This study investigates the effects of recombinant TNF-�� on intervertebral disc (IVD) degeneration under mechanical stress. By creating a model of early-stage IVD disease, the research highlights the inflammatory and apoptotic pathways activated by TNF-��.
Take an intervertebral disc (IVD), a cushion-like structure located between the vertebrae, from a bovine tail.
Position it vertically and inject recombinant TNF-α, a pro-inflammatory cytokine, into the nucleus pulposus (NP), the gel-like core.
Withdraw the syringe halfway and pull the plunger to create a vacuum, preventing TNF-α backflow. Then, remove the syringe.
Place the IVD in a bioreactor chamber containing media, and apply an intense pressure load daily.
TNF-α binds to its receptors on NP cells, triggering a signaling cascade.
This activates initiator and executioner caspases, which degrade cellular proteins and induce apoptosis.
TNF-α also activates the transcription factor NF-κB, which induces inflammatory gene expression, leading to the release of enzymes that degrade the IVD's extracellular matrix.
Additionally, the intense pressure load induces mechanical stress, further degrading the IVD.
This reduces the IVD's height in comparison to untreated controls, establishing a degenerative and proinflammatory model of early-stage IVD disease.
After the first dynamic loading cycle on day one, directly place the IVDs in a Petri dish in a vertical position and stabilize them with a tweezer. Inject recombinant TNF alpha with a 30-gauge insulin needle, slowly at a speed of approximately 70 microliters per minute into the IVDs of the pathological group. After injecting, pull the syringe halfway back and pull the plunger to create a vacuum that prevents the injected solution from leaking. Then, remove the syringe completely from the IVD.
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