discs (IVD) and thus is involved in the pathogenesis of low back pain (LBP). We aimed to delineate the COX-2 working mechanism and its interacting molecules, and to explore the effect of NSAIDs and selective COX-2 inhibitor on degenerative spinal diseases. Here, lumbar spinal samples harvested from Cox-2 mutant (Cox-2-/-) and wild type (WT) mice were used for histological examinations. Nucleus pulposus (NP) cells isolated from rat were treated with PGE-2. Mouse endplate chondrocytes (mEC) isolated from mice were treated with a recombinant sonic hedgehog (Shh) protein. A mouse IVD organ culture system was established and treated COX-2 inhibitor Celecoxib. Human lumbar endplate chondrocytes were cultured and treated with Celecoxib. Immunohistochemical (IHC) studies were done in the human and mouse VEP samples. Radiographic and histological examinations revealed delayed VEP ossification in Cox-2-/- mice compared to WT ones. Decreased PGE-2 expression was found to promote Shh expression in rat NP cells, while Shh increased noggin expression in mEC. IHC showed that noggin expression was increased while pSmad1 expression decreased in the VEP of Cox-2-/- mice. Human VEP samples from patients with severe IVD degeneration showed decreased expression of Shh and noggin and increased expression of COX-2 and pSmad1 compared with milder cases. In cultured mouse IVDs and human endplate chondrocytes, Celecoxib enhanced expression of Shh and noggin and decreased Smad1 phosphorylation. In conclusion, COX-2/PGE-2 axis plays an important role in VEP ossification and IVD degeneration through crosstalk with Shh and BMP signaling pathways. These findings may facilitate clinical use of COX-2 inhibitor to prevent LBP progression.