Evidence of spinal stiffening following fusionless bipolar fixation for neuromuscular scoliosis: a shear wave elastography assessment of lumbar annulus fibrosus.

OBJECTIVES: There are no established criteria for stiffness after fusionless surgery for neuromuscular scoliosis (NMS). As a result, there is no consensus regarding the surgical strategy to propose at long-term follow-up. This study reports the first use of shear wave elastography for assessing the mechanical response of lumbar intervertebral discs (IVDs) after fusionless bipolar fixation (FBF) for NMS and compares them with healthy controls. The aim was to acquire evidence from the stiffness of the spine following FBF. PATIENTS AND METHODS: Nineteen NMS operated on with FBF (18 ± 2y at last follow-up, 6 ± 1 y after surgery) were included prospectively. Preoperative Cobb was 89 ± 20° and 35 ± 1° at latest follow-up. All patients had reached skeletal maturity. Eighteen healthy patients (20 ± 4 y) were also included. Shear wave speed (SWS) was measured in the annulus fibrosus of L3L4, L4L5 and L5S1 IVDs and compared between the two groups. A measurement reliability was performed. RESULTS: In healthy subjects, average SWS (all disc levels pooled) was 7.5 ± 2.6 m/s. In NMS patients, SWS was significantly higher at 9.9 ± 1.4 m/s (p < 0.05). Differences were significant between L3L4 (9.3 ± 1.8 m/s vs. 7.0 ± 2.5 m/s, p = 0.004) and L4L5 (10.3 ± 2.3 m/s vs. 7.1 ± 1.1 m/s, p = 0.0006). No difference was observed for L5S1 (p = 0.2). No correlation was found with age at surgery, Cobb angle correction and age at the SWE measurement. CONCLUSIONS: This study shows a significant increase in disc stiffness at the end of growth for NMS patients treated by FBF. These findings are a useful adjunct to CT-scan in assessing stiffness of the spine allowing the avoidance of surgical final fusion at skeletal maturity.

In-vivo characterization of the lumbar annulus fibrosus in adults with ultrasonography and shear wave elastography.

In vivo characterization of intervertebral disc (IVD) mechanical properties and microstructure could give an insight into the onset and progression of disc pathologies. Ultrasound shearwave elastography provided promising results in children, but feasibility in adult lumbar discs, which are deep in the abdomen, was never proved. The aim of this work was to determine the feasibility and reliability of ultrasound assessment of lumbar IVD in adults. Thirty asymptomatic adults were included (22 to 67 years old). Subjects were lying supine, and the annulus fibrosus of the L3-L4 IVD was imaged by conventional ultrasonography and shearwave elastography. Shear wave speed (SWS) and lamellar thickness were measured. Reliability was determined through repeated measurements acquired by three operators. Average SWS in AF at the L3L4 level was 4.0 ± 0.9 m/s, with an inter-operator uncertainty of 8.7%, while lamellar thickness was 255 ± 27 µm with an uncertainty of 9.6%. Measurement was not feasible in one out of four subjects with BMI > 24 kg/m² (overweight). Ultrasound assessment of annulus fibrosus revealed feasible, within certain limitations, and reproducible. This method gives an insight into disc microstructure and mechanical properties, and it could be applied for the early detection or follow-up of disc pathologies.

A Minimally Invasive Annulus Fibrosus Needle Puncture Model of Intervertebral Disc Degeneration in Rats.

BACKGROUND: The needle puncture model in rats has been accepted as an ordinary model to induce intervertebral disc degeneration (IVDD). However, the model primarily penetrated the whole intervertebral disc, resulting in injury to the nucleus pulposus (NP) and annulus fibrosus (AF). The intention of this research was to explore a minimally invasive approach through needle puncture of the AF percutaneously in rats. METHODS: Twenty SD rats underwent puncture at Co8/9 via a 20 G percutaneous needle. The needle was slowly advanced perpendicular to the tail skin to penetrate the whole AF without damaging the NP limited by a hand-made sheath. The X-rays and magnetic resonance imaging T2 relaxation was evaluated at 1, 2, and 3 weeks to assess the disc height index and signal changes. Histological and immunohistochemical staining of the IVD were obtained under a light microscope. RESULTS: X-rays showed that the disc height had progressively narrowed to 49% of baseline 3 weeks after injury. magnetic resonance imaging evaluation demonstrated that the mean T2-weighted signal intensity at 3 weeks was 43% of that in the uninjured control group at the Co8/9 level. Histological staining demonstrated disorganized lamellae in the AF and decreased proteoglycan content and cellularity within the NP in the injured discs. CONCLUSIONS: The present research demonstrates a reliable and convenient approach to induce an AF tear in rats through percutaneous needle puncture. This model reduces harm to the experimental animals significantly while imitating the progressive degeneration process. More importantly, the model confirmed that AF damage alone could lead to IVDD and provided a research method for AF degeneration in IVDD.

Contrast-enhanced microCT evaluation of degeneration following partial and full width injuries to the mouse lumbar intervertebral disc.

A targeted injury to the mouse intervertebral disc (IVD) is often used to recapitulate the degenerative cascade of the human pathology. Since injuries can vary in magnitude and localization, it is critical to examine the effects of different injuries on IVD degeneration. We thus evaluated the degenerative progression resulting from either a partial- or full-width injury to the mouse lumbar IVD using contrast-enhanced micro-computed tomography and histological analyses. A lateral-retroperitoneal surgical approach was used to access the lumbar IVD, and the injuries to the IVD were produced by either incising one side of the annulus fibrosus or puncturing both sides of the annulus fibrosus. Female C57BL/6J mice of 3-4 months age were used in this study. They were divided into three groups to undergo partial-width, full-width, or sham injuries. The L5/6 and L6/S1 lumbar IVDs were surgically exposed, and then the L6/S1 IVDs were injured using either a surgical scalpel (partial-width) or a 33G needle (full-width), with the L5/6 serving as an internal control. These animals recovered and then euthanized at either 2-, 4-, or 8-weeks after surgery for evaluation. The IVDs were assessed for degeneration using contrast-enhanced microCT (CEµCT) and histological analysis. The high-resolution 3D CEµCT evaluation of the IVD confirmed that the respective injuries were localized within one side of the annulus fibrosus or spanned the full width of the IVD. The full-width injury caused significant deteriorations in the nucleus pulposus, annulus fibrous and at the interfaces after 2 weeks, which was sustained through the 8 weeks, while the partial width injury caused localized disruptions that remained limited to the annulus fibrosus. The use of CEµCT revealed distinct IVD degeneration profiles resulting from partial- and full-width injuries. The partial width injury may serve as an alternative model for IVD degeneration resulting from localized annulus fibrosus injuries.

Delineation of Inner Annulus Fibrosus and Nucleus Pulposus on Routine T2-weighted MR Images.

STUDY DESIGN: Retrospective study of 150 IVDs. OBJECTIVE: Assessment of costume algorithm ability to delineate the IAF and NP on routine T2 images. SUMMARY OF BACKGROUND DATA: Central hyperintense region on T2-weighted MR images of normal lumbar IVDs represents a combination of IAF and NP. Ability to identify NP as distinct from IAF can help improve our understanding of IVD morphology in-vivo. METHODS: Sagittal T2-weighted TSE MR images of 150 lumbar IVDs from 25 patients were analyzed. MR images were processed using a custom algorithm that markedly increased the signal intensity of structures with inherent signal intensity within 2 defined intensity thresholds. Signal intensity and contrast-to- noise ratio between outer annulus fibrosus, IAF, and NP were assessed at baseline and after processing. To assess consistency of underlying T2 differences, similar analysis was done on 108 discs from 18 patients in whom additional sagittal T2-weighted STIR images were available. RESULTS: Following image processing, apparent IAF and NP were rendered visible in 86% and 84.3% IVDs on T2-weighted TSE and STIR images respectively. While signal intensity of these 2 regions was inherently different (P< 0.001) before processing on TSE and STIR images, their visualization was facilitated by a significant increase (P<0.001) in contrast-to-noise ratio after processing. Nonvisualization of NP was associated with disc degeneration (P<0.001). CONCLUSION: Inherent differences exist in signal intensities of normal NP and IAF on T2-weighted MR images. Accentuating these differences using image postprocessing techniques can render these 2 structures visible.

Regenerating and repairing degenerative intervertebral discs by regulating the micro/nano environment of degenerative bony endplates based on low-tension mechanics.

BACKGROUND: Conservative treatment is the recommended first-line treatment for degenerative disc diseases. Traction therapy has historically been one of the most common clinical methods to address this, but the clinical effect remains controversial. METHODS: Forty-two six-month-old male Sprague-Dawley rats were randomly divided into six groups: the model group (Group A, four coccyx vertebrae (Co7-Co10) were fixed with customized external fixators, and the vertebral disc degeneration model was constructed by axial compression of the target segment Co8 - Co9 for 4 weeks), the experimental control group (Group B, after successful modeling, the external fixation device was removed and self-rehabilitation was performed) and four intervention groups (Groups C to F): Groups C and E: Co8 - Co9 vertebrae compressed for 4 weeks followed by two or 4 weeks of high tension traction (HTT), respectively, and Groups D and F: vertebrae compressed for 4 weeks followed by two or 4 weeks of low-tension traction (LTT), respectively. Imaging tests (X-ray and MRI) were performed to assess disc height and T2 signal intensity at each time point. After the experiment, the animals were euthanized, and the caudal vertebrae were collected for analysis of intervertebral disc histopathology, proteoglycan content, and micronanostructure of the annulus fibrosus, nucleus pulposus and bony endplate. RESULTS: Signs of tissue regeneration were apparent in all four intervention groups. After two to 4 weeks of intervention (HTT and LTT), the morphology of pores in the bony endplate, their number, and diameter had recovered significantly compared with those in Group A. The LTT group was superior to the HTT group, and the 4w in situ group was significantly superior to the 2w group. Meanwhile, the histological scores of discs, the mean fibril diameter and modulus of annulus fibrosus were significantly improved compared with the control groups, and the LTT group was superior to HTT group. CONCLUSIONS: Low-tension traction better promotes active reconstruction of bony endplates and improves the elastic modulus and micro/nanostructure of the disc. Thus, it further promotes the regeneration and repair of intervertebral discs.

Selective Retention of Bone Marrow Stromal Cells with Gelatin Sponge for Repair of Intervertebral Disc Defects after Microendoscopic Discectomy: A Prospective Controlled Study and 2-Year Follow-Up.

OBJECTIVE: Discectomy remains the classic procedure for treating lumbar intervertebral disc (IVD) herniation, but the occurrence of defects after discectomy is thought to be an important cause generating recurrent and accelerated IVD degeneration. Previous studies attempted suture of the annulus fissure, but the validity of this technique on restraining the degenerative process is controversial. On the other hand, cell therapies have been shown in multiple clinical and basic studies. Our purpose was to investigate the effectiveness of selective retention of autologous Bone Marrow Stromal Cells (BMSCs) with gelatin sponge in combination with annulus fibrosus suture (AFS) for the repair of IVD defects following mobile microendoscopic discectomy (MMED). METHODS: This prospective, two-armed, and controlled clinical study was conducted from December 2016 to December 2018. Written informed consent was obtained from each patient. Forty-five patients with typical symptoms, positive signs of radiculopathy, and obvious lumbar disc herniation observed by MRI were enrolled. Patients were divided into 3 groups with different treating methods: MMED (n = 15), MMED+AFS (n = 15), and MMED+AFS+BMSCs (n = 15). A postoperative 2-year follow-up was performed to evaluate the patient-reported outcomes of VAS, ODI, and SF-36. The improvement rate of VAS and ODI was calculated as [(latest-preoperative)/preoperative] to evaluate the therapeutic effect of the three groups. Assessment parameters included Pfirrmann grade, intervertebral disc height (IDH), and disc protrusion size (DPS), as measured by MRI to evaluate the morphological changes. RESULTS: All patients enrolled had a postoperative follow-up at 3, 6, 12, and 24 months. VAS and ODI scores were significantly improved compared to the preoperative status in all three groups with a mean DPS reduction rate over 50%. At the final follow-up, the improvement rate of the VAS score in the MMED+AFS+BMSCs group was significantly higher than the MMED+AFS and MMED groups (80.1% ± 7.6% vs. 71.3% ± 7.0% vs. 70.1% ± 7.8%), while ODI improvement showed a significant change (65.6% ± 8.8% vs. 59.9% ± 5.5% vs. 57.8% ± 8.1%). All participants showed significant improvement in SF-36 PCS and MCS; the differences between each group were not significant. The mean IDH loss rate of the MMED+AFS+BMSCs group was also significantly lower than other groups (-17.2% ± 1.3% vs. -27.6% ± 0.7% vs. -29.3% ± 2.2%). The Pfirrmann grade was aggravated in the MMED and MMED+AFS groups while maintained at the preoperative grade in the MMED+AFS+BMSCs group. No adverse events of cell transplantation or recurrence were found in all patients during the postoperative follow-up period. CONCLUSIONS: It is feasible and effective to repair lumbar IVD defects using SCR-enriched BMSCs with gelatin sponges, which warrants further study and development as a cell-based therapy for IVD repair.

A microstructure-based model for a full lamellar-interlamellar displacement and shear strain mapping inside human intervertebral disc core.

The determinant role of the annulus fibrosus interlamellar zones in the intervertebral disc transversal and volumetric responses and hence on their corresponding three-dimensional conducts have been only revealed and appreciated recently. Their consideration in disc modeling strategies has been proven to be essential for the reproduction of correct local strain and displacement fields inside the disc especially in the unconstrained directions of the disc. In addition, these zones are known to be the starting areas of annulus fibrosus circumferential tears and disc delamination failure mode, which is often judged as one of the most dangerous disc failure modes that could evolve with time leading to disc hernia. For this latter reason, the main goal of the current contribution is to incorporate physically for the first time, the interlamellar zones, at the scale of a complete human lumbar intervertebral disc, in order to allow a correct local vision and replication of the different lamellar-interlamellar interactions and an identification of the interlamellar critical zones. By means of a fully tridimensional chemo-viscoelastic constitutive model, which we implemented into a finite element code, the physical, mechanical and chemical contribution of the interlamellar zones is added to the disc. The chemical-induced volumetric response is accounted by the model for both the interlamellar zones and the lamellae using experimentally-based fluid kinetics. Computational simulations are performed and critically discussed upon different simple and complex physiological movements. The disc core and the interlamellar zones are numerically accessed, allowing the observation of the displacement and shear strain fields that are compared to direct MRI experiments from the literature. Important conclusions about the correct lamellar-interlamellar-nucleus interactions are provided thanks to the developed model. The critical interlamellar spots with the highest delamination potentials are defined, analyzed and related to the local kinetics and microstructure.

Direct Quantification of Intervertebral Disc Water Content Using MRI.

BACKGROUND: Water content is a key parameter for simulating tissue swelling and nutrient diffusion. Accurately measuring water content throughout the intervertebral disc (NP = nucleus pulposus; AF = annulus fibrosus) is important for developing patient-specific models. Water content is measured using destructive techniques, Quantitative MRI has been used to estimate water content and detect early degeneration, but it is dependent on scan parameters, concentration of free water molecules, and fiber architecture. PURPOSE: To directly measure disc-tissue water content using quantitative MRI and compare MRI-based measurements with biochemical assays, and to quantify changes in disc geometry due to compression. STUDY TYPE: Basic science, controlled. SPECIMEN: Twenty bone-disc-bone motion segments from skeletally mature bovines. FIELD STRENGTH/SEQUENCE: 7T/3D fast low angle shot (FLASH) pulse sequence and a T2 rapid imaging with refocused echoes (RARE) sequence. ASSESSMENT: Disc volumes, NP and AF volumetric water content, and T2 relaxation times were measured through MRI; NP and AF tissue gravimetric water content, mass density, and glycosaminoglycan content were measured through a biochemical assay. STATISTICAL TESTS: Correlations between MRI-based measurement and biochemical composition were evaluated using Pearson's linear regression. RESULTS: Mechanical dehydration resulted in a decrease in disc volume by up to 20% and a decrease in disc height by up to 35%. Direct water content measurements for the NP was achieved by normalizing MRI-based spin density by NP mass density (1.10 ± 0.03 g/cm3 ). However, the same approach underestimated water content in the AF by ~10%, which may be due to a higher concentration of collagen fibers and bound water molecules. DATA CONCLUSION: Spin density or spin density normalized by mass density to estimate NP and AF water content was more accurate than correlations between water content and relaxation times. Mechanical dehydration decreased disc volume and disc height, and increased maximum cross-sectional area. LEVEL OF EVIDENCE: TECHNICAL EFFICACY STAGE: J. Magn. Reson. Imaging 2020;52:1152-1162.

The application of key feature extraction algorithm based on Gabor wavelet transformation in the diagnosis of lumbar intervertebral disc degenerative changes.

OBJECTIVE: Based on the theoretical basis of Gabor wavelet transformation, the application effects of feature extraction algorithm in Magnetic Resonance Imaging (MRI) and the role of feature extraction algorithm in the diagnosis of lumbar vertebra degenerative diseases were explored. METHOD: The structure of lumbar vertebra and degenerative changes were respectively introduced to clarify the onset mechanism and pathological changes of lumbar vertebra degenerative changes. Most importantly, the theoretical basis of Gabor wavelet transformation and the extraction effect of feature information in lumbar vertebra MRI images were introduced. The differentiation effects of feature information extraction algorithm on annulus fibrosus and nucleus pulposus were analyzed. In this study, the data of lumbar spine MRI was randomly selected from the Wenzhou Lumbar Spine Research Database as research objects. A total of 130 discs were successfully fitted, and 109 images were graded by a doctor after observation, which was compared with the results of the artificial diagnosis. Through the comparison with the results of observation and diagnosis by professional doctors, the accuracy of feature extraction algorithm based on Gabor wavelet transformation in the diagnosis of lumbar vertebra degenerative changes was analyzed. RESULTS: 1. Compared with the results of the manual diagnosis, the accuracy of the classification method was 88.3%. In addition, the specificity (SPE), accuracy (ACC), and sensitivity (SEN) of the classification method were respectively 89.5%, 92.4%, and 87.6%. 2. The mutual information method and the KLT algorithm were utilized for vertebral body tracking. The maximum mutual information method was more effective in the case of fewer image sequences; however, with the increase of image frames, the accumulation of errors would make the tracking effects of images get worse. Based on the KLT algorithm, the enhanced vertebral boundary information was selected; the soft tissues showed in the obtained images were smooth, the boundary information of vertebral body was enhanced, and the results were more accurate. CONCLUSION: The feature extraction algorithm based on Gabor wavelet transformation could easily and quickly realize the localization of the lumbar intervertebral disc, and the accuracy of the results was ensured. In addition, from the aspect of vertebral body tracking, the tracking effects based on the KLT algorithm were better and faster than those based on the maximum mutual information method.

Development, validation, and application of ligamentous cervical spinal segment C6-C7 of a six-year-old child and an adult.

BACKGROUND AND OBJECTIVE: The cervical spine is one of the primary regions that is easily injured in traffic accidents. Although adult cervical spine finite element models have been widely adopted to investigate the cervical injury, few efforts have been made with respect to the development and application of FE models of the pediatric cervical spine, especially that of a six-year-old child. The objective of this study is to develop and validate high quality cervical spinal segment C6-C7 FE models of a six-year-old child and an adult, and to further investigate the differences of C6-C7 between the child and adult under different loading conditions. METHODS: The cervical spinal segment C6-C7 FE models were developed by a structured multiblock method, and were verified under flexion, extension, axial rotation, and lateral bending conditions. The validated models were used to investigate the differences of C6-C7 between the child and adult under different loading conditions. RESULTS: The global angular displacement of C6-C7, the ligament elongation ratio, and the maximum effective strain of annulus fibrosus of the child were obviously larger than those of the adult under the same loading conditions. Regarding the loading forms, the flexion angular displacement of C6-C7 of the child was obviously larger than those of the extension and lateral bending, while for the adult cervical segment C6-C7, no obvious differences existed. The elongation ratio of different ligaments was highly dependent on the types of loadings. The maximum effective strain of annulus fibrosus under flexion, extension and lateral bending loads occurred at the compressive region of the front, rear, and one compressive lateral side, in which the annulus fibrosus was more susceptible to injury under the lateral bending condition, compared with those of the flexion and extension conditions. CONCLUSIONS: Both the developed child and adult cervical spinal segment C6-C7 FE models exhibited high biofidelity. The responses (angular displacement, the ligament elongation ratio, and the maximum effective strain of annulus fibrosus) of the child and adult are dependent on the loading types, and the responses of the child were obviously larger than those of the adult under the same loading conditions.

Longitudinal characterization of intervertebral disc remodeling following acute annular injury in a rat model of degenerative disc disease.

Objective: Characterize 3D remodeling of the rat intervertebral disc (IVD) following acute annular injury via in vivo micro-computed tomography (µCT), ex vivo contrast-enhanced (CE)-µCT, and histology. Design: Female Lewis rats (N = 4/group) underwent either sham surgery or anterior annular puncture to L3-L4 and L5-L6 (n = 8 IVDs/group) to induce IVD degeneration. Rats were allowed ad libidum cage activity before and after surgery and underwent in vivo µCT scanning at baseline and every 2 weeks post-op for 12 weeks to characterize longitudinal changes in IVD height. At 12 weeks, lumbar spines were dissected and underwent CE-µCT scanning to characterize endpoint glycosaminoglycan distribution and nucleus pulposus (NP) volume ratio. Spines were processed for safranin-O-stained sagittal histology, and IVD degeneration was graded via the Rutges scale. Results: Puncture IVDs exhibited loss of IVD height at all time points from 4 weeks onward compared to Sham-the most severe height loss occurred posteriorly, with significant changes also occurring in the NP and laterally. Puncture IVDs exhibited higher CE-µCT attenuation, indicative of lower glycosaminoglycan content, and reduced NP volume ratio compared to Sham. Histologically, Puncture IVDs had higher Rutges damage scores and exhibited reduced NP cellularity and hydration, disorganized annulus fibrosus (AF) lamellae with evidence of the stab tract, and indistinct AF-NP border compared to Sham. Conclusions: Characterization of the complex, 3D alterations involved in the onset and early progression of IVD degeneration can foster greater understanding of the pathoetiology of IVD degeneration and may inform future studies assessing more sensitive diagnostic techniques or novel therapies.

Shear wave elastography of lumbar annulus fibrosus in adolescent idiopathic scoliosis before and after surgical intervention.

OBJECTIVES: To determine lumbar intervertebral disc properties with shear wave elastography in adolescent idiopathic scoliotic (AIS) patients before and after surgery, and compare them with asymptomatic controls. METHODS: Twenty-five severe AIS patients with an indication for fusion surgery (15 ± 1.5 years old, the Cobb angle ranging between 40 and 93°) and fifty-nine asymptomatic adolescents (13 ± 2 years old) were included prospectively. Shear wave speed (SWS) was measured in the annulus fibrosus of L3-L4, L4-L5, and L5-S1 discs of each subject. In AIS patients, measurements took place before surgery, and 3 months (N = 13) or 1 year after (N = 12). RESULTS: No difference was observed between disc levels in any group. When pooling disc levels, SWS was significantly higher in preop AIS patients (4.0 ± 0.5 m/s) than in asymptomatic subjects (3.1 ± 0.5 m/s, p < 0.0001). SWS decreased 3 months postop (3.5 ± 0.3 m/s), and it decreased further towards normal values 1 year after (3.3 ± 0.4 m/s). SWS in preop AIS patients tended to decrease with the Cobb angle (Spearman's rho = - 0.4, p = 0.05). CONCLUSION: Shear wave elastography measurements showed that discs in AIS patients were altered relative to asymptomatic ones, and this alteration tended to normalize 1 year post fusion surgery. Further studies should aim at determining if bracing of mild scoliosis has an effect on disc properties. KEY POINTS: • Shear wave elastography shows alteration of annulus fibrosus in adolescent idiopathic scoliosis. • Disc elastography tends to normalize 1 year after surgery.

Aberrant mechanosensing in injured intervertebral discs as a result of boundary-constraint disruption and residual-strain loss.

In fibrous tissues, prestressed boundary constraints at bone interfaces instil residual strain throughout the tissue, even when unloaded. For example, internal swelling pressures in the central nucleus pulposus of the intervertebral disc generate prestrain in the outer annulus fibrosus. With injury and depressurization, these residual strains are lost. Here we show that the loss of residual strains in the intervertebral disc alters the microenvironment and instigates aberrant tissue remodelling and the adoption of atypical cellular phenotypes. By using puncture surgery of the annulus fibrosus in rabbits, ex vivo puncture experiments and electrospun nanofibrous scaffolds recapitulating these evolving boundary constraints, we show that the loss of residual strain promotes short-term apoptosis and the emergence of a fibrotic phenotype. We also show that local fibre organization and cellular contractility mediate this process and that the aberrant cellular changes could be abrogated by targeting the cell-mechanosensing machinery with small molecules. Our findings indicate that injury to dense connective tissues under prestrain alters boundary constraints and residual strain; this leads to aberrant mechanosensing, which in turn promotes disease progression.

Differences in IVD characteristics between low back pain patients and controls associated with HIZ as revealed with quantitative MRI.

BACKGROUND: Magnetic resonance imaging (MRI) can provide objective continuous intervertebral disc (IVD) measures in low back pain (LBP) patients. However, there are limited studies comparing quantitative IVD measures of symptomatic and asymptomatic individuals. PURPOSE: This study aimed to investigate possible differences in IVD tissue composition in patients with chronic LBP and controls using quantitative MRI and correlate IVD measures with the phenotype High-Intensity Zone (HIZ). METHODS: The lumbar spine of 25 LBP-patients (25-69y, mean 38y, 11 males) and 12 controls (25-59y, mean 38y, 7 males) was examined with T2-mapping on a 1.5T MRI scanner. The mean T2-map value and standard deviation were determined in three midsagittal IVD slices and five sub-regions dividing each IVD in the sagittal plane. The distribution of T2-map values over the IVD was also determined with histogram analysis (Δµ = distribution width). RESULTS: When compared to controls, patient IVDs displayed lower values for all metrics, with significant differences for the T2-map value, standard deviation (p = 0.026) and Δµ (p = 0.048). Significantly different T2-map values were found between cohorts in the region representing nucleus pulposus and the border zone between nucleus pulposus and posterior annulus fibrosus (p = 0.047-0.050). Excluding all IVDs with HIZs resulted in no significant difference between the cohorts for any of the analyzed metrics (p = 0.053-0.995). Additionally, the T2-map values were lower in patients with HIZ in comparison without HIZ. CONCLUSIONS: Differences in IVD characteristics, measured with quantitative MRI, between LBP patients and controls were found. The T2-differences may reflect altered IVD function associated with HIZ. Future studies are recommended to explore IVD functionality in relation to HIZ and LBP.

Characterization of the microstructure of the intervertebral disc in patients with chronic low back pain by diffusion kurtosis imaging.

PURPOSE: Multivariate analysis of T2-weighted signal, diffusion ADC, and DKI parameters and tractography were used to differentiate chronic non-specific low back pain (CLBP) patients and asymptomatic controls (AC). METHODS: A total of 30 patients with CLBP and 23 AC underwent diffusion kurtosis imaging (DKI) of lumbar spine with a 3T MRI scanner to get the ADC values and seven parameters of DKI in the nucleus pulposus (NP) of the intervertebral disc. The tractography and the tract-related parameters as other parameters were also generated to indicate the intactness of annulus fibrosus (AF). T2-grades of the discs were also quantified based on an eight-grade degeneration grading system. ADC and T2-grades were compared with DKI parameters for the differentiation of CLBP and AC groups. RESULTS: There was no difference in the T2 grades, ADC value, and multiple parameters in DKI of NP between CLBP and AC groups (P > 0.05). The average FA values in NP in AC group were found significantly higher than in the CLBP group (P < 0.05). The scores for the intactness of AF of the intervertebral discs were significantly different in CLBP and AC groups, with 90% of sensitivity and 70% specificity (P < 0.05). Additionally, there were significantly differences in the length and volume values of the AF in CLBP and AC groups (P < 0.05). CONCLUSION: DKI is a good noninvasive method, and it might help to differentiate CLBP from AC. Particularly, the continuation of DKI tractography reflects the presence of annulus fibrosus fissures, an important character in the generation of the low back pain. These slides can be retrieved under Electronic Supplementary Material.

Multiscale and multimodal structure-function analysis of intervertebral disc degeneration in a rabbit model.

OBJECTIVES: The objective of this study was to perform a quantitative analysis of the structural and functional alterations in the intervertebral disc during in vivo degeneration, using emerging tools that enable rigorous assessment from the microscale to the macroscale, as well as to correlate these outcomes with noninvasive, clinically relevant imaging parameters. DESIGN: Degeneration was induced in a rabbit model by puncturing the annulus fibrosus (AF) with a 16-gauge needle. 2, 4, 8, and 12 weeks following puncture, degenerative changes in the discs were evaluated via magnetic resonance imaging (MRI), whole motion segment biomechanics, atomic force microscopy, histology and polarized light microscopy, immunohistochemistry, biochemical content, and second harmonic generation imaging. RESULTS: Following puncture, degeneration was evident through marked changes in whole disc structure and mechanics. Puncture acutely compromised disc macro and microscale mechanics, followed by progressive stiffening and remodeling. Histological analysis showed substantial anterior fibrotic remodeling and osteophyte formation, as well as an overall reduction in disc height, and disorganization and infolding of the AF lamellae into the NP space. Increases in NP collagen content and aggrecan breakdown products were also noted within 4 weeks. On MRI, NP T2 was reduced at all post-puncture time points and correlated significantly with microscale indentation modulus. CONCLUSION: This study defined the time dependent changes in disc structure-function relationships during IVD degeneration in a rabbit annular injury model and correlated degeneration severity with clinical imaging parameters. Our findings identified AF infolding and occupancy of the space as a principle mechanism of disc degeneration in response to needle puncture, and provide new insights to direct the development of novel therapeutics.

Microstructural characterization of annulus fibrosus by ultrasonography: a feasibility study with an in vivo and in vitro approach.

The main function of the intervertebral disc is biomechanical function, since it must resist repetitive high loadings, while giving the spine its flexibility and protecting the spinal cord from over-straining. It partially owes its mechanical characteristics to the lamellar architecture of its outer layer, the annulus fibrosus. Today, no non-invasive means exist to characterize annulus lamellar structure in vivo. The aim of this work was to test the feasibility of imaging annulus fibrosus microstructure in vivo with ultrasonography. Twenty-nine healthy adolescents were included. Ultrasonographies of L3-L4 disc were acquired with a frontal approach. Annulus fibrosus was segmented in the images to measure the thickness of the lamellae. To validate lamellar appearance in ultrasonographies, multimodality images of two cow tail discs were compared: ultrasonography, magnetic resonance and optical microscopy. In vivo average lamellar thickness was 229.7 ± 91.5 µm, and it correlated with patient body mass index and age. Lamellar appearance in the three imaging modalities in vitro was consistent. Lamellar measurement uncertainty was 7%, with good agreement between two operators. Feasibility of ultrasonography for the analysis of lumbar annulus fibrosus structure was confirmed. Further work should aim at validating measurement reliability, and to assess the relevance of the method to characterize annulus alterations, for instance in disc degeneration or scoliosis.

In vitro and in vivo evaluation of an electrospun-aligned microfibrous implant for Annulus fibrosus repair.

Annulus fibrosus (AF) impairment is associated with reherniation, discogenic pain, and disc degeneration after surgical partial discectomy. Due to a limited intrinsic healing capacity, defects in the AF persist over time and it is hence necessary to adopt an appropriate strategy to close and repair the damaged AF. In this study, a cell-free biodegradable scaffold made of polycaprolactone (PCL), electrospun, aligned microfibers exhibited high levels of cell colonization, alignment, and AF-like extracellular matrix deposition when evaluated in an explant culture model. The biomimetic multilayer fibrous scaffold was then assessed in an ovine model of AF impairment. After 4 weeks, no dislocation of the implants was detected, and only one sample out of six showed a partial delamination. Histological and immunohistochemical analyses revealed integration of the implant with the surrounding tissue as well as homogeneously aligned collagen fiber organization within each lamella compared to the disorganized and scarcer fibrous tissue in a randomly organized control fibrous scaffold. In conclusion, this biomimetic electrospun implant exhibited promising properties in terms of AF defect closure, with AF-like neotissue formation that fully integrated with the surrounding ovine tissue.

Assessment of changes in the micro-nano environment of intervertebral disc degeneration based on Pfirrmann grade.

BACKGROUND CONTEXT: Pfirrmann grading can be used to assess intervertebral disc degeneration (IVDD). There is growing evidence that IVDD is not simply a structural disorder but also involves changes to the substructural characteristics of the disc. Whether Pfirrmann grade can accurately represent these micro-nano environmental changes remains unclear. PURPOSE: We aimed to assess the micro-nano structural characteristics of the degenerative disc to provide more specific biomechanical information than the Pfirrmann score. STUDY DESIGN: A micro- and nano-level structural analysis of degenerative discs of rat tails. METHODS: In this study, 12-week-old adult male Sprague-Dawley rats were divided randomly into five groups: control (no intervention to the intervertebral disc of the tail) and four intervention groups that all had caudal vertebrae immobilized using a custom-made external device to fix four caudal vertebrae (Co7-Co10) but with variable subsequent compression of Co8 and Co9 for 2, 4, 6, or 8 weeks. Magnetic resonance imaging detection of rat coccygeal vertebrae was conducted at each time node of the experiment, and the T2 signal intensity and disc space were evaluated. Animals were euthanized and the caudal vertebrae were harvested for further analysis. Histopathology, glycosaminoglycan (GAG) content, histologic score, end plate structure, and elastic modulus of the intervertebral discs were evaluated. RESULTS: IVDD was observed at an earlier Pfirrmann grade (Pfirrmann II) under the microscope. With an increase in Pfirrmann grade to III-V, the pore structure of the bony end plate changed significantly and the number of pores decreased gradually. Furthermore, the total GAG content of the nucleus pulposus decreased from an average of 640.33 µg GAG/ng DNA in Pfirrmann grade I to 271.33 µg GAG/ng DNA in Pfirrmann grade V (p < .0001). At the early stage of clinical degeneration of intervertebral discs (Pfirrmann grades II and III), there were significant changes in mechanical properties of the outer annulus fibrosus compared with the inner layer (p < .05). Further, the fibril diameters exhibited significant changes compared with the control group (p < .05). CONCLUSIONS: Our study found that the Pfirrmann grading system combined with intervertebral disc micro-nano structural changes more comprehensively reflected the extent of disc degeneration. These data may help improve our understanding of the pathogenesis and process of clinical disc degeneration.