Assessing collagen alterations in enzymatic degradation models of osteoarthritis via second harmonic generation microscopy.

INTRODUCTION: Structural changes in the collagen II architecture of osteoarthritis (OA) are poorly understood, which is a large shortcoming in the early diagnosis of this disease. Though degradation can be simulated by enzymes including trypsin and bacterial collagenase, the specific structural features of each digestion and their relationship to naturally occurring OA remain unclear. EXPERIMENTAL DESIGN: We used collagen sensitive/specific Second Harmonic Generation (SHG) microscopy in conjunction with optical scattering measurements to probe the resulting architecture changes in bovine knee cartilage upon trypsin and collagenase degradation. Image features extracted from SHG images were used to train a linear discriminant (LD) model capable of classifying enzymatic degradation, which was then applied to human cartilage with varied modified Mankin histological scores. RESULTS: The treatment of cartilage with these enzymes resulted in more disorganized collagen structure, where this effect was greatest with collagenase treatment. Using the LD model, we classified the control and degraded tissues in the three zones with >92% accuracy, showing that these enzymes have distinct activity on the collagen assembly. Application of the LD model to human cartilage indicated that collagenase effects were more representative of in vivo degeneration and were also consistent with damage beginning at the articular surface and progressing into deeper zones. CONCLUSIONS: SHG and optical scattering measurements successfully delineate trypsin and collagenase degradation and suggest that collagen alterations in human OA are better simulated by the latter mechanism. These results lay the groundwork for using high-resolution SHG and optical scattering as an earlier diagnostic tool than is currently available.

Relaxation capacity of cartilage is a critical factor in rate- and integrity-dependent fracture.

Articular cartilage heals poorly but experiences mechanically induced damage across a broad range of loading rates and matrix integrity. Because loading rates and matrix integrity affect cartilage mechanical responses due to poroviscoelastic relaxation mechanisms, their effects on cartilage failure are important for assessing and preventing failure. This paper investigated rate- and integrity-dependent crack nucleation in cartilage from pre- to post-relaxation timescales. Rate-dependent crack nucleation and relaxation responses were obtained as a function of matrix integrity through microindentation. Total work for crack nucleation increased with decreased matrix integrity, and with decreased loading rates. Critical energy release rate of intact cartilage was estimated as 2.39 ± 1.39 to 2.48 ± 1.26 kJ m-2 in a pre-relaxation timescale. These findings showed that crack nucleation is delayed when cartilage can accommodate localized loading through poroviscoelastic relaxation mechanisms before fracture at a given loading rate and integrity state.

Evaluation of patient tissue selection methods for deriving equivalent density calibration for femoral bone quantitative CT analyses.

Osteoporosis affects an increasing number of people every year and patient specific finite element analysis of the femur has been proposed to identify patients that could benefit from preventative treatment. The aim of this study was to demonstrate, verify, and validate an objective process for selecting tissues for use as the basis of phantomless calibration to enable patient specific finite element analysis derived hip fracture risk prediction. Retrospective reanalysis of patient computed tomography (CT) scans has the potential to yield insights into more accurate prediction of osteoporotic fracture. Bone mineral density (BMD) specific calibration scans are not typically captured during routine clinical practice. Tissue-based BMD calibration can therefore empower the retrospective study of patient CT scans captured during routine clinical practice. Together the method for selecting tissues as the basis for phantomless calibration coupled with the post-processing steps for deriving a calibration equation using the selected tissues provide an estimation of quantitative equivalent density results derived using calibration phantoms. Patient tissues from a retrospective cohort of 211 patients were evaluated. The best phantomless calibration resulted in a femoral strength (FS) [N] bias of 0.069 ± 0.07% over FS derived from inline calibration and a BMD [kg/cm3] bias of 0.038 ± 0.037% over BMD derived from inline calibration. The phantomless calibration slope for the best method presented was within the range of patient specific calibration curves available for comparison and demonstrated a small bias of 0.028 ± 0.054 HU/(mg/cm3), assuming the Mindways Model 3 BMD inline calibration phantom as the gold standard. The presented method of estimating a calibration equation from tissues showed promise for CT-based femoral fracture analyses of retrospective cohorts without readily available calibration data.

Real-time optical redox imaging of cartilage metabolic response to mechanical loading.

OBJECTIVE: Metabolic dysregulation has recently been identified as a key feature of osteoarthritis. Mechanical overloading has been postulated as a primary cause of this metabolic response. Current methods of real-time metabolic activity analysis in cartilage are limited and challenging. However, optical redox imaging leverages the autofluorescence of co-enzymes NAD(P)H and FAD to provide dye-free real-time analysis of metabolic activity. This technique has not yet been applied to cartilage. This study aimed to assess the effects of a compressive load on cartilage using optical redox imaging. METHOD: Cartilage samples were excised from porcine femoral condyles. To validate this imaging modality in cartilage, glycolysis was inhibited via 2-deoxy-D-glucose (2DG) and oxidative phosphorylation was inhibited by rotenone. Optical redox images were collected pre- and post-inhibition. To assess the effects of mechanical loading, samples were subjected to a compressive load and imaged for approximately 30 min. Load and strain parameters were determined using high-speed camera images in Matlab. A range of loading magnitudes and rates were applied across samples. RESULTS: 2DG and rotenone demonstrated the expected inhibitory effects on fluorescence intensity in the channels corresponding to NAD(P)H and FAD, respectively. Mechanical loading induced an increase in NAD(P)H channel fluorescence which subsided by 30 min post-loading. Magnitude of loading parameters had mixed effects on metabolites. CONCLUSIONS: Optical redox imaging provides an opportunity to assess real-time metabolic activity in cartilage. This approach revealed a metabolic response to a single load and can be used to provide insight into the role of metabolism in mechanically-mediated cartilage degradation.

Computed tomography arthrography with traction in the human hip for three-dimensional reconstruction of cartilage and the acetabular labrum.

AIM: To develop and demonstrate the efficacy of a computed tomography arthrography (CTA) protocol for the hip that enables accurate three-dimensional reconstructions of cartilage and excellent visualization of the acetabular labrum. MATERIALS AND METHODS: Ninety-three subjects were imaged (104 scans); 68 subjects with abnormal anatomy, 11 patients after periacetabular osteotomy surgery, and 25 subjects with normal anatomy. Fifteen to 25 ml of contrast agent diluted with lidocaine was injected using a lateral oblique approach. A Hare traction splint applied traction during CT. The association between traction force and intra-articular joint space was assessed qualitatively under fluoroscopy. Cartilage geometry was reconstructed from the CTA images for 30 subjects; the maximum joint space under traction was measured. RESULTS: Using the Hare traction splint, the intra-articular space and boundaries of cartilage could be clearly delineated throughout the joint; the acetabular labrum was also visible. Dysplastic hips required less traction (∼5 kg) than normal and retroverted hips required (>10 kg) to separate the cartilage. An increase in traction force produced a corresponding widening of the intra-articular joint space. Under traction, the maximum width of the intra-articular joint space during CT ranged from 0.98-6.7 mm (2.46 ± 1.16 mm). CONCLUSIONS: When applied to subjects with normal and abnormal hip anatomy, the CTA protocol presented yields clear delineation of the cartilage and the acetabular labrum. Use of a Hare traction splint provides a simple, cost-effective method to widen the intra-articular joint space during CT, and provides flexibility to vary the traction as required.

A new sensor for measurement of dynamic contact stress in the hip.

Various techniques exist for quantifying articular contact stress distributions, an important class of measurements in the field of orthopaedic biomechanics. In situations where the need for dynamic recording has been paramount, the approach of preference has involved thin-sheet multiplexed grid-array transducers. To date, these sensors have been used to study contact stresses in the knee, shoulder, ankle, wrist, and spinal facet joints. Until now, however, no such sensor had been available for the human hip joint due to difficulties posed by the deep, bi-curvilinear geometry of the acetabulum. We report here the design and development of a novel sensor capable of measuring dynamic contact stress in human cadaveric hip joints (maximum contact stress of 20 MPa and maximum sampling rate 100 readings/s). Particular emphasis is placed on issues concerning calibration, and on the effect of joint curvature on the sensor's performance. The active pressure-sensing regions of the sensors have the shape of a segment of an annulus with a 150-deg circumferential span, and employ a polar/circumferential "ring-and-spoke" sensel grid layout. There are two sensor sizes, having outside radii of 44 and 48 mm, respectively. The new design was evaluated in human cadaver hip joints using two methods. The stress magnitudes and spatial distribution measured by the sensor were compared to contact stresses measured by pressure sensitive film during static loading conditions that simulated heel strike during walking and stair climbing. Additionally, the forces obtained by spatial integration of the sensor contact stresses were compared to the forces measured by load cells during the static simulations and for loading applied by a dynamic hip simulator. Stress magnitudes and spatial distribution patterns obtained from the sensor versus from pressure sensitive film exhibited good agreement. The joint forces obtained during both static and dynamic loading were within ±10% and ±26%, respectively, of the forces measured by the load cells. These results provide confidence in the measurements obtained by the sensor. The new sensor's real-time output and dynamic measurement capabilities hold significant advantages over static measurements from pressure sensitive film.

Patient-specific analysis of cartilage and labrum mechanics in human hips with acetabular dysplasia.

BACKGROUND: Acetabular dysplasia is a major predisposing factor for development of hip osteoarthritis (OA), and may result from alterations to chondrolabral loading. Subject-specific finite element (FE) modeling can be used to evaluate chondrolabral mechanics in the dysplastic hip, thereby providing insight into mechanics that precede OA. OBJECTIVE: To evaluate chondrolabral contact mechanics and congruency in dysplastic hips and normal hips using a validated approach to subject-specific FE modeling. METHODS: FE models of ten subjects with normal acetabula and ten subjects with dysplasia were constructed using a previously validated protocol. Labrum load support, and labrum and acetabular cartilage contact stress and contact area were compared between groups. Local congruency was determined at the articular surface for two simulated activities. RESULTS: The labrum in dysplastic hips supported 2.8-4.0 times more of the load transferred across the joint than in normal hips. Dysplastic hips did not have significantly different congruency in the primary load-bearing regions than normal hips, but were less congruent in some unloaded regions. Normal hips had larger cartilage contact stress than dysplastic hips in the few regions that had significant differences. CONCLUSIONS: The labrum in dysplastic hips has a far more significant role in hip mechanics than it does in normal hips. The dysplastic hip is neither less congruent than the normal hip, nor subjected to elevated cartilage contact stresses. This study supports the concept of an outside-in pathogenesis of OA in dysplastic hips and that the labrum in dysplastic hips should be preserved during surgery.

Finite element predictions of cartilage contact mechanics in hips with retroverted acetabula.

BACKGROUND: A contributory factor to hip osteoarthritis (OA) is abnormal cartilage mechanics. Acetabular retroversion, a version deformity of the acetabulum, has been postulated to cause OA via decreased posterior contact area and increased posterior contact stress. Although cartilage mechanics cannot be measured directly in vivo to evaluate the causes of OA, they can be predicted using finite element (FE) modeling. OBJECTIVE: The objective of this study was to compare cartilage contact mechanics between hips with normal and retroverted acetabula using subject-specific FE modeling. METHODS: Twenty subjects were recruited and imaged: 10 with normal acetabula and 10 with retroverted acetabula. FE models were constructed using a validated protocol. Walking, stair ascent, stair descent and rising from a chair were simulated. Acetabular cartilage contact stress and contact area were compared between groups. RESULTS: Retroverted acetabula had superomedial cartilage contact patterns, while normal acetabula had widely distributed cartilage contact patterns. In the posterolateral acetabulum, average contact stress and contact area during walking and stair descent were 2.6-7.6 times larger in normal than retroverted acetabula (P ≤ 0.017). Conversely, in the superomedial acetabulum, peak contact stress during walking was 1.2-1.6 times larger in retroverted than normal acetabula (P ≤ 0.044). Further differences varied by region and activity. CONCLUSIONS: This study demonstrated superomedial contact patterns in retroverted acetabula vs widely distributed contact patterns in normal acetabula. Smaller posterolateral contact stress in retroverted acetabula than in normal acetabula suggests that increased posterior contact stress alone may not be the link between retroversion and OA.