The goal of the investigation was to test the hypothesis that the PI3K/AKT signaling pathway regulates the expression of the major extracellular matrix component of the intervertebral disc, aggrecan, in nucleus pulposus cells. Primary rat nucleus pulposus cells were treated with PI3K inhibitor to measure changes in gene and protein expression. In addition, cells were transfected with various luciferase reporter plasmids to investigate mechanisms of regulation of aggrecan gene expression. We found that treatment of nucleus pulposus cells with a PI3K inhibitor, LY294002 resulted in decreased expression of aggrecan and a reduction in deposition of sulfated glycosaminoglycans. Moreover, pharmacological suppression or co-expression of dominant negative (DN)-PI3K or DN-AKT resulted in downregulation of aggrecan promoter activity. Expression of constitutively active (CA)-PI3K significantly induced aggrecan promoter activity. We observed that PI3K maintained Sox9 gene expression and activity: inhibition of PI3K/AKT resulted in decreased Sox9 expression, lowered promoter activity and mediated a reduction in Sox9 transcriptional activity. PI3K effects were independent of phosphorylation status of Cterminus transactivation domain (TAD) of Sox9. Finally, we noted that in nucleus pulposus cells, PI3K signaling controlled transactivation of p300 (p300-TAD activity), an important transcriptional coactivator of Sox9. Results of these studies demonstrate for the first time that PI3K/AKT signaling controls aggrecan gene expression, in part by modulating Sox9 expression and activity in cells of the nucleus pulposus.

Cervical interbody device subsidence can result in screw breakage, plate dislodgement, and/or kyphosis. Preoperative bone density measurement may be helpful in predicting the complications associated with anterior cervical surgery. This is especially important when a motion preserving device is implanted given the detrimental effect of subsidence on the postoperative segmental motion following disc replacement. To evaluate the structural properties of the cervical endplate and examine the correlation with CT measured trabecular bone density. Eight fresh human cadaver cervical spines (C2-T1) were CT scanned and the average trabecular bone densities of the vertebral bodies (C3-C7) were measured. Each endplate surface was biomechanically tested for regional yield load and stiffness using an indentation test method. Overall average density of the cervical vertebral body trabecular bone was 270 +/- 74 mg/cm3. There was no significant difference between levels. The yield load and stiffness from the indentation test of the endplate averaged 139 +/- 99 N and 156 +/- 52 N/mm across all cervical levels, endplate surfaces, and regional locations. The posterior aspect of the endplate had significantly higher yield load and stiffness in comparison to the anterior aspect and the lateral aspect had significantly higher yield load in comparison to the midline aspect. There was a significant correlation between the average yield load and stiffness of the cervical endplate and the trabecular bone density on regression analysis. Although there are significant regional variations in the endplate structural properties, the average of the endplate yield loads and stiffnesses correlated with the trabecular bone density. Given the morbidity associated with subsidence of interbody devices, a reliable and predictive method of measuring endplate strength in the cervical spine is required. Bone density measures may be used preoperatively to assist in the prediction of the strength of the vertebral endplate. A threshold density measure has yet to be established where the probability of endplate fracture outweighs the benefit of anterior cervical procedure.

There is a significant loss of endplate integrity when 1 mm of endplate (44% loss) or 2 mm of endplate (52% loss) is removed. Although the implant interface plays an important role in the magnitude of the subsidence of a device, this study in general shows that the endplate is important in terms of maximizing the strength of a construct.