Development of a whole organ culture model for intervertebral disc disease

Abstract

Background/Objective: Whole organ in vitro intervertebral disc models have been associated with poor maintenance of cell viability. No previous studies have used a rotating wall vessel bioreactor for intervertebral disc explants culture. The purpose of this study was to develop and validate an in vitro model for the assessment of biological and biomechanical measures of intervertebral disc health and disease. Methods: To this end, endplate-intervertebral disc-endplate whole organ explants were harvested from the tails of rats. For the injured group, the annulus fibrosus was penetrated with a 20G needle to the nucleus pulposus and aspirated. Explants were cultured in a rotating wall vessel bioreactor for 14 days. Results: Cell viability and histologic assessments were performed at Day 0, Day 1, Day 7, and Day 14. Compressive mechanical properties of the intervertebral disc were assessed at Day 0 and Day 14. In the annulus fibrosus and nucleus pulposus cells, the uninjured group maintained high viability through 14 days of culture, whereas cell viability in annulus fibrosus and nucleus pulposus of the injured intervertebral discs was markedly lower at Day 7 and Day 14. Histologically, the uninjured intervertebral discs maintained cell viability and tissue morphology and architecture through 14 days, whereas the injured intervertebral discs showed areas of cell death, loss of extracellular matrix integrity, and architecture by Day 14. Stiffness values for uninjured intervertebral discs were similar at Day 0 and Day 14, whereas the stiffness for the injured intervertebral discs was approximately 2.5 times greater at Day 14. Conclusion: These results suggest that whole organ intervertebral discs explants can be successfully cultured in a rotating wall vessel bioreactor to maintain cell viability and tissue architecture in both annulus fibrosus and nucleus pulposus for at least 14 days. In addition, the injury used produced pathologic changes consistent with those seen in degenerative intervertebral disc disease in humans. This model will permit further study into potential future treatments and other mechanisms of addressing intervertebral disc disease.

Keywords

• biomechanics
• degeneration
• intervertebral disc
• rat tail
• whole organ culture