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Background: Articular cartilage is a fiber reinforced hydrated solid that serves a largely mechanical role of supporting load and enabling low friction joint articulation. Daily activities that load cartilage, lead to fluid exudation and compressive axial strain. To date, the only mechanism shown to recover this cartilage strain in vivo is unloading (e.g., lying supine). Based on recent work in cartilage explants, we hypothesized that loaded joint activity (walking) would also be capable of strain recovery in cartilage. Methods: Eight asymptomatic young adults performed a fixed series of tasks, each of which was followed by magnetic resonance imaging to track changes in their knee cartilage thickness. The order of tasks was as follows: 1) stand for 30 ​min, 2) walk for 10 ​min, 3) stand for 30 ​min, and 4) lie supine for 50 ​min. The change in cartilage thickness was used to compute the axial cartilage strain. Results: Standing produced an average axial strain of −5.1 ​% (compressive) in the tibiofemoral knee cartilage, while lying supine led to strain recovery. In agreement with our hypothesis, walking also led to cartilage strain recovery. Interestingly, the recovery rate during walking (0.19 ​% strain/min) was nearly 3-fold faster than lying supine (0.07 ​% strain/min). Conclusions: This study represents the first in vivo demonstration that joint activity is capable of recovering compressive strain in cartilage. These findings indicate that joint activities such as walking may play a key role in maintaining and recovering cartilage strain, with implications for maintaining cartilage health and preventing or delaying cartilage degeneration.

Abstract Background Magnetic resonance imaging (MRI) noninvasively quantifies disc structure but requires segmentation that is both time intensive and susceptible to human error. Recent advances in neural networks can improve on manual segmentation. The aim of this study was to establish a method for automatic slice‐wise segmentation of 3D disc volumes from subjects with a wide range of age and degrees of disc degeneration. A U‐Net convolutional neural network was trained to segment 3D T1‐weighted spine MRI. Methods Lumbar spine MRIs were acquired from 43 subjects (23–83 years old) and manually segmented. A U‐Net architecture was trained using the TensorFlow framework. Two rounds of model tuning were performed. The performance of the model was measured using a validation set that did not cross over from the training set. The model version with the best Dice similarity coefficient (DSC) was selected in each tuning round. After model development was complete and a final U‐Net model was selected, performance of this model was compared between disc levels and degeneration grades. Results Performance of the final model was equivalent to manual segmentation, with a mean DSC = 0.935 ± 0.014 for degeneration grades I–IV. Neither the manual segmentation nor the U‐Net model performed as well for grade V disc segmentation. Compared with the baseline model at the beginning of round 1, the best model had fewer filters/parameters (75%), was trained using only slices with at least one disc‐labeled pixel, applied contrast stretching to its input images, and used a greater dropout rate. Conclusion This study successfully trained a U‐Net model for automatic slice‐wise segmentation of 3D disc volumes from populations with a wide range of ages and disc degeneration. The final trained model is available to support scientific use.

Abstract Background Cadaveric intervertebral discs are often studied for a variety of research questions, and outcomes are interpreted in the in vivo context. Unfortunately, the cadaveric disc does not inherently represent the LIVE condition, such that the disc structure (geometry), composition (T2 relaxation time), and mechanical function (opening pressure, OP) measured in the cadaver do not necessarily represent the in vivo disc. Methods We conducted serial evaluations in the Yucatan minipig of disc geometry, T2 relaxation time, and OP to quantify the changes that occur with progressive dissection and used axial loading to restore the in vivo condition. Results We found no difference in any parameter from LIVE to TORSO; thus, within 2 h of sacrifice, the TORSO disc can represent the LIVE condition. With serial dissection and sample preparation the disc height increased (SEGMENT height 18% higher than TORSO), OP decreased (POTTED was 67% lower than TORSO), and T2 time was unchanged. With axial loading, an imposed stress of 0.20–0.33 MPa returned the disc to in vivo, LIVE disc geometry and OP, although T2 time was decreased. There was a linear correlation between applied stress and OP, and this was conserved across multiple studies and species. Conclusion To restore the LIVE disc state in human studies or other animal models, we recommend measuring the OP/stress relationship and using this relationship to select the applied stress necessary to recover the in vivo condition.

The menisci increase the contact area of load bearing in the knee and thus disperse the mechanical stress via their circumferential tensile fibers. Traumatic meniscus injuries cause mechanical symptoms in the knee, and are more prevalent amongst younger, more active patients, compared to degenerative tears amongst the elderly population. Traumatic meniscus tears typically result from the load-and-shear mechanism in the knee joint. The treatment depends on the size, location, and pattern of the tear. For non-repairable tears, partial or total meniscal resection decreases its tensile stress and increases joint contact stress, thus potentiating the risk of arthritis. A longitudinal vertical tear pattern at the peripheral third red-red zone leads to higher healing potential after repair. The postoperative rehabilitation protocols after repair range from immediate weight-bearing with no range of motion restrictions to non-weight bearing and delayed mobilization for weeks. Pediatric and adolescent patients may require special considerations due to their activity levels, or distinct pathologies such as a discoid meniscus. Further biomechanical and biologic evidence is needed to guide surgical management, postoperative rehabilitation protocols, and future technology applications for traumatic meniscus injuries.

Abstract Objectives Mechanical loading is crucial for tendon matrix homeostasis. Under-stimulation of tendon tissue promotes matrix degradation and ultimately tendon failure. In this study, we examined the expression of tendon matrix molecules and matrix-degrading enzymes (matrix metalloproteinases) in stress-deprived tail tendons and compared to tendons that were mechanically loaded by a simple restraining method. Data description Isolated mouse tail fascicles were either floated or restrained by magnets in cell culture media for 24 h. The gene expression of tendon matrix molecules and matrix metalloproteinases in the tendon fascicles of mouse tails were examined by real-time RT-PCR. Stress deprivation of tail tendons increase Mmp3 mRNA levels. Restraining tendons represses these increases in Mmp3. The gene expression response to restraining was specific to Mmp3 at 24 h as we did not observe mRNA level changes in other matrix related genes that we examined (Col1, Col3, Tnc, Acan, and Mmp13). To elucidate, the mechanisms that may regulate load transmission in tendon tissue, we examined filamentous (F-)actin staining and nuclear morphology. As compared to stress deprived tendons, restrained tendons had greater staining for F-actin. The nuclei of restrained tendons are smaller and more elongated. These results indicate that mechanical loading regulates specific gene expression potentially through F-actin regulation of nuclear morphology. A further understanding on the mechanisms involved in regulating Mmp3 gene expression may lead to new strategies to prevent tendon degeneration.

Abstract Background Intervertebral disc degeneration is often implicated in low back pain; however, discs with structural degeneration often do not cause pain. It may be that disc mechanics can provide better diagnosis and identification of the pain source. In cadaveric testing, the degenerated disc has altered mechanics, but in vivo, disc mechanics remain unknown. To measure in vivo disc mechanics, noninvasive methods must be developed to apply and measure physiological deformations. Aim Thus, this study aimed to develop methods to measure disc mechanical function via noninvasive MRI during flexion and extension and after diurnal loading in a young population. This data will serve as baseline disc mechanics to later compare across ages and in patients. Materials & Methods To accomplish this, subjects were imaged in the morning in a reference supine position, in flexion, in extension, and at the end of the day in a supine position. Disc deformations and vertebral motions were used to quantify disc axial strain, changes in wedge angle, and anterior–posterior (A‐P) shear displacement. T2 weighted MRI was also used to evaluate disc degeneration via Pfirrmann grading and T2 time. All measures were then tested for effect of sex and disc level. Results We found that flexion and extension caused level‐dependent strains in the anterior and posterior of the disc, changes in wedge angle, and A‐P shear displacements. Flexion had higher magnitude changes overall. Diurnal loading did not cause level‐dependent strains but did cause small level‐dependent changes in wedge angle and A‐P shear displacements. Discussion Correlations between disc degeneration and mechanics were largest in flexion, likely due to the smaller contribution of the facet joints in this condition. Conclusion In summary, this study established methods to measure in vivo disc mechanical function via noninvasive MRI and established a baseline in a young population that may be compared to older subjects and clinical disorders in the future.

Abstract Finite element models of the intervertebral disc are used to address research questions that cannot be tested through typical experimentation. A disc model requires complex geometry and tissue properties to be accurately defined to mimic the physiological disc. The physiological disc possesses residual strain in the annulus fibrosus (AF) due to osmotic swelling and due to inherently pre‐strained fibers. We developed a disc model with residual contributions due to swelling‐only, and a multigeneration model with residual contributions due to both swelling and AF fiber pre‐strain and validated it against organ‐scale uniaxial, quasi‐static and multiaxial, dynamic mechanical tests. In addition, we demonstrated the models' ability to mimic the opening angle observed following radial incision of bovine discs. Both models were validated against organ‐scale experimental data. While the swelling only model responses were within the experimental 95% confidence interval, the multigeneration model offered outcomes closer to the experimental mean and had a bovine model opening angle within one SD of the experimental mean. The better outcomes for the multigeneration model, which allowed for the inclusion of inherently pre‐strained fibers in AF, is likely due to its uniform fiber contribution throughout the AF. We conclude that the residual contribution of pre‐strained fibers in the AF should be included to best simulate the physiological disc and its behaviors.

Abstract Noninvasive assessments of intervertebral disc health and degeneration are critical for addressing disc degeneration and low back pain. Magnetic resonance imaging (MRI) is exceptionally sensitive to tissue with high water content, and measurement of the MR transverse relaxation time, T2, has been applied as a quantitative, continuous, and objective measure of disc degeneration that is linked to the water and matrix composition of the disc. However, T2 measurement is susceptible to inaccuracies due to Rician noise, T1 contamination, and stimulated echo effects. These error generators can all be controlled for with proper data collection and fitting methods. The objective of this study was to identify sequence parameters to appropriately acquire MR data and to establish curve fitting methods to accurately calculate disc T2 in the presence of noise by correcting for Rician noise. To do so, we compared T2 calculated from the typical monoexponential (MONO) fits and noise corrected exponential (NCEXP) fits. We examined how the selected sequence parameters altered the calculated T2 in silico and in vivo. Typical MONO fits were frequently poor due to Rician noise, and NCEXP fits were more likely to provide accurate T2 calculations. NCEXP is particularly less biased and less uncertain at low SNR. This study showed that the NCEXP using sequences with data from 20 echoes out to echo times of ~300 ms is the best method for calculating T2 of discs. By acquiring signal data out to longer echo times and accounting for Rician noise, the curve fitting is more robust in calculating T2 despite the noise in the data. This is particularly important when considering degenerate discs or AF tissue because the SNR of these regions is lower.

The kinematics of the intervertebral disc are defined by six degrees of freedom (DOF): three translations (Tz: axial compression, Tx: lateral shear, and Ty: anterior‐posterior shear) and three rotations (Rz: torsion, Rx: flexion‐extension, and Ry: lateral bending). There is some evidence that the six DOFs are mechanically coupled, such that loading in one DOF affects the mechanics of the other five “off‐axis” DOFs, however, most studies have not controlled and/or measured all six DOFs simultaneously. Additionally, the relationships between disc geometry and disc mechanics are important for evaluation of data from different sized donor and patient discs. The objectives of this study were to quantify the mechanical behavior of the intervertebral disc in all six degrees of freedom (DOFs), measure the coupling between the applied motion in each DOF with the resulting off‐axis motions, and test the hypothesis that disc geometry influences these mechanical behaviors. All off‐axis displacements and rotations were significantly correlated with the applied DOF and were of similar magnitude as physiologically relevant motion, confirming that off‐axis coupling is an important mechanical response. Interestingly, there were pairs of DOFs that were especially strongly coupled: lateral shear (Tx) and lateral bending (Ry), anterior‐posterior shear (Ty) and flexion‐extension (Rx), and compression (Tz) and torsion (Rz). Large off‐axis shears may contribute to injury risk in bending and flexion. In addition, the disc responded to shear (Tx, Ty) and rotational loading (Rx, Ry, and Rz) by increasing in disc height in order to maintain the applied compressive load. Quantifying these mechanical behaviors across all six DOF are critical for designing and testing disc therapies, such as implants and tissue engineered constructs, and also for validating finite element models.

Tears are central to knee meniscus pathology and, from a mechanical perspective, are crack-like defects (cracks). In many materials, cracks create stress concentrations that cause progressive local rupture and reduce effective strength. It is currently unknown if cracks in meniscus have these consequences; if they do, this would have repercussions for management of meniscus pathology. The objective of this study was to determine if a short crack in meniscus tissue, which mimics a preclinical meniscus tear, (a) causes crack growth and reduces effective strength, (b) creates a near-tip strain concentration and (c) creates unloaded regions on either side of the crack. Specimens with and without cracks were tested in uniaxial tension and compared in terms of macroscopic stress–strain curves and digital image correlation strain fields. The strain fields were used as an indicator of stress concentrations and unloaded regions. Effective strength was found to be insensitive to the presence of a crack (potential effect < 0.86 s.d.; β = 0.2), but significant strain concentrations, which have the potential to lead to long-term accumulation of tissue or cell damage, were observed near the crack tip.

Degeneration of the intervertebral discs, a process characterized by a cascade of cellular, biochemical, structural and functional changes, is strongly implicated as a cause of low back pain. Current treatment strategies for disc degeneration typically address the symptoms of low back pain without treating the underlying cause or restoring mechanical function. A more in-depth understanding of disc degeneration, as well as opportunities for therapeutic intervention, can be obtained by considering aspects of intervertebral disc development. Development of the intervertebral disc involves the coalescence of several different cell types through highly orchestrated and complex molecular interactions. The resulting structures must function synergistically in an environment that is subjected to continuous mechanical perturbation throughout the life of an individual. Early postnatal changes, including altered cellularity, vascular regression and altered extracellular matrix composition, might set the disc on a slow course towards symptomatic degeneration. In this Perspective, we review the pathogenesis and treatment of intervertebral disc degeneration in the context of disc development. Within this scope, we examine how model systems have advanced our understanding of embryonic morphogenesis and associated molecular signaling pathways, in addition to the postnatal changes to the cellular, nutritional and mechanical microenvironment. We also discuss the current status of biological therapeutic strategies that promote disc regeneration and repair, and how lessons from development might provide clues for their refinement.

Abstract Background Inflammatory cytokines such as interleukin‐1 beta (IL‐1β) contribute to the progression of intervertebral disc degeneration. Previously we demonstrated, in vitro, that by delivering interleukin‐1 receptor antagonist (IL‐1ra) from poly(lactic co‐glycolic acid) (PLGA) microspheres, we could attenuate the degradative effects of IL‐1β on the nucleus pulposus (NP) for up to 20 days. The objective of this study was to undertake a preliminary investigation into whether microspheres could be successfully delivered to and retained in the disc in vivo, and whether IL‐1ra released from those microspheres remained biologically active. For retention studies, fluorescently‐labeled microspheres were delivered to the NPs of rat caudal discs. Rats were sacrificed at time points up to 56 days, and microspheres were localized using fluorescent microscopy. To investigate whether IL‐1ra microspheres could effectively inhibit the effects of IL‐1β in vivo, four disc levels were allocated to the following treatment groups: intact; saline; IL‐1β; or IL‐1β + IL‐1ra microspheres. Rats were sacrificed after seven days and NP glycosaminoglycan content was measured. Findings Microspheres were visible in the disc at all time points up to 28 days, and localized to the NP, the annulus fibrosus (AF), or both. Glycosaminoglycan content for discs injected with IL‐1β alone was significantly lower than for intact controls. For discs injected with IL‐1β along with IL‐1ra microspheres, glycosaminoglycan content was not significantly different from intact controls. Conclusions Microspheres can successfully be delivered to the disc in vivo and retained for a clinically relevant time frame. IL‐1ra released from microspheres can effectively prevent IL‐1β‐induced NP glycosaminoglycan loss in vivo.

Identification of the deformation mechanisms and specific components underlying the mechanical function of biological tissues requires mechanical testing at multiple levels within the tissue hierarchical structure. Dichlorotriazinylaminofluorescein (DTAF) is a fluorescent dye that is used to visualize microscale deformations of the extracellular matrix in soft collagenous tissues. However, the DTAF concentrations commonly employed in previous multiscale experiments (≥2000 µg/ml) may alter tissue mechanics. The objective of this study was to determine whether DTAF affects tendon fascicle mechanics and if a concentration threshold exists below which any observed effects are negligible. This information is valuable for guiding the continued use of this fluorescent dye in future experiments and for interpreting the results of previous work. Incremental strain testing demonstrated that high DTAF concentrations (≥100 µg/ml) increase the quasi-static modulus and yield strength of rat tail tendon fascicles while reducing their viscoelastic behavior. Subsequent multiscale testing and modeling suggests that these effects are due to a stiffening of the collagen fibrils and strengthening of the interfibrillar matrix. Despite these changes in tissue behavior, the fundamental deformation mechanisms underlying fascicle mechanics appear to remain intact, which suggests that conclusions from previous multiscale investigations of strain transfer are still valid. The effects of lower DTAF concentrations (≤10 µg/ml) on tendon mechanics were substantially smaller and potentially negligible; nevertheless, no concentration was found that did not at least slightly alter the tissue behavior. Therefore, future studies should either reduce DTAF concentrations as much as possible or use other dyes/techniques for measuring microscale deformations.