In vitro mechanical testing of intervertebral discs is crucial for basic science and preclinical testing. Generally, these tests aim to replicate in vivo conditions, but simplifications are necessary in specimen preparation and mechanical testing due to complexities in both structure and the loading conditions required to replicate in vivo conditions. There has been a growing interest in developing a consensus of testing protocols within the spine community to improve comparison of results between studies. The objective of this study was to perform axial compression experiments on bovine bone-disc-bone specimens at three institutions. No differences were observed between testing environment being air, with PBS soaked gauze, or a PBS bath (P > .206). A 100-fold increase in loading rate resulted in a small (2%) but significant increase in compressive mechanics (P < .017). A 7% difference in compressive stiffness between Labs B and C was eliminated when values were adjusted for test system compliance. Specimens tested at Lab A, however, were found to be stiffer than specimens from Lab B and C. Even after normalizing for disc geometry and adjusting for system compliance, an 35% difference was observed between UK based labs (B and C) and the USA based lab (A). Large differences in specimen stiffness may be due to genetic differences between breeds or in agricultural feed and use of growth hormones; highlighting significant challenges in comparing mechanics data across studies. This research provides a standardized test protocol for the comparison of spinal specimens and provides steps towards understanding how location and test setup may affect biomechanical results.
A comprehensive understanding of multiscale and multiphasic intervertebral disc mechanics is crucial for designing advanced tissue engineered structures aiming to recapitulate native tissue behavior. The bovine caudal disc is a commonly used human disc analog due to its availability, large disc height and area, and similarities in biochemical and mechanical properties to the human disc. Because of challenges in directly measuring subtissue-level mechanics, such as in situ fiber mechanics, finite element models have been widely employed in spinal biomechanics research. However, many previous models use homogenization theory and describe each model element as a homogenized combination of fibers and the extrafibrillar matrix while ignoring the role of water content or osmotic behavior. Thus, these models are limited in their ability in investigating subtissue-level mechanics and stress-bearing mechanisms through fluid pressure. The objective of this study was to develop and validate a structure-based bovine caudal disc model, and to evaluate multiscale and multiphasic intervertebral disc mechanics under different loading conditions and with degeneration. The structure-based model was developed based on native disc structure, where fibers and matrix in the annulus fibrosus were described as distinct materials occupying separate volumes. Model parameters were directly obtained from experimental studies without calibration. Under the multiscale validation framework, the model was validated across the joint-, tissue-, and subtissue-levels. Our model accurately predicted multiscale disc responses for 15 of 16 cases, emphasizing the accuracy of the model, as well as the effectiveness and robustness of the multiscale structure-based modeling-validation framework. The model also demonstrated the rim as a weak link for disc failure, highlighting the importance of keeping the cartilage endplate intact when evaluating disc failure mechanisms in vitro. Importantly, results from this study elucidated important fluid-based load-bearing mechanisms and fiber-matrix interactions that are important for understanding disease progression and regeneration in intervertebral discs. In conclusion, the methods presented in this study can be used in conjunction with experimental work to simultaneously investigate disc joint-, tissue-, and subtissue-level mechanics with degeneration, disease, and injury.
Mechanical testing is a valuable tool for assessing intervertebral disc health, but the wide range of testing protocols makes it difficult to compare results from different studies. Normalizing mechanical properties by disc geometry allows for such comparisons, but there is little consistency in the methods by which disc geometry is measured. As such, we hypothesized that methods used to measure disc geometry would impact reported mechanical properties. Disc height and area were measured using computed tomography (CT), digital calipers, and ImageJ to yield three different measurements for disc height and six for disc area. Disc heights measured by digital calipers ex situ were >30% less than disc heights measured in situ by CT, and disc areas measured ex situ using ImageJ were >30% larger than those measured by CT. This significantly affected reported mechanical properties, leading to a 65% reduction in normalized compressive stiffness in the most extreme case. Though we cannot quantitatively correct between methods, results presented in this study suggest that disc geometry measurement methods have a significant impact on normalized mechanical properties and should be accounted for when comparing results.
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