Polyphenol-modified biomimetic bioadhesives for the therapy of annulus fibrosus defect and nucleus pulposus degeneration after discectomy.

Discectomy is the surgical standard of care to relieve low back pain caused by intervertebral disc (IVD) herniation. However, there remains annulus fibrosus (AF) defect and nucleus pulposus (NP) degeneration, which often result in recurrent herniation (re-herniation). Herein, we develop a polyphenol-modified waterborne polyurethane bioadhesives (PPU-glues) to promote therapy prognosis after discectomy. Being composed of tannic acid (TA) mixed cationic waterborne polyurethane nanodispersions (TA/WPU+) and curcumin (Cur) embedded anionic waterborne polyurethane nanodispersions (Cur-WPU-), PPU-glue gels rapidly (<10 s) and exhibits low swelling ratios, tunable degradation rates and good biocompatibility. Due to the application of an adhesion strategy combing English ivy mechanism and particle packing theory, PPU-glue also shows considerable lap shear strength against wet porcine skin (≈58 kPa) and burst pressure (≈26 kPa). The mismatched particle sizes and the opposite charges of TA/WPU+ and Cur-WPU- in PPU-glue bring electrostatic interaction and enhance particle packing density. PPU-glue possesses superior reactive oxygen species (ROS)-scavenging capacity derived from polyphenols. PPU-glue can regulate extracellular matrix (ECM) metabolism in degenerated NP cells, and it can promote therapy biologically and mechanically in degenerated rat caudal discs. In summary, this study highlights the therapeutic approach that combines AF seal and NP augmentation, and PPU-glue holds great application potentials for post discectomy therapy. STATEMENT OF SIGNIFICANCE: Currently, there is no established method for the therapy of annulus fibrosus (AF) defect and nucleus pulposus (NP) degeneration after discectomy. Herein, we developed a polyphenol-modified biomimetic polyurethane bioadhesives (PPU-glue) with strong adhesive strength and superior bioactive property. The adhesion strategy that combined a particle packing theory and an English ivy mechanism was firstly applied to the intervertebral disc repair field, which benefited AF seal. The modified method of incorporating polyphenols was utilized to confer with ROS-scavenging capacity, ECM metabolism regulation ability and anti-inflammatory property, which promoted NP augmentation. Thus, PPU-glue attained the synergy effect for post discectomy therapy, and the design principle could be universally expanded to the bioadhesives for other surgical uses.

3D bioprinting of an intervertebral disc tissue analogue with a highly aligned annulus fibrosus via suspended layer additive manufacture.

Intervertebral disc (IVD) function is achieved through integration of its two component regions: the nucleus pulposus (NP) and the annulus fibrosus (AF). The NP is soft (0.3-5 kPa), gelatinous and populated by spherical NP cells in a polysaccharide-rich extracellular matrix (ECM). The AF is much stiffer (~100 kPa) and contains layers of elongated AF cells in an aligned, fibrous ECM. Degeneration of the disc is a common problem with age being a major risk factor. Progression of IVD degeneration leads to chronic pain and can result in permanent disability. The development of therapeutic solutions for IVD degeneration is impaired by a lack of in vitro models of the disc that are capable of replicating the fundamental structure and biology of the tissue. This study aims to investigate if a newly developed suspended hydrogel bioprinting system (termed SLAM) could be employed to fabricate IVD analogues with integrated structural and compositional features similar to native tissue. Bioprinted IVD analogues were fabricated to recapitulate structural, morphological and biological components present in the native tissue. The constructs replicated key structural components of native tissue with the presence of a central, polysaccharide-rich NP surrounded by organised, aligned collagen fibres in the AF. Cell tracking, actin and matrix staining demonstrated that embedded NP and AF cells exhibited morphologies and phenotypes analogous to what is observed in vivo with elongated, aligned AF cells and spherical NP cells that deposited HA into the surrounding environment. Critically, it was also observed that the NP and AF regions contained a defined cellular and material interface and segregated regions of the two cell types, thus mimicking the highly regulated structure of the IVD.

Intervertebral disc deformation in the lower lumbar spine during object lifting measured in vivo using indwelling bone pins.

Object lifting is often categorized into squat and stoop techniques, with the former believed to protect the back by maintaining a neutral spine, and the latter considered harmful due to spinal flexion. Despite the widespread promotion of these beliefs, there is no evidence to support such dichotomy, as spinal flexion is not conclusively linked to low back pain. This study aimed to investigate intervertebral disc deformation in the lower lumbar spine during squat and stoop lifting using indwelling bone pins. Five healthy males underwent insertion of Kirschner wires into the L3, L4, and L5 spinous processes, followed by biomechanical data collection using magnetic and optical tracking systems during upright standing, isolated flexion/extension, and object lifting with both squat and stoop techniques. Except for one subject, stoop lifting resulted in up to 90 % greater disc wedging compared to squat lifting, with a significant difference at L4/L5 (p = 0.042). The anterior annulus fibrosus experienced 10 % to 40 % more compression during stoop lifting, but no significant differences were found in posterior annulus fibrosus expansion between techniques. Lever arms were about 35 % longer during stoop compared to squat lifting. These results indicate that even though stoop lifting generally led to greater disc deformation, significant deformation was also observed during squat lifting, challenging the notion of maintaining a neutral spine with this technique. Moreover, the considerable variability observed among participants raises concerns about the suitability of current one-size-fits-all lifting guidelines.

Stress relaxation behavior of the transition zone in the intervertebral disc.

The stress relaxation of the TZ region, located at the interface of the Annulus Fibrosus (AF) and Nucleus Pulposus (NP) of the disc, and how its stress is relaxed compared to the adjacent regions is unknown. The current study aimed to identify the TZ stress relaxation properties under different strain magnitudes (0.2, 0.4, and 0.6 mm/mm) and compared the TZ stress relaxation characteristics to the NP and inner AF (IAF) regions at a specific strain magnitude (0.6 mm/mm). The results of the current study revealed that the TZ region exhibited different stress relaxation properties under various strain magnitudes with significantly higher initial (p < 0.008) and reduced stresses (marginally; p = 0.06) at higher strains. Our experimental stress relaxation data revealed a significantly higher equilibrium stress for the IAF compared to the TZ and NP regions (p < 0.001) but not between the TZ and NP regions (p = 0.7). We found that NP radial stress relaxed significantly faster (p < 0.04) than the TZ and NP. Additionally, the current study proposed a simple mathematical model and identified that, consistent with experimental data, the overall effect of region on both the level of decayed stress and the rate at which stress is relaxed was significant (p < 0.006). The current study found a similar stress relaxation characteristic between the NP and TZ regions, while IAF exhibited different stress relaxation properties. It is possible that this mismatch in stress relaxation acts as a shape transformation mechanism triggered by viscoelastic behavior. STATEMENT OF SIGNIFICANCE: Our understanding of the biomechanical properties of the transition zone (TZ) in the IVD, a region at the interface of the Nucleus Pulposus (NP) and Annulus Fibrosus (AF), is sparse. Unfortunately, there are no current studies that investigate the TZ stress relaxation properties and how stress is relaxed in the TZ compared to the adjacent regions. For the first time, the current study characterized the stress relaxation properties of the TZ and described how the TZ stress is relaxed compared to its adjacent regions.

The mechanical properties measurement could be affected by forceps: a technical note on sampling human annulus fibrosus.

PURPOSE: To compare the mechanical properties of human annulus fibrosus obtained by forceps versus bistoury and observe whether the measurement could be affected by forceps sampling method. METHODS: In this study, the mechanical properties of the the extracellular matrix (ECM) of human annulus fibrosus, including elastic modulus and stiffness, were investigated using atomic force microscope (AFM). Tissue was obtained from patients during operation using a bistoury or nucleus pulposus forceps. Tissues obtained with the nucleus pulposus forceps were considered as the forceps group and those obtained with a bistoury were considered as the bistoury group. RESULTS: There was no significant difference observed between the forceps and bistoury group according to histological staining. The elastic modulus of the forceps group was 0.41 ± 0.08 MPa, and that of bistoury group was 0.53 ± 0.13 MPa, and the difference between the two groups was statistically significant (p < 0.05). The stiffness of the forceps group was 0.024 ± 0.003 N/m, and that of the bistoury group was 0.037 ± 0.003 N/m, and the difference between the two groups was statistically significant (p < 0.05). CONCLUSION: The results indicate that the forceps sampling method has a substantial negative effect on the micromechanical properties of the ECM of the annulus fibrosus. Bistoury sampling method is recommended as the experimental subject for exploring the micromechanics mechanisms of cervical degenerative disease.

Comprehensive analysis of damage evolution in human annulus fibrosus: Numerical exploration of mechanical sensitivity to biological age-dependent alteration.

BACKGROUND AND OBJECTIVE: The annulus fibrosus is an essential part of the intervertebral disc, critical for its structural integrity. Mechanical deterioration in this component can lead to complete disc failure, particularly through tears development, with radial tears being the most common. These tears are often the result of both mechanical and biological factors. This study aims to numerically investigate the mechanisms of radial failure in the annulus tissue, taking into account the mechanical and age-dependent biological damage origins. A newly developed microstructure-based model was upgraded to predict damage evolution in the different annulus regions. METHODS: The study employs a computational model to predict mechanical failures in various annulus regions, using experimental data for comparison. The model incorporates age-dependent microstructural changes to evaluate the effects of biological aging on the mechanical behavior. It specifically includes a detailed analysis of the temporal changes in circumferential rigidity and failure strain of the annulus. RESULTS: The model demonstrated a strong ability to replicate the experimental responses of the different annulus regions to failure. It revealed that age-related microstructural changes significantly impact the rigidity and failure response of the annulus, particularly in the posterior regions and as well the anterior inner side. These changes increase susceptibility to rupture with aging. A correlation was also observed between the composition of collagen fibers, water content, and the annulus transversal response in both radial and axial directions. CONCLUSION: The findings challenge previous assumptions, showing that age-dependent microstructural changes have a notable effect on the annulus mechanical properties. The computational model closely aligns with experimental observations, underscoring the determinant role of oriented collagen fibers in radial failure. This study enhances the understanding of annulus failure and provides a foundation for further research on the impact of aging on disc mechanical integrity and failure.

New complex cell junctions in and around the intervertebral discs discovered using autoantibodies from patients affected by endemic pemphigus foliaceus in El Bagre, Colombia, South America.

BACKGROUND: Traditionally, the intervertebral disks' (IVD) nucleus pulposus (NP) and annulus fibrosus (AF) are considered to have few cellular components and cell junctions. Patients affected by a new variant of endemic pemphigus foliaceus in El Bagre, Colombia, experience back pain in the spinal areas of the lower and upper back. Here, we investigate the reactivity of the patient's autoantibodies to structures in and around the IVDs at the cellular level. METHODS: We first administered a questionnaire and performed a medical examination. We then tested for autoreactivity against IVDs by indirect immunofluorescence, confocal microscopy, and reflectance confocal microscopy using bovine and human tissues as antigen sources. We tested 45 sera from patients affected by the disease and 45 control sera from the endemic area matched by age, gender, demographics, and work activity. RESULTS: Most of the patient sera revealed polyclonal antibodies against newly discovered cell junctions in the NP and AF, including their translamellar cross-bridges. Additional reactivities were detected against cell junctions in the spinal cord neurons, paraspinal nerves, blood vessels, anterior and posterior longitudinal ligaments, and paraspinal skeletal muscles. The reactivities of the patient's autoantibodies co-localized with those of commercially available antibodies to desmoplakins I-II, armadillo repeat gene deleted in velo-cardio-facial syndrome, plakophilin-4, and myocardium-enriched zonula occludens-1-associated protein (p < 0.001). CONCLUSIONS: We discovered novel complex cell junctions in the IVDs using patients' autoantibodies. These discoveries open a new chapter in the knowledge of IVD, representing a breakthrough pertinent to many diseases.

Repair of annulus fibrosus defects using decellularized annulus fibrosus matrix/chitosan hybrid hydrogels.

Degenerative disc disease is the leading cause of lower back and leg pain, considerably impacting daily life and incurring substantial medical expenses for those affected. The development of annulus fibrosus tissue engineering offers hope for treating this condition. However, the current annulus fibrosus tissue engineering scaffolds fail to accurately mimic the natural biological environment of the annulus fibrosus, resulting in limited secretion of extracellular matrix produced by the seeded cells and poor biomechanical properties of the constructed biomimetic annulus fibrosus tissue. This inability to match the biomechanical performance of the natural annulus fibrosus hinders the successful treatment of annulus fibrosus defects. In this study, we fabricated decellularized annulus fibrosus matrix (DAFM)/chitosan hydrogel-1 (DAFM: Chitosan 6:2) and DAFM/chitosan hydrogel-2 (DAFM: Chitosan 4:4) by varying the ratio of DAFM to chitosan. Rat annulus fibrosus (AF)-derived stem cells were cultured on these hydrogel scaffolds, and the cell morphology, AF-related gene expression, and Interleukin-6 (IL-6) levels were investigated. Additionally, magnetic resonance imaging, Hematoxylin and eosin staining, and Safranine and Fast Green staining were performed to evaluate the repair effect of the DAFM/chitosan hydrogels in vivo. The gene expression results showed that the expression of Collagen type I (Col-I), Collagen type I (Col-II), and aggrecan by annulus fibrosus stem cells (AFSCs) cultured on the DAFM/chitosan-1 hydrogel was higher compared with the DAFM/chitosan-2 hydrogel. Conversely, the expression of metalloproteinase-9 (MMP-9) and IL-6 was lower on the DAFM/chitosan-1 hydrogel compared with the DAFM/chitosan-2 hydrogel. In vivo, both the DAFM/chitosan-1 and DAFM/chitosan-2 hydrogels could partially repair large defects of the annulus fibrosus in rat tail vertebrae. In conclusion, the DAFM/chitosan-1 hydrogel could be regarded as a candidate scaffold material for the repair of annulus fibrosus defects, offering the potential for improved treatment outcomes.

Role of intra-lamellar collagen and hyaluronan nanostructures in annulus fibrosus on lumbar spine biomechanics: insights from molecular mechanics-finite element-based multiscale analyses.

Annulus fibrosus' (AF) ability to transmit multi-directional spinal motion is contributed by a combination of chemical interactions among biomolecular constituents-collagen type I (COL-I), collagen type II (COL-II), and proteoglycans (aggrecan and hyaluronan)-and mechanical interactions at multiple length scales. However, the mechanistic role of such interactions on spinal motion is unclear. The present work employs a molecular mechanics-finite element (FE) multiscale approach to investigate the mechanistic role of molecular-scale collagen and hyaluronan nanostructures in AF, on spinal motion. For this, an FE model of the lumbar segment is developed wherein a multiscale model of AF collagen fiber, developed from COL-I, COL-II, and hyaluronan using the molecular dynamics-cohesive finite element multiscale method, is incorporated. Analyses show AF collagen fibers primarily contribute to axial rotation (AR) motion, owing to angle-ply orientation. Maximum fiber strain values of 2.45% in AR, observed at the outer annulus, are 25% lower than the reported values. This indicates native collagen fibers are softer, attributed to the softer non-fibrillar matrix and higher interfibrillar sliding. Additionally, elastic zone stiffness of 8.61 Nm/° is observed to be 20% higher than the reported range, suggesting native AF lamellae exhibit lower stiffness, resulting from inter-collagen fiber bundle sliding. The presented study has further implications towards the hierarchy-driven designing of AF-substitute materials.

A novel technique for posterior lumbar interbody fusion to obtain a good local lordosis angle: anterior-release posterior lumbar interbody fusion.

Herein, we describe a novel posterior lumbar interbody fusion (PLIF) technique with annulus fibrosus (AF) release and the use of expandable cages (called "anterior-release PLIF" [ARPLIF]). In this technique, posterior column osteotomy (PCO) and AF release provide excellent intervertebral mobility. AF release involves circumferentially peeling off the AF above or below the endplate between the fixed vertebrae under radiographic guidance without cutting the AF and anterior longitudinal ligament. Subsequently, high-angle variable-angle expandable cages are used to simultaneously expand both sides before inserting the percutaneous pedicle screws and correcting to achieve good local lumbar lordosis. PCO and AF release achieve excellent intervertebral mobility. Intervertebral mobility and simultaneous expansion of both cages disperse the force on the endplates, reducing cage subsidence, and the high-angle cages facilitate high intervertebral angle creation. The novel ARPLIF intervertebral manipulation technique can promote good local lumbar lordosis formation.

Collagen integrity of the annulus fibrosus in degenerative disc disease individuals quantified with collagen hybridizing peptide.

Introduction: Degenerative disc disease (DDD) is accompanied by structural changes in the intervertebral discs (IVD). Extra-cellular matrix degradation of the annulus fibrosus (AF) has been linked with degeneration of the IVD. Collagen is a vital component of the IVD. Collagen hybridizing peptide (CHP) is an engineered protein that binds to degraded collagen, which we used to quantify collagen damage in AF. This method was used to compare AF samples obtained from donors with no DDD to AF samples from patients undergoing surgery for symptomatic DDD. Methods: Fresh AF tissue was embedded in an optimal cutting temperature compound and cryosectioned at a thickness of 8 µm. Hematoxylin and Eosin staining was performed on sections for general histomorphological assessment. Serial sections were stained with Cy3-conjugated CHP and the mean fluorescence intensity and areal fraction of Cy3-positive staining were averaged for three regions of interest (ROI) on each CHP-stained section. Results: Increases in mean fluorescence intensity (p = 0.0004) and percentage of positively stained area (p = 0.00008) with CHP were detected in DDD samples compared to the non-DDD samples. Significant correlations were observed between mean fluorescence intensity and percentage of positively stained area for both non-DDD (R = 0.98, p = 5E-8) and DDD (R = 0.79, p = 0.0012) samples. No significant differences were detected between sex and the lumbar disc level subgroups of the non-DDD and DDD groups. Only tissue pathology (non-DDD versus DDD) influenced the measured parameters. No three-way interactions between tissue pathology, sex, and lumbar disc level were observed. Discussion and Conclusions: These findings suggest that AF collagen degradation is greater in DDD samples compared to non-DDD samples, as evidenced by the increased CHP staining. Strong positive correlations between the two measured parameters suggest that when collagen degradation occurs, it is detected by this technique and is widespread throughout the tissue. This study provides new insights into the structural alterations associated with collagen degradation in the AF that occur during DDD.

The Role of Microvascular Variations in the Process of Intervertebral Disk Degeneration and Its Regulatory Mechanisms: A Literature Review.

Microvascular changes are considered key factors in the process of intervertebral disk degeneration (IDD). Microvascular invasion and growth into the nucleus pulposus (NP) and cartilaginous endplates are unfavorable factors that trigger IDD. In contrast, the rich distribution of microvessels in the bony endplates and outer layers of the annulus fibrosus is an important safeguard for the nutrient supply and metabolism of the intervertebral disk (IVD). In particular, the adequate supply of microvessels in the bony endplates is the main source of the nutritional supply for the entire IVD. Microvessels can affect the progression of IDD through a variety of pathways. Many studies have explored the effects of microvessel alterations in the NP, annulus fibrosus, cartilaginous endplates, and bony endplates on the local microenvironment through inflammation, apoptosis, and senescence. Studies also elucidated the important roles of microvessel alterations in the process of IDD, as well as conducted in-depth explorations of cytokines and biologics that can inhibit or promote the ingrowth of microvessels. Therefore, the present manuscript reviews the published literature on the effects of microvascular changes on IVD to summarize the roles of microvessels in IVD and elaborate on the mechanisms of action that promote or inhibit de novo microvessel formation in IVD.

A multifunctional scaffold that promotes the scaffold-tissue interface integration and rescues the ROS microenvironment for repair of annulus fibrosus defects.

Due to the limited self-repair ability of the annulus fibrosus (AF), current tissue engineering strategies tend to use structurally biomimetic scaffolds for AF defect repair. However, the poor integration between implanted scaffolds and tissue severely affects their therapeutic effects. To solve this issue, we prepared a multifunctional scaffold containing loaded lysyl oxidase (LOX) plasmid DNA exosomes and manganese dioxide nanoparticles (MnO2 NPs). LOX facilitates extracellular matrix (ECM) cross-linking, while MnO2 NPs inhibit excessive reactive oxygen species (ROS)-induced ECM degradation at the injury site, enhancing the crosslinking effect of LOX. Our results revealed that this multifunctional scaffold significantly facilitated the integration between the scaffold and AF tissue. Cells were able to migrate into the scaffold, indicating that the scaffold was not encapsulated as a foreign body by fibrous tissue. The functional scaffold was closely integrated with the tissue, effectively enhancing the mechanical properties, and preventing vascular invasion, which emphasized the importance of scaffold-tissue integration in AF repair.

Composite Hydrogel Sealants for Annulus Fibrosus Repair.

Intervertebral disc (IVD) herniation is a leading cause of disability and lower back pain, causing enormous socioeconomic burdens. The standard of care for disc herniation is nucleotomy, which alleviates pain but does not repair the annulus fibrosus (AF) defect nor recover the biomechanical function of the disc. Existing bioadhesives for AF repair are limited by insufficient adhesion and significant mechanical and geometrical mismatch with the AF tissue, resulting in the recurrence of protrusion or detachment of bioadhesives. Here, we report a composite hydrogel sealant constructed from a composite of a three-dimensional (3D)-printed thermoplastic polyurethane (TPU) mesh and tough hydrogel. We tailored the fiber angle and volume fraction of the TPU mesh design to match the angle-ply structure and mechanical properties of native AF. Also, we proposed and tested three types of geometrical design of the composite hydrogel sealant to match the defect shape and size. Our results show that the sealant could mimic native AF in terms of the elastic modulus, flexural modulus, and fracture toughness and form strong adhesion with the human AF tissue. The bovine IVD tests show the effectiveness of the composite hydrogel sealant for AF repair and biomechanics recovery and for preventing herniation with its heightened stiffness and superior adhesion. By harnessing the combined capabilities of 3D printing and bioadhesives, these composite hydrogel sealants demonstrate promising potential for diverse applications in tissue repair and regeneration.

Innovative hydrogel-patch combination for large annulus fibrosus defects: a prospective approach to address herniation recurrence.

BACKGROUND CONTEXT: Large annulus fibrosus (AF) defects often lead to a high rate of reherniation, particularly in the medial AF region, which has limited self-healing capabilities. The increasing prevalence of herniated discs underscores the need for effective repair strategies. PURPOSE: The objectives of this study were to design an AF repair technique to reduce solve the current problems of insufficient mechanical properties and poor sealing capacity. STUDY DESIGN: In vitro biomechanical experiments and finite element analysis. METHODS: The materials used in this study were patches and hydrogels with good biocompatibility and sufficient mechanical properties to withstand loading in the lumbar spine. Five repair techniques were assessed in this study: hydrogel filler (HF), AF patch medial barrier (MB), AF patch medial barrier and hydrogel filler (MB&HF), AF patch medial-lateral barrier (MLB), and AF patch medial-lateral barrier and hydrogel filler (MLB&HF). The repair techniques were subjected to in vitro testing (400 N axial compression and 0-500 N fatigue loading at 5Hz) and finite element analysis (400 N axial compression) to evaluate the effectiveness at repairing large AF defects. The evaluation included repair tightness, spinal stability, and fatigue resistance. RESULTS: From the in vitro testing, the failure load of the repair techniques was in the following order HF MLB >MB&HF >MLB&HF. CONCLUSIONS: The combined use of patches and hydrogels exhibited promising mechanical properties postdiscectomy, providing a promising solution for addressing large AF defects and improving disc stability. CLINICAL SIGNIFICANCE: This study introduces a promising method for repairing large annular fissure (AF) defects after disc herniation, combining patch repair with a hydrogel filler. These techniques hold potential for developing clinical AF repair products to address this challenging issue.

Biomechanics of annulus fibrosus: Elastic fiber simplification and degenerative impact on damage initiation and propagation.

This study addresses three primary objectives related to lumbar intervertebral disc (IVD) biomechanics under ramping quasi-static loading conditions. First, we explore the conditions justifying the simplification of axisymmetric elastic fiber families into single fiber bundles through discretized strain energy functions. Simulations reveal that a concentration factor exceeding 10 allows for a consistent deviation below 10% between simplified and non-simplified responses. Second, we investigate the impact of elastic fibers on the physiological stiffness in IVDs, revealing minimal influence on biological motions but significant effects on degeneration. Lastly, we examine the initiation and progression of annulus fibrosus (AF) damage. Our findings confirm the validity of simplifying elastic fiber families and underscore the necessity of considering elastic fiber damage in biomechanical studies of AF tissues. Elastic fibers contribute to increased biaxial stretch stiffness, and their damage significantly affects the loading capacity of the inner AF. Additionally, degeneration significantly alters the susceptibility to damage in the AF, with specific regions exhibiting higher vulnerability. Damage tends to extend circumferentially and radially, emphasizing the regional variations in collagen and elastic fiber properties. This study offers useful insights for refining biomechanical models, paving the way for a more comprehensive understanding of IVD responses and potential clinical implications.