Effects of Asymmetric Dynamic Loading on Intervertebral Disc – towards a Scoliosis Mimicking Organ Culture Model

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Introduction The etiology of spinal deformity in idiopathic scoliosis is unclear to date. One of the suspected influences is the asymmetric loading condition involved in the disorder. The aim of this project is to test the hypothesis that asymmetric dynamic loading influences the morphological and biological characteristics of the intervertebral discs in scoliosis. The study is performed with organ cultured discs by using a custom-designed asymmetrical loading device. Material and Methods Bovine caudal discs (6–10 months) were used in current study. For symmetric dynamic loading (Parallel), discs were placed in custom-designed chambers, and compressed by parallel metal plates in a Bose mechanical testing device. For asymmetric dynamic loading (Wedge), a 10° wedge was placed underneath the discs to mimic the load bearing condition of discs in scoliotic patients. The discs were submitted to 2 different load regimes: (1) 1 hour dynamic loading (0.02–0.4 MPa, 1Hz) and 23 hours free swelling culture for 7 days; (2) 1 hour dynamic loading (0.02–0.4 MPa, 1Hz) and 23 hours static loading (0.2 MPa) for 7 days. Disc heights were measured with caliper before and after each loading. After 7 days of culture, gene expression levels of aggrecan (ACAN), type I and II collagen (COL1 and COL2), IL1, IL6, and MMP1 in the annulus fibrosus was analyzed by real-time PCR. Genes that have been found dysregulated in human scoliotic discs compared with healthy controls were also measured in the organ cultured discs, including MMP13, type X collagen (COL10), CXCR4, BMP3, S100A12, and S100A8 ( n = 8). Results Disc height showed a constant drop in load regime 2, while a temporary decrease after 1h dynamic loading followed by free swelling recovery was noted in load regime 1. After 7th dynamic loading, the change in shape was greater in load regime 2 (disc height ratio wedged to non-wedged side of 0.81), than that in load regime 1 (height ratio of 0.87, p < 0.05). Under load regime 2, MMP13 gene expression level increased 6.1-fold in Wedge disc compared with Parallel disc, while gene expression levels of COL10, CXCR4, BMP3, S100A12, and S100A8 were not affected. Gene expression levels of ACAN, COL1 and COL2 under load regime 1 were significantly higher compared with load regime 2. Moreover, discs under load regime 2 showed a trend in higher IL1, IL6, and MMP1 gene expression compared with regime 1. Conclusion Diurnal dynamic loading and free swelling recovery could maintain the gene expression of organ cultured discs at their physiological level. Diurnal dynamic loading followed by static loading mimicked a degenerative condition, as indicated by lower anabolic and higher catabolic gene expression. These results suggest that recovery of disc height and morphology after dynamic load may help to prevent degeneration of discs under constant loading. Asymmetric dynamic and static loading regime induces an increase in MMP13 gene expression compared with symmetric loading, which was also observed in a human scoliosis sample dataset. These results indicate that short-term asymmetric loading may be used to mimic early changes associated with the onset of scoliosis. Acknowledgment This study is supported by AOSpine International.