Dual inversion recovery ultrashort echo time (DIR-UTE) imaging and quantification of the zone of calcified cartilage (ZCC).

OBJECTIVE: To develop ultrashort echo time (UTE) magnetic resonance imaging (MRI) techniques to image the zone of calcified cartilage (ZCC), and quantify its T2*, T1 and T1ρ. DESIGN: In this feasibility study a dual inversion recovery UTE (DIR-UTE) sequence was developed for high contrast imaging of the ZCC. T2* of the ZCC was measured with DIR-UTE acquisitions at progressively increasing TEs. T1 of the ZCC was measured with saturation recovery UTE acquisitions at progressively increasing saturation recovery times. T1ρ of the ZCC was measured with spin-locking prepared DIR-UTE acquisitions at progressively increasing spin-locking times. RESULTS: The feasibility of the qualitative and quantitative DIR-UTE techniques was demonstrated on phantoms and in six cadaveric patellae using a clinical 3 T scanner. On average the ZCC has a short T2* ranging from 1.0 to 3.3 ms (mean ± standard deviation = 2.0 ± 1.2 ms), a short T1 ranging from 256 to 389 ms (mean ± standard deviation = 305 ± 45 ms), and a short T1ρ ranging from 2.2 to 4.6 ms (mean ± standard deviation = 3.6 ± 1.2 ms). CONCLUSION: UTE MR based techniques have been developed for high resolution imaging of the ZCC and quantitative evaluation of its T2*, T1 and T1ρ relaxation times, providing non-invasive assessment of collagen orientation and proteoglycan content at the ZCC and the bone cartilage interface. These measurements may be useful for non-invasive assessment of the ZCC, including understanding the involvement of this tissue component in osteoarthritis.

Biomechanical, structural, and biochemical indices of degenerative and osteoarthritic deterioration of adult human articular cartilage of the femoral condyle.

OBJECTIVE: To compare the tensile biomechanical properties of age-matched adult human knee articular cartilage exhibiting distinct stages of degenerative or osteoarthritic deterioration and to determine the relationships between tensile properties and biochemical and structural properties hypothesized to underlie functional biomechanical deterioration. METHODS: Age-matched articular cartilage samples, obtained from the lateral and medial femoral condyles (LFC and MFC), exhibited (1) minimal fibrillation, characteristic of normal aging (NLA), (2) overt fibrillation associated with degeneration (DGN), or (3) overt fibrillation associated with osteoarthritis (OA). DGN samples were from knees that exhibited degeneration but not osteophytes while OA samples were from fragments removed during total knee arthroplasty. Cartilage samples were analyzed for tensile properties, cell and matrix composition, and histopathological structure. RESULTS: Differences in tensile, compositional and surface structural properties were indicative of distinct stages of cartilage degeneration, early (OA) advanced (DGN) and late (OA) with early degenerative changes in NLA samples being more advanced in the MFC than the LFC, including higher surface fibrillation, lower intrinsic fluorescence, and lower mechanical integrity. The transition from early to advanced degeneration involved a diminution in mechanical function, surface integrity, and intrinsic fluorescence. The transition from advanced to late degeneration involved an increase in cartilage water content, an increase in degraded collagen, and loss of collagen. CONCLUSIONS: These results provide evidence of coordinated mechanical dysfunction, collagen network remodeling, and surface fibrillation. Even in the cartilage of knees exhibiting overt fibrillation but not extensive erosions characteristic of clinical osteoarthritis, most features of advanced cartilage degeneration were present.

Wear-lines and split-lines of human patellar cartilage: relation to tensile biomechanical properties.

BACKGROUND: Articular cartilage undergoes age-associated degeneration, resulting in both structural and functional biomechanical changes. At early stages of degeneration, wear-lines develop in the general direction of joint movement. With aging, cartilage exhibits a decrease in tensile modulus. The tensile modulus of cartilage has also been related to the orientation of the collagen network, as revealed by split-lines. OBJECTIVE: To determine the relative contribution of wear-line and split-line orientation on the tensile biomechanical properties of human patellar cartilage from different depths. METHODS: In human patellar cartilage, wear- and split-lines are aligned parallel to each other at the proximal facet, and perpendicular to each other at the medial facet. Using superficial, middle, and deep cartilage sections from these two sites, tensile samples were prepared in two orthogonal orientations. Thus, for each depth, there were four groups of samples, with their long axes were aligned either parallel or perpendicular to wear-line direction and also aligned parallel or perpendicular to split-line direction. Uniaxial tensile tests were performed to assess equilibrium and ramp moduli. RESULTS: Tensile equilibrium moduli varied with wear-line orientation (P<0.05) and depth (P<0.001), in an interactive manner (P<0.05), and tended to vary with split-line orientation (P=0.16). In the superficial layer, equilibrium and ramp modulus were higher when the samples were loaded parallel to wear-lines (P<0.05). CONCLUSION: These results indicate that mild wear (i.e., wear-line formation) at the articular surface has deleterious functional effects on articular cartilage and represent an early aging-associated degenerative change. The identification and recognition of functional biomechanical consequences of wear-lines are useful for planning and interpreting tensile biomechanical tests in human articular cartilage.

Age- and site-associated biomechanical weakening of human articular cartilage of the femoral condyle.

OBJECTIVE: To determine the time sequence of biochemical and structural events associated with, and hypothesized to underlie, age-associated tensile weakening of macroscopically normal adult human articular cartilage of the knee. METHODS: Macroscopically normal human articular cartilage of the lateral and medial femoral condyles (LFC and MFC) from Young (21-39 yrs), Middle (40-59 yrs), and Old (>/=60 yrs) age donors were analyzed for tensile properties, surface wear, and cell and matrix composition. RESULTS: Variations in tensile, compositional, and surface structural properties were indicative of early, intermediate, and late stages of age-associated cartilage deterioration, occurring at an earlier age in the MFC than the LFC. Differences between Young and Middle age groups (indicative of early-to-intermediate stage changes) included decreased mechanical function in the superficial zone, with a loss of (or low) tensile integrity, and surface wear, with faint striations and mild staining on the articular surface after application of India ink. Differences between Middle and Old age groups (indicative of intermediate-to-late stage changes) included maintenance of moderate level biomechanical function, a decrease in cellularity, and a decrease in matrix glycosaminoglycan content. Tissue fluorescence increased steadily with age. CONCLUSIONS: Many of these age-associated differences are identical to those regarded as pathological features of cartilage degeneration in early osteoarthritis. These findings provide evidence for the roles of mechanical wear, cell death, and enzymatic degradation in mediating the progression through successive and distinguishable stages of early cartilage deterioration.

Indentation probing of human articular cartilage: Effect on chondrocyte viability.

BACKGROUND: Clinical arthroscopic probes based on indentation testing are being developed. However, the biological effects of certain design parameters (i.e., tip geometry and size) and loading protocols (i.e., indentation depth, rate, and repetition) on human articular cartilage are unclear. OBJECTIVE: Determine if indenter design and indentation protocol modulate mechanical injury of probed cartilage samples. METHODS: The objectives of this study were to determine the effects of indentation testing using clinically applicable tips (0.4mm radius, plane- or sphere-ended) and protocols (indentation depths of 100, 200, or 300 microm, applied at a rate of 50 or 500 microm/s) on the extent and the pattern of chondrocyte death, should it occur. Grossly normal osteochondral blocks were harvested from human talar dome, indented, stained with live/dead dyes, and imaged en face on a fluorescence microscope. RESULTS: The occurrence and the extent of cell death generally increased with indentation depth, being undetected at an indentation depth of 100 microm but marked at 300 microm. In addition, tip geometry affected the pattern of cell death: ring- and solid circle-shaped areas of cell deaths were apparent when compressed to 300 microm using plane- and sphere-ended indenters. CONCLUSION: Indenter design and indentation protocol modulated the extent and the pattern of chondrocyte death. These results have implications for designing indentation probes and protocols, as well as clinicians performing arthroscopic probing.

Continuous passive motion applied to whole joints stimulates chondrocyte biosynthesis of PRG4.

UNLABELLED: Continuous passive motion (CPM) is currently a part of patient rehabilitation regimens after a variety of orthopedic surgical procedures. While CPM can enhance the joint healing process, the direct effects of CPM on cartilage metabolism remain unknown. Recent in vivo and in vitro observations suggest that mechanical stimuli can regulate articular cartilage metabolism of proteoglycan 4 (PRG4), a putative lubricating and chondroprotective molecule found in synovial fluid and at the articular cartilage surface. OBJECTIVES: (1) Determine the topographical variation in intrinsic cartilage PRG4 secretion. (2) Apply a CPM device to whole joints in bioreactors and assess effects of CPM on PRG4 biosynthesis. METHODS: A bioreactor was developed to apply CPM to bovine stifle joints in vitro. Effects of 24h of CPM on PRG4 biosynthesis were determined. RESULTS: PRG4 secretion rate varied markedly over the joint surface. Rehabilitative joint motion applied in the form of CPM regulated PRG4 biosynthesis, in a manner dependent on the duty cycle of cartilage sliding against opposing tissues. Specifically, in certain regions of the femoral condyle that were continuously or intermittently sliding against meniscus and tibial cartilage during CPM, chondrocyte PRG4 synthesis was higher with CPM than without. CONCLUSIONS: Rehabilitative joint motion, applied in the form of CPM, stimulates chondrocyte PRG4 metabolism. The stimulation of PRG4 synthesis is one mechanism by which CPM may benefit cartilage and joint health in post-operative rehabilitation.

Static and dynamic compression regulate cartilage metabolism of PRoteoGlycan 4 (PRG4).

The boundary lubrication function of articular cartilage is mediated in part by molecules at the articular surface and in synovial fluid, encoded by Prg4. The objective of this study was to determine whether static and dynamic compression regulate PRG4 biosynthesis by cartilage explants. Articular cartilage disks were harvested to include the articular surface from immature bovines. Some disks were subjected to 24 h (day 1) of loading, followed by 72 h (days 2-4) of free-swelling culture to assess chondrocyte responses following unloading. Loading consisted of 6 or 100 kPa of static compression, with or without superimposed dynamic compression (10 or 300 kPa peak amplitude, 0.01 Hz). Other disks were cultured free-swelling as controls. PRG4 secretion into culture medium was inhibited by all compression protocols during day 1. Following unloading, cartilage previously subjected to dynamic compression to 300 kPa exhibited a rebound effect, secreting more PRG4 than did controls, while cartilage previously subjected to 100 kPa static loading secreted less PRG4. Immunohistochemistry revealed that all compression protocols also affected the number of cells expressing PRG4. The paradigm that mechanical stimuli regulate biosynthesis in cartilage appears operative not only for load bearing matrix constituents, but also for PRG4 molecules mediating lubrication.

A morphologic, biochemical, and biomechanical assessment of short-term effects of osteochondral autograft plug transfer in an animal model.

PURPOSE: The objective of this study was to assess the short-term changes that occur after an osteochondral autograft plug transfer from the femoral trochlea to the medial femoral condyle in a goat model. TYPE OF STUDY: Articular cartilage repair animal study. METHODS: Six adult male goats were used in this study. Two 4.5-mm osteochondral plugs were transferred from the superolateral femoral trochlea to 2 recipient sites in the central portion of the medial femoral condyle for a survival period of 12 weeks. Postmortem, the global effects of the procedure were assessed by gross morphologic inspection and by analyzing the synovial DNA for inflammatory response. The recipient sites were also evaluated histologically and biomechanically. Metabolic activity was determined by (35)SO(4) uptake, and viability was assessed using a live/dead stain and by confocal laser microscopy. RESULTS: There was no evidence of significant gross morphologic or histologic changes in the operative knee as a result of the osteochondral donor or recipient sites. The patella, tibial plateau, and medial meniscus did not show any increased degenerative changes as a result of articulating against the donor or recipient sites of the osteochondral autografts. Analysis of synovial DNA revealed no inflammatory response. Biomechanically, 6- to 7-fold greater stiffness was noted in the cartilage of the transferred plugs compared with the control medial femoral condyle. Furthermore, on histologic examination, the healing subchondral bone interface at the recipient site had increased density. Glycosaminoglycan synthesis as determined by (35)SO(4) uptake was upregulated in the transplanted cartilage plug relative to the contralateral control, showing a repair response at the site of implantation. And finally, confocal microscopy showed 95% viability of the transferred plugs in the medial femoral condyle region. CONCLUSIONS: Our findings demonstrate the ability to successfully transfer an osteochondral autograft plug with maintenance of chondrocyte cellular viability. The transferred cartilage is stiffer than the control medial femoral condyle cartilage, and there is concern regarding the increased trabecular mass in the healing subchondral plate, but these do not result in increased degenerative changes of the opposing articular surfaces in the short term.

Depth- and strain-dependent mechanical and electromechanical properties of full-thickness bovine articular cartilage in confined compression.

Compression tests have often been performed to assess the biomechanical properties of full-thickness articular cartilage. We tested whether the apparent homogeneous strain-dependent properties, deduced from such tests, reflect both strain- and depth-dependent material properties. Full-thickness bovine articular cartilage was tested by oscillatory confined compression superimposed on a static offset up to 45%. and the data fit to estimate modulus, permeability, and electrokinetic coefficient assuming homogeneity. Additional tests on partial-thickness cartilage were then performed to assess depth- and strain-dependent properties in an inhomogeneous model, assuming three discrete layers (i = 1 starting from the articular surface, to i = 3 up to the subchondral bone). Estimates of the zero-strain equilibrium confined compression modulus (H(A0)), the zero-strain permeability (kp0) and deformation dependence constant (M), and the deformation-dependent electrokinetic coefficient (ke) differed among individual layers of cartilage and full-thickness cartilage. HiA0 increased from layer 1 to 3 (0.27 to 0.71 MPa), and bracketed the apparent homogeneous value (0.47 MPa). ki(p0) decreased from layer 1 to 3 (4.6 x 10(-15) to 0.50 x 10(-15) m2/Pa s) and was less than the homogeneous value (7.3 x 10(-15) m2/Pa s), while Mi increased from layer 1 to 3 (5.5 to 7.4) and became similar to the homogeneous value (8.4). The amplitude of ki(e) increased markedly with compressive strain, as did the homogeneous value: at low strain, it was lowest near the articular surface and increased to a peak in the middle-deep region. These results help to interpret the biomechanical assessment of full-thickness articular cartilage.

Cartilage repair with autogenic perichondrium cell and polylactic acid grafts.

The repair of articular cartilage injuries remains a challenge, with many of the current therapeutic strategies based on the grafting or recruitment of chondrogenic tissues or cells. This 1-year study compared the repair of a 3.7-mm diameter by 3-mm deep osteochondral defect in the medial femoral condyle of 24 New Zealand White rabbits; the defect was obtained using an autogenic perichondrium cell polylactic acid composite graft with a contralateral control in which the osteochondral defect remained empty. To elucidate the effect of host immune responses on the repair process after perichondrium cell transplantation, the results of the autogenic perichondrium cell polylactic acid graft group were compared with those obtained in the authors' previous 1-year study of allogenic perichondrium cell polylactic acid composite grafts implanted in a similar model. One year after surgery, the repair site underwent gross inspection and histologic, histomorphometric, biochemical, and biomechanical analyses. The autogenic perichondrium cell polylactic acid graft group (92%) and the control group in which the osteochondral defect remained empty (88%) resulted in a high percentage of grossly acceptable repairs. The autogenic grafts appeared to augment the intrinsic healing capacity of the animals (as compared with the animals in the No Implant Group). The autogenic perichondrium cell polylactic and grafts improved the histologic appearance and percentage of Type II collagen of the cartilaginous repair tissue. Compared with allogenic grafts, the autogenic grafts had better reconstitution of the subchondral bone. However, the results of this experimental model suggest a suboptimal concentration of glycosaminoglycans in the neocartilage matrix, a depressed surface of the repair tissue, a histologic appearance that was not equivalent to that of normal articular cartilage, and reduced biomechanical properties for the repair tissue. The future application of growth factors to this model may yield a treatment that can be applied in the clinical arena.