Immunohistochemistry was performed to assess protein expression of IL-1β, IL-6, IL-8, and collagen type II in the IVD tissue. Cell viability in the IVD was assessed using lactate dehydrogenase and ethidium homodimer-1 staining. Release of nitric oxide (NO), interleukin 8 (IL-8) and glycosaminoglycan (GAG) into the IVD conditioned medium were analyzed. Gene expression in the IVD tissue was measured by RT-qPCR. Etanercept was injected intradiscally while Tofacitinib was supplemented into the culture medium. Bovine caudal IVDs were cultured in a bioreactor system for 4 days to simulate physiological or degenerative conditions: (1) Phy-physiological loading (0.02–0.2 MPa 0.2 Hz 2 h/day) and high glucose DMEM medium (4.5 g/L) (2) Deg+Tumor necrosis factor α (TNF-α)-degenerative loading (0.32–0.5 MPa 5 Hz 2 h/day) and low glucose DMEM medium (2 g/L), with TNF-α injection. The present study sought to investigate the potential of Etanercept and Tofacitinib for maintaining disc homeostasis in a preclinical intervertebral disc (IVD) organ culture model within IVD bioreactors allowing for dynamic loading and nutrient exchange. However, their underlying anti-inflammatory and regenerative activity is poorly explored. Anti-inflammatory agents are currently under investigation as they demonstrated to alleviate symptoms in patients having IDD.
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