Compressive nanomechanics of opposing aggrecan macromolecules

In this study, we have measured the nanoscale compressive interactions between opposing aggrecan macromolecules in near-physiological conditions, in order to elucidate the molecular origins of tissue-level cartilage biomechanical behavior. Aggrecan molecules from fetal bovine epiphyseal cartilage were chemically end-grafted to planar substrates, standard nanosized atomic force microscopy (AFM) probe tips (R(tip) approximately 50 nm), and larger colloidal probe tips (R(tip) approximately 2.5 microm). To assess normal nanomechanical interaction forces between opposing aggrecan layers, substrates with microcontact printed aggrecan were imaged using contact mode AFM, and aggrecan layer height (and hence deformation) was measured as a function of solution ionic strength (IS) and applied normal load. Then, using high-resolution force spectroscopy, nanoscale compressive forces between opposing aggrecan on the tip and substrate were measured versus tip-substrate separation distance in 0.001-1M NaCl. Nanosized tips enabled measurement of the molecular stiffness of 2-4 aggrecan while colloidal tips probed the nanomechanical properties of larger assemblies (approximately 10(4) molecules). The compressive stiffness of aggrecan was much higher when using a densely packed colloidal tip than the stiffness measured for using the nanosized tip with a few aggrecan, demonstrating the importance of lateral interactions to the normal nanomechanical properties. The measured stress at 0.1M NaCl (near-physiological ionic strength) increased sharply at aggrecan densities under the tip of approximately 40 mg/ml (physiological densities are approximately 20-80 mg/ml), corresponding to an average inter-GAG spacing of 4-5 Debye lengths (4-5 nm); this characteristic spacing is consistent with the onset of significant electrostatic interactions between GAG chains of opposing aggrecan molecules. Comparison of nanomechanical data to the predictions of Poisson-Boltzmann-based models further elucidated the regimes over which electrostatic and nonelectrostatic interactions affect aggrecan stiffness in compression. The most important aspects of this study include: the incorporation of experiments at two different length scales, the use of microcontact printing to enable quantification of aggrecan deformation and the corresponding nanoscale compressive stress vs. strain curve, the use of tips of differing functionality to provide insights into the molecular mechanisms of deformation, and the comparison of experimental data to the predictions of three increasingly refined Poisson-Boltzmann (P-B)-based theoretical models for the electrostatic double layer component of the interaction.     

Nanoscale shear deformation mechanisms of opposing cartilage aggrecan macromolecules

The nanoscale shear deformation behavior of two opposing end-grafted aggrecan layers was studied in aqueous solutions using atomic force microscopy, and was observed to depend markedly on bath ionic strength, the presence of calcium ions, and the applied lateral displacement rate. These results provide molecular-level insights into the contribution of aggrecan deformation mechanisms to cartilage tissue-level material properties.     

Mapping the depth dependence of shear properties in articular cartilage

Determining the depth dependence of the shear properties of articular cartilage is essential for understanding the structure-function relation in this tissue. Here, we measured spatial variations in the shear modulus G of bovine articular cartilage using a novel technique that combines shear testing, confocal imaging and force measurement. We found that G varied by up to two orders of magnitude across a single sample, exhibited a global minimum 50-250 microm below the articular surface in a region just below the superficial zone and was roughly constant at depths > 1000 microm (the "plateau region"). For plateau strains gamma(plateau) approximately 0.75% and overall compressive strains epsilon approximately 5%, G(min) and G(plateau) were approximately 70 and approximately 650 kPa, respectively. In addition, we found that the shear modulus profile depended strongly on the applied shear and axial strains. The greatest change in G occurred at the global minimum where the tissue was highly nonlinear, stiffening under increased shear strain, and weakening under increased compressive strain. Our results can be explained through a simple thought model describing the observed nonlinear behavior in terms of localized buckling of collagen fibers and suggest that compression may decrease the vulnerability of articular cartilage to shear-induced damage by lowering the effective strain on individual collagen fibrils.     

Age-related nanostructural and nanomechanical changes of individual human cartilage aggrecan monomers and their glycosaminoglycan side chains

The nanostructure and nanomechanical properties of aggrecan monomers extracted and purified from human articular cartilage from donors of different ages (newborn, 29 and 38 year old) were directly visualized and quantified via atomic force microscopy (AFM)-based imaging and force spectroscopy. AFM imaging enabled direct comparison of full length monomers at different ages. The higher proportion of aggrecan fragments observed in adult versus newborn populations is consistent with the cumulative proteolysis of aggrecan known to occur in vivo. The decreased dimensions of adult full length aggrecan (including core protein and glycosaminoglycan (GAG) chain trace length, end-to-end distance and extension ratio) reflect altered aggrecan biosynthesis. The demonstrably shorter GAG chains observed in adult full length aggrecan monomers, compared to newborn monomers, also reflects markedly altered biosynthesis with age. Direct visualization of aggrecan subjected to chondroitinase and/or keratanase treatment revealed conformational properties of aggrecan monomers associated with chondroitin sulfate (CS) and keratan sulfate (KS) GAG chains. Furthermore, compressive stiffness of chemically end-attached layers of adult and newborn aggrecan was measured in various ionic strength aqueous solutions. Adult aggrecan was significantly weaker in compression than newborn aggrecan even at the same total GAG density and bath ionic strength, suggesting the importance of both electrostatic and non-electrostatic interactions in nanomechanical stiffness. These results provide molecular-level evidence of the effects of age on the conformational and nanomechanical properties of aggrecan, with direct implications for the effects of aggrecan nanostructure on the age-dependence of cartilage tissue biomechanical and osmotic properties.     

Cartilage aggrecan can undergo self-adhesion

Here it is reported that aggrecan, the highly negatively charged macromolecule in the cartilage extracellular matrix, undergoes Ca²⁺-mediated self-adhesion after static compression even in the presence of strong electrostatic repulsion in physiological-like solution conditions. Aggrecan was chemically end-attached onto gold-coated planar silicon substrates and gold-coated microspherical atomic force microscope probe tips (end radius R ≈ 2.5 μm) at a density (∼40 mg/mL) that simulates physiological conditions in the tissue (∼20-80 mg/mL). Colloidal force spectroscopy was employed to measure the adhesion between opposing aggrecan monolayers in NaCl (0.001-1.0 M) and NaCl + CaCl₂ ([Cl⁻] = 0.15 M, [Ca²⁺] = 0 - 75 mM) aqueous electrolyte solutions. Aggrecan self-adhesion was found to increase with increasing surface equilibration time upon compression (0-30 s). Hydrogen bonding and physical entanglements between the chondroitin sulfate-glycosaminoglycan side chains are proposed as important factors contributing to aggrecan self-adhesion. Self-adhesion was found to significantly increase with decreasing bath ionic strength (and hence, electrostatic double-layer repulsion), as well as increasing Ca²⁺ concentration due to the additional ion-bridging effects. It is hypothesized that aggrecan self-adhesion, and the macromolecular energy dissipation that results from this self-adhesion, could be important factors contributing to the self-assembled architecture and integrity of the cartilage extracellular matrix in vivo.     

Mechanisms of anterior-posterior stability of the knee joint under load-bearing

The anterior-posterior (AP) stability of the knee is an important aspect of functional performance. Studies have shown that the stability increases when compressive loads are applied, as indicated by reduced laxity, but the mechanism has not been fully explained. A test rig was designed which applied combinations of AP shear and compressive forces, and measured the AP displacements relative to the neutral position. Five knees were evaluated at compressive loads of 0, 250, 500, and 750 N, with the knee at 15° flexion. At each load, three cycles of shear force at ±100 N were applied. For the intact knee under load, the posterior tibial displacement was close to zero, due to the upward slope of the anterior medial tibial surface. The soft tissues were then resected in sequence to determine their role in AP laxity. After anterior cruciate ligament (ACL) resection, the anterior tibial displacement increased significantly even under load, highlighting its importance in stability. Meniscal resection further increased displacement but also the vertical displacement increased, implying the meniscus was providing a buffering effect. The PCL had no effect on any of the displacements under load. Plowing cartilage deformation and surface friction were negligible. This work highlighted the particular importance of the upward slope of the anterior medial tibial surface and the ACL to AP knee stability under load. The results are relevant to the design of total knees which reproduce anatomic knee stability behavior.     

Effect of indenter size on elastic modulus of cartilage measured by indentation

Our preliminary indentation experiments showed that the equilibrium elastic modulus of murine tibial cartilage increased with decreasing indenter size: flat-ended 60 deg conical tips with end diameters of 15 microm and 90 microm gave 1.50+/-0.82 MPa (mean+/-standard deviation) and 0.55+/-0.11 MPa, respectively (p<0.01). The goal of this paper is to determine if the dependence on tip size is an inherent feature of the equilibrium elastic modulus of cartilage as measured by indentation. Since modulus values from nonindentation tests are not available for comparison for murine cartilage, bovine cartilage was used. Flat-ended conical or cylindrical tips with end diameters ranging from 5 microm to 4 mm were used to measure the equilibrium elastic modulus of bovine patellar cartilage. The same tips were used to test urethane rubber for comparison. The equilibrium modulus of the bovine patellar cartilage increased monotonically with decreasing tip size. The modulus obtained from the 2 mm and 4 mm tips (0.63+/-0.21 MPa) agreed with values reported in the literature; however, the modulus measured by the 90 microm tip was over two and a half times larger than the value obtained from the 1000 microm tip. In contrast, the elastic modulus of urethane rubber obtained using the same 5 microm-4 mm tips was independent of tip size. The equilibrium elastic modulus of bovine patellar cartilage measured by indentation depends on tip size. This appears to be an inherent feature of indentation of cartilage, perhaps due to its inhomogeneous structure.     

Force-mediated dissociation of proteoglycan aggregate in articular cartilage

Proteoglycan aggregate is the primary component in articular cartilage responsible for resisting compressive loading. It consists of a core molecule of hyaluronan and a number of side chains of aggrecan bound to hyaluronan non-covalently. The loss of aggrecan from articular cartilage is considered to be a major factor in the development of osteoarthritis. Though enzymatic digestion of aggrecan is believed to be responsible for the release of aggrecan from osteoarthritic cartilage, other mechanisms, such as direct force-mediated detachment of aggrecan from hyaluronan may also be involved. In this study, the rupture force of the single bond between hyaluronan and aggrecan in articular cartilage was directly quantified using experimental measurement and Monte Carlo simulation. Low rupture force of this bond, as determined in this study suggested a possible direct force-mediated detachment of aggrecan from proteoglycan aggregate in osteoarthritic cartilage.     

Microscale frictional response of bovine articular cartilage from atomic force microscopy

The objective of this study was to compare micro- and macroscale friction coefficients of bovine articular cartilage. Microscale measurements were performed using standard atomic force microscopy (AFM) techniques, using a 5 microm spherical probe tip. Twenty-four cylindrical osteochondral plugs were harvested in pairs from adjacent positions in six fresh bovine humeral heads (4-6 months old), and divided into two groups for AFM and macroscopic friction measurements. AFM measurements of friction were observed to be time-independent, whereas macroscale measurements demonstrated the well-documented time-dependent increase from a minimum to an equilibrium value. The microscale AFM friction coefficient (mu(AFM), 0.152+/-0.079) and macroscale equilibrium friction coefficient (mu(eq), 0.138+/-0.036) exhibited no statistical differences (p=0.50), while the macroscale minimum friction coefficient (mu(min), 0.004+/-0.001) was significantly smaller than mu(eq) and mu(AFM) (p<0.0001). Variations in articular surface roughness (Rq= 462+/-216 nm) did not correlate significantly with mu(AFM), mu(eq) or mu(min). The effective compressive modulus determined from AFM indentation tests using a Hertz contact analysis was E*=45.8+/-18.8 kPa. The main finding of this study is that mu(AFM) is more representative of the macroscale equilibrium friction coefficient, which represents the frictional response in the absence of cartilage interstitial fluid pressurization. These results suggest that AFM measurements may be highly suited for exploring the role of boundary lubricants in diarthrodial joint lubrication independently of the confounding effect of fluid pressurization to provide greater insight into articular cartilage lubrication.     

Repeated application of incremental landing impact loads to intact knee joints induces anterior cruciate ligament failure and tibiofemoral cartilage deformation and damage: A preliminary cadaveric investigation

Anterior cruciate ligament (ACL) injury is a major problem worldwide and prevails during high-impact activities. It is not well-understood how the extent and distribution of cartilage damage will arise from repetitive landing impact loads that can lead to ACL failure. This study seeks to investigate the sole effect of repetitive incremental landing impact loads on the induction of ACL failure, and extent and distribution of tibiofemoral cartilage damage in cadaveric knees. Five cadaveric knees were mounted onto a material testing system at 70° flexion to simulate landing posture. A motion-capture system was used to track rotational and translational motions of the tibia and femur, respectively. Each specimen was compressed at a single 10 Hz haversine to simulate landing impact. The compression trial was successively repeated with increasing actuator displacement till a significant compressive force drop was observed. All specimens underwent ACL failure, which was confirmed via magnetic resonance scans and dissection. Volume analysis, thickness measurement and histological techniques were employed to assess cartilage lesion status. For each specimen, the highest peak compressive force (1.9–7.8 kN) was at the final trial in which ACL failure occurred; corresponding posterior femoral displacement (7.6–18.0 mm) and internal tibial rotation (0.6°–4.7°) were observed. Significant compressive force drop (79.8–90.9%) was noted upon ACL failure. Considerable cartilage deformation and damage were found in exterior, posterior and interior femoral regions with substantial volume reduction in lateral compartments. Repeated application of incremental landing impact loads can induce both ACL failure and cartilage damage, which may accelerate the risk of developing osteoarthritis.     

Dynamic unconfined compression of articular cartilage under a cyclic compressive load

To study the effect of dynamic mechanical force on cartilage metabolism, many investigators have applied a cyclic compressive load to cartilage disc explants in vitro. The most frequently used in vitro testing protocol has been the cyclic unconfined compression of articular cartilage in a bath of culture medium. Cyclic compression has been achieved by applying either a prescribed cyclic displacement or a prescribed cyclic force on a loading platen placed on the top surface of a cylindrical cartilage disc. It was found that the separation of the loading platen from the tissue surface was likely when a prescribed cyclic displacement was applied at a high frequency.The purpose of the present study was to simulate mathematically the dynamic behavior of a cylindrical cartilage disc subjected to cyclic unconfined compression under a dynamic force boundary condition protocol, and to provide a parametric analysis of mechanical deformations within the extracellular matrix. The frequency-dependent dynamic characteristics of dilatation, hydrostatic pressure and interstitial fluid velocity were analyzed over a wide range of loading frequencies without the separation of the loading platen. The result predicted that a cyclic compressive force created an oscillating positive-negative hydrostatic pressure together with a forced circulation of interstitial fluid within the tissue matrix. It was also found that the load partitioning mechanism between the solid and fluid phases was a function of loading frequency. At a relatively high loading frequency, a localized dynamic zone was developed near the peripheral free surface of the cartilage disc, where a large dynamic pressure gradient exists, causing vigorous interstitial fluid flow.     

Shear deformation kinematics during cartilage articulation: effect of lubrication, degeneration, and stress relaxation

During joint articulation, the biomechanical behavior of cartilage not only facilitates load-bearing and low-friction, but also provides regulatory cues to chondrocytes. Elucidation of cartilage kinematics under combined compression and shearing conditions clarifies these cues in health and disease. The objectives of this study were to elucidate the effects of lubricant, tissue degeneration, and stress relaxation duration on cartilage shear kinematics during articulation. Human osteochondral cores with normal and mildly degenerate surface structures were isolated. Paired blocks from each core were apposed, compressed, allowed to stress relax for 5 or 60 min, and shear tested with a micro-scale video microscopy system using phosphate-buffered saline (PBS) or synovial fluid as lubricant. During applied lateral motion, local and overall shear strain (Exz) of articular cartilage were determined. The applied lateral displacement at which Exz reached 50% of the peak (Deltax(1/2)) was also determined. Quantitatively, surface Exz increased at the onset of lateral motion and peaked just as surfaces detached and slid. With continued lateral motion, surface Exz was maintained. After short stress relaxation, effects of lubrication on Exz and Deltax(1/2) were not apparent. With prolonged stress relaxation, Exz and Deltax(1/2) near the articular surface increased markedly when PBS was used as lubricant. Similar patterns were observed for overall Exz and Deltax(1/2). With degeneration, surface Exz was consistently higher for all cases after the onset of lateral motion. Thus, cartilage shear kinematics is markedly affected by lubricant, cartilage degeneration, and loading duration. Changes in these factors may be involved in the pathogenesis of osteoarthritis.     

Structure and interactions of aggrecans: statistical thermodynamic approach

Weak polyelectrolytes tethered to cylindrical surfaces are investigated using a molecular theory. These polymers form a model system to describe the properties of aggrecan molecules, which is one of the main components of cartilage. We have studied the structural and thermodynamical properties of two interacting aggrecans with a molecular density functional theory that incorporates the acid-base equilibrium as well as the molecular properties: including conformations, size, shape, and charge distribution of all molecular species. The effect of acidity and salt concentration on the behavior is explored in detail. The repulsive interactions between two cylindrical-shaped aggrecans are strongly influenced by both the salt concentration and the pH. With increasing acidity, the polyelectrolytes of the aggrecan acquire charge and with decreasing salt concentration those charges become less screened. Consequently the interactions increase in size and range with increasing acidity and decreasing salt concentration. The size and range of the forces offers a possible explanation to the aggregation behavior of aggrecans and for their ability to resist compressive forces in cartilage. Likewise, the interdigitation of two aggrecan molecules is strongly affected by the salt concentration as well as the pH. With increasing pH, the number of charges increases, causing the repulsions between the polymers to increase, leading to a lower interdigitation of the two cylindrical polymer layers of the aggrecan molecules. The low interdigitation in charged polyelectrolytes layers provides an explanation for the good lubrication properties of polyelectrolyte layers in general and cartilage in particular.     

Biphasic poroviscoelastic simulation of the unconfined compression of articular cartilage: I--Simultaneous prediction of reaction force and lateral displacement

This study investigated the ability of the linear biphasic poroelastic (BPE) model and the linear biphasic poroviscoelastic (BPVE) model to simultaneously predict the reaction force and lateral displacement exhibited by articular cartilage during stress relaxation in unconfined compression. Both models consider articular cartilage as a binary mixture of a porous incompressible solid phase and an incompressible inviscid fluid phase. The BPE model assumes the solid phase is elastic, while the BPVE model assumes the solid phase is viscoelastic. In addition, the efficacy of two additional models was also examined, i.e., the transversely isotropic BPE (TIBPE) model, which considers transverse isotropy of the solid matrix within the framework of the linear BPE model assumptions, and a linear viscoelastic solid (LVE) model, which assumes that the viscoelastic behavior of articular cartilage is solely governed by the intrinsic viscoelastic nature of the solid matrix, independent of the interstitial fluid flow. It was found that the BPE model was able to accurately account for the lateral displacement, but unable to fit the short-term reaction force data of all specimens tested. The TIBPE model was able to account for either the lateral displacement or the reaction force, but not both simultaneously. The LVE model was able to account for the complete reaction force, but unable to fit the lateral displacement measured experimentally. The BPVE model was able to completely account for both lateral displacement and reaction force for all specimens tested. These results suggest that both the fluid flow-dependent and fluid flow-independent viscoelastic mechanisms are essential for a complete simulation of the viscoelastic phenomena of articular cartilage.     

Effect of dynamic loading on the frictional response of bovine articular cartilage

The objective of this study was to test the hypotheses that (1) the steady-state friction coefficient of articular cartilage is significantly smaller under cyclical compressive loading than the equilibrium friction coefficient under static loading, and decreases as a function of loading frequency; (2) the steady-state cartilage interstitial fluid load support remains significantly greater than zero under cyclical compressive loading and increases as a function of loading frequency. Unconfined compression tests with sliding of bovine shoulder cartilage against glass in saline were carried out on fresh cylindrical plugs (n=12), under three sinusoidal loading frequencies (0.05, 0.5 and 1 Hz) and under static loading; the time-dependent friction coefficient mu(eff) was measured. The interstitial fluid load support was also predicted theoretically. Under static loading mu(eff) increased from a minimum value (mu(min)=0.005+/-0.003) to an equilibrium value (mu(eq)=0.153+/-0.032). In cyclical compressive loading tests mu(eff) similarly rose from a minimum value (mu(min)=0.004+/-0.002, 0.003+/-0.001 and 0.003+/-0.001 at 0.05, 0.5 and 1 Hz) and reached a steady-state response oscillating between a lower-bound (mu(lb)=0.092+/-0.016, 0.083+/-0.019 and 0.084+/-0.020) and upper bound (mu(ub)=0.382+/-0.057, 0.358+/-0.059, and 0.298+/-0.061). For all frequencies it was found that mu(ub)>mu(eq) and mu(lb)相似文献   

Articular cartilage proteoglycans as boundary lubricants: structure and frictional interaction of surface-attached hyaluronan and hyaluronan--aggrecan complexes

Mammalian synovial joints are extremely efficient lubrication systems reaching friction coefficient μ as low as 0.001 at high pressures (up to 100 atm) and shear rates (up to 10(6) to 10(7) Hz); however, despite much previous work, the exact mechanism responsible for this behavior is still unknown. In this work, we study the molecular mechanism of synovial joint lubrication by emulating the articular cartilage superficial zone structure. Macromolecules extracted and purified from bovine hip joints using well-known biochemical techniques and characterized with atomic force microscope (AFM) have been used to reconstruct a hyaluronan (HA)--aggrecan layer on the surface of molecularly smooth mica. Aggrecan forms, with the help of link protein, supramolecular complexes with the surface-attached HA similar to those at the cartilage/synovial fluid interface. Using a surface force balance (SFB), normal and shear interactions between a HA--aggrecan-coated mica surface and bare mica have been examined, focusing, in particular, on the frictional forces. In each stage, control studies have been performed to ensure careful monitoring of the macromolecular surface layers. We found the aggrecan--HA complex to be a much better boundary lubricant than the HA alone, an effect attributed largely to the fluid hydration sheath bound to the highly charged glycosaminoglycan (GAG) segments on the aggrecan core protein. A semiquantitative model of the osmotic pressure is used to describe the normal force profiles between the surfaces and interpret the boundary lubrication mechanism of such layers.     

In situ friction measurement on murine cartilage by atomic force microscopy

Articular cartilage provides a low-friction, wear-resistant surface for the motion of diarthrodial joints. The objective of this study was to develop a method for in situ friction measurement of murine cartilage using a colloidal probe attached to the cantilever of an atomic force microscope. Sliding friction was measured between a chemically functionalized microsphere and the cartilage of the murine femoral head. Friction was measured at normal loads ranging incrementally from 20 to 100 nN with a sliding speed of 40 microm/s and sliding distance of 64 microm. Under these test conditions, hydrostatic pressurization and biphasic load support in the cartilage were minimized, providing frictional measurements that predominantly reflect boundary lubrication properties. Friction coefficients measured on murine tissue (0.25+/-0.11) were similar to those measured on porcine tissue (0.23+/-0.09) and were in general agreement with measurements of boundary friction on cartilage by other researchers. Using the colloidal probe as an indenter, the elastic mechanical properties and surface roughness were measured in the same configuration. Interfacial shear was found to be the principal mechanism of friction generation, with little to no friction resulting from plowing forces, collision forces, or energy losses due to normal deformation. This measurement technique can be applied to future studies of cartilage friction and mechanical properties on genetically altered mice or other small animals.     

Regional structural and viscoelastic properties of fibrocartilage upon dynamic nanoindentation of the articular condyle

Fibrocartilage,a tissue with macromaterial properties between dense fibrous tissue and hyaline cartilage, is not well understood in its ultrastructure and regional viscoelastic properties. Here nanoindentation with atomic force microscopy was performed on fresh fibrocartilage samples of rabbit jaw joint condyles. Each sample was divided into anteromedial, anterolateral, posteromedial, and posterolateral regions for probing and topographic imaging in 2 x 2 microm and 10 x 10 microm scan sizes. Young's moduli differed significantly among these regions in a descending gradient from the anteromedial (2.34 +/- 0.26 MPa) to the posterolateral (0.95 +/- 0.06 MPa). The Poisson ratio, defined as lateral strain over axial strain, had the same gradient distribution: highest for the anteromedial region (0.46 +/- 0.05) and lowest for the posterolateral region (0.31 +/- 0.05). The same four regions showed a descending gradient of surface roughness: highest for the anteromedial (321.6 +/- 13.8 nm) and lowest for the posterolateral (155.6 +/- 12.6 nm). Thus, the regional ultrastructural and viscoelastic properties of fibrocartilage appear to be coregulated. Based on these region-specific gradient distributions, fibrocartilage is constructed to withstand tissue-borne shear stresses, which likely propagate across its different regions. A model of shear gradient and concentric gradient is proposed to describe the region-specific capacity of fibrocartilage to sustain shear stresses in tendons, ligaments, joints, and the healing bone across species.     

Micropipette aspiration on the outer hair cell lateral wall

The mechanical properties of the lateral wall of the guinea pig cochlear outer hair cell were studied using the micropipette aspiration technique. A fire-polished micropipette with an inner diameter of approximately 4 microm was brought into contact with the lateral wall and negative pressure was applied. The resulting deformation of the lateral wall was recorded on videotape and subjected to morphometric analysis. The relation between the length of the aspirated portion of the cell and aspiration pressure is characterized by the stiffness parameter, K(s) = 1.07 +/- 0.24 (SD) dyn/cm (n = 14). Values of K(s) do not correlate with the original cell length, which ranges from 29 to 74 microm. Theoretical analysis based on elastic shell theory applied to the experimental data yields an estimate of the effective elastic shear modulus, mu = 15.4 +/- 3.3 dyn/cm. These data were obtained at subcritical aspiration pressures, typically less than 10 cm H2O. After reaching a critical (vesiculation) pressure, the cytoplasmic membrane appeared to separate from the underlying structures, a vesicle with a length of 10-20 microm was formed, and the cytoplasmic membrane resealed. This vesiculation process was repeated until a cell-specific limit was reached and no more vesicles were formed. Over 20 vesicles were formed from the longest cells in the experiment.     

Direct contribution of axial impact compressive load to anterior tibial load during simulated ski landing impact

Anterior tibial loading is a major factor involved in the anterior cruciate ligament (ACL) injury mechanism during ski impact landing. We sought to investigate the direct contribution of axial impact compressive load to anterior tibial load during simulated ski landing impact of intact knee joints without quadriceps activation. Twelve porcine knee specimens were procured. Four specimens were used as non-impact control while the remaining eight were mounted onto a material-testing system at 70° flexion and subjected to simulated landing impact, which was successively repeated with incremental actuator displacement. Four specimens from the impacted group underwent pre-impact MRI for tibial plateau angle measurements while the other four were subjected to histology and microCT for cartilage morphology and volume assessment. The tibial plateau angles ranged from 29.4 to 38.8°. There was a moderate linear relationship (Y=0.16X; R²=0.64; p<0.001) between peak axial impact compressive load (Y) and peak anterior tibial load (X). The anterior and posterior regions in the impacted group sustained surface cartilage fraying, superficial clefts and tidemark disruption, compared to the control group. MicroCT scans displayed visible cartilage deformation for both anterior and posterior regions in the impacted group. Due to the tibial plateau angle, increased axial impact compressive load can directly elevate anterior tibial load and hence contribute to ACL failure during simulated landing impact. Axial impact compressive load resulted in shear cartilage damage along anterior–posterior tibial plateau regions, due to its contribution to anterior tibial loading. This mechanism plays an important role in elevating ACL stress and cartilage deformation during impact landing.