Analysis of cell death and vertebral end plate bone mineral density in the annulus of the aging sand rat

Abstract

Background context: The relationship between disc degeneration and end plate sclerosis is poorly understood. The sand rat is an excellent, economical small-animal model in which disc degeneration is age related, spontaneous, reliable, and well characterized. This model is used here to evaluate disc degeneration, disc cell viability, and vertebral end plate bone mineral density (BMD) in lumbar sites.

Purpose: To determine the proportion of live and dead cells and end plate bone mineral density in the aging sand rat annulus.

Study design: Young and old sand rats were used in work approved by the Institutional Animal Care and Use Committee. Outcome measures were the percentage of live/dead annulus cells in the disc and the BMD of cranial and caudal end plates of lumbar vertebrae.

Methods: Bone densitometry was used to obtain endplate BMD on lumbar spines of 16 young sand rats aged 2 to 6 months and 26 older animals aged 22 to 46 months. X-ray films were analyzed for wedging, end plate calcification, and disc-space narrowing. Additional discs were also harvested and incubated with fluorochromes, and the percentage of live or dead cells were determined for the outer, inner annulus, and entire annulus.

Results: Radiographically old animals had significantly greater incidence of lumbar wedging (p<0.004) and a significantly greater incidence of end plate calcification and disc-space narrowing (p<0.01). In the live-dead study, the mean percentage of dead annulus cells for the three age groups were significantly different for the outer annulus (p<0.001), inner annulus (p=0.005), and total annulus (p<0.0001). The percentages of dead cells for the entire annulus were 46.14%+/-7.99% (age 2-6 months), 48.13%+/-17.32% (age, 13-19 months), and 76.80%+/-7.27% (age 26-38 months). The percentage of dead disc cells correlated significantly with age for outer annulus, inner annulus, and total annulus (p<0.006). The percentage of dead cells in the entire annulus and the inner annulus correlated significantly with end plate BMD (p<0.02).

Conclusions: Data are novel and show that in very aged sand rats, end plate BMD is significantly greater than that of young animals. Live/dead cell analyses showed increasing cell death in both outer and inner annulus, which correlated significantly with age and with end plate BMD.