Effect of Articular Surface Compression on Cartilage Extracellular Matrix Deformation.

Early stage osteoarthritis is characterized by disruption of the superficial zone (SZ) of articular cartilage, including collagen damage and proteoglycan loss, resulting in "mechanical softening" of the extracellular matrix (ECM). The role of the SZ in controlling fluid exudation and imbibition during loading and unloading, respectively, was studied using confined creep compression tests. Bovine osteochondral (OC) plugs were subjected to either a static (88 kPa) or cyclic (0-125 kPa at 1 Hz) compressive stress for five minutes, and the cartilage deformation and recovery were measured during tissue loading and unloading, respectively. During unloading, the articular surface of the cartilage was either loaded with a small 1% tare load (∼1 kPa) applied through a porous load platen (covered), or completely unloaded (uncovered). Then the SZ (∼10%) of the cartilage was removed and the creep tests were repeated. Randomized tests were performed on each OC specimen to assess variability within and between plugs. Static creep strain was always greater than cyclic creep strain except at the beginning of loading (10-20 cycles). Uncovering the articular surface after creep deformation resulted in faster thickness recovery compared to the covered recovery. Removal of the SZ resulted in increased static and cyclic creep strains, as well as an increase in the cyclic peak-to-peak strain envelope. Our results indicate that an intact SZ is essential for normal cartilage mechanical function during joint motion by controlling fluid exudation and imbibition, and concomitantly ECM deformation and recovery, when loaded and unloaded, respectively.

A new paradigm for mechanobiological mechanisms in tumor metastasis.

Tumor metastases and epithelial to mesenchymal transition (EMT) involve tumor cell invasion and migration through the dense collagen-rich extracellular matrix surrounding the tumor. Little is neither known about the mechanobiological mechanisms involved in this process, nor the role of the mechanical forces generated by the cells in their effort to invade and migrate through the stroma. In this paper we propose a new fundamental mechanobiological mechanism involved in cancer growth and metastasis, which can be both protective or destructive depending on the magnitude of the forces generated by the cells. This new mechanobiological mechanism directly challenges current paradigms that are focused mainly on biological and biochemical mechanisms associated with tumor metastasis. Our new mechanobiological mechanism describes how tumor expansion generates mechanical forces within the stroma to not only resist tumor expansion but also inhibit or enhance tumor invasion by, respectively, inhibiting or enhancing matrix metalloproteinase (MMP) degradation of the tensed interstitial collagen. While this mechanobiological mechanism has not been previously applied to the study of tumor metastasis and EMT, it may have the potential to broaden our understanding of the tumor invasive process and assist in developing new strategies for preventing or treating cancer metastasis.

Differential cross-linking and radio-protective effects of genipin on mature bovine and human patella tendons.

Gamma irradiation is a proven sterilization method, but is not widely used on allografts for anterior cruciate ligament reconstruction (e.g., patella tendon) due to radiation-induced decreases in mechanical strength. Addressing this drawback would improve the safety and supply of allografts to meet current and future demand. It was hypothesized that genipin-induced collagen cross-linking would increase the tensile modulus of patella tendon tissue such that 5 MRad gamma irradiation would not reduce the tissue mechanical strength below the original untreated values. Optimized genipin treatment increased the tensile modulus of bovine tendons by ~2.4-fold. After irradiation, genipin treated tissue did not significantly differ from native tissue, proving the hypothesis. Optimized genipin treatment of human tendons increased the tensile modulus by ~1.3-fold. After irradiation, both control and genipin-treated tissues possessed ~50-60% of their native tendon modulus, disproving the hypothesis. These results highlight possible age- and species- dependent effects of genipin cross-linking on tendon tissue. Cross-linking of human allografts may be beneficial only in younger donor tissues. Future research is warranted to better understand the mechanisms and applications of collagen cross-linking for clinical use.

A novel device to apply controlled flexion and extension to the rat knee following anterior cruciate ligament reconstruction.

We designed and validated a novel device for applying flexion-extension cycles to a rat knee in an in vivo model of anterior cruciate ligament reconstruction (ACL-R). Our device is intended to simulate rehabilitation motion and exercise post ACL-R to optimize physical rehabilitation treatments for the improved healing of tendon graft ligament reconstructions. The device was validated for repeatability of the knee kinematic motion by measuring the force versus angular rotation response from repeated trials using cadaver rats. The average maximum force required for rotating an ACL reconstructed rat knee through 100 degrees of flexion-extension was 0.4 N with 95% variability for all trials within ±0.1 N.

Role of coracoacromial ligament and related structures in glenohumeral stability: a cadaveric study.

This study sought to determine the role of the coracoacromial ligament and related arch structures in glenohumeral joint stabilization. Eight fresh-frozen cadaver specimens were tested at multiple angles of glenohumeral abduction and rotation for translations (in the direction of and perpendicular to a 50-N force) in intact, vented shoulders and after three interventions: coracoacromial veil release, coracoacromial ligament release, and anterior acromioplasty. After releasing the veil, an inferior force significantly increased inferior translation at lower angles of abduction with no additional increase after coracoacromial ligament section or acromioplasty. After ligament release or acromioplasty, a superior force increased superior translation at all angles. Few increases in anterior or posterior translations were observed. The coracoacromial veil interacts with the structures of the coracoacromial arch and glenohumeral capsule to limit inferior humeral translation. Likewise, the coracoacromial ligament and the acromian serve to limit superior translation. Attempts to preserve these structures may help improve surgical outcomes.

ACL transection results in a posterior shift and increased velocity of contact on the medial tibial plateau.

Our objective was to quantify the effect of ACL transection on dynamic knee joint contact force distributions during simulated gait. Given the prevalence of medial compartment osteoarthritis in un-reconstructed ACL ruptured knees, we hypothesized that changes in contact mechanics after ACL transection would be most prevalent in the medial compartment. Twelve human cadaveric knees were tested using a dynamic knee gait simulator which was programmed to mimic a clinical Lachman exam and gait. An electronic pressure sensor was placed on the medial and lateral tibial plateaus under the menisci to quantify dynamic contact forces before and after ACL transection. Tibial translations and rotations, medial and lateral plateau peak contact stress, and position and velocity of the Weighted Center of Contact (WCoC) were computed. After ACL transection, the tibia translated more anteriorly in the Lachman examination and at heel strike during gait. Changes in contact mechanics across the medial tibial plateau during simulated gait were: an increase in the velocity of WCoC and a posterior shift in the WCoC, both of which occurred at heel strike; increased peak contact forces in the posterior-peripheral quadrant of the tibial plateau at 45% of the gait cycle; and an additional posterior shift in WCoC from 25 to 55% of the gait cycle. The only change in contact mechanics in the lateral plateau was a decrease in WCoC velocity in late stance. This data is suggested to further the study of biomechanical pathways (biomechanical biomarkers) in the relationship between altered knee contact mechanics and chondrocyte metabolic responses after ACL transection.

Squeeze-film properties of synovial fluid and hyaluronate-based viscosupplements.

The rheological properties of synovial fluid and hyaluronate (HA) solutions have been studied using a variety of viscometers and rheometers. These devices measure the viscosity of the fluid's resistance to shearing forces, which is useful when studying the lubrication and frictional properties of movable joints. Less commonly used is a squeeze-film fluid test, mechanistically similar to when two joint surfaces squeeze interposed fluid. In our study, we used squeeze-film tests to determine the rheological response of normal bovine synovial fluid and 10 mg/ml HA-based solutions, Hyalgan/Hyalovet, commercially available 500-700 kDa HA viscosupplements, and a 1000 kDa sodium hyaluronate (NaHy) solution. We found similar rheological responses (fluid thickness, viscosity, viscosity-pressure relationship) for all three fluids, though synovial fluid's minimum squeeze-film thickness was slightly thicker. Squeeze-film loading speed did not affect these results. Different HA concentrations and molecular weights also did not have a significant or consistent effect on the squeeze-film responses. An unexpected result for the HA-solutions was a linear increase in minimum fluid-film thickness with increasing initial fluid-film thickness. This result was attributed to faster gelling of thicker HA-solutions, which formed at a lower squeeze-film strain and higher squeeze-film strain rate compared to thinner layers. Also included is a review of the literature on viscosity measurements of synovial fluid and HA solutions.

Ultraviolet light (365 nm) transmission properties of articular cartilage as a function of depth, extracellular matrix, and swelling.

Current tissue engineering approaches for treatment of injured or diseased articular cartilage use ultraviolet light (UV) for in situ photopolymerization of biomaterials to fill chondral and osteochondral defects as well as resurfacing, stiffening and bonding the extracellular matrix and tissue interfaces. The most commonly used UV light wavelength is UVA 365 nm, the least cytotoxic and deepest penetrating. However, little information is available on the transmission of UVA 365 nm light through the cartilage matrix. In the present study, 365 nm UV light transmission was measured as a function of depth through 100 µm thick slices of healthy articular cartilage removed from mature bovine knees. Transmission properties were measured in normal (Native) cartilage and after swelling equilibration in phosphate-buffered saline (Swollen). Single-factor and multiple linear regression analyses were performed to determine depth-dependencies between the effective attenuation coefficients and proteoglycan, collagen and water contents. For both cartilages, a significant depth-dependency was found for the effective attenuation coefficients, being highest at the articular surface (superficial zone) and decreasing with depth. The effective attenuation coefficients for full-thickness cartilages were approximately a third lower than the total attenuation coefficients calculated from the individual slices. Analysis of absorption and scattering effects due to the ECM and chondrocytes found that UV light scatter coefficients were ∼10 times greater than absorption coefficients. The greater transmittance of UV light through the thicker cartilage was attributed to the collagen within the ECM causing significant backscatter forward reflectance.

Collagen peptide simulated bending after applied axial deformation.

Structural proteins in the extracellular matrix are subjected to a range of mechanical loading conditions, including varied directions of force application. Molecular modeling suggests that these mechanical forces directly affect collagen's conformation and the subsequent mechanical response at the molecular level is complex. For example, tensile forces in the axial direction result in collagen triple helix elongation and unwinding, while perpendicular forces can cause local triple helix disruption. However, the effects of more complicated mechanical loading, such as the effect of axial pretension on collagen bending and triple helix microunfolding are unknown. In this study we used steered molecular dynamics to first model a collagen peptide under axial tension and then apply a perpendicular bending force. Axial tension causes molecular elongation and increased the subsequent perpendicular bending stiffness, but surprisingly did not increase the predicted collagen triple helix microunfolding threshold. We believe these results elucidate a key potential mechanism by which microscale mechanical loads translate from cellular and micro scales down to the nano and atomistic. Further, these data predict that cryptic force-induced collagen triple helix unwinding is axial-deformation independent, supporting the possibility that cell traction forces could be a key molecular mechanism to alter the cellular matrix microenvironment to facilitate collagen enzymatic degradation and subsequent cellular migration, such as in tumor extravasation.

Light Absorptive Properties of Articular Cartilage, ECM Molecules, Synovial Fluid, and Photoinitiators as Potential Barriers to Light-Initiated Polymer Scaffolding Procedures.

OBJECTIVE: Many in vivo procedures to repair chondral defects use ultraviolet (UV)-photoinitiated in situ polymerization within the cartilage matrix. Chemical species that absorb UV light might reduce the effectiveness of these procedures by acting as light absorption barriers. This study evaluated whether any of the individual native biochemical components in cartilage and synovial fluid interfered with the absorption of light by common scaffolding photosensitizers. MATERIALS: UV-visible spectroscopy was performed on each major component of cartilage in solution, on bovine synovial fluid, and on four photosensitizers, riboflavin, Irgacure 2959, quinine, and riboflavin-5'-phosphate. Molar extinction and absorption coefficients were calculated at wavelengths of maximum absorbance and 365 nm. Intact articular cartilage was also examined. RESULTS: The individual major biochemical components of cartilage, Irgacure 2959, and quinine did not exhibit a significant absorption at 365 nm. Riboflavin and riboflavin-5'-phosphate were more effectual light absorbers at 365 nm, compared with the individual native species. Intact cartilage absorbed a significantly greater amount of UV light in comparison with the native species. CONCLUSION: Our results indicate that none of the individual native species in cartilage will interfere with the absorption of UV light at 365 nm by these commonly used photoinitiators. Intact cartilage slices exhibited significant light absorption at 365 nm, while also having distinct absorbance peaks at wavelengths less than 300 nm. Determining the UV absorptive properties of the biomolecules native to articular cartilage and synovial fluid will aid in optimizing scaffolding procedures to ensure sufficient scaffold polymerization at a minimum UV intensity.

Effect of interface mechanical discontinuities on scaffold-cartilage integration.

A consistent lack of lateral integration between scaffolds and adjacent articular cartilage has been exhibited in vitro and in vivo. Given the mismatch in mechanical properties between scaffolds and articular cartilage, the mechanical discontinuity that occurs at the interface has been implicated as a key factor, but remains inadequately studied. Our objective was to investigate how the mechanical environment within a mechanically loaded scaffold-cartilage construct might affect integration. We hypothesized that the magnitude of the mechanical discontinuity at the scaffold-cartilage interface would be related to decreased integration. To test this hypothesis, chondrocyte seeded scaffolds were embedded into cartilage explants, pre-cultured for 14 days, and then mechanically loaded for 28 days at either 1N or 6N of applied load. Constructs were kept either peripherally confined or unconfined throughout the duration of the experiment. Stress, strain, fluid flow, and relative displacements at the cartilage-scaffold interface and within the scaffold were quantified using biphasic, inhomogeneous finite element models (bFEMs). The bFEMs indicated compressive and shear stress discontinuities occurred at the scaffold-cartilage interface for the confined and unconfined groups. The mechanical strength of the scaffold-cartilage interface and scaffold GAG content were higher in the radially confined 1N loaded groups. Multivariate regression analyses identified the strength of the interface prior to the commencement of loading and fluid flow within the scaffold as the main factors associated with scaffold-cartilage integration. Our study suggests a minimum level of scaffold-cartilage integration is needed prior to the commencement of loading, although the exact threshold has yet to be identified. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

Anatomy of the coracoacromial veil.

The coracoacromial (CA) ligament plays an important role in the stability of the shoulder joint by limiting superior translation of the glenohumeral joint. This ligament is further divided into anterolateral and posteromedial bands. Attached to the CA ligament, a supportive structure was noted in some previous studies. The purpose of this study was to learn more about the anatomy of this structure. Twenty-eight shoulders were obtained. Deltoid and trapezius muscles were removed without damaging the rotator cuff and coracoacromial arch. The CA ligament was dissected further to reveal two constituent bands, an anterolateral and a posteromedial band. A connective tissue structure was noted between the posteromedial band, CA ligament, and rotator interval capsule. This structure was oriented as an L-shaped curtain, which the authors termed the "coracoacromial veil." Anatomical position of this veil provides a stabilizing link between the CA ligament and the rotator interval capsule. This structure potentially limits inferior translation of the glenohumeral joint.

Influence of the pericellular and extracellular matrix structural properties on chondrocyte mechanics.

Understanding the mechanical factors that drive the biological responses of chondrocytes is central to our interpretation of the cascade of events that lead to osteoarthritic changes in articular cartilage. Chondrocyte mechanics is complicated by changes in tissue properties that can occur as osteoarthritis (OA) progresses and by the interaction between macro-scale, tissue level, properties, and micro-scale pericellular matrix (PCM) and local extracellular matrix (ECM) properties, both of which cannot be easily studied using in vitro systems. Our objective was to study the influence of macro- and micro-scale OA-associated structural changes on chondrocyte strains. We developed a multi-scale finite element model of articular cartilage subjected to unconfined loading, for the following three conditions: (i) normal articular cartilage, (ii) OA cartilage (where macro and micro-scale changes in collagen content, matrix modulus, and permeability were modeled), and (iii) early-stage OA cartilage (where only micro-scale changes in matrix modulus were modeled). In the macro-scale model, we found that a depth-dependent strain field was induced in both healthy and OA cartilage and that the middle and superficial zones of OA cartilage had increased tensile and compressive strains. At the micro-scale, chondrocyte shear strains were sensitive to PCM and local ECM properties. In the early-OA model, micro-scale spatial softening of PCM and ECM resulted in a substantial increase (30%) of chondrocyte shear strain, even with no structural changes in macro-scale tissue properties. Our study provides evidence that micromechanical changes at the cellular level may affect chondrocyte activities before macro-scale degradations at the tissue level become apparent. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:721-729, 2018.

Articular cartilage response to a sliding load using two different-sized spherical indenters1.

BACKGROUND: Cartilage surface contact geometry influences the deformational behavior and stress distribution throughout the extracellular matrix (ECM) under load. OBJECTIVE: To test the correlation between the mechanical and cellular response of articular cartilage when loaded with two different-sized spherical indenters under dynamic reciprocating sliding motion. METHODS: Articular cartilage explants were subjected to a reciprocating sliding load using a 17.6 mm or 30.2 mm spherical ball for 2000 cycles at 10 mm/s and 4 kg axial load. Deformation of the cartilage was recorded and contact parameters were calculated according to Hertzian theory. After mechanical loading cartilage samples were collected and analyzed for ECM collagen damage, gene regulation and proteoglycan (PG) loss. RESULTS: Significantly higher ECM deformation and strain and lower dynamic effective modulus were found for explants loaded with the smaller diameter indenter whereas contact radius and stress remained unaffected. Also, the 17.6 mm indenter increased PG loss and significantly upregulated genes for ECM proteins and enzymes as compared to the 30.2 mm indenter. CONCLUSION: Sliding loads that increase ECM deformation/strain were found to induce enzyme-mediated catabolic processes in articular cartilage explants. These observations provide further understanding of how changes in cartilage contact mechanics under dynamic conditions can affect the cellular response.

Photocrosslinked tyramine-substituted hyaluronate hydrogels with tunable mechanical properties improve immediate tissue-hydrogel interfacial strength in articular cartilage.

Articular cartilage lacks the ability to self-repair and a permanent solution for cartilage repair remains elusive. Hydrogel implantation is a promising technique for cartilage repair; however for the technique to be successful hydrogels must interface with the surrounding tissue. The objective of this study was to investigate the tunability of mechanical properties in a hydrogel system using a phenol-substituted polymer, tyramine-substituted hyaluronate (TA-HA), and to determine if the hydrogels could form an interface with cartilage. We hypothesized that tyramine moieties on hyaluronate could crosslink to aromatic amino acids in the cartilage extracellular matrix. Ultraviolet (UV) light and a riboflavin photosensitizer were used to create a hydrogel by tyramine self-crosslinking. The gel mechanical properties were tuned by varying riboflavin concentration, TA-HA concentration, and UV exposure time. Hydrogels formed with a minimum of 2.5 min of UV exposure. The compressive modulus varied from 5 to 16 kPa. Fluorescence spectroscopy analysis found differences in dityramine content. Cyanine-3 labelled tyramide reactivity at the surface of cartilage was dependent on the presence of riboflavin and UV exposure time. Hydrogels fabricated within articular cartilage defects had increasing peak interfacial shear stress at the cartilage-hydrogel interface with increasing UV exposure time, reaching a maximum shear stress 3.5× greater than a press-fit control. Our results found that phenol-substituted polymer/riboflavin systems can be used to fabricate hydrogels with tunable mechanical properties and can interface with the surface tissue, such as articular cartilage.

Shape of chondrocytes within articular cartilage affects the solid but not the fluid microenvironment under unconfined compression.

Metabolic activity of the chondrocytes in articular cartilage is strongly related to their zone-specific shape and the composition and mechanical properties of their surrounding extracellular matrix (ECM). However the mechanisms by which cell shape influences the response of the ECM microenvironment to mechanical loading is yet to be elucidated. This relationship was studied using a biphasic multiscale finite element model of different shaped chondrocytes in the superficial and deep zones of the ECM during unconfined stress relaxation. For chondrocytes in the superficial zone, increasing the cell's initial aspect ratio (length/height) increased the deformation and solid stresses of the chondrocyte and pericellular matrix (PCM) during the loading phase; for chondrocytes in the deep zone the effect of the cell shape on the solid microenvironment was time and variable dependent. However, for superficial and deep zone chondrocytes the cell shape did not affect the fluid pressure and fluid shear stress. These results suggest that mechanotransduction of chondrocytes in articular cartilage may be regulated through the solid phase rather than the fluid phase, and that high stresses and deformations in the solid microenvironment in the superficial zone may be essential for the zone-specific biosynthetic activity of the chondrocyte. The biphasic multiscale computational analysis suggests that maintaining the cell shape is critical for regulating the microenvironment and metabolic activity of the chondrocyte in tissue engineering constructs. STATEMENT OF SIGNIFICANCE: We investigated the effect of chondrocyte shape on the cellular microenvironment using a biphasic multiscale finite element analysis. Our study showed that cell shapes affects the solid but not the fluid microenvironment of the chondrocyte, and that maintaining the cell shape is critical for regulating the microenvironment and metabolic activity of the chondrocyte in native cartilage and tissue engineering constructs. As far as we know, this is the first study on the mechanotransduction mechanisms by which cell shape influences the response of the microenvironment to mechanical loading. This study is important for understanding cell mechanobiology, not only for regulation of cell phenotype in tissue engineered constructs but, as important, for understanding changes in normal chondrocyte function after post-traumatic injury and in the initiation and progression of osteoarthritis.

A Model to Study Articular Cartilage Mechanical and Biological Responses to Sliding Loads.

In physiological conditions, joint function involves continuously moving contact areas over the tissue surface. Such moving contacts play an important role for the durability of the tissue. It is known that in pathological joints these motion paths and contact mechanics change. Nevertheless, limited information exists on the impact of such physiological and pathophysiological dynamic loads on cartilage mechanics and its subsequent biological response. We designed and validated a mechanical device capable of applying simultaneous compression and sliding forces onto cartilage explants to simulate moving joint contact. Tests with varying axial loads (1-4 kg) and sliding speeds (1-20 mm/s) were performed on mature viable bovine femoral condyles to investigate cartilage mechanobiological responses. High loads and slow sliding speeds resulted in highest cartilage deformations. Contact stress and effective cartilage moduli increased with increasing load and increasing speed. In a pilot study, changes in gene expression of extracellular matrix proteins were correlated with strain, contact stress and dynamic effective modulus. This study describes a mechanical test system to study the cartilage response to reciprocating sliding motion and will be helpful in identifying mechanical and biological mechanisms leading to the initiation and development of cartilage degeneration.

Use of novel chitosan hydrogels for chemical tissue bonding of autologous chondral transplants.

The objective of this study was to evaluate the effect of chemical tissue bonding (CTB) on adhesion strength, fluid permeability, and cell viability across a cartilaginous graft-host interface in an in vitro autologous chondral transplant (ACT) model. Chitosan-based cross-linkers; Chitosan-Rose Bengal [Chi-RB (Ch-ABC)], Chitosan-Genipin [Chi-GP (Ch-ABC)], and Chitosan-Rose Bengal-Genipin [Chi-RB-GP (Ch-ABC)] were applied to bovine immature cartilage explants after pre-treatment with surface degrading enzyme, Chondroitinase-ABC (Ch-ABC). Adhesion strength, fluid permeability and cell viability were assessed via mechanical push-out shear testing, fluid transport and live/dead cell staining, respectively. All three chitosan-based cross-linkers significantly increased the adhesion strength at the graft-host interface, however, only a statistically significant decrease in fluid permeability was noted in Chi-GP (Ch-ABC) specimen compared to untreated controls. Cell viability was maintained for 7 days of culture across all three treatment groups. These results show the potential clinical relevance of novel chitosan-based hydrogels in enhancing tissue integration and reducing synovial fluid penetration after ACT procedures in diarthoidal joints such as the knee and ankle. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1139-1146, 2016.

Influence of stress rate on water loss, matrix deformation and chondrocyte viability in impacted articular cartilage.

The biomechanical response of articular cartilage to a wide range of impact loading rates was investigated for stress magnitudes that exist during joint trauma. Viable, intact bovine cartilage explants were impacted in confined compression with stress rates of 25, 50, 130 and 1000 MPa/s and stress magnitudes of 10, 20, 30 and 40 MPa. Water loss, cell viability, dynamic impact modulus (DIM) and matrix deformation were measured. Under all loading conditions the water loss was small (approximately 15%); water loss increased linearly with increasing peak stress and decreased exponentially with increasing stress rate. Cell death was localized within the superficial zone (< or =12% of total tissue thickness); the depth of cell death from the articular surface increased with peak stress and decreased with increasing stress rate. The DIM increased (200-700 MPa) and matrix deformation decreased with increasing stress rate. Initial water and proteoglycan (PG) content had a weak, yet significant influence on water loss, cell death and DIM. However, the significance of the inhomogeneous structure and composition of the cartilage matrix was accentuated when explants impacted on the deep zone had less water loss and matrix deformation, higher DIM, and no cell death compared to explants impacted on the articular surface. The mechano-biological response of articular cartilage depended on magnitude and rate of impact loading.

An in vivo rabbit model for cartilage trauma: a preliminary study of the influence of impact stress magnitude on chondrocyte death and matrix damage.

OBJECTIVE: This study was designed to evaluate the postimpact response of the articular cartilage in the rabbit knee after a single traumatic episode. DESIGN: A novel servo-controlled Rabbit Impact Test System (RITS) was developed to apply a well-defined trauma to the femoral condyle in the rabbit knee. The RITS was first used in an in vitro study to determine an appropriate stress to cause cartilage damage without bone fracture. Viable rabbit knees (n = 18) were impacted with stresses of 15 to 50 MPa at a stress rate of 420 MPa/s, the latter corresponding to joint impact rates commonly seen in sports injuries and vehicular accidents. Based on the in vitro study, we performed an in vivo study by impacting the knees of rabbits (n = 9) with a 35 MPa peak stress at a stress rate of 420 MPa/s. The articular cartilage in these knees was analyzed at 0 and 3 weeks after impaction. SETTING: Center for Laboratory Animal Services, Hospital for Special Surgery. SUBJECTS: A total of 27 New Zealand White rabbits were used in this study. INTERVENTION: A rabbit's knee was rigidly immobilized in the adjustable frame of the RITS. A small incision on the knee exposed the lateral condyle and the impactor was positioned perpendicular to the surface of the condyle. The lateral femoral condyle of the left knee was impacted, whereas the right knee was sham operated and used as a control. MAIN OUTCOME MEASUREMENTS: Visual matrix damage, cell viability, and microscopic matrix damage was assessed. RESULTS: In the in vitro study, matrix damage was observed at stress magnitudes > or =30 MPa. However, cell death was initiated at approximately 20 MPa at the articular surface and increased in depth with increasing stress magnitude (2.8 +/- 2% thickness/MPa,). In the in vivo study, visible surface damage was observed immediately after impaction but not at 3 weeks after impaction. At 3 weeks, the articular cartilage showed significant arthritic changes (matrix damage, chondrocyte death, and proteoglycan loss) typical of late-stage osteoarthritis. CONCLUSIONS: Our novel impact test system was able to accurately apply a quantifiable stress magnitude at a constant stress rate to rabbit femoral condyles in the in vitro and in vivo settings. At the time of impaction, the extent of cell death depended with the intensity of trauma (stress magnitude) in which complete cell death was observed in the impacted site at >40 MPa. Under in vivo conditions, the test system was able to consistently produce superficial matrix damage and cell death at 35 MPa stress magnitude at the time of impaction. This resulted in cartilage "arthritic" changes by 3 weeks postinjury.