Generating a Pro-Inflammatory Organ Culture Model to Simulate an Early Intervertebral Disc Disease
Transcription
Take a bovine tail.
Remove the overlying tissue and vertebral processes, then cut transversely to obtain segments.
Identify the intervertebral disc or IVD, located between the growth plates and vertebral end plates.
Cut on either side of the IVD and place it in a dish containing gauze soaked in buffer to maintain tissue viability.
Scrape off any attached vertebral tissue and measure the IVD height.
Wash with buffer to remove blood remnants and disinfect the IVD in a buffer containing antibiotics.
Place the IVDs in bioreactor chambers containing media.
The media in the physiological group has high glucose, while in the pathological group, a low-glucose medium induces nutrient deprivation.
Place the chambers in the bioreactor.
For the physiological IVD, apply a gentle load daily. For the pathological IVD, apply an intense load at a higher rate daily.
Between the daily loading tests, allow the IVDs to recover in media.
After all loading procedures, measure the IVD heights.
A reduced height in the pathological IVDs indicates a degenerative and pro-inflammatory condition.
