Mechanical response of bovine articular cartilage under dynamic unconfined compression loading at physiological stress levels

OBJECTIVE: The objective of this study was to characterize the dynamic modulus and compressive strain magnitudes of bovine articular cartilage at physiological compressive stress levels and loading frequencies. DESIGN: Twelve distal femoral cartilage plugs (3mm in diameter) were loaded in a custom apparatus under load control, with a load amplitude up to 40 N and loading frequencies of 0.1, 1, 10 and 40 Hz, resulting in peak Cauchy stress amplitudes of 4.8 MPa (engineering stress 5.7 MPa). RESULTS: The equilibrium Young's modulus under a tare load of 0.4N was 0.49+/-0.10 MPa. In the limit of zero applied stress, the incremental dynamic modulus derived from the slope of the stress-strain curve increased from 14.6+/-6.9 MPa at 0.1 Hz to 28.7+/-7.8 MPa at 40 Hz. At 4 MPa of applied stress, the corresponding increase was from 48.2+/-13.5 MPa at 0.1 Hz to 64.8+/-13.0 MPa at 40 Hz. Peak compressive strain amplitudes varied from 15.8+/-3.4% at 0.1 Hz to 8.7+/-1.8% at 40 Hz. The phase angle decreased from 28.8 degrees +/-6.7 degrees at 0.1 Hz to-0.5 degrees +/-3.8 degrees at 40 Hz. DISCUSSION: These results are representative of the functional properties of articular cartilage under physiological load magnitudes and frequencies. The viscoelasticity and nonlinearity of the tissue helps to maintain the compressive strains below 20% under the physiological compressive stresses achieved in this study. These findings have implications for our understanding of cartilage metabolism and chondrocyte viability under various loading regimes. They also help establish guidelines for cartilage functional tissue engineering studies.     

Deformation of loaded articular cartilage prepared for scanning electron microscopy with rapid freezing and freeze-substitution fixation

To investigate the effect of joint loading on collagen fibers in articular cartilage, 45 knees of adult rabbits were examined by scaning electron microscopy. The knees were loaded at the patella with a simulated “quadriceps force” of 0.5-4 times body weight for 0.5 or 25 minutes, plunge-frozen, and fixed by freezesubstitution with aldehydes. Six knees were loaded for 3 hours and then fixed conventionally. Fixed tibial plateaus were examined and then freeze-fractured through the area of tibiofemoral contact, dried, coated, and examined by scanning electron microscopy to assess the overall deformation of the tibial articular surface and matrix collagen fibers. With tissue prepared by conventional fixation used as a standard, the quality of fixation was graded by light and transmission electron microscopy of patellar cartilage taken from half of the freeze-fixed knees. In loaded specimens, an indentation was present where the femur contacted the tibial plateau. The diameter and apparent depth of the dent were proportional to the magnitude and duration of the load; no dent was seen in the controls. The thickness of the cartilage at the center of the indentation was reduced 15-80%. Meniscectomy always produced larger deformations in otherwise equivalent conditions. Icecrystal damage to cells was evident by transmission electron microscopy and scanning electron microscopy, but at magnifications as high as ×30,000 the collagen fibrils prepared by freeze-substitution and conventional aqueous methods were identical. In loaded regions, the collagen matrix of the tibial cartilage was deformed in two ways: (a) radial collagen fibers exhibited a periodic crimp, and (b) in regions where an indentation was created by the femoral condyle, the radial fibers were bent, in effect creating tangential zone where none had existed before. The radial fibers apparently are loaded axially and buckle under normal loads.     

How severe must repetitive loading be to kill chondrocytes in articular cartilage?

OBJECTIVE: Little is known about the effects of severe repetitive loading on articular cartilage chondrocytes, even though epidemiological studies associate this type of loading with osteoarthritis. We hypothesize that repetitive loading can kill cartilage chondrocytes in a dose-related manner. DESIGN: Large cartilage-on-bone specimens were cut from the patella groove of bovine knees obtained directly from a slaughterhouse. Cartilage was loaded using a flat impermeable indenter in such a manner that the loaded region was supported naturally by surrounding cartilage and subchondral bone. Specimens received 3600 cycles of compressive loading at 1 Hz, with the peak load lying in the range 1-70% of the force required to damage cartilage in a single loading cycle (35 MPa). Cell viability was assessed in thick sections of loaded and control cartilage using a paravital staining method: fluorescein diacetate stained live cells green, and propidium iodide stained dead cells red. The assay was validated on cartilage which had been subjected to repeated freeze-thaw cycles to kill the chondrocytes. RESULTS: Paravital staining revealed 100% cell death after one freeze-thaw cycle at -196 degrees C and three cycles at -20 degrees C. Baseline chondrocyte viability was 80% in unloaded cartilage, and viability decreased when applied compressive loading exceeded 6 MPa. Above this threshold, cell viability was inversely proportional to applied stress. When gross damage to the cartilage surface first became evident, above 14 MPa, 40% of cells remained viable. Load-induced chondrocyte death was greatest in the surface zone, and extended beyond the loaded area. Electron micrographs indicated that some cells were dying by apoptosis. CONCLUSIONS: Some chondrocytes are much more vulnerable to repetitive mechanical loading than others, suggesting that vigorous activity may lead to cell death in articular cartilage.     

Articular cartilage preservation and storage. III. Quantitative zonal analysis of cytoplasmic components of stored versus in vivo articular cartilage chondrocytes

The cytoplasmic components of chondrocytes in the various zones of articular cartilage of the medial femoral condyle of six-week-old male New Zealand white rabbits stored in tissue culture medium at 37 degrees in 5% CO2 and air were quantitated from electron micrographs, and the results were compared statistically with the cytoplasmic components of chondrocytes in the corresponding zones of in vivo articular cartilage. The major changes that occurred during storage were: (1) an increase in the amount of lipid bodies in chondrocytes in the tangential, transitional, and calcified zones; (2) a decrease in the number of holes in the cytoplasm of chondrocytes in the radial and calcified zones; (3) a decrease in the amount of endoplasmic reticulum in the radial and calcified zones; and (4) an increase in cell size and cytosol area in the tangential and transitional zones but a significant decrease in cell size and cytosol area in the calcified zone. Stored articular cartilage chondrocytes demonstrated cellular changes associated with aging, whereas in vivo articular cartilage chondrocytes demonstrated changes associated with degeneration. The matrix of stored articular cartilage in the tangential, transitional, and upper part of the radial zones showed a decrease in opacity due to a decrease in the number of collagen fibers per unit area of matrix, a condition termed "chondroporosis." This study demonstrates that articular cartilage stored in standard tissue culture medium under ideal physiological conditions is morphologically abnormal. Based on these findings, one would not expect such stored cartilage to remain functionally intact when transplanted to replace articular cartilage loss.     

The acute structural changes of loaded articular cartilage following meniscectomy or ACL-transection

Objective Meniscectomy and anterior cruciate ligament (ACL) rupture have been identified as precursors of osteoarthrosis (OA) in clinical reviews and animal experiments. In this study, the acute effects of these injuries on articular cartilage matrix deformation, preserved in a loaded state using a cryopreservation technique, were studied by scanning electron microscopy (SEM).Method Whole knee joints from adult White New Zealand rabbits (N=87) were loaded ex vivo, using a simulated quadriceps pull under static and cyclic loading conditions, following medial meniscectomy or transection of the ACL. Specimens were plunge-frozen while under load, or following a recovery period, and prepared for SEM by cryofixation. Using SEM and photographic images, the medial tibial plateau cartilage was assessed both qualitatively and quantitatively.Results After meniscectomy, significantly increased bending and crimping of radial collagen fibers occurred with static loading. Compared to intact knees, the area of tibial cartilage showing an indentation was increased by 80% (P< 0.05), the articular cartilage thickness was significantly more reduced when under load (for high force long duration static loading, intact joints had 53%±3 reduction in cartilage thickness compared to 39%±4 after meniscectomy, P< 0.05), and it took nearly twice as long for the cartilage thickness to recover following loading. These post-meniscectomy differences were either not present or were minimal when the joint was allowed to extend when loaded. ACL-transection slightly increased collagen deformation in the deeper zones, but only with cyclic loading.Conclusion The findings indicate that, with static loading, significantly increased deformation of articular cartilage collagen structure can occur following meniscectomy, but is minimized by joint motion. This increased deformation may be relevant to the etiology and progression of joint degeneration.     

Articular cartilage functional histomorphology and mechanobiology: a research perspective

The histomorphogenesis of articular cartilage is regulated during skeletal development by the intermittent forces and motions imposed at diarthrodial joints. A key feature in this development is the formation of the superficial, transitional, radial, and calcified cartilage zones through the cartilage thickness. The histomorphological, biological, and mechanical characteristics of these zones can be correlated with the distributions of pressures, deformations, and pressure-induced fluid flow that are created in vivo. In a mature joint, cyclic loads produce cyclic hydrostatic fluid pressure through the entire cartilage thickness that is comparable in magnitude to the applied joint pressure. Prolonged physical activity can cause the total cartilage thickness to decrease about 5%, although the consolidation strains vary tremendously in the different zones. The superficial zone can experience significant fluid exudation and consolidation (compressive strains) in the range of 60% while the radial zone experiences relatively little fluid flow and consolidation. The topological variation in the histomorphologic appearance of articular cartilage is influenced by the local mechanical loading of chondrocytes in the different zones. Patterns of stress, strain, and fluid flow created in the joint result in spatial and temporal changes in the rates of synthesis and degradation of matrix proteins. When viewed over the course of a lifetime, even subtle difference in these cellular processes can affect the micro- and macro-morphology of articular cartilage. This hypothesis is supported by in vivo and ex vivo experiments where load-induced changes in matrix synthesis and catabolism, gene expression, and signal transduction pathways have been observed.     

Cartilage viability after repetitive loading: a preliminary report.

OBJECTIVE: To assess matrix changes and chondrocyte viability during static and continuous repetitive mechanical loading in mature bovine articular cartilage explants. METHODS: Cartilage explants were continuously loaded either statically or cyclically (0.5 Hz) for 1-72 h (max. stress 1 megapascal). Cell death was assessed using fluorescent probes and detection of DNA strand breakage characteristic of apoptosis. Cell morphology and matrix integrity were evaluated using histology and transmission electron microscopy. RESULTS: Repetitive loading of articular cartilage at physiological levels of stress (1 megapascal) was found to be harmful to only the chondrocytes in the superficial tangential zone (STZ) and depended on the characteristics (static vs cyclic) and duration (1-72 h) of the applied load. The chondrocytes in the middle and deep zone remained viable at all times. Static loads caused cell death at an early time (3 h) as compared with cyclic loads (sinusoidal, 0.5 cycles per s for 6 h). The amount and extent of cell death peaked at 6 h of cyclic loading, and did not change in subsequent experiments run for longer periods of time (up to 72 h). There was no indication of fragmented nuclear DNA but there was evidence of injurious cell death (necrosis) by electron microscopy. Morphological analysis of cartilage repetitively loaded for 24 h showed matrix damage only in the uppermost superficial layer at the articular surface, reminiscent of the early stages of osteoarthritis. CONCLUSIONS: Cell death in mature cartilage explants occurred after 6 hours of continuous repetitive load or 3 h of static load. Cell death was directly related to the mechanical load, as control (free-swelling) explants remained viable at all times. The excessive, repetitive loading conditions imposed are not physiological, and demonstrate the deleterious effects of mechanical overload resulting in morphological and cellular damage similar to that seen in degenerative joint disease.     

Increased stromelysin-1 (MMP-3), proteoglycan degradation (3B3- and 7D4) and collagen damage in cyclically load-injured articular cartilage

OBJECTIVE: To determine whether load-induced injury causes alterations in proteoglycan (PG), stromelysin-1 (MMP-3) and collagen in articular cartilage. METHODS: Mature bovine cartilage was cyclically loaded at 0.5 Hz with 1 and 5 MPa for 1, 6 and 24h. Immediately after loading explants were evaluated for cell viability. Alterations in matrix integrity were determined by measuring PG content, PG degradation using 7D4 and 3B3(-) antibodies, broken collagen using COL2-3/4m antibody, and stromelysin-1 content using a MMP-3 antibody. RESULTS: Mechanical load caused cell death and PG loss starting from the articular surface and increasing in depth with loading time. There was a decrease in the 7D4 epitope (native chondroitin sulfate) in the superficial zone of cartilage loaded for longer than 1h, but an increase around chondrocytes in the deep zone. The 3B3(-) staining for degraded/abnormal chondroitin-4-sulfate neoepitope appeared only in cartilage loaded under the most severe condition (5 MPa, 24 h). The elevation of stromelysin-1 was co-localized with broken collagen (COL2-3/4m) at the articular surface in explants loaded with 1 and 5 MPa for 24 h. CONCLUSIONS: Cell death and PG loss occurred within 6h of cyclic loading. The elevation of MMP-3 following cell death was consistently found in the superficial zone of loaded cartilage. Since MMP-3 can degrade PG and super-activate procollagenase, the increase of MMP-3 can therefore induce matrix degradation and PG depletion in mechanically injured articular cartilage, both of which are important to the development of osteoarthritis.     

Unfolding of membrane ruffles of in situ chondrocytes under compressive loads

Impact loading results in chondrocyte death. Previous studies implicated high tensile strain rates in chondrocyte membranes as the cause of impact‐induced cell deaths. However, this hypothesis relies on the untested assumption that chondrocyte membranes unfold in vivo during physiological tissue compression, but do not unfold during impact loading. Although membrane unfolding has been observed in isolated chondrocytes during osmotically induced swelling and mechanical compression, it is not known if membrane unfolding also occurs in chondrocytes embedded in their natural extracellular matrix. This study was aimed at quantifying changes in membrane morphology of in situ superficial zone chondrocytes during slow physiological cartilage compression. Bovine cartilage‐bone explants were loaded at 5 μm/s to nominal compressive strains ranging from 0% to 50%. After holding the final strains for 45 min, the loaded cartilage was chemically pre‐fixed for 12 h. The cartilage layer was post‐processed for visualization of cell ultrastructure using electron microscopy. The changes in membrane morphology in superficial zone cells were quantified from planar electron micrographs by measuring the roughness and the complexity of the cell surfaces. Qualitatively, the cell surface ruffles that existed before loading disappeared when cartilage was loaded. Quantitatively, the roughness and complexity of cell surfaces decreased with increasing load magnitudes, suggesting a load‐dependent use of membrane reservoirs. Chondrocyte membranes unfold in a load‐dependent manner when cartilage is compressed. Under physiologically meaningful loading conditions, the cells likely expand their surface through unfolding of the membrane ruffles, and therefore avoid direct stretch of the cell membrane. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:304–310, 2017.

     

Deformation of the articular cartilage and joint space of the human knee joint under static load

In order to define the deformation of articular cartilage and the joint space under statically loaded knee joints, we studied serial sections in the frontal plane of knee joints which were frozen under the application of a load. Normal articular cartilage surfaces did not make contact even under loads of 100kg or more, leaving always a space between. The compressive deformation of articular cartilage mainly occurred at the central area near to the intercondylar notch or prominence, but the deformation of cartilage in the area covered by menisci was negligible.     

Articular cartilage MR imaging and thickness mapping of a loaded knee joint before and after meniscectomy

OBJECTIVE: We describe a technique to axially compress a sheep knee joint in an MRI scanner and measure articular cartilage deformation. As an initial application, tibial articular cartilage deformation patterns after 2 h of static loading before and after medial meniscectomy are compared. METHODS: Precision was established for repeated scans and repeated segmentations. Accuracy was established by comparing to micro-CT measurements. Four sheep knees were then imaged unloaded, and while statically loaded for 2 h at 1.5 times body weight before and after medial meniscectomy. Images were obtained using a 3D gradient echo sequence in a 4.7 T MRI. Corresponding 3D cartilage thickness models were created. Nominal strain patterns for the intact and meniscectomized conditions were compared. RESULTS: Coefficients of variation were all 2% or less. Root mean squared errors of MR cartilage thickness measurements averaged less than 0.09 mm. Meniscectomy resulted in a 60% decrease in the contact area (P=0.001) and a 13% increase in maximum cartilage deformation (P=0.01). Following meniscectomy, there were greater areas of articular cartilage experiencing abnormally high and low nominal strains. Areas of moderate nominal strain were reduced. CONCLUSIONS: Medial meniscectomy resulted in increased medial tibial cartilage nominal strains centrally and decreased strains peripherally. Areas of abnormally high nominal strain following meniscectomy correlated with areas that are known to develop fibrillation and softening 16 weeks after medial meniscectomy. Areas of abnormally low nominal strain correlated with areas of osteophyte formation. Studies of articular cartilage deformation may prove useful in elucidating the mechanical etiology of osteoarthritis.     

Deformation of articular cartilage collagen structure under static and cyclic loading

Relatively little is known about the morphology of articuar cartilage under conditions of normal use. yet a more profound knowledge is both critical to the understanding of cartilage function and helpful for the validation of tissue-engineered cartilage. In this study, the deformation of the articular cartilage of the tibial plateau under compressive static and cyclic loading is characterized. Whole knee joints of rabbits were loaded ex vivo while the knee was held statically or allowed to move against resistance. Load magnitudes of quadriceps were maintained at either three (high) or one (low) times body weight for 30 minutes. For cyclic loading, the tibia was flexed between 70 and 150° relative to the femur at 1 Hz with either a cyclic or constant force. The recovery of cartilage after unloading was examined for each loading condition. At the end of the loading, specimens were cryofixed while under load, freeze-substituted, and prepared for scanning electron microscopy. Morphological examination demonstrated significantly higher deformation of the collagen structure throughout all cartilage zones under static loading conditions compared with cyclic loading condition's in which deformation was limited to the superficial regions. The minimum thickness of the cartilage that remained after loading was dependent on the magnitude of load and was significantly smaller with static loads (54% of the thickness of the unloaded controls) than after cyclic loading or constant-force cyclic loading (78 or 66% of the thickness of the unloaded controls, p < 0.05). Acute bending of the collagen fibers was observed under both loading conditions: in the superficial half of the articular cartilage after static loading and in the superficial quarter after cyclic loading. Complete recovery of all deformation occurred within 30 minutes but was significantly faster after cyclic loading. These data suggest that the structure of the collagen of articular cartilage exhibits a zone-specific deformation that is dependent on the magnitude and type of load.     

Effect of compressive loading and unloading on the synthesis of total protein,proteoglycan, and fibronectin by canine cartilage explants

Full-thickness canine articular cartilage explants were subjected to compressive loads equivalent to a uniaxial stress of 0.025–1.2 MPa. A single cycle (18 h) of unconfined compression resulted in inhibition of total protein, proteoglycan, and fibronectin synthesis. The inhibition of fibronectin synthesis followed that of total protein synthesis. The magnitude of inhibition increased nonlinearly with increasing load levels. The signal that depressed synthesis remained effective for several hours after removal of load, but by 24 h proteoglycan synthesis had partially recovered and fibronectin and protein synthesis had fully recovered and sometimes exceeded the rate of synthesis in free-swelling controls. Forty-eight hours after five cycles of intermittent unconfined compression with similar loads, proteoglycan content and synthesis did not differ in loaded disks and in disks that were never loaded in vitro. Interestingly, the percentage of water in disks that had never been loaded in vitro increased significantly after 10 days in culture, relative to the percentage of water in free-swelling disks on the day of harvest. Intermittent compressive loading in the range of 0.5–1.2 MPa partially prevented this increase. Our results confirmed the previously reported inhibition of biosynthesis with static loading but also suggested that exposure to intermittent compressive loading may help to maintain the normal ratio of dry to wet weight in the explant.     

Subchondral damage after acute transarticular loading: an in vitro model of joint injury.

Intact canine metacarpophalangeal and metatarsophalangeal joints were subjected to a variety of loads in vitro. Intraarticular fracture occurred in 19 joints loaded to an average force of 2.4 +/- 0.4 kN with a corresponding loading rate of 88 +/- 23 kN/s. The remaining 29 joints were without gross evidence of fracture with an average load and loading rate of 1.7 +/- 0.9 kN and 64 +/- 32 kN/s, respectively. In the fractured specimens, damage to the zone of calcified cartilage and subchondral bone was much more extensive than was initially evident by gross inspection when assessed by scanning electron microscopy. Cracks with associated step-off displacement at the zone of calcified cartilage were found distant to the gross fractures. These findings were confirmed histologically. In addition, cracks localized to the zone of calcified cartilage were commonly identified histologically in specimens loaded in the range of 1.9-2.8 kN, but were not grossly fractured. The contact area determined with pressure-sensitive film increased with increasing load up to the point of fracture. The average pressure generated at the articular cartilage surface at the time of fracture in this model is > or = 40 MPa, and the fracture occurred at the contact site. Our findings suggest that failure in acute transarticular loading begins in the zone of calcified cartilage and subsequently involves the subchondral bone and overlying cartilage. This type of injury may contribute to the development of osteoarthritis after intraarticular fracture, or at high loads that do not result in gross fracture.     

In vitro measurement of articular cartilage deformations in the intact human hip joint under load.

Using a new roentgenographic technique for measuring cartilage deformation in intact joint specimens, twenty-eight normal human hip joints from subjects twenty-five to eighty-five years old were loaded with a force of five times body weight in a testing machine. The initial unloaded thickness of the articular cartilage of the femoral head and the changes in thickness of this cartilage under load were measured roentgenographically at seven to twelve sites on each femoral head. These measurements showed that the deformations of femiral-head articular cartilage under load in the intact joint are non-uniform and increase greatly with age. In twelve specimens measurements were also made of the increase in cartilage deformation with time when the load of five times body weight was maintained on the joint. A single osteoarthrotic joint was also studied. The experimental findings imply changes in the fundamental mechanical properties of the cartilage with age, which probably result from age-related alterations in cartilage microstructure and chemical composition.     

Changes induced by chronic in vivo load alteration in the tibiofemoral joint of mature rabbits

We investigated the relationship between the magnitude and duration of chronic compressive load alteration and the development and progression of degenerative changes in the rabbit tibiofemoral joint. Varus loading devices were attached to the hind limb of mature NZW rabbits. Altered compressive loads of 0%, 50%, and 80% body weight (BW) were applied to the tibiofemoral joint for 12 h per day for 12 and 24 weeks (n = 4 animals/group). Compartment‐specific assessment of the tibial plateau included histological assessments (articular cartilage, calcified cartilage, and subchondral bone thicknesses, degeneration score, and articular cartilage cellularity) and biomechanical measures (aggregate modulus, permeability, Poisson's ratio). Analyses of variance techniques were used to examine the relationship between each outcome measure with load magnitude and duration as independent variables in the model. Degenerative changes developed in the medial compartment with increased magnitude of compressive loading and included fibrillation, increased degeneration score, and reduced cellularity of the articular cartilage. Increased calcified cartilage thickness was observed in both the medial and lateral compartments following exposure to altered loading of 80% BW for 24 weeks. This work demonstrates that in vivo chronic compressive load alteration to the tibiofemoral joint can initiate progressive macroscopic and histological‐based degenerative changes analogous to the early changes occurring in OA. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1413–1422, 2012     

The distribution of surface strain in the cadaveric lumbar spine

The fourth lumbar vertebrae and L4-5 discs from six cadaveric lumbar spines were subjected to detailed strain gauge analysis under conditions of controlled loading. With central compression loads, maximal compressive strain was found to occur near the bases of the pedicles and on both superficial and deep surfaces of the pars interarticularis, which emphasises the importance of the posterior elements of lumbar vertebrae in transmitting load. Radial bulge and tangential strain of the disc wall were maximal at the posterolateral surface, in agreement with the fact that disc degeneration and prolapse commonly occur there. Under posterior offset loads simulating extension, both compressive and tensile strains were found to be increased on both surfaces of the pars interarticularis, which suggests that hyperextension may lead to stress fractures and spondylolisthesis. Posterior offset loads also increased the radial bulge of the posterior disc wall and tangential strain at the anterior surface of the disc. Anterior offset loads simulating flexion increased the radial bulge of the anterior disc wall and tangential strain at the posterior surface of the disc. These findings are compatible with movement of the nucleus pulposus within the disc during flexion and extension. This hypothesis was supported by post-mortem discography.     

Articular cartilage adjacent to experimental defects is subject to atypical strains

We tested the hypothesis that articular cartilage adjacent to experimental osteochondral defects is not subject to unusual strains under load. A 2.5-mm drill hole was made in the medial femoral condyle of 15 knees from 10 adult rabbits. Experimental joints were loaded with simulated quadriceps force, then frozen under load and preserved by freeze-substitution fixation. Deformation in the region of the defect was evaluated by scanning electron and light microscopy and compared with nondrilled and nonloaded control knees. To simulate blood clot, alginate was placed into some defects before loading. In loaded knees, articular cartilage at the edge of the drill hole was abnormally flattened and folded into the defect. Opposing tibial cartilage or meniscus intruded into the femoral defect beyond the cement line. Alginate did not prevent incursion of opposing cartilage. In this standard drill-hole model, the articular cartilage defect is occupied by the opposing surface when a joint is loaded. Any tissue growing or surgically implanted in the defect is subject to loading and displacement, therefore complicating attempts to characterize the healing or regenerative potential in similar drill-hole models. Deformation of cartilage at the defect edge suggests load concentration or increased compliance. Either phenomenon would contribute to subsequent degeneration of the cartilage adjacent to defects.     

Cartilage induction by controlled mechanical stimulation in vivo.

To study mechanical control of tissue differentiation, we designed a new version of the previously described bone conduction chamber. The bone conduction chamber consists of a cylindrical titanium chamber for implantation in the rat tibia. It has tissue ingrowth openings at one end, located subcortically, and the other end protrudes into the subcutis. The newly developed load chamber has a mobile piston so that an external compressive load can be transferred to the tissue within the chamber. Sprague-Dawley rats had a regular bone conduction chamber implanted in one tibia and a load chamber implanted in the other. Mesenchymal tissue was allowed to grow into the chamber for 3 weeks before the mechanical loading was started. Thereafter, twice a day, 20 cycles of compressive load were applied with a frequency of 0.17 Hz to the load chamber. This was estimated to produce a compressive hydrostatic stress of 2 MPa. The chambers, harvested after 7 weeks of loading, all contained newly formed bone. The bone ingrowth distance into the chamber was decreased in the loaded specimens compared with the contralateral unloaded controls (p = 0.01). Instead, cartilage was found in the loaded chambers next to the piston. Beneath the cartilage was a dense bone plate under which a marrow cavity had formed. No cartilage was found in the unloaded controls, but the architecture of the bone and marrow cavity was similar to that of the loaded specimens. We conclude that this model allows load to be transmitted onto the ingrowing tissue and that the load parameters used cause this tissue to differentiate into cartilage close to the piston.