Subchondral bone plate thickness is associated with micromechanical and microstructural changes in the bovine patella osteochondral junction with different levels of cartilage degeneration.

The influence of joint degeneration on the biomechanical properties of calcified cartilage and subchondral bone plate at the osteochondral junction is relatively unknown. Common experimental difficulties include accessibility to and visualization of the osteochondral junction, application of mechanical testing at the appropriate length scale, and availability of tissue that provides a consistent range of degenerative changes. This study addresses these challenges. A well-established bovine patella model of early joint degeneration was employed, in which micromechanical testing of fully hydrated osteochondral sections was carried out in conjunction with high-resolution imaging using differential interference contrast (DIC) optical light microscopy. A total of forty-two bovine patellae with different grades of tissue health ranging from healthy to mild, moderate, and severe cartilage degeneration, were selected. From the distal-lateral region of each patella, two adjacent osteochondral sections were obtained for the mechanical testing and the DIC imaging, respectively. Mechanical testing was carried out using a robotic micro-force acquisition system, applying compression tests over an array (area: 200 µm × 1000 µm, step size: 50 µm) across the osteochondral junction to obtain a stiffness map. Morphometric analysis was performed for the DIC images of fully hydrated cryo-sections. The levels of cartilage degeneration, DIC images, and the stiffness maps were used to associate the mechanical properties onto the specific tissue regions of cartilage, calcified cartilage, and subchondral bone plate. The results showed that there were up to 20% and 24% decreases (p < 0.05) in the stiffness of calcified cartilage and subchondral bone plate, respectively, in the severely degenerated group compared to the healthy group. Furthermore, there were increases (p < 0.05) in the number of tidemarks, bone spicules at the cement line, and the mean thickness of the subchondral bone plate with increasing levels of degeneration. The decreasing stiffness in the subchondral bone plate coupled with the presence of bone spicules may be indicative of a subchondral remodeling process involving new bone formation. Moreover, the mean thickness of the subchondral bone plate was found to be the strongest indicator of mechanical and associated structural changes in the osteochondral joint tissues.

Age-related differences in hamstring tendon used as autograft in reconstructive anterior cruciate ligament surgery.

PURPOSE: The hamstring tendon is the most commonly used autograft material in reconstructive surgeries of anterior cruciate ligament (ACL) tears. Younger patients have worse surgical outcomes, with a higher risk of re-rupture. We hypothesized that age-related changes in hamstring tendon properties affect the tendon's propensity to rupture when used as an autograft in ACL reconstructions. The purpose of this study was to compare hamstring tendon samples obtained from people aged 20 years or younger to samples obtained from older people. METHODS: Superfluous hamstring tendon material was collected from 13 young donors (aged 16-20 years) and 17 older donors undergoing ACL reconstructive surgery. Sections of the tendon samples were used for biomechanical testing, structural analysis of collagen fibrils by electron microscopy, and global analysis of gene expression by microarrays. RESULTS: We found that tendon samples from the older group had lower Young's modulus than the younger group (P = 0.015), whereas the stress to failure was similar in the two groups. We found no difference in the average diameter of collagen fibrils between the two groups. Microarray analysis identified 162 differentially expressed genes (fold change ≥ 1.5, P < 0.05), with overrepresentation of several biological processes, including regulation of adhesion, migration, inflammation, and differentiation (fold enrichment > 2.0, false discovery rate P < 0.05). CONCLUSION: The hamstring tendon from younger people has higher stiffness than tendon from older people, and the profile of gene expression in tendon varies with age. These differences may negatively affect the performance of the hamstring tendon in ACL reconstructions in younger people.

The ultrastructure of cartilage tissue and its swelling response in relation to matrix health.

This multi-length scale anatomical study explores the influence of mild cartilage structural degeneration on the tissue swelling response. While the swelling response of cartilage has been studied extensively, this is the first study to reveal and correlate tissue microstructure and ultrastructure, with the swelling induced cartilage tissue strains. Cartilage sample strips (n = 30) were obtained from the distal-lateral quadrant of thirty mildly degenerate bovine patellae and, following excision from the bone, the cartilage strips were allowed to swell freely for 2 h in solutions of physiological saline and distilled water successively. The swelling response of this group of samples were compared with that of healthy cartilage, with (n = 20) and without the surface layer (n = 20). The subsequent curling response of cartilage showed that in healthy tissue it was highly variable, and with the surface removed some samples curved in the opposite direction, while in the mildly degenerate tissue group, virtually all tissue strips curved in a consistent upward manner. A significant difference in strain was observed between healthy samples with surface layer removed and mildly degenerate samples, illustrating how excision of the surface zone from pristine cartilage is insufficient to model the swelling response of tissue which has undergone natural degenerative changes. On average, total tissue thickness increased from 940 µm (healthy) to 1079 µm (mildly degenerate), however, looking at the zonal strata, surface and transition zone thicknesses both decreased while deep zone thickness increased from healthy to mildly degenerate tissue. Morphologically, changes to the surface zone integrity were correlated with a diminished surface layer which, at the ultrastructural scale, correlated with a decreased fibrillar density. Similarly, fibrosity of the general matrix visible at the microscale was associated with a loss of later interconnectivity resulting in large, aggregated fibril bundles. The microstructural and ultrastructural investigation revealed that the key differences influencing the tissue swelling strain response was (1) the thickness and extent of disruption to the surface layer and (2) the amount of fibrillar network destructuring, highlighting the importance of the collagen and tissue matrix structure in restraining cartilage swelling.

The micro and ultrastructural anatomy of bone spicules found in the osteochondral junction of bovine patellae with early joint degeneration.

The structural changes in the tissues of the osteochondral junction are a topic of interest, especially considering how bone changes are involved in the initiation and progression of osteoarthritis (OA). Our research group has previously demonstrated that at the cement line boundary between the zone of calcified cartilage (ZCC) and the subchondral bone, in mature bovine patellae with early OA, there are numerous bone spicules that have emerged from the underlying bone. These spicules contain a central vascular canal and a bone cuff. In this study, we use high-resolution differential interference contrast optical microscopy and scanning electron microscopy to compare the cartilage-bone junction of three groups of mature bovine patellae showing healthy to mild to moderately degenerate cartilage. The ZCC and bone junction was carefully examined to estimate the frequency of marrow spaces, bone spicules and fully formed bone bulges. The results reveal that bone spicules are associated with all grades of cartilage tissue studied, with the most occurring in the intermediate stages of tissue health. The micro and ultrastructure of the bone spicule are consistent with that of an osteon, especially those found in compression zones in long bones. Also considering the coexistence of marrow spaces and fully formed bone, this study suggests that these bone spicules arise similar to the formation of osteons in the bone remodelling process. The significance of this conclusion is in the way researchers approach the bone formation issue in the early degenerative joint. Instead of endochondral ossification, we propose that bone formation in OA is more akin to a combination of primary bone remodelling and de novo bone formation.

The dynamic impact behavior of the human neurocranium.

Realistic biomechanical models of the human head should accurately reflect the mechanical properties of all neurocranial bones. Previous studies predominantly focused on static testing setups, males, restricted age ranges and scarcely investigated the temporal area. This given study determined the biomechanical properties of 64 human neurocranial samples (age range of 3 weeks to 94 years) using testing velocities of 2.5, 3.0 and 3.5 m/s in a three-point bending setup. Maximum forces were higher with increasing testing velocities (p ≤ 0.031) but bending strengths only revealed insignificant increases (p ≥ 0.052). The maximum force positively correlated with the sample thickness (p ≤ 0.012 at 2.0 m/s and 3.0 m/s) and bending strength negatively correlated with both age (p ≤ 0.041) and sample thickness (p ≤ 0.036). All parameters were independent of sex (p ≥ 0.120) apart from a higher bending strength of females (p = 0.040) for the 3.5 -m/s group. All parameters were independent of the post mortem interval (p ≥ 0.061). This study provides novel insights into the dynamic mechanical properties of distinct neurocranial bones over an age range spanning almost one century. It is concluded that the former are age-, site- and thickness-dependent, whereas sex dependence needs further investigation.

Impact-induced cartilage damage assessed using polarisation-sensitive optical coherence tomography.

Non-invasive determination of structural changes in articular cartilage immediately after impact and rehydration provides insight into the response and recovery of the soft tissue, as well as provides a potential methodology for clinicians to quantify early degenerative changes. In this study, we use polarisation-sensitive optical coherence tomography (PS-OCT) to examine subtle alterations of the optical properties in healthy and early-stage degenerate articular cartilage immediately after impact loading to identify structurally relevant metrics required for understanding the mechanical factors of osteoarthritic initiation and progression. A custom-designed impact testing rig was used to deliver 0.9 J and 1.4 J impact energies to bovine articular cartilage. A total of 52 (n=26 healthy, n=26 mildly degenerate) cartilage-on-bone samples were imaged before, immediately after, and 3 h after impact. PS-OCT images were analyzed to assess changes relating to surface irregularity, optical attenuation, and birefringence. Mildly degenerate cartilage exhibits a significant change in birefringence following 1.4 J impact energies compared to healthy samples which is believed to be attributable to degenerate cartilage being unable to fully utilise the fluid phase to distribute and dampen the energy. After rehydration, the polarisation-sensitive images appear to 'optically-recover' reducing the reliability of birefringence as an absolute metric. Surface irregularity and optical attenuation encode diagnostically relevant information and may serve as markers to predict the mechanical response of articular cartilage. PS-OCT with its ability to non-invasively image the sub-surface microstructural abnormalities of cartilage presents as an ideal modality for cartilage degeneration assessment and identification of mechanically vulnerable tissue.

Protein Levels and Microstructural Changes in Localized Regions of Early Cartilage Degeneration Compared with Adjacent Intact Cartilage.

OBJECTIVE: It was hypothesized that the respective protein profiles of bovine cartilage from sites of localized mild to moderate (GI to GII) degeneration versus adjacent sites of intact tissue would vary in accordance with the tissue microstructural changes associated with a pre-osteoarthritic state. METHODS: A total of 15 bovine patellae were obtained for this study. Paired samples of tissue were collected from the lateral region of each patella. If the patella contained a site of degeneration, a paired tissue set involved taking one sample each from the degenerated site and the intact tissue adjacent to it. Sufficient tissue was collected to facilitate 2 arms of investigation: microstructural imaging and proteome analysis. The microstructural analysis used a bespoke tissue preparation technique imaged with differential interference contrast optical microscopy to assess fibrillar scale destructuring and underlying bone spicule formation. An iTRAQ-based proteome analysis was performed using liquid chromatography-tandem mass spectrometry to identify the differential levels of proteins across the intact and degenerated cartilage and further, the results were validated with multiple reaction monitoring assay. RESULTS: In the healthy cartilage pairs, there was no significant variation in protein profiles between 2 adjacent sample sites. In pairs of tissue that contained a sample of GI/GII tissue, there were both significant microstructural changes as well as the difference in abundance levels of 24 proteins. CONCLUSIONS: From the known functions of the 24 proteins, found to be strongly aligned with the specific microstructural changes observed, a unique "proteins ensemble" involved in the initiation and progression of early cartilage degeneration is proposed.

Raman microspectroscopic analysis of the tissue-specific composition of the human osteochondral junction in osteoarthritis: A pilot study.

This study investigates the influence of osteoarthritis (OA) disease severity on the bio-composition of the osteochondral junction at the human tibial plateau using Raman microspectroscopy. We specifically aim to analyze the spatial composition of mineralized osteochondral tissues, i.e., calcified cartilage (CC) and subchondral bone plate (SBP) from unfixed, hydrated specimens. We hypothesize that the mineralization of CC and SBP decreases in advanced OA. Twenty-eight cylindrical osteochondral samples (d = 4 mm) from tibial plateaus of seven cadaveric donors were harvested and sorted into three groups following histopathological grading: healthy (n = 5), early OA (n = 8), and advanced OA (n = 15). Raman spectra were subjected to multivariate cluster analyses to identify different tissues. Finally, the tissue-specific composition was analyzed, and the impact of OA was statistically evaluated with linear mixed models. Cluster analyses of Raman spectra successfully distinguished CC and SBP as well as a tidemark region and uncalcified cartilage. CC was found to be more mineralized and the mineral was more crystalline compared with SBP. Both tissues exhibited similar compositional changes as a function of histopathological OA severity. In early OA, the mineralization tends to increase, and the mineral contains fewer carbonate substitutions. Compared with early OA, mineral crystals are rich in carbonate while the overall mineralization decreases in advanced OA. This Raman spectroscopic study advances the methodology for investigating the complex osteochondral junction from native tissue. The developed methodology can be used to elucidate detailed tissue-specific changes in the chemical composition with advancing OA. STATEMENT OF SIGNIFICANCE: In this study, Raman microspectroscopy was utilized to investigate the influence of osteoarthritic degeneration on the tissue-specific biochemical composition of the human osteochondral junction. Multivariate cluster analyses allowed us to characterize subtle compositional changes in the calcified cartilage and subchondral bone plate as well as in the tidemark region. The compositional differences found between the calcified cartilage and subchondral bone plate in both organic and mineral phases will serve as critical benchmark parameters when designing biomaterials for osteochondral repair. We found tissue-specific changes in the mineralization and carbonate substitution as a function of histopathological OA severity. Our developed methodology can be used to investigate the metabolic changes in the osteochondral junction associated with osteoarthritis.

The mechanical significance of the zonally differentiated collagen network of articular cartilage in relation to tissue swelling.

BACKGROUND: We hypothesise that the Benninghoff arcade fibril structure motif of cartilage is able to predict the swelling response of cartilage. METHODS: A total of ten healthy adult bovine patellae were used for this study, yielding 20 paired full depth cartilage samples (half with surface layer intact and half with surface layer removed). Following excision from the bone, samples were allowed to equilibrate first in physiological saline for 2 h, and then in distilled water for another 2 h to maximise the swelling response. Images were captured using a stereomicroscope to measure strain and the fully-swollen samples were fixed in 10% formalin to retain shape for microscopic and ultrastructural imaging. FINDINGS: We expected all swelling samples with an intact 'strain-limiting' surface layer to curl upwards, instead only 70% of them did. For samples without a surface layer, we expected the swelling to be evenly distributed and to remain relatively uncurled; but in 40% of the samples there was a downward curvature (i.e. opposite to that of the previous group). Micro-to-ultrastructural imaging, to determine fibrillar structure and organisation, revealed the deep zone cartilage was an additional counter layer limiting swelling strain, and was the likely cause of the unexpected swelling responses. INTERPRETATION: Our expectations that the surface layer alone will influence the swelling response, was based on the assumptions of the Benninghoff arcade model. This study highlights the additional importance of sub-micron scale fibrillar interconnectivity and the role of the deep zone.

Differential Response of Bovine Mature Nucleus Pulposus and Notochordal Cells to Hydrostatic Pressure and Glucose Restriction.

OBJECTIVE: The nucleus pulposus of the human intervertebral disc contains 2 cell types: notochordal (NC) and mature nucleus pulposus (MNP) cells. NC cell loss is associated with disc degeneration and this process may be initiated by mechanical stress and/or nutrient deprivation. This study aimed to investigate the functional responses of NC and MNP cells to hydrostatic pressures and glucose restriction. DESIGN: Bovine MNP and NC cells were cultured in 3-dimensional alginate beads under low (0.4-0.8 MPa) and high (1.6-2.4 MPa) dynamic pressure for 24 hours. Cells were cultured in either physiological (5.5 mM) glucose media or glucose-restriction (0.55 mM) media. Finally, the combined effect of glucose restriction and high pressure was examined. RESULTS: Cell viability and notochordal phenotypic markers were not significantly altered in response to pressure or glucose restriction. MNP cells responded to low pressure with an increase in glycosaminoglycan (GAG) production while high pressure significantly decreased ACAN gene expression compared with atmospheric controls. NC cells showed no response in matrix gene expression or GAG production with either loading regime. Glucose restriction decreased NC cell TIMP-1 expression but had no effect on MNP cells. The combination of glucose restriction and high pressure only affected MNP cell gene expression, with decreased ACAN, Col2α1, and ADAMTS-5 expression. CONCLUSION: This study shows that NC cells are more resistant to acute mechanical stresses than MNP cells and provides a strong rationale for future studies to further our understanding the role of NC cells within the disc, and the effects of long-term exposure to physical stresses.

Sagittal Alignment With Downward Slope of the Lower Lumbar Motion Segment Influences Its Modes of Failure in Direct Compression: A Mechanical and Microstructural Investigation.

STUDY DESIGN: Microstructural investigation of compression-induced herniation of ovine lumbar discs with and without added component of anterior-inferior slope. OBJECTIVE: Does increased shear arising from a simulated component of motion segment slope imitating sacral slope weaken the lateral annulus and increase risk of overt herniation at this same region. SUMMARY OF BACKGROUND DATA: An increase in sacral slope secondary to lordosis and pelvic incidence increases shear stresses at the lumbosacral junction and has been associated with an increase in spondylolisthetic disorders and back injury. The small component of forward shear induced when a segment is compressed in flexion is suggested to cause differential recruitment of the lateral annular fibers leading to its early disruption followed by intra-annular nuclear tracking to the posterolateral/posterior regions. However, the influence of even greater forward shear arising from the added component of slope seen where pelvic incidence and lumbar lordosis are increased in the lower lumbar spine is less understood. METHODS: Ovine motion segments were compressed at 40 mm/min up to failure; 9 with a horizontal disc alignment and 26 with a segment slope of 15° and then analyzed structurally. RESULTS: All the horizontal discs failed (11.8 ±â€Š2.4 kN) via vertebral fracture without any evidence of soft tissue failure even in the lateral aspects of the discs. The increased forward shear resulting from the slope decreased the failure load (6.4 ±â€Š1.6 kN). The sloping discs mostly suffered mid-span, noncontinuous disruption of the lateral annulus with some extruding nuclear material directly from these same lateral regions. CONCLUSION: The increased level of forward shear generated in moderately sloping lumbar segments when compressed was abnormally damaging to the lateral regions of the disc annulus. This is consistent with the view that shear differentially loads the oblique-counter oblique fiber sets in the lateral annulus, increasing its vulnerability to early disruption and overt herniation. LEVEL OF EVIDENCE: N/A.

New evidence for structural integration across the cartilage-vertebral endplate junction and its relation to herniation.

BACKGROUND CONTEXT: The cartilaginous and bony material that can be present in herniated tissue suggests that failure can involve both cartilaginous and vertebral-endplates. How structural integration is achieved across the junction between these two distinct tissue regions via its fibril and mineral components is clearly relevant to the modes of endplate failure that occur. PURPOSE: To understand how structural integration is achieved across the cartilaginous-vertebral endplate junction. STUDY DESIGN: A micro- and fibril-level structural analysis of the cartilage-vertebral endplate region was carried out using healthy, mature ovine motion segments. METHODS: Oblique vertebra-annulus-vertebra samples were prepared such that alternate layers of lamellar fibers extended from vertebra to vertebra. The endplate region of each sample was then decalcified in a targeted manner before being loaded in tension along the fiber direction to achieve incomplete rupture within the region of the endplate. The failure regions were then analyzed with differential interference contrast microscopy and scanning electron microscopy. RESULTS: Microstructural analysis revealed that failure within the endplate region was not confined to the cement line. Instead, rupture continued into the underlying vertebral endplate with bony material still attached to the now unanchored annular bundles. Ultrastructural analysis of the partially ruptured regions of the cement line revealed clear evidence of blending/interweaving relationships between the fibrils of the annular bundles, the calcified cartilage and the bone with no one pattern of association appearing dominant. These findings suggest that fibril-based structural cohesion exists across the cement line at the site of annular insertion, with strengthening via a mechanism somewhat analogous to steel-reinforced concrete. The fibrils are brought into a close intermingling association with interfibril forces mediated via the mineral component. CONCLUSIONS: This study provides clear evidence of structural connectivity across the cartilaginous-vertebral endplate junction by the intermingling of their fibrillar components and mediated by the mineral phase. This is consistent with the clinical observation that in some disc herniations bony material can be still attached to the extruded soft tissue.

Quantifying birefringence in the bovine model of early osteoarthritis using polarisation-sensitive optical coherence tomography and mechanical indentation.

Recent studies have shown potential for using polarisation sensitive optical coherence tomography (PS-OCT) to study cartilage morphology, and to be potentially used as an in vivo, non-invasive tool for detecting osteoarthritic changes. However, there has been relatively limited ability of this method to quantify the subtle changes that occur in the early stages of cartilage degeneration. An established mechanical indenting technique that has previously been used to examine the microstructural response of articular cartilage was employed to fix the bovine samples in an indented state. The samples were subject to creep loading with a constant compressive stress of 4.5 MPa and, when imaged using PS-OCT, enabled birefringent banding patterns to be observed. The magnitude of the birefringence was quantified using the birefringence coefficient (BRC) and statistical analysis revealed that PS-OCT is able to detect and quantify significant changes between healthy and early osteoarthritic cartilage (p < 0.001). This presents a novel utilization of PS-OCT for future development as an in vivo assessment tool.

The Influence of Concordant Complex Posture and Loading Rate on Motion Segment Failure: A Mechanical and Microstructural Investigation.

STUDY DESIGN: Microstructural investigation of compression-induced herniation of a lumbar disc held in a concordant complex posture. OBJECTIVE: To explore the significance of loading rate in a highly asymmetric concordant posture, comparing the mechanisms of failure to an earlier study using a nonconcordant complex posture. SUMMARY OF BACKGROUND DATA: A recent study with a nonconcordant complex posture (turning in the opposite direction to that which the load is applied) demonstrated the vulnerability of the disc to loading that is borne by one set of oblique-counter oblique fiber sets in the alternating lamellae of the annulus, and aggravated by an elevated loading rate. Given the strain rate-dependent properties of the disc it might be expected that the outcome differs if the posture is reversed. METHODS: Forty-one motion segments from ovine 16 spines were split into two cohorts; adopting the previously employed low rate (40 mm/min) and surprise rate (400 mm/min) of loading. Both groups of damaged discs were then analyzed microstructurally. RESULTS: With the lower rate loading the concordant posture significantly reduced the load required to cause disc failure than earlier described for nonconcordant posture (6.9 vs. 8.4 kN), with more direct tears and alternate lamella damage extending to the anterior disc. Contrary to this result, with a surprise rate, the load at failure was significantly increased with the concordant posture (8.08 vs. 6.96 kN), although remaining significantly less than that from a simple flexed posture (9.6 kN). Analysis of the damage modes and postures suggest facet engagement plays a significant role. CONCLUSION: This study confirms that adding shear to the posture lowers the load at failure, and causes alternate lamella rupture. Load at failure in a complex posture is not determined by loading rate alone. Rather, the strain rate-dependent properties of the disc influence which elements of the system are brought into play. LEVEL OF EVIDENCE: N/A.

A Microstructural Investigation of Disc Disruption Induced by Low Frequency Cyclic Loading.

STUDY DESIGN: Microstructural investigation of low frequency cyclic loading and flexing of the lumbar disc. OBJECTIVE: To explore micro-level structural damage in motion segments subjected to low frequency repetitive loading and flexing at sub-acute loads. SUMMARY OF BACKGROUND DATA: Cumulative exposure to mechanical load has been implicated in low back pain and injury. The mechanical pathways by which cyclic loading physically affects spine tissues remain unclear, in part due to the absence of high quality microstructural evidence. METHODS: The study utilized seven intact ovine lumbar spines and from each spine one motion segment was used as a control, two others were cyclically loaded. Ten motion segments were subjected to 5000 cycles at 0.5 Hz with a peak load corresponding to ∼30% of that required to achieve failure. An additional small group of segments subjected to 10,000 or 30,000 cycles was similarly analyzed. Following chemical fixation and decalcification samples were cryosectioned along one of the oblique fiber angles and imaged in their fully hydrated state using differential interference contrast optical microscopy. Structural damage obtained from the images was organized into an algebraic shell for analysis. RESULTS: At 5000 cycles the disc damage was limited to inner wall distortions, evidence of stress concentrations at bridging-lamellae attachments, and small delaminations. The high-cycle discs tested exhibited significant mid-wall damage. There was no evidence of nuclear material being displaced. CONCLUSION: At this low frequency and without the application of sustained loading or a more severe loading regime, or maintaining a constant flexion with repetitive loading, it seems unlikely that actual nuclear migration occurs. It is possible that the inner-annular damage shown in the low dose group could disrupt pathways for nutrient diffusion leading to earlier cell death and matrix degradation, thus contributing to a cascade of degeneration. LEVEL OF EVIDENCE: N/A.

Does an Annular Puncture Influence the Herniation Path?: An In Vitro Mechanical and Structural Investigation.

STUDY DESIGN: A study of mechanically induced herniation in punctured ovine discs followed by structural analysis. OBJECTIVE: To investigate whether an annular puncture influences the path that herniation takes by providing direct passage for nucleus through the annulus and therefore whether it increases the risk of acute herniation from overload at the site of damage independent of any longer-term degeneration. SUMMARY OF BACKGROUND DATA: Ten years after treatment with discography both degenerative changes and frequency of herniation have been shown to increase compared to untreated discs. Although the effect of an annular puncture over time has been widely investigated the question of whether it increases the risk of acute herniation has not been resolved. METHODS: The posterolateral annuli of healthy ovine lumbar discs were punctured with either a 25-gauge (n = 8) or a larger 18-gauge (n = 8) needle and then compressed in a flexed posture of 10° until initial indications of failure. The entire volume of the disc was visually assessed for structural damage by obtaining progressive, full transverse cross-sections of its entire height thus exposing all regions of the disc. RESULTS: There was no association between the 25-gauge puncture and disc disruption and herniation. In contrast, nuclear material was observed to migrate through the 18-gauge needle puncture. Disruption of the lateral inner annulus was observed in 12 out of the 16 discs tested. CONCLUSION: The risk of acute herniation through the puncture site is dependent on the needle diameter used. Under the conditions employed the lateral inner annulus remains the site most vulnerable to disruption independent of the presence of a posterolateral puncture. LEVEL OF EVIDENCE: N /A.

How a decreased fibrillar interconnectivity influences stiffness and swelling properties during early cartilage degeneration.

OBJECTIVE: The functional coupling between the fibrillar network and the high-swelling proteoglycans largely determines the mechanical properties of the articular cartilage matrix. The objective of this new study was to show specifically how changes in fibrillar interconnectivity arising from early cartilage degeneration influence transverse stiffness and swelling properties at the tissue level. DESIGN: Radial zone transverse layers of cartilage matrix were obtained from intact and mildly degenerate bovine patellae. Each layer was then subdivided to assess tensile stiffness, free-swelling response, glycosaminoglycan (GAG) content, and micro- and ultra-structural features. RESULTS: The tensile modulus was significantly lower and the degree of swelling significantly higher for the degenerate matrix compared to the intact. Scanning electron microscopy revealed a homogeneous response to transverse strain in the intact cartilage, whereas large non-fibrillar spaces between fibril aggregates were visible in the degenerate matrix. Although there were no significant differences in GAG content it did correlate significantly with stiffness and swelling in the intact samples but not in the degenerate. CONCLUSIONS: The lower degree of fibril network interconnectivity in the degenerate matrix led to both a decreased transverse stiffness and reduced resistance to osmotic swelling. This network 'de-structuring' also resulted in a reduced functional interaction between the fibrillar network and the proteoglycans. The study provides new insights into the role of the fibrillar network and how changes in the network arising from the degenerative cascade will influence tissue level behaviour.

A more realistic disc herniation model incorporating compression, flexion and facet-constrained shear: a mechanical and microstructural analysis. Part II: high rate or 'surprise' loading.

PURPOSE: Part I of this study explored mechanisms of disc failure in a complex posture incorporating physiological amounts of flexion and shear at a loading rate considerably lower than likely to occur in a typical in vivo manual handling situation. Given the strain-rate-dependent mechanical properties of the heavily hydrated disc, loading rate will likely influence the mechanisms of disc failure. Part II investigates the mechanisms of failure in healthy discs subjected to surprise-rate compression while held in the same complex posture. METHODS: 37 motion segments from 13 healthy mature ovine lumbar spines were compressed in a complex posture intended to simulate the situation arising when bending and twisting while lifting a heavy object at a displacement rate of 400 mm/min. Seven of the 37 samples reached the predetermined displacement prior to a reduction in load and were classified as early stage failures, providing insight to initial areas of disc disruption. Both groups of damaged discs were then analysed microstructurally using light microscopy. RESULTS: The average failure load under high rate complex loading was 6.96 kN (STD 1.48 kN), significantly lower statistically than for low rate complex loading [8.42 kN (STD 1.22 kN)]. Also, unlike simple flexion or low rate complex loading, direct radial ruptures and non-continuous mid-wall tearing in the posterior and posterolateral regions were commonly accompanied by disruption extending to the lateral and anterior disc. CONCLUSION: This study has again shown that multiple modes of damage are common when compressing a segment in a complex posture, and the load bearing ability, already less than in a neutral or flexed posture, is further compromised with high rate complex loading.

A more realistic disc herniation model incorporating compression, flexion and facet-constrained shear: a mechanical and microstructural analysis. Part I: Low rate loading.

PURPOSE: To date, the mechanisms of disc failure have been explored at a microstructural level in relatively simple postures. However, in vivo the disc is known to be subjected to complex loading in compression, bending and shear, and the influence of these factors on the mechanisms of disc failure is yet to be described at a microstructural level. The purpose of this study was to provide a microstructural analysis of the mechanisms of failure in healthy discs subjected to compression while held in a complex posture incorporating physiological amounts of flexion and facet-constrained shear. METHODS: 30 motion segments from 10 healthy mature ovine lumbar spines were compressed in a complex posture intended to simulate the situation arising when bending and twisting while lifting a heavy object, and at a displacement rate of 40 mm/min. Nine of the 30 samples reached the predetermined displacement prior to a reduction in load and were classified as early-stage failures, providing insight into initial areas of disc disruption. Both groups of damaged discs were then analysed microstructurally using light microscopy. RESULTS: Complex postures significantly reduced the load required to cause disc failure than earlier described for flexed postures [8.42 kN (STD 1.22 kN) compared to 9.69 kN (STD 2.56 kN)] and resulted in a very different failure morphology to that observed in either simple flexion or direct compression, involving infiltration of nucleus material in a circuitous path to the annular periphery. CONCLUSION: The complex posture as used in this study significantly reduced the load required to cause disc failure, providing further evidence that asymmetric postures while lifting should be avoided if possible.

Microstructure Variations in the Soft-Hard Tissue Junction of the Human Anterior Cruciate Ligament.

The role of the sub-bundles in the anterior cruciate ligament (ACL) has been defined, such that the anterior-medial bundle directly resists anterior tibial translation while the posterior lateral bundle is involved in rotational stability. With regards to this biomechanical function, much of the previous work on bundle-specific morphology has been carried out on the macroscale, with much less attention given to the micro-to-ultrastructural scalar levels. This is especially true of the enthesis and its microstructure, a biomechanically significant region that has been largely neglected in the published literature dealing with ACL sub-bundle anatomy. In this study, the human ACL tibial enthesis was investigated at multiple scalar levels using differential interference contrast and scanning electron microscopies with the aim of determining whether the sub-bundle ligament structure, and its known macroscale function, is consistent with its micro-architecture at the ligament-bone junction. The investigation found that different ligament insertion morphologies exist between the two bundles, where the AM bundle has more intense interdigitation with the bone matrix than that of the PL bundle. The results suggest that such structure-function relationships, especially across scalar-levels, provide new insight into the significance of the sub-bundle anatomy of the ACL. Anat Rec, 300:1547-1559, 2017. © 2017 Wiley Periodicals, Inc.