ABSTRACT
OBJECTIVE: Notochordal cells in the intervertebral disc interact with nucleus pulposus (NP) cells and support the maintenance of disc homeostasis by regulation of matrix production. However, the influence of notochordal cells has not been evaluated in the annulus fibrosus (AF), which is the primary pain generator in the disc. We hypothesized that the notochordal cell has the capacity to modulate inflammatory mediators secreted by AF cells secondary to stimulation. METHODS: Notochordal and AF cells were isolated from adult New Zealand white rabbits. AF pellets were cultured with notochordal cell clusters or in notochordal cell-conditioned media (NCCM) for 24 or 48 hours with proinflammatory cytokines at varying concentrations. Gene expression in AF pellets were assayed for nitric oxide synthase (iNOS), cyclo-oxygenase (COX)-2, and interleukin (IL)-6 by real time reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: AF pellet in NCCM significantly decreased the iNOS and COX-2 messenger ribonucleic acid (mRNA) levels compared to AF pellets alone and AF pellets with notochordal cells (p < 0.05). AF pellet resulted in dose-dependent iNOS and COX-2 expression in response to IL-1beta, stimulation, demonstrating that 1 ng/ml for 24 hours yielded a maximal response. AF pellet in NCCM significantly decreased the expression of iNOS and COX-2 in response to 1ng/ml IL-1beta, stimulation at 24 hours (p < 0.05). There was no difference in IL-6 expression compared to AF pellets alone or AF pellets with notochordal cell clusters. CONCLUSION: We conclude that soluble factors from notochordal cells mitigate the gene expression of inflammatory mediators in stimulated AF, as expected after annular injury, suggesting that notochordal cells could serve as a novel therapeutic approach in symptomatic disc development.
Subject(s)
Adult , Humans , Rabbits , Cytokines , Gene Expression , Homeostasis , Interleukin-6 , Interleukins , Intervertebral Disc , Nitric Oxide Synthase , Notochord , Prostaglandin-Endoperoxide Synthases , Reverse Transcriptase Polymerase Chain Reaction , RNAABSTRACT
BACKGROUND: Endobronchial tuberculosis often complicates bronchostenosis, which can cause dyspnea due to an airway obstruction, and can be misdiagnosed as bronchial asthma or lung cancer. This study investigated the possible correlation between the interferon-gamma(IFN-gamma) and transforming growth factor-beta(TGF-beta) levels in the serum and bronchial washing fluid and the treatment results in endobronchial tuberculosis patients. METHODS: Sixteen patients, who were diagnosed as endobronchial tuberculosis using bronchoscopy, and 10 healthy control subjects were enrolled in this study. The IFN-gamma and TGF-beta levels were measured in the serum and bronchial washing fluid of 16 endobronchial tuberculosis patients before and after treatment using the ELISA method. The endobronchial tuberculosis patients were divided into those who showed bronchial fibrostenosis after treatment and those who did not. RESULTS: The IFN-gamma and TGF-beta levels in the bronchial washing fluid in endobronchial tuberculosis patients were elevated comparing to the control (p < 0.05). After treatment, 7 of the 16 endobronchial tuberculosis patients showed bronchial fibrostenosis and the other 9 cases healed without this sequela. In the patients with fibrostenosis after treatment, the initial serum TGF-beta level was lower than the patients without fibrostenosis after treatment (p < 0.05). Moreover, the serum TGF-beta level after treatment further decreased comparing to the patients without fibrostenosis after treatment(p < 0.05). CONCLUSION: Elevated IFN-gamma and TGF-beta levels in the bronchial washing fluid in endobronchial tuberculosis patients are believed to be related to the pathogenesis of endobronchial tuberculosis. The decreased initial serum TGF-beta level and the change in the serum TGF-beta level after treatment are believed to be involved in bronchial fibrostenosis during the course of the disease.