Effects of Cyclic Mechanical Stress on the Production of Inflammatory Agents by Nucleus Pulposus and Anulus Fibrosus Derived Cells In Vitro

Abstract
Cyclic mechanical stress (CMS) was applied to cultured nucleus pulposus and anulus fibrosus cells, and the production of inflammatory agents by these cells was evaluated. To investigate the involvement of CMS in the production of inflammatory agents by disc cells. It has been reported that CMS affects degeneration of the disc. However, little is known about the effect of CMS on the production of inflammatory agents by both cell types in vitro. Cells derived from nucleus pulposus and anulus fibrosus of Sprague-Dawley rat tails were cultured with or without CMS applied by the Flexercell Strain Unit (Flexcell International Corp., Hillsborough, NC) in the presence or absence of inflammatory stimulus. Doses of prostaglandin-E2 (PGE2) were measured in the culture supernatants. Semiquantitative evaluations of the expressions of cyclooxygenase (COX)-2 and phospholipase-A2 IIA messenger ribonucleic acids (mRNAs) were also examined. Sole application of CMS on nucleus pulposus and anulus fibrosus cells increased PGE2 synthesis. Coincidence of CMS and inflammatory stimulus synergistically enhanced PGE2 synthesis of both cell types. Anulus fibrosus cells showed a stronger reactivity to these stimuli than nucleus pulposus cells. The expression of COX-2 mRNA of anulus fibrosus cells tended to correlate to the amount of PGE2, whereas COX-2 mRNA was constitutively expressed in nucleus pulposus cells, suggesting that the roles of COX-2 might be different between nucleus pulposus and anulus fibrosus. Phospholipase-A2 IIA mRNA was constitutively expressed in both cell types. The results of this study suggested that CMS might be involved in the pathomechanism of pain induction of lumbar disc diseases.