Femoral Head Articular Cartilage Research Articles

Characterization of the Age-Related Differences in Porcine Acetabulum and Femoral Head Articular Cartilage.

The use of porcine animal models for cartilage injury has increased recently due to their similarity with humans with regard to cartilage thickness, limited intrinsic healing of chondral defects, and joint loading biomechanics. However, variations in the mechanical and biochemical properties of porcine hip articular cartilage among various tissue ages and weightbearing (WB) regions are still unknown. This study's aim was to characterize the mechanical and biochemical properties of porcine hip articular cartilage across various ages and WB regions. Articular cartilage explants were harvested from WB and non-weightbearing (NWB) surfaces of the femoral head and acetabulum of domesticated pigs (Sus scrofa domesticus) at fetal (gestational age: 80 days), juvenile (6 months), and adult (2 years) ages. Explants underwent compressive stress-relaxation mechanical testing, biochemical analysis for total collagen and glycosaminoglycan (GAG) content, and histological staining. Juvenile animals consistently had the highest mechanical properties, with 2.2- to 7.6-time increases in relaxation modulus, 1.3- to 2.3-time increases in instantaneous modulus, and 4.1- to 14.2-time increases in viscosity compared with fetal cartilage. Mechanical properties did not significantly differ between the WB and NWB regions. Collagen content was highest in the NWB regions of the juvenile acetabulum (65.3%/dry weight [DW]) and femoral head (75.4%/DW) cartilages. GAG content was highest in the WB region of the juvenile acetabulum (23.7%/DW) and the WB region of the fetal femoral head (27.5%/DW) cartilages. Histological staining for GAG and total collagen content followed the trends from the quantitative biochemical assays. This study provides a benchmark for the development and validation of preclinical porcine models for hip cartilage pathologies.

The surgical destabilization ofthe abductor muscle leads to development ofinstability-associated hip osteoarthritis inmice.

Osteoarthritis (OA) of the hip is a common and debilitating painful joint disease. However, there is paucity of surgically induced hip OA models in small animals that allow scientists to study the onset and progression of the disease. A growing body of evidence indicates a positive association between periarticular myotendinous pathology and the development of hip OA. Thus, in the current study, we aimed to establish a novel mouse instability-associated hip OA model via selective injury of the abductor complex around the hip joint. C57BL6/J mice were randomized to sham surgery or abductor injury, in which the myotendinous insertion at the third trochanter and greater trochanter were surgically detached. Mice were allowed free active movement until they were sacrificed at either 3 weeks or 20 weeks post-injury. Histologic analyses and immunohistochemical staining of the femoral head articular cartilage were performed, along with microCT (µCT) analysis to assess subchondral bone remodeling. We observed that mice receiving abductor injury exhibited significantly increased instability-associated OA severity with loss of proteoglycan and type II collagen staining compared to sham control mice at 20 weeks post-surgery, while comparable matrix metalloproteinase 13 expression was observed between injury and sham groups. No significant differences in subchondral bone remodeling were found after 3 or 20 weeks following injury. Our study further supports the link between abductor dysfunction and the development of instability-associated hip OA. Importantly, this novel surgically induced hip OA mouse model may provide a valuable tool for future investigations into the pathogenesis and treatment of hip OA.

Whole-genome RNAsequencing identifies distinct transcriptomic profiles in impingement cartilage between patients with femoroacetabular impingement and hip osteoarthritis.

Femoroacetabular impingement (FAI) has a strong clinical association with the development of hip osteoarthritis (OA); however, the pathobiological mechanisms underlying the transition from focal impingement to global joint degeneration remain poorly understood. The purpose of this study is to use whole-genome RNA sequencing to identify and subsequently validate differentially expressed genes (DEGs) in femoral head articular cartilage samples from patients with FAI and hip OA secondary to FAI. Thirty-seven patients were included in the study with whole-genome RNA sequencing performed on 10 gender-matched patients in the FAI and OA cohorts and the remaining specimens were used for validation analyses. We identified a total of 3531 DEGs between the FAI and OA cohorts with multiple targets for genes implicated in canonical OA pathways.Quantitative reverse transcription-polymerase chain reaction validation confirmed increased expression of FGF18 and WNT16 in the FAI samples, while there was increased expression of MMP13 and ADAMTS4 in the OA samples. Expression levels of FGF18 and WNT16 were also higher in FAI samples with mild cartilage damage compared to FAI samples with severe cartilage damage or OA cartilage. Our study further expands the knowledge regarding distinct genetic reprogramming in the cartilage between FAI and hip OA patients. We independently validated the results of the sequencing analysis and found increased expression of anabolic markers in patients with FAI and minimal histologic cartilage damage, suggesting that anabolic signaling may be increased in early FAI with a transition to catabolic and inflammatory gene expression as FAI progresses towards more severe hip OA.Clinical significance:Cam-typeFAI has a strong clinical association with hip OA; however, the cellular pathophysiology of disease progression remains poorly understood. Several previous studies have demonstrated increased expression of inflammatory markers in FAI cartilage samples, suggesting the involvement of these inflammatory pathways in the disease progression. Our study further expands the knowledge regarding distinct genetic reprogramming in the cartilage between FAI and hip OA patients. In addition to differences in inflammatory gene expression, we also identified differential expression in multiple pathways involved in hip OA progression.

Correlation of Delayed Gadolinium-Enhanced MRI of Cartilage (dGEMRIC) Value With Hip Arthroscopy Intraoperative Findings and Midterm Periacetabular Osteotomy Outcomes.

Background:Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) is an advanced imaging technique that is purported to quantify cartilage damage in acute and chronic joint disease and predict periacetabular osteotomy (PAO) outcomes. There is a paucity of literature relating dGEMRIC values to arthroscopic findings before PAO and postoperative outcomes after PAO.Purpose:To assess the utility and validity of dGEMRIC as a preoperative and prognostic assessment tool of cartilage status and integrity as it relates to intraoperative findings and midterm postoperative outcomes after PAO.Study Design:Case series; Level of evidence, 4.Methods:We analyzed a cohort of 58 patients (70 hips) with a median age of 30.1 years (range, 15-50) with hip dysplasia who underwent hip arthroscopy, followed by a PAO with preoperative dGEMRIC. The primary outcome measures were intraoperative assessment and correlation with cartilage damage (presence of cartilage flap, Outerbridge grade of the acetabulum and femoral head). Secondary outcome measures were postoperative patient-reported outcome (PRO) scores, including the International Hip Outcome Tool and Non-arthritic Hip Score. Correlation analyses were performed to determine the relationship between dGEMRIC values and (1) PROs and (2) intraoperative assessment of cartilage damage.Results:There were significant negative linear relationships between dGEMRIC values and the primary outcome measures: presence of a cartilage flap (coronal, P = .004; sagittal, P < .001), Outerbridge grade of acetabular articular cartilage lesion (coronal, P = .002; sagittal, P = .003), and Outerbridge grade of femoral head articular cartilage lesion (coronal, P = .001; sagittal, P < .001). Despite significant overall improvement in all patients, there was no significant correlation between preoperative dGEMRIC values and improvement in PROs from presurgery to latest postoperative follow-up (median, 2.2 years; range, 1.0-5.0 years).Conclusion:Although dGEMRIC values (sagittal and coronal) were significant predictors of the intraoperative presence of cartilage flaps and overall cartilage integrity, they were not associated with midterm outcomes after PAO.

Quantitative Magnetic Resonance Imaging of Femoral Head Articular Cartilage Change in Patients with Hip Osteonecrosis Treated with Extracorporeal Shock Wave Therapy.

Background Multiple reports have demonstrated the therapeutic potential of extracorporeal shock wave (ESWT) in osteonecrosis of the femoral head (ONFH). However, few studies reported the changes in hip articular cartilage after the intervention. This study aimed to investigate the effect of ESWT on femoral head cartilage using a novel technique, quantitative T2-mapping magnetic resonance imaging. Methods A total of 143 eligible patients with unilateral early-stage ONFH were randomized into the ESWT group and control group. Seventy-three patients in the ESWT group received two sessions of ESWT with oral drug treatment, while seventy patients in the control group received oral drug treatment only. The visual analog pain scale (VAS) and Harris hip score (HHS) at 3-month, 6-month, and 12-month follow-up were used as the clinical evaluation index. The radiological evaluation index used the T2 mapping values, necrotic size, and China-Japan Friendship Hospital (CJFH) classification. Results A total of 143 patients (62 females and 81 males) were finally included, and the characteristics before treatment were comparable between the two groups. At the last follow-up (12 months), the T2 values and ΔT2 changes in the ESWT group were all smaller than those in the control group (p=0.042; p=0.039), while the CJFH classification of ONFH and necrotic lesion size were not statistically significant. At 3 months and 6 months, the VAS in the ESWT group was lower than that in the control group (p=0.021; p=0.046) and the HHS in the ESWT group was higher (p=0.028; p=0.039). However, there were no significant differences in the VAS and HHS at 12 months between the ESWT and control groups. Conclusions The results of the current study indicated that, based on drug treatment, ESWT is an effective treatment method for nontraumatic ONFH, which could result in significant pain relief and function restoration. Furthermore, it could delay the injury of femoral head cartilage during the progression of ONFH.

Morphology of the articular cartilage of the femoral head broiler chickens cross Ross-308 in the age aspect and against the background of the use of dietary supplements

In modern broiler poultry farming, the problem of different rates of formation of muscle and connective tissue has become increasingly common. To a greater extent, this problem is affected by highly productive crosses of broiler chickens (Ross-308, Cobb500), whose selection is aimed at accelerated recruitment of live weight, without taking into account the development of the backbone and joints. According to Russian and foreign authors, during the period of broiler cultivation, the rate of poultry culling is on average 10%. Of these, more than half are pathologies of the musculoskeletal system, both non-infectious (injuries, unbalanced diet in terms of macro-and microelement composition) and infectious (reovirus tenosynovitis) etiology.Our study presents the results of an experiment to study the effect of silicon dioxide and dihydroquercetin on the zonal structure of the articular cartilage of the femoral head of broiler chickens in the age aspect. The object of the study was broiler chickens of the Ross-308 cross. 160 head of chickens on the whole were examined. During the experiment, four groups of chickens with 40 chicks each were formed according to the principle of analogues. The first group was a control one and received the main diet until the age of 38 days, the chickens of the second, third and fourth groups were fed with feed additives "Covelos-Sorb" in the amount of 0.1 g, 0.14 g and 0.18 g/ 1 kg of live weight per day respectively and "Ecostimul2" in the amount of 1 mg/1 kg of live weight per day. As a result of the study, it was found that the change in the thickness of the structural zones of the articular cartilage of the broiler chicken femoral head is periodic and asynchronous, depending on the age, individual characteristics and dosage of biologically active supplements. There was also an increase in the proportion of isogenic groups in the intermediate zone of the articular cartilage of the femoral head in experimental chickens, which may indicate an increase in the activity of chondrogenesis.

Systemic administration of a pharmacologic iron chelator reduces cartilage lesion development in the Dunkin-Hartley model of primary osteoarthritis

Iron has been emerging as a key contributor to aging-associated, chronic disorders due to the propensity for generating reactive oxygen species. To date, there are a limited number of publications exploring the role of iron in the pathogenesis of primary/age-related osteoarthritis (OA). The objective of this study was to determine whether reduced iron via pharmacologic iron chelation with deferoxamine (DFO) affected the development and/or severity of cartilage lesions in a primary OA model. At 12-weeks-of-age, 15 male Dunkin-Hartley guinea pigs received either 46 mg/kg DFO (n = 8) or vehicle control (n = 7) injected subcutaneously twice daily for five days each week. Movement changes, captured via overhead enclosure monitoring, were also determined. Termination occurred at 30-weeks-of-age. Iron was quantified in serum, urine, liver, and femoral head articular cartilage. Left knees were evaluated for: structural changes using histopathology guidelines; and immunohistochemistry. Gene expression analysis was conducted on right knee articular cartilage. DFO reduced iron levels in femoral head articular cartilage (p = 0.0006) and liver (p = 0.02), and increased iron within urine (p = 0.04) and serum (p = 0.0009). Mobility of control animals declined, while the DFO group maintained activity levels similar to the first month of treatment (p = 0.05). OA-associated cartilage lesions were reduced in knees of DFO animals (p = 0.0001), with chondrocyte hypocellularity a key histologic difference between groups (p < 0.0001). DFO-receiving animals had increased immunostaining for phosphorylated adenosine monophosphate activated protein kinase alpha within knee articular cartilage; lower transcript counts of several proapoptotic genes (p = 0.04–0.0004) and matrix-degrading enzymes (p = 0.02-<0.0001), and increased expression of the anti-apoptotic gene Bcl-2 (p < 0.0001) and a tissue inhibitor of matrix-metalloproteinases (p = 0.03) were also observed. These results suggest that iron chelation delayed the progression of primary OA in an animal model and could hold potential as a translational intervention. These findings provide expanded insight into factors that may contribute to the pathogenesis of primary OA.

Minimally invasive medial hip approach for femoral head fixation: How much can we see and what is the anatomical at risk? – A cadaveric injection study

IntroductionMost Pipkin I and II femoral head fractures are treated with either an anterior or a posterior approach. A medial hip approach is commonly used in children, and some surgeons have suggested it for femoral head fixation. The objectives of this study were to identify the structures at risk with the medial hip approach and to demonstrate the areas of the femoral head exposed using this approach. MethodsThe first part of this study involved vascular injection conducted in four fresh human cadavers using the medial hip approach. The surgical technique was described and the structures at risk, mainly arteries, were identified. The second part was done in 14 hips to identify and measured the maximum exposure area of the femoral head with the medial hip approach. ResultsThe structures at risk with the medial hip approach were the medial femoral circumflex artery (MFCA) after it branches from the deep femoral artery and runs posteromedially across the femoral neck medial to the iliopsoas tendon and the deep branch of the MFCA lies over the posterior hip capsule. The femoral head exposure area in anterior view with anteroinferior articular cartilage exposure was 4.57 ± 1.09 cm2. In the medial view with the hip in internal rotation and in external rotation, the inferomedial articular cartilage exposure was 2.50 ± 1.52 cm2 and 6.48 ± 2.29 cm2, respectively, and the combined area of exposure was 9.49 ± 2.86 cm2 . In the posterior view, the posteroinferior articular exposure was 1.69 ± 1.16 cm2. The medial hip approach allowed visualization of the small area of the posteroinferior articular cartilage in all specimens. The percentages of visualization of the femoral head articular cartilage in anterior view, medial view and posterior view were 50.07% ± 13.53, 45.56% ± 12.76 and 18.07% ± 11.27%, respectively. ConclusionsThe structures at risk with the medial hip approach is the MFCA along the anterior acetabular rim and the deep branch on the posteromedial aspect of the femoral neck. It is an alternative which provides excellent access in Pipkin I and some part of Pipkin II, but it requires that the MFCA be protected by the use of meticulous surgical techniques.

Circadian time series proteomics reveals daily dynamics in cartilage physiology

SummaryObjectiveCartilage in joints such as the hip and knee experiences repeated phases of heavy loading and low load recovery during the 24-h day/night cycle. Our previous work has shown 24 h rhythmic changes in gene expression at transcript level between night and day in wild type mouse cartilage which is lost in a circadian clock knock-out mouse model. However, it remains unknown to what extent circadian rhythms also regulate protein level gene expression in this matrix rich tissue.MethodsWe investigated daily changes of protein abundance in mouse femoral head articular cartilage by performing a 48-h time-series LC-MS/MS analysis.ResultsOut of the 1,177 proteins we identified across all time points, 145 proteins showed rhythmic changes in their abundance within the femoral head cartilage. Among these were molecules that have been implicated in key cartilage functions, including CTGF, MATN1, PAI-1 and PLOD1 & 2. Pathway analysis revealed that protein synthesis, cytoskeleton and glucose metabolism exhibited time-of-day dependent functions. Analysis of published cartilage proteomics datasets revealed that a significant portion of rhythmic proteins were dysregulated in osteoarthritis and/or ageing.ConclusionsOur circadian proteomics study reveals that articular cartilage is a much more dynamic tissue than previously thought, with chondrocytes driving circadian rhythms not only in gene transcription but also in protein abundance. Our results clearly call for the consideration of circadian timing mechanisms not only in cartilage biology, but also in the pathogenesis, treatment strategies and biomarker detection in osteoarthritis.

Systemic iron overload exacerbates osteoarthritis in the strain 13 guinea pig

Iron is emerging as a key player in aging-associated diseases due to its propensity for driving free radical formation. Studies examining the role of iron in the pathogenesis of primary osteoarthritis (OA) are limited. Our objective was to establish a direct relationship between excess iron and OA by administering iron dextran to a guinea pig strain with decreased propensity for developing this disease. Twenty, 12-week-old Strain 13 guinea pigs received either iron dextran or dextran control intraperitoneally once weekly for 4 weeks; termination occurred at 16 weeks of age. Iron levels were determined systemically (serum and liver) and within diarthrodial joints [femoral head articular cartilage and infrapatellar fat pads (IFPs) of knee joints]. One knee was collected to score structural changes associated with OA via microcomputed tomography (microCT) and histology using published grading schemes. Articular cartilage and IFPs were harvested from contralateral knees for gene expression analyses. Iron overload was confirmed systemically via increased serum iron and liver iron concentration. Articular cartilage and IFPs in the iron dextran group also had higher levels of iron. Excess iron worsened knee OA using both microCT and histologic scoring systems. Gene analyses revealed that exogenous iron altered the expression of iron trafficking proteins, select cytokines, and structural components of cartilage. These results demonstrate that systemic iron overload caused cellular iron accumulation in the knee joint. This excess iron is associated with increased expression of local inflammatory mediators and early onset and progression of knee joint OA in Strain 13 animals.

Histopathological Findings of Failed Free Vascularized Fibular Grafting for Osteonecrosis of the Femoral Head after Long-Term Follow-Up.

Purpose The aim of this study was to report the histopathology of failed free vascularized fibular grafting (FVFG) for osteonecrosis of the femoral head (ONFH) after a mean follow-up of 11.5 years (ranged from 10.6 to 14.2 years). Methods Six hips of 5 patients with a history of steroid use, aged 34–67 years, were in stage II of ONFH as classified by the Ficat and Arlet classification at the time of FVFG treatment. Grafting failure led to osteoarthritis of the hip joint during a mean of 11.5 years of follow-up. Femoral head specimens were first evaluated macroscopically. Bone specimens were sectioned into long strips, divided into necrotic, transitional, and healthy zones, and then prepared for nondecalcified and decalcified histopathological examination using hematoxylin and eosin (HE) staining, Goldner's trichrome staining, and immunofluorescence (IF) staining. Results Femoral head articular cartilage surfaces appeared thin, opaque, and partially cartilaginous missing, with gradual collapse detected in weight-bearing areas. The interface with the fibular graft showed well union, with no obvious gaps between graft and host bone, as observed macroscopically. The necrotic area was filled with fibular graft, cancellous bone, and cartilaginous or soft tissue invasion. Histopathology results revealed well integration between fibular graft and host bone, with thickened trabecular bone. Gaps occurred in transitional and healthy zones. In the necrotic zone, cartilaginous or soft tissue invasion occurred, while thin or missing articular cartilage exposed subchondral bone to hip joint surfaces. By IF counterstaining with CD-31 and α-SMA, blood vessel transplanted during fibular grafting could be clearly observed along the graft from healthy to necrotic zones. In the necrotic zone, blood vessels presented obviously and spread into the surrounding area of the graft tip. Conclusion After FVFG procedure with a mean follow-up of 11.5 years, fibular grafts retained their integrity as viable, vascularized, cortical bone that fused well with host bone and formed thickened trabecular bone surrounding the surface of the graft. However, the revascularization of FVFG's blood vessels spreading from the tip of the fibular graft into subchondral area of necrotic lesion did not improve significantly in these failure cases. The local necrotic lesion failed to be repaired as healthy trabecular bone to buttress articular surface and was occupied by soft tissues.

Evaluation of musculoskeletal phenotype of the G608G progeria mouse model with lonafarnib, pravastatin, and zoledronic acid as treatment groups

Hutchinson-Gilford progeria syndrome (HGPS) is a uniformly fatal condition that is especially prevalent in skin, cardiovascular, and musculoskeletal systems. A wide gap exists between our knowledge of the disease and a promising treatment or cure. The aim of this study was to first characterize the musculoskeletal phenotype of the homozygous G608G BAC-transgenic progeria mouse model, and to determine the phenotype changes of HGPS mice after a five-arm preclinical trial of different treatment combinations with lonafarnib, pravastatin, and zoledronic acid. Microcomputed tomography and CT-based rigidity analyses were performed to assess cortical and trabecular bone structure, density, and rigidity. Bones were loaded to failure with three-point bending to assess strength. Contrast-enhanced µCT imaging of mouse femurs was performed to measure glycosaminoglycan content, thickness, and volume of the femoral head articular cartilage. Advanced glycation end products were assessed with a fluorometric assay. The changes demonstrated in the cortical bone structure, rigidity, stiffness, and modulus of the HGPS G608G mouse model may increase the risk for bending and deformation, which could result in the skeletal dysplasia characteristic of HGPS. Cartilage abnormalities seen in this HGPS model resemble changes observed in the age-matched WT controls, including early loss of glycosaminoglycans, and decreased cartilage thickness and volume. Such changes might mimic prevalent degenerative joint diseases in the elderly. Lonafarnib monotherapy did not improve bone or cartilage parameters, but treatment combinations with pravastatin and zoledronic acid significantly improved bone structure and mechanical properties and cartilage structural parameters, which ameliorate the musculoskeletal phenotype of the disease.

Enoxolone suppresses apoptosis in chondrocytes and progression of osteoarthritis via modulating the ERK1/2 signaling pathway.

Osteoarthritis (OA) is an inflammatory disorder of synovial joints which is mainly treated with therapeutic agents showing side effects associated with the gastrointestinal (GI) and metabolic system. Consequently, there is urgent need for a potent, safe and novel agent for treating OA and related disorders. Enoxolone is a pentacyclic triterpenoid obtained from the herb liquorice. Based on earlier findings, we postulated that enoxolone may produce chondroprotective activity by exerting anti-inflammatory, anti-catabolic and oxidative stress-decreasing effects. The chondrocytes were extracted from the femoral head articular cartilage of healthy rats. Immunofluorescence staining was done for identification of chondrocytes. Cell viability and proliferation studies were done using Cell Counting Kit-8. Apoptotic cells were identified by TUNEL assay and flow cytometry analysis. Autophagy was assessed by monodansylcadaverine assay. Western blot analysis was done for expression of proteins. In the present study we investigated the protective effect of enoxolone on interleukin 1β (IL-1β) treated Iry chondrocytes in vitro. Treatment with IL-1β resulted in a significant reduction in cell viability of cells in increasing dose and time. Treatment with enoxolone along with IL-1β caused a significant decrease in growth inhibition. Also, enoxolone inhibited the IL-1β mediated apoptosis and activation of caspase-3 in cells. We also observed that enoxolone elevated the levels of p-ERK1/2, light chain 3 (LC3)-II and Beclin-1 (autophagy markers) in chondrocytes. The expression of (LC3)-II and Beclin-1 was decreased when the cells were treated with U0126 (ERK1/2 inhibitor). Our findings demonstrate that enoxolone could suppress inflammatory signaling and apoptosis via the ERK1/2 pathway in chondrocytes.

GC–MS method for metabolic profiling of mouse femoral head articular cartilage reveals distinct effects of tissue culture and development

The metabolic profile of cartilage is important to define as it relates to both normal and pathophysiological conditions. Our aim was to develop a precise, high-throughput method for gas/chromatography-mass/spectrometry (GC-MS) semi-targeted metabolic profiling of mouse cartilage. Femoral head (hip) cartilage was isolated from 5- and 15-week-old male C57BL/6J mice immediately after death for invivo analyses. In vitro conditions were evaluated in 5-week-old samples cultured ±10% fetal bovine serum (FBS). We optimized cartilage processing for GC-MS analysis and evaluated group-specific differences by multivariate and parametric statistical analyses. 55 metabolites were identified in pooled cartilage (4 animals per sample), with 29 metabolites shared between invivo and invitro conditions. Multivariate analysis of these common metabolites demonstrated that culturing explants was the strongest factor altering cartilage metabolism, followed by age and serum starvation. In vitro culture altered the relative abundance of specific metabolites; whereas, cartilage development between five and 15-weeks of age reduced the levels of 36 out of 43 metabolites >2-fold, especially in TCA cycle and alanine, aspartate, and glutamate pathways. In vitro serum starvation depleted six out of 41 metabolites. This study describes the first GC-MS method for mouse cartilage metabolite identification and quantification. We observed fundamental differences in femoral head cartilage metabolic profiles between invivo and in vitro conditions, suggesting opportunities to optimize in vitro conditions for studying cartilage metabolism. In addition, the reductions in TCA cycle and amino acid metabolites during cartilage maturation illustrate the plasticity of chondrocyte metabolism during development.

Viscoelasticity of articular cartilage: Analysing the effect of induced stress and the restraint of bone in a dynamic environment.

The aim of this study was to determine the effect of the induced stress and restraint provided by the underlying bone on the frequency-dependent storage and loss stiffness (for bone restraint) or modulus (for induced stress) of articular cartilage, which characterise its viscoelasticity. Dynamic mechanical analysis has been used to determine the frequency-dependent viscoelastic properties of bovine femoral and humeral head articular cartilage. A sinusoidal load was applied to the specimens and out-of-phase displacement response was measured to determine the phase angle, the storage and loss stiffness or modulus. As induced stress increased, the storage modulus significantly increased (p < 0.05). The phase angle decreased significantly (p < 0.05) as the induced stress increased; reducing from 13.1° to 3.5°. The median storage stiffness ranged from 548 N/mm to 707 N/mm for cartilage tested on-bone and 544 N/mm to 732 N/mm for cartilage tested off-bone. On-bone articular cartilage loss stiffness was frequency independent (p > 0.05); however, off-bone, articular cartilage loss stiffness demonstrated a logarithmic frequency-dependency (p < 0.05). In conclusion, the frequency-dependent trends of storage and loss moduli of articular cartilage are dependent on the induced stress, while the restraint provided by the underlying bone removes the frequency-dependency of the loss stiffness.

A genome-wide transcriptomic analysis of articular cartilage during normal maturation in pigs

ObjectiveThe articular cartilage undergoes dramatic changes in structure and composition during post-natal maturation, but the associated transcriptional changes are not well characterized. Compared to a mature stage, the immature articular cartilage shows developmental features such as increased thickness, presence of blood vessels, and the presence of a deep layer of growth cartilage which undergoes endochondral ossification. These features decrease during normal development. Following maturation, the articular cartilage is known to undergo few minor modifications. Since mature articular cartilage has poor regenerative and repair capacity compared to the immature articular cartilage, a better understanding of the molecular changes during the normal postnatal articular cartilage development might reveal insights on the molecular adaptation. It may also provide new therapeutic strategies. The purpose of this study was to determine the differential expression of genes in the femoral head articular cartilage of 6-weeks old and 6-months old pigs using a genome-wide transcriptomic analysis. MethodsThe articular cartilage of the femoral head of 6-weeks and 6-months old normal pigs was assessed for thickness and vascularity (number of vascular canals) using Safranin O/Fast Green staining of paraffin sections (n=4 pigs/age group). The measurements were determined using Image J software. RNA was isolated from the femoral head articular cartilage from 6-weeks and 6-months old pigs (n=8 pigs/age group). A microarray analysis was performed using an Affymetrix Porcine GeneChip Array. A gene enrichment analysis and a functional clustering analysis were performed by DAVID and STRING software, respectively. The differential expression of selected genes was confirmed by a quantitative RTPCR analysis. ResultsThe femoral head articular cartilage showed a significant decrease in thickness and number of vascular canals in 6-months old compared to 6-weeks old pigs. A microarray analysis revealed a differential gene expression of 576 genes, with 206 genes that were significantly upregulated and 370 genes that were significantly downregulated (>2-fold change, p<0.05) at 6-months compared to 6-weeks of age. Among the upregulated genes, DAVID analysis revealed that a significant number of genes represented the biological processes of responses to external stimuli, and wounding and inflammation at 6-months of age. These processes involved genes representing secretory and signaling proteins such as MMP-1, MMP-3, IL-8 and STAT3 suggesting increased inflammatory activity. In addition, an assessment of the downregulated genes indicated a decrease in the expression of genes representing the biological processes of developmental processes (e.g. BMPR1A, BMPR2, ACVR2, periostin, SFRP2, COL5A3) and regulation of blood vessel size (e.g. alpha adrenergic receptor 1B, alpha-SMA) at 6-months of age. A real-time qRTPCR analysis of selected upregulated genes, fibronectin, MMP-3, IL-8 and downregulated genes, BMPR2, PECAM, CCL2, TLR4 confirmed the differential gene expression in the microarray analysis. ConclusionDuring the process of articular cartilage maturation from 6-weeks to 6-months of age in normal pigs, genes associated with inflammatory responses to injury were upregulated and genes involved in the development and vascular responses were downregulated. These findings suggest that during articular cartilage maturation, the transcriptional changes might increase the susceptibility of cartilage to inflammatory damage and decrease the regenerative capacity.

Osteoprotegerin deficiency leads to deformation of the articular cartilage in femoral head.

Osteoarthritis (OA) was a degenerative joint disease characterized by articular cartilage degradation and extensive remodeling of the subchondral bone. Multiple lines of evidence indicated that Osteoprotegerin (OPG), a member of TNF receptor superfamily that was expressed in the chondrocytes of articular cartilage and adjacent locations in the physiological setting, was involved in maintaining integrity of articular cartilage. OPG could prevent subchondral bone from resorption, and also protect cartilage from degradation. In this study, we used Osteoprotegerin-knockout mice (Opg-KO mice) to find out the role of OPG in articular cartilage. We examined articular cartilage in the femoral head of Opg-KO mice began in early adulthood using modern molecular and imaging methods. We found cartilage changes starting from adulthood and progressively with age, reminiscent of pathological changes in OA. Deficiency of OPG caused thinned articular cartilage and extensive remodeling of the subchondral bone in femoral head in comparison with wild-type mice (WT mice). Also, the articular cartilage of femoral head expressed significantly less of Aggrecan, Col-II and Col-X, but more Col-I and Matrix Metalloproteinases-13 (Mmp-13) than WT mice both at gene and protein level. Moreover, increased chondrocyte apoptosis and decreased chondrocyte proliferation were observed in femoral head of Opg-KO mice compared to WT mice. These data suggested that OPG played an important role in maintaining the homeostasis of articular cartilage of femoral head.

Chondrogenic capability of osteoarthritic chondrocytes from the trapeziometacarpal and hip joints.

Osteoarthritis is the most common degenerative disease of joints like the hip and the trapeziometacarpal joint (rhizarthrosis). In this in vitro study, we compared the chondrogenesis of chondrocytes derived from the trapezium and the femoral head cartilage of osteoarthritic patients to have a deeper insight on trapezium chondrocyte behavior as autologous cell source for the repair of cartilage lesions in rhizarthrosis. Chondrocytes collected from trapezium and femoral head articular cartilage were cultured in pellets and analyzed for chondrogenic differentiation, cell proliferation, glycosaminoglycan production, gene expression of chondrogenic and fibrous markers, histological and immunohistochemical analyses. Our results showed a higher cartilaginous matrix deposition and a lower fibrocartilaginous phenotype of the femoral chondrocytes with respect to the trapezium chondrocytes assessed by a higher absolute glycosaminoglycan and type II collagen production, thus demonstrating a superior chondrogenic potential of the femoral with respect to the trapezium chondrocytes. The differences in chondrogenic potential between trapezium and femoral head chondrocytes confirmed a lower regenerative capability in the trapezium than in the femoral head cartilage due to the different environment and loading acting on these joints that affects the metabolism of the resident cells. This could represent a limitation to apply the cell therapy for rhizoarthrosis.

DEVELOPING A TECHNIQUE FOR ULTRASOUND‐GUIDED INJECTION OF THE ADULT CANINE HIP

An accurate method for guiding injections into the canine hip would facilitate diagnostic localization of lameness and targeted treatments. Ultrasound-guided hip injections are commonly used in humans and large animals. Aims of this prospective study were to describe ultrasound (US) anatomy of the adult canine hip and determine the feasibility and accuracy of intra-articular placement of injectate using US-guidance. Seven adult dogs were used to describe US anatomy, five dog cadavers were used to assess the feasibility of the injection technique and 11 dog cadavers were used to assess accuracy of injections. For the accuracy test, 22 joints were injected with iodinated contrast medium by three operators with different experience. With dogs in lateral recumbency, the hyperechoic femoral head surface was identified by following the femoral neck from the greater trochanter or the acetabular rim was localized by following caudally the ilium from the iliac wing. An anechoic gap between the femoral head and acetabular surface represented the joint. The capsule was visible as a triangular echoic structure and the femoral head articular cartilage appeared as an anechoic band. The needle was inserted axial to the greater trochanter and directed in a dorsolateral-ventromedial direction toward the joint space and then pushed through the capsule. Based on postinjection radiography, accuracy was 81.8% at first attempt and 100% at second attempt. This study indicated that US-guided injection is a feasible and accurate technique for injecting the adult canine hip. Future studies in live dogs are needed to assess safety and efficacy.

Arthroscopic Treatment of Cam Type Femoroacetabular Impingement: Short Term Results

Purpose: We evaluated the short term results after treatment of cam type femoroacetabular impingement (FAI) by arthroscopy. Materials and Methods: We evaluated the clinical and radiological results of arthroscopically treated cam type FAI in patients who had failed conservative treatment with hip pain, with at least 12 months follow-up, from November 2010 to December 2012. There were 19 males and six females. Mean age of patients was 32.9 years (19-57 years) and mean follow up period was 17.2 months (13-31 months). We analyzed the alpha angle, head neck offset, visual analogue scale (VAS), and modified Harris hip score (MHHS). Results: Mean alpha angle improved from 64.8。to 39.9。and mean head neck offset also improved from 0.8 to 7.6 mm. Peripheral longitudinal and radial fibrillated labral tear was the most common in the anterosuperior quadrant. Damage to acetabular cartilage was identified in 14 patients. Mean VAS improved from 6.3 to 0.9 and mean MHHS improved from 51.7 to 73.6. Complications associated with the operation included three cases of femoral head articular cartilage injury, two cases of pudendal nerve injury, and two cases of lateral femoral cutaneous nerve injury. Conclusion: Although the short term results for arthroscopically treated cam type FAI were satisfactory, care must be taken to reduce the complications associated with arthroscopy and long term follow is needed in order to determine whether or not it can reduce osteoarthritis of the hip.