Proteoglycan Depletion Research Articles

Computational simulation of the multiphasic degeneration of the bone-cartilage unit during osteoarthritis via indentation and unconfined compression tests

It has been experimentally proposed that the discrete regions of articular cartilage, along with different subchondral bone tissues, known as the bone-cartilage unit, are biomechanically altered during osteoarthritis degeneration. However, a computational framework capturing all of the dominant changes in the multiphasic parameters has not yet been developed. This article proposes a new finite element model of the bone-cartilage unit by combining several validated, nonlinear, depth-dependent, fibril-reinforced, and swelling models, which can computationally simulate the variations in the dominant parameters during osteoarthritis degeneration by indentation and unconfined compression tests. The mentioned dominant parameters include the proteoglycan depletion, collagen fibrillar softening, permeability, and fluid fraction increase for approximately non-advanced osteoarthritis. The results depict the importance of subchondral bone tissues in fluid distribution within the bone-cartilage units by decreasing the fluid permeation and pressure (up to a maximum of 100 kPa) during osteoarthritis, supporting the notion that subchondral bones might play a role in the pathogenesis of osteoarthritis. Furthermore, the osteoarthritis composition-based studies shed light on the significant biomechanical role of the calcified cartilage, which experienced a maximum change of 70 kPa in stress, together with relative load contributions of articular cartilage constituents during osteoarthritis, in which the osmotic pressure bore around 70% of the loads after degeneration. To conclude, the new insights provided by the results reveal the significance of the multiphasic osteoarthritis simulation and demonstrate the functionality of the proposed bone-cartilage unit model.

Combined enzymatic degradation of proteoglycans and collagen significantly alters intratissue strains in articular cartilage during cyclic compression

As degenerative joint diseases such as osteoarthritis (OA) progress, the matrix constituents, particularly collagen fibrils and proteoglycans, become damaged, therefore deteriorating the tissue's mechanical properties. This study aims to further the understanding of the effect of degradation of the different cartilage constituents on the mechanical loading environment in early stage OA. To this end, intact, collagen- and proteoglycan-depleted cartilage plugs were cyclically loaded in axial compression using an experimental model simulating in vivo cartilage-on-cartilage contact conditions in a micro-MRI scanner. Depletion of collagen and proteoglycans was achieved through enzymatic degradation with collagenase and chondroitinase ABC, respectively. Using a displacement-encoded imaging sequence (DENSE), strains were computed and compared in intact and degraded samples. The results revealed that, while degradation with one or the other enzyme had little effect on the contact strains, degradation with a combination of both enzymes caused an increase in the means and variance of the transverse, axial and shear strains, particularly in the superficial zone of the cartilage. This effect indicates that the balance between cartilage matrix constituents plays an essential role in maintaining the mechanical properties of the tissue, and a disturbance in this balance leads to a decrease of the load bearing capacity associated with degenerative joint diseases such as OA.

Detection of proteoglycan loss from articular cartilage using Brillouin microscopy, with applications to osteoarthritis.

The degeneration of articular cartilage (AC) occurs in osteoarthritis (OA), which is a leading cause of pain and disability in middle-aged and older people. The early disease-related changes in cartilage extra-cellular matrix (ECM) start with depletion of proteoglycan (PG), leading to an increase in tissue hydration and permeability. These early compositional changes are small (<10%) and hence difficult to register with conventional non-invasive imaging technologies (magnetic resonance and ultrasound imaging). Here we apply Brillouin microscopy for detecting changes in the mechanical properties and composition of porcine AC. OA-like degradation is mimicked by enzymatic tissue digestion, and we compare Brillouin microscopy measurements against histological staining of PG depletion over varying digestion times and enzyme concentrations. The non-destructive nature of Brillouin imaging technology opens new avenues for creating minimally invasive arthroscopic devices for OA diagnostics and therapeutic monitoring.

GagCEST MRI at 3T can detect cartilage differences between healthy and osteoarthritic subjects

Purpose: Evaluation of osteoarthritis (OA) is hampered by a lack of non-invasive tools to effectively detect early and reversible changes associated with the disease. Early OA is characterized by cartilage proteoglycan depletion related to an increase in enzymatic degradation. GagCEST is a promising quantitative MR technique for assessing content of Glycosaminoglycan (GAG) side chains attached to proteoglycans at 7T and is of particular interest due to its specificity to GAG content without the use of exogenous contrast agents or specialized hardware. While gagCEST has been shown to be a useful biomarker for cartilage composition at 7T, there is limited evidence of its utility at 3T. In this study, we assess the utility of an optimized gagCEST protocol at 3T by comparing gagCEST asymmetry in subjects with established OA to healthy controls. Methods: Ten healthy subjects (5 female and 5 male, ages 20-30) and twenty-four subjects with medial compartment OA were scanned on a 3T MRI system. The OA cohort (ages 60-70) was grouped by Kellgren-Lawrence (KL) grade (assessed using radiography) with five subjects with a KL grade of 1 (4 female and 1 male), twelve with a KL grade of 2 (8 female and 4 male), and five with a KL grade of 3 (4 female and 3 male). Other inclusion criteria included no previous ACL reconstruction surgery and BMI < 35. GagCEST images were acquired with a magnetization prepared 3D spoiled gradient-echo sequence (imaging parameters in Table 1). After correcting for magnetic field inhomogeneities, gagCEST asymmetry (%) was calculated using the B0-field corrected signal intensity at ±1.0 ppm, and the chemical shift of GAG hydroxyl protons, using the equation: gagCESTasym= [S(-1.0 ppm)-S(+1.0 ppm)]/ S0]. Femoral cartilage was manually segmented and subdivided into medial and lateral subregions. Statistical comparisons between cartilage compartments and healthy vs OA subjects were made using an ANOVA 2-way test with a Tukey HSD Test for multiple comparisons. Results: Subjects with OA showed a reduced gagCEST asymmetry compared to healthy controls. Further, there was a trend of an inverse relationship between GagCEST asymmetry and KL grade. There was a statistically significant difference in gagCEST asymmetry among KL grade values (p = 0.019). Further analysis comparing healthy subjects to the different OA KL grade subjects, showed a significant difference between healthy and OA KL-2 subjects and OA KL-3 subjects (p = 0.022 and p = 0.046 respectively). The comparison between healthy and KL-1 subjects did not show a significant difference (p = 0.536), which may be due to insufficient power in this cohort. There was no statistically significant difference between medial and lateral compartments in the femoral cartilage (p = 0.977). Additionally, a negative correlation of GagCEST with KL grade was observed (r= -0.404, p=0.01). The results show that optimized gagCEST methods at 3T were able to detect a statistically significant reduction in gagCEST asymmetry in osteoarthritic subjects, known to have cartilage degeneration and lower biochemical GAG composition, compared to healthy controls. Further analysis showed a significant difference in gagCEST asymmetry between the healthy and OA KL-2 and KL-3 subjects. This decrease can be seen in the unrolled gagCEST asymmetry maps, especially in the medial compartment of the knee, which corresponds well with the diagnosis in our cohort of medial compartment OA. Further, at a total acquisition time of 8:76 minutes for whole joint coverage, these methods offer translational potential. Conclusions: Our results demonstrate promise for the application of gagCEST at 3T to detect differences between healthy cartilage and cartilage in subjects with OA. Taken together, our results show that GagCEST could be a viable tool to assess early degenerative changes in cartilage in clinical trials. A strength of MRI over X-ray is that it is more sensitive to early changes in cartilage composition that occur due to OA. GagCEST has the potential for distinguishing these changes that are unseen in X-rays because it may be sensitive to signs of changes in GAG content that radiographs are not able to visualize.View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Maximum shear strain-based algorithm can predict proteoglycan loss in damaged articular cartilage

Post-traumatic osteoarthritis (PTOA) is a common disease, where the mechanical integrity of articular cartilage is compromised. PTOA can be a result of chondral defects formed due to injurious loading. One of the first changes around defects is proteoglycan depletion. Since there are no methods to restore injured cartilage fully back to its healthy state, preventing the onset and progression of the disease is advisable. However, this is problematic if the disease progression cannot be predicted. Thus, we developed an algorithm to predict proteoglycan loss of injured cartilage by decreasing the fixed charge density (FCD) concentration. We tested several mechanisms based on the local strains or stresses in the tissue for the FCD loss. By choosing the degeneration threshold suggested for inducing chondrocyte apoptosis and cartilage matrix damage, the algorithm driven by the maximum shear strain showed the most substantial FCD losses around the lesion. This is consistent with experimental findings in the literature. We also observed that by using coordinate system-independent strain measures and selecting the degeneration threshold in an ad hoc manner, all the resulting FCD distributions would appear qualitatively similar, i.e., the greatest FCD losses are found at the tissue adjacent to the lesion. The proposed strain-based FCD degeneration algorithm shows a great potential for predicting the progression of PTOA via biomechanical stimuli. This could allow identification of high-risk defects with an increased risk of PTOA progression.

Intra-articular effects of combined xenogenous serum rich in growth factors (SRGF) and vitamin C on histopathology grading and staging of osteoarthritis in rat model

ObjectiveOsteoarthritis (OA) is one of the most common degenerative diseases especially in the knee joint. The definitive method for the treatment of this disease is not known. In recent years, use of platelet-derived products has been considered as a new therapeutic approach because of its low cost, easy to use, and minimum side effects. Serum rich of growth factors (SRGF) is one of the biological compounds used to healing and regeneration. Its effects may improve in combination with antioxidants such as vitamin C. This vitamin increases the synthesis of proteoglycans by chondrocytes. The present study investigated effect of xenogenous SRGF in combination with vitamin C on the monosodium iodoacetate-induction osteoarthritis in rats. MethodsAnimals were randomly categorized into three groups including OA, SRGF, and vitamin C+SRGF. Treatments were performed with 3 time intra-articular injection in weekly intervals. Knee samples were taken after two weeks of the last treatment for histopathologic investigations. ResultsIn the OA group, surface fibrillation and irregularity, multiple clefts, loss of chondrocytes, proteoglycan depletion with Toluidine blue staining were detected. In the treated group with SRGF/vitamin C, the severity of degenerative lesions was decreased. Chondrocytes had proliferated and matrix proteoglycan increased in compared to the SRGF and OA groups. Also, osteoarthritis stage was markedly reduced in this group rather than two other groups. ConclusionThe results of this study show the synergic effect of vitamin C and growth factors on accelerating articular repair.

Nanostructural and mechanical changes in the sclera following proteoglycan depletion

The mechanical properties of ocular tissues, such as the sclera, have a major impact on healthy eye function, and are governed by the properties and composition of the microstructural components. For example, biomechanical degradation associated with myopia occurs alongside a reduction of proteoglycans (PGs). In this study, the role of PG degradation in the nanomechanical properties of the porcine sclera is explored. In-vitro enzymatic degradation of PGs was conducted with α-amylase and chondroitinase ABC enzymes. Collagen fibril morphology and nanomechanical stiffness were measured with atomic force microscopy (AFM). The elastic modulus of the tissue was reduced in all enzyme-treated samples relative to controls. In addition, collagen fibril organization was disrupted by PG depletion. Our data demonstrate that PGs play an important role in determining not only the mechanical properties at these length scales, but also collagen fibril arrangement.

Development of an in vivo mouse model of discogenic low back pain.

Discogenic low back pain (DLBP) is extremely common and costly. Effective treatments are lacking due to DLBP's unknown pathogenesis. Currently, there are no in vivo mouse models of DLBP, which restricts research in this field. The aim of this study was to establish a reliable DLBP model in mouse that captures the pathological changes in the disc and allows longitudinal pain testing. The model was generated by puncturing the mouse lumbar discs (L4/5, L5/6, and L6/S1) and removing the nucleus pulposus using a microscalpel under the microscope. Histology, molecular pathways, and pain-related behaviors were examined. Over 12 weeks post-surgery, animals displayed the mechanical, heat, and cold hyperalgesia along with decreased burrowing and rearing. Histology showed progressive disc degeneration with loss of disc height, nucleus pulposus reduction, proteoglycan depletion, and annular fibrotic disorganization. Immunohistochemistry revealed a substantial increase in inflammatory mediators at 2 and 4 weeks. Nerve growth factor was upregulated from 2 weeks to the end of the experiment. Nerve fiber ingrowth was induced in the injured discs after 4 weeks. Disc-puncture also produced an upregulation of neuropeptides in dorsal root ganglia neurons and an activation of glial cells in the spinal cord dorsal horn. These findings indicate that the cellular and structural changes in discs, as well as peripheral and central nervous system plasticity, paralleled persistent, and robust behavioral pain responses. Therefore, this mouse DLBP model could be used to investigate mechanisms underlying discogenic pain, thereby facilitating effective drug screening and development of treatments for DLBP.

Progression of Post-Traumatic Osteoarthritis in rat meniscectomy models: Comprehensive monitoring using MRI

Knee injury often triggers post-traumatic osteoarthritis (PTOA) that affects articular cartilage (AC), subchondral bone, meniscus and the synovial membrane. The available treatments for PTOA are largely ineffective due to late diagnosis past the “treatment window”. This study aimed to develop a detailed understanding of the time line of the progression of PTOA in murine models through longitudinal observation of the femorotibial joint from the onset of the disease to the advanced stage. Quantitative magnetic resonance microimaging (µMRI) and histology were used to evaluate PTOA-associated changes in the knee joints of rats subjected to knee meniscectomy. Systematic longitudinal changes in the articular cartilage thickness, cartilage T2 and the T2 of epiphysis within medial condyles of the tibia were all found to be associated with the development of PTOA in the animals. The following pathogenesis cascade was found to precede advanced PTOA: meniscal injury → AC swelling → subchondral bone remodelling → proteoglycan depletion → free water influx → cartilage erosion. Importantly, the imaging protocol used was entirely MRI-based. This protocol is potentially suitable for whole-knee longitudinal, non-invasive assessment of the development of OA. The results of this work will inform the improvement of the imaging methods for early diagnosis of PTOA.

Critical-sized cartilage defects in the equine carpus

ABSTRACTAim: The horse joint, due to its similarity with the human joint, is the ultimate model for translational articular cartilage repair studies. This study was designed to determine the critical size of cartilage defects in the equine carpus and serve as a benchmark for the evaluation of new cartilage treatment options. Material and Methods: Circular full-thickness cartilage defects with a diameter of 2, 4, and 8 mm were created in the left middle carpal joint and similar osteochondral (3.5 mm in depth) defects in the right middle carpal joint of 5 horses. Spontaneously formed repair tissue was examined macroscopically, with MR and µCT imaging, polarized light microscopy, standard histology, and immunohistochemistry at 12 months. Results: Filling of 2 mm chondral defects was good (77.8 ± 8.5%), but proteoglycan depletion was evident in Safranin-O staining and gadolinium-enhanced MRI (T1Gd). Larger chondral defects showed poor filling (50.6 ± 2.7% in 4 mm and 31.9 ± 7.3% in 8 mm defects). Lesion filling in 2, 4, and 8 mm osteochondral defects was 82.3 ± 3.0%, 68.0 ± 4.6% and 70.8 ± 15.4%, respectively. Type II collagen staining was seen in 9/15 osteochondral defects but only in 1/15 chondral defects. Subchondral bone pathologies were evident in 14/15 osteochondral samples but only in 5/15 chondral samples. Although osteochondral lesions showed better neotissue quality than chondral lesions, the overall repair was deemed unsatisfactory because of the subchondral bone pathologies. Conclusion: We recommend classifying 4 mm as critical osteochondral lesion size and 2 mm as critical chondral lesion size for cartilage repair research in the equine carpal joint model.

A method for characterising human intervertebral disc glycosaminoglycan disaccharides using liquid chromatography-mass spectrometry with multiple reaction monitoring.

Intervertebral disc (IVD) degeneration results in the depletion of proteoglycans and glycosaminoglycans (GAGs), which can lead to structural and mechanical loss of IVD function, ingrowth of nociceptive nerve fbres and eventually discogenic pain. Specifc GAG types as well as their disaccharide paterns can be predictive of disease and degeneration in several tissues but have not been comprehensively studied within the IVD. A highly sensitive mass spectrometry based technique with multiple reaction monitoring (MRM) was used to provide characterisation of chondroitin sulphate (CS), hyaluronic acid (HA), heparan sulphate (HS) and their disaccharide sulphation paterns across diferent anatomical regions of human IVDs. Principal component analysis further distinguished important regional variations and proposed potential ageing variations in GAG profles. CS was the GAG in greatest abundance in the IVD followed by HA and HS. Principal component analysis identifIed clear separation of GAG profiles between nucleus pulposus and annulus fibrosus in young and old specimens. Distinct patterns of predominantly expressed disaccharides of CS and HS between young and old IVD samples, provided preliminary evidence that important alterations in disaccharides occur within IVDs during ageing. This technique offered a novel approach to identify and quantify specific GAG disaccharides in human IVDs and the data presented were the first to offer insight into the spatial distribution as well as association with ageing of GAGs and GAG disaccharide sulphation patterns across the human IVD.

Intra-articular delivery of tetramethylpyrazine microspheres with enhanced articular cavity retention for treating osteoarthritis

Tetramethylpyrazine (TMP) is a traditional Chinese herbal medicine with strong anti-inflammatory and cartilage protection activities, and thus a promising candidate for treating osteoarthritis. However, TMP is rapidly cleared from the joint cavity after intra-articular injection and requires multiple injections to maintain efficacy. The aim of this study was to encapsulate TMP into poly (lactic-co-glycolic acid) (PLGA) microspheres to enhance the TMP retention in the joint, reducing injection frequencies and decreasing dosage. TMP microspheres were prepared by emulsion/solvent evaporation method. The intra-articular retention of the drug was assessed by detecting the drug concentration distributed in the joint tissue at different time points. The therapeutic effect of TMP microspheres was evaluated by the swelling of knee joints and histologic analysis in papain-induced OA rat model. The prepared freeze-dried microspheres with a particle size of about 10 µm can effectively prolong the retention time of the drug in the articular cavity to 30 d, which is 4.7 times that of the TMP solution. Intra-articular injection of TMP microspheres efficiently relieved inflammatory symptoms, improved joint lesions and decreased the depletion of proteoglycan. In conclusion, intra-articular injection of TMP loaded microspheres was a promising therapeutic method in the treatment of OA.

New algorithm for simulation of proteoglycan loss and collagen degeneration in the knee joint: Data from the osteoarthritis initiative.

Osteoarthritis is a harmful joint disease but prediction of disease progression is problematic. Currently, there is only one modeling framework which can be applied to predict the progression of knee osteoarthritis but it only considers degenerative changes in the collagen fibril network. Here, we have developed the framework further by considering all of the major tissue changes (proteoglycan content, fluid flow, and collagen fibril network) occurring in osteoarthritis. While excessive levels of tissue stresses controlled degeneration of the collagen fibril network, excessive levels of tissue strains controlled the decrease in proteoglycan content and the increase in permeability. We created four knee joint models with increasing degrees of complexity based on the depth-wise composition. Models were tested for normal and abnormal, physiologically relevant, loading conditions in the knee. Finally, the predicted depth-wise compositional changes from each model were compared against experimentally observed compositional changes in vitro. The model incorporating the typical depth-wise composition of cartilage produced the best match with experimental observations. Consistent with earlier in vitro experiments, this model simulated the greatest proteoglycan depletion in the superficial and middle zones, while the collagen fibril degeneration was located mostly in the superficial zone. The presented algorithm can be used for predicting simultaneous collagen degeneration and proteoglycan loss during the development of knee osteoarthritis. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1673-1683, 2018.

Anti-citrullinated protein antibodies cause arthritis by cross-reactivity to joint cartilage.

Today, it is known that autoimmune diseases start a long time before clinical symptoms appear. Anti-citrullinated protein antibodies (ACPAs) appear many years before the clinical onset of rheumatoid arthritis (RA). However, it is still unclear if and how ACPAs are arthritogenic. To better understand the molecular basis of pathogenicity of ACPAs, we investigated autoantibodies reactive against the C1 epitope of collagen type II (CII) and its citrullinated variants. We found that these antibodies are commonly occurring in RA. A mAb (ACC1) against citrullinated C1 was found to cross-react with several noncitrullinated epitopes on native CII, causing proteoglycan depletion of cartilage and severe arthritis in mice. Structural studies by X-ray crystallography showed that such recognition is governed by a shared structural motif "RG-TG" within all the epitopes, including electrostatic potential-controlled citrulline specificity. Overall, we have demonstrated a molecular mechanism that explains how ACPAs trigger arthritis.

A Histopathological Scheme for the Quantitative Scoring of Intervertebral Disc Degeneration and the Therapeutic Utility of Adult Mesenchymal Stem Cells for Intervertebral Disc Regeneration.

The purpose of this study was to develop a quantitative histopathological scoring scheme to evaluate disc degeneration and regeneration using an ovine annular lesion model of experimental disc degeneration. Toluidine blue and Haematoxylin and Eosin (H&E) staining were used to evaluate cellular morphology: (i) disc structure/lesion morphology; (ii) proteoglycan depletion; (iii) cellular morphology; (iv) blood vessel in-growth; (v) cell influx into lesion; and (vi) cystic degeneration/chondroid metaplasia. Three study groups were examined: 5 × 5 mm lesion; 6 × 20 mm lesion; and 6 × 20 mm lesion plus mesenchymal stem cell (MSC) treatment. Lumbar intervertebral discs (IVDs) were scored under categories (i–vi) to provide a cumulative score, which underwent statistical analysis using STATA software. Focal proteoglycan depletion was associated with 5 × 5 mm annular rim lesions, bifurcations, annular delamellation, concentric and radial annular tears and an early influx of blood vessels and cells around remodeling lesions but the inner lesion did not heal. Similar features in 6 × 20 mm lesions occurred over a 3–6-month post operative period. MSCs induced a strong recovery in discal pathology with a reduction in cumulative histopathology degeneracy score from 15.2 to 2.7 (p = 0.001) over a three-month recovery period but no recovery in carrier injected discs.

Are axial intervertebral disc biomechanics determined by osmosis?

The intervertebral disc faces high compressive forces during daily activities. Axial compression induces creeping fluid loss and reduction in disc height. With degeneration, disc fluids and height are progressively lost, altering biomechanics. It is assumed that this reduction of fluids is caused by a decline of osmolality within the disc due to proteoglycan depletion. Here we investigate the isolated effect of a reduction in osmosis on the biomechanical properties of the intervertebral disc. Continuous diurnal loading was applied to healthy caprine intervertebral discs in a loaded disc culture system for a total of 6days. We increased testing bath osmolality with two doses of polyethylene-glycol (PEG), thereby reducing the osmotic gradient between the disc and the surrounding fluid. This way we could study the isolated effect of reduced osmosis on axial creep, without damaging the disc. We evaluated: daily creep and recovery, recovery time-constants and compressive stiffness. Additionally, we investigated water content. There was a strong dose-dependent effect of PEG concentration on water content and axial creep behaviour: disc height, amplitude and rate of creep and recovery were all significantly reduced. Axial compressive stiffness of the disc was not affected. Reduction of water content and amplitude of creep and recovery showed similarity to degenerative disc biomechanics. However, the time-constants increased, indicating that the hydraulic permeability was reduced, in contrast to what happens with degeneration. This suggests that besides the osmotic gradient, the permeability of the tissues determines healthy intervertebral disc biomechanics.

A new light on Alkaptonuria: A Fourier-transform infrared microscopy (FTIRM) and low energy X-ray fluorescence (LEXRF) microscopy correlative study on a rare disease

BackgroundAlkaptonuria (AKU) is an ultra-rare disease associated to the lack of an enzyme involved in tyrosine catabolism. This deficiency results in the accumulation of homogentisic acid (HGA) in the form of ochronotic pigment in joint cartilage, leading to a severe arthropathy. Secondary amyloidosis has been also unequivocally assessed as a comorbidity of AKU arthropathy. Composition of ochronotic pigment and how it is structurally related to amyloid is still unknown. MethodsWe exploited Synchrotron Radiation Infrared and X-Ray Fluorescence microscopies in combination with conventional bio-assays and analytical tools to characterize chemical composition and morphology of AKU cartilage. ResultsWe evinced that AKU cartilage is characterized by proteoglycans depletion, increased Sodium levels, accumulation of lipids in the peri-lacunar regions and amyloid formation. We also highlighted an increase of aromatic compounds and oxygen-containing species, depletion in overall Magnesium content (although localized in the peri-lacunar region) and the presence of calcium carbonate fragments in proximity of cartilage lacunae. ConclusionsWe highlighted common features between AKU and arthropathy, but also specific signatures of the disease, like presence of amyloids and peculiar calcifications. Our analyses provide a unified picture of AKU cartilage, shedding a new light on the disease and opening new perspectives. General significanceOchronotic pigment is a hallmark of AKU and responsible of tissue degeneration. Conventional bio-assays have not yet clarified its composition and its structural relationship with amyloids. The present work proposes new strategies for filling the aforementioned gap that encompass the integration of new analytical approaches with standardized analyses.

T1ρ/T2 mapping and histopathology of degenerative cartilage in advanced knee osteoarthritis.

AIMTo investigate whether normal thickness cartilage in osteoarthritic knees demonstrate depletion of proteoglycan or collagen content compared to healthy knees.METHODSMagnetic resonance (MR) images were acquired from 5 subjects scheduled for total knee arthroplasty (TKA) (mean age 70 years) and 20 young healthy control subjects without knee pain (mean age 28.9 years). MR images of T1ρ mapping, T2 mapping, and fat suppressed proton-density weighted sequences were obtained. Following TKA each condyle was divided into 4 parts (distal medial, posterior medial, distal lateral, posterior lateral) for cartilage analysis. Twenty specimens (bone and cartilage blocks) were examined. For each joint, the degree and extent of cartilage destruction was determined using the Osteoarthritis Research Society International cartilage histopathology assessment system. In magnetic resonance imaging (MRI) analysis, 2 readers performed cartilage segmentation for T1ρ/T2 values and cartilage thickness measurement.RESULTSEleven areas in MRI including normal or near normal cartilage thickness were selected. The corresponding histopathological sections demonstrated mild to moderate osteoarthritis (OA). There was no significant difference in cartilage thickness in MRI between control and advanced OA samples [medial distal condyle, P = 0.461; medial posterior condyle (MPC), P = 0.352; lateral distal condyle, P = 0.654; lateral posterior condyle, P = 0.550], suggesting arthritic specimens were morphologically similar to normal or early staged degenerative cartilage. Cartilage T2 and T1ρ values from the MPC were significantly higher among the patients with advanced OA (P = 0.043). For remaining condylar samples there was no statistical difference in T2 and T1ρ values between cases and controls but there was a trend towards higher values in advanced OA patients.CONCLUSIONThough cartilage is morphologically normal or near normal, degenerative changes exist in advanced OA patients. These changes can be detected with T2 and T1ρ MRI techniques.

Magnetic Resonance Imaging Findings in the Knee Before and After Long-Distance Running—Documentation of Irreversible Structural Damage? A Systematic Review

Background: Various studies have investigated structural knee changes after running, with conflicting results. Purpose: To perform a systematic review of acute changes in knee structures as detected by magnetic resonance imaging (MRI) after running and assess the reversibility of these changes. Study Design: Systematic review. Methods: A systematic literature search in Medline, Cochrane, Embase, and Scopus was performed. Articles that fulfilled predefined inclusion criteria were included and systematically reviewed according to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) statement. Results: A total of 19 studies were included in this review. All studies performed standard knee MRI; 6 studies additionally performed T1ρ and T2 mapping. Sixteen studies assessed cartilage or meniscal morphological changes. Ten found no significant morphological changes after running. Six studies showed significant changes at the first follow-up. Six performed a second follow-up. Five showed no change compared with baseline and the first follow-up, and 1 showed a significant recovery compared with the first follow-up and no significant difference compared with baseline. Five of the 6 studies performing T1ρ and T2 mapping found significant changes in T2 and T1ρ values at the first follow-up. Three performed a second follow-up. Two found a significant recovery of T2 but no recovery of T1ρ. One study did not find a significant change compared with baseline. Ten studies assessed the patellar tendon, ligaments, synovial fluid, or subchondral bone. Changes at the first follow-up were not significant. A second follow-up was performed in 5 studies. All studies discovered recovery from the first follow-up. Conclusion: These data suggest that healthy athletes who have no risk factors for degenerative joint disease may present fleeting quantitative alterations after running. No irreversible, qualitative harmful effects seemed to occur, with the exception of persistent T1ρ elevation representing a proteoglycan depletion. Whether T1ρ changes need more than 3 months to recover or represent permanent structural damage remains to be investigated.

Effects of BIS076 in a model of osteoarthritis induced by anterior cruciate ligament transection in ovariectomised rats.

BackgroundOsteoarthritis (OA) is the most frequent articular disease and a leading cause of disability. There is a need for effective treatments able to slow the progression of disease. Some of the available treatments are dietary supplements providing natural components. Recent studies have shown that estrogen deficiency contributes to the pathophysiological events of OA progression.MethodsWe have used the anterior cruciate ligament transection model of OA in ovariectomised rats to study the effects of BIS076, a new formulation of a natural porcine cartilage extract associated with hydroxyapatite (as a source of calcium) and vitamin D3. Cartilage degradation, proteoglycan depletion and synovitis were followed by histochemistry. Effects on bone microstructure were determined by μCT. The levels of biomarkers in serum and inflammatory mediators in knee homogenates were measured by luminex or ELISA.ResultsOral administration of BIS076 reduced articular cartilage damage and serum levels of cartilage degradation markers C-telopeptide of type II collagen and cartilage oligomeric matrix protein, as well as matrix metalloproteinase-3. The local inflammatory response was down-regulated by BIS076 with lower production of pro-inflammatory cytokines and prostaglandin E2 in joint tissues. In addition, BIS076 was effective on metaphyseal bone alterations as this formulation increased volumetric bone mineral density and improved bone micro-architecture. These effects were related to the modification of bone metabolism reflected by changes in bone biomarkers with reductions in the ratio receptor activator of nuclear factor κB ligand/osteoprotegerin and the levels of tartrate-resistant acid phosphatase-5b, suggesting an inhibitory activity of BIS076 on trabecular bone resorption.ConclusionsWe have demonstrated the protective properties of a new formulation (BIS076) on joint lesion and bone alterations in an experimental model of OA in ovariectomised rats. This study supports the interest of BIS076 in OA treatments.