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The primary purpose of this study is to determine whether recreational runners with patellofemoral pain (PFP) exhibit greater peak patella cartilage stress compared with pain-free runners. A secondary purpose was to determine the kinematic and/or kinetic predictors of peak patella cartilage stress during running. A total of 22 female recreational runners (12 with PFP and 10 pain-free controls) participated in this study. Patella cartilage stress profiles were quantified using subject-specific finite element models simulating the maximum knee flexion angle during the stance phase of running. Input parameters to the finite element model included subject-specific patellofemoral joint geometry, quadriceps muscle forces, and lower-extremity kinematics in the frontal and transverse planes. Tibiofemoral joint kinematics and kinetics were quantified to determine the best predictor of stress using stepwise regression analysis. Compared with the pain-free runners, those with PFP exhibited greater peak hydrostatic pressure (PFP vs control: 21.2 [5.6] MPa vs 16.5 [4.6] MPa) and maximum shear stress (PFP vs control: 11.3 [4.6] MPa vs 8.7 [2.3] MPa). Knee external rotation was the best predictor of peak hydrostatic pressure and peak maximum shear stress (38% and 25% of variances, respectively), followed by the knee extensor moment (21% and 25% of variances, respectively). Runners with PFP exhibit greater peak patella cartilage stress during running compared with pain-free individuals. The combination of knee external rotation and a high knee extensor moment best predicted the elevated peak stress during running. [ABSTRACT FROM AUTHOR]

Cartilage dysplasia is one of the important reasons for the weakness of pig limbs and hooves. Porcine rickets with weak limbs and hooves bring huge economic losses to the pig industry. However, research on the development of pig cartilage is lacking. This study investigated the key genes and molecular mechanisms involved in cartilage development via an RNA-seq technique. Samples of proximal tibia cartilage were collected from three normal piglets with 1 day, 14 days, and 28 days of age, respectively, and then these samples were divided into two comparison groups (1-day vs. 14-day group, 14-day vs. 28-day group). Through the transcriptome analysis, 108 differentially expressed genes (DEGs), such as FORL2, were obtained from 1-day vs. 14-day comparison group, and 3602 DEGs were obtained from 14-day vs. 28-day comparison group, including SOX9, BMP6, and MMP13. The gene ontology (GO) functional and KEGG pathway enrichment revealed that many functions of DEGs were related to bone development. The pathways of DEGs from Day 1 vs. Day 14 were mainly enriched in mineral absorption, but the DEGs of Day 14 vs. Day 28 were enriched in osteoclast differentiation. Then, the expression patterns of six candidate genes were verified via qPCR. In conclusion, candidate genes affecting cartilage development in Yorkshire pigs were obtained by transcriptome analysis, and the clues showed that Day 14 to Day 28 is a more active and extensive period in cartilage developments, which played a key role in revealing the molecular mechanism of pig cartilage development basis, also compensating for vacancies in cartilage research. [ABSTRACT FROM AUTHOR]

Cartilage failure in diarthrodial joints results in pain and a reduction in quality of life. The goal of cartilage tissue engineering is to replace or regenerate these mechanically loaded tissues to restore function to the joint. Recent advances in the authors' laboratory have resulted in the production of cartilage and fibrocartilage with clinically relevant properties. A review of salient results will constitute the bulk of this manuscript. After providing a brief background of the clinical problem, this review will highlight several specific tissue engineering tools. The approaches used to produce mechanically functional cartilage through tissue engineering have several parallels to the problems faced in osseointegration, eg, the need for mechanically appropriate tissues at the implantation site. The discussion that follows will focus on how approaches developed in identifying alternative cell sources and various exogenous stimuli for producing new cartilage may be applicable to osseointegration. [ABSTRACT FROM AUTHOR]

Introduction. Computational modeling of the human pelvis using the finite elements (FE) method has become increasingly important to understand the mechanisms of load distribution under both healthy and pathologically altered conditions and to develop and assess novel treatment strategies. The number of accurate and validated FE models is however small, and given models fail resembling the physiologic joint motion in particular of the sacroiliac joint. This study is aimed at using an inverted validation approach, using in vitro load deformation data to refine an existing FE model under the same mode of load application and to parametrically assess the influence of altered morphology and mechanical data on the kinematics of the model. Materials and Methods. An osteoligamentous FE model of the pelvis including the fifth lumbar vertebra was used, with highly accurate representations of ligament orientations. Material properties were altered parametrically for bone, cartilage, and ligaments, followed by changes in bone geometry (solid versus 3 and 2 mm shell) and material models (linear elastic, viscoelastic, and hyperelastic isotropic), and the effects of varying ligament fiber orientations were assessed. Results. Elastic modulus changes were more decisive in both linear elastic and viscoelastic bone, cartilage, and ligaments models, especially if shell geometries were used for the pelvic bones. Viscoelastic material properties gave more realistic results. Surprisingly little change was observed as a consequence of altering SIJ ligament orientations. Validation with in vitro experiments using cadavers showed close correlations for movements especially for 3 mm shell viscoelastic model. Discussion. This study has used an inverted validation approach to refine an existing FE model, to give realistic and accurate load deformation data of the osteoligamentous pelvis and showed which variation in the outcomes of the models are attributed to altered material properties and models. The given approach furthermore shows the value of accurate validation and of using the validation data to fine tune FE models. [ABSTRACT FROM AUTHOR]

The varus knee has been defined as a Hip-Knee-Ankle alignment of less than 180 degrees. Varus knee alignment increases the load on the medial knee and also the risk of osteoarthritis. High tibial osteotomy has been designed to modify the malalignment of varus knee. The aim of this study was to investigate the osteotomy effects on knee adduction moment (KAM) and contact forces using a musculoskeletal and subject-specific knee model. A patient with varus knee and no symptoms of any other disease or disability participated in this study. The geometry of the multibody knee model has been modified using MR images. The solutions of its finite element model have been used to determine the parameters of the multibody model. The motion data, ground reaction force and kinetic data have been applied to run the subject-specific musculoskeletal model during the stance phase of gait. After osteotomy, the adduction moment decreased, where the maximum values are comparable to other studies. The pattern of KAM did not witness any significant changes. The total and medial contact forces reduced considerably after surgery, but the lateral contact force did not significantly change. The changes in total and medial contact forces and lack of change in lateral contact force could be explained by modification of the gait pattern after surgery. [ABSTRACT FROM AUTHOR]

Objective Secondary cartilage develops from osteochondral progenitor cells. Hypertrophic chondrocytes in secondary cartilage increase within a very short time and then ossify rapidly. In the present study, we investigated the sequential development process of osteochondral progenitor cells, and the morphology and size of hypertrophic chondrocytes in secondary cartilage. Design ICR mice at embryonic days (E) 14.5–17.5 were used. The mandibular condyle and the medial pterygoid process of the sphenoid bone were observed as secondary cartilage, and the cranial base and the lateral pterygoid process of the sphenoid bone, which is primary cartilage, were observed as a control. Thin sections were subjected to immunostaining and alkaline phosphatase (ALP) staining. Using a confocal laser microscope, 3D stereoscopic reconstruction of hypertrophic cells was performed. To evaluate the size of hypertrophic chondrocytes objectively, the cell size was measured in each cartilage. Results Hypertrophic chondrocytes of secondary cartilage first expressed type X collagen (Col X) at E15.5. SRY-box 9 (Sox 9) and ALP were co-expressed in the fibroblastic/polymorphic tissue layer of secondary cartilage. This layer was very thick at E15.5, and then rapidly became thin. Hypertrophic cells in secondary cartilage were markedly smaller than those in primary cartilage. Conclusions The small hypertrophic cells present in secondary cartilage may have been a characteristic acquired in order for the cartilage to smoothly promote a marked increase in hypertrophic cells and rapid calcification. [ABSTRACT FROM AUTHOR]

Purpose: Many studies have shown that local anesthetics may impede chondrocyte metabolism. However, the influence of a single-dose local anesthetics is controversial. The aim of this metaanalysis was to review the literature for studies investigating the cytotoxic effects of single-dose local anesthetics on chondrocytes and cartilage.Methods: A comprehensive literature search was performed using established search engines (Medline, Embase) to identify studies, investigating the influence of single-dose local anesthetics on cartilage. The systematic analysis included the influence on histology, cell viability, morphology, and matrix production depending upon dose, exposure time, and type of local anesthetics.Results: Twelve studies with four different local anesthetics were included in this metaanalysis. Bupivacaine and lidocaine were found to be more chondrotoxic than mepivacaine and ropivacaine. The amount of dead cells increased in a substance-, dose-, and time-dependent process. Osteoarthritic cartilage seems to be more vulnerable compared to intact cartilage. The toxic effects occur first in the superficial cartilage layers and include damage to membrane integrity, mitochondrial DNA, and nuclear changes. There is no study that could show a significant chondrotoxic effect with low concentrations of bupivacaine (0.0625%), ropivacaine (0.1 and 0.2%), and mepivacaine (0.5%).Conclusions: The cytotoxicity of local anesthetics on chondrocytes is dependent on dose, time, and type of local anesthetics. Single-dose intra-articular administration of local anesthetics impede chondrocyte metabolism and should be performed only with low concentrations for selected diagnostic purposes and painful joints. The use of lidocaine should be avoided.Level Of Evidence: II. [ABSTRACT FROM AUTHOR]

Intermuscular bones (IBs) only exist in the myosepta of lower teleosts and its molecular mechanism remains to be clarified. Bone morphogenetic proteins (BMPs) have been demonstrated to be involved in various physiological processes, including bone and cartilage formation. In this study, we firstly obtained and characterized nine bmp genes for Megalobrama amblycephala , which belongs to Cyprinidae and have a certain amount of IBs. Sequence alignment and phylogenetic analysis both documented that the mature proteins of M. amblycephala bmp genes were highly conserved with other corresponding homologs, respectively, indicating that the function of each bmp gene has been conserved throughout evolution. As a step to characterize potential involvement of bmp genes in IB formation and development, spatiotemporal expressions of nine bmp genes ( bmp2a , bmp2b , bmp3 , bmp4 , bmp5 , bmp7b , bmp8a , bmp14 and bmp16 ) were investigated during the key development stages of IBs. During the ossification process from stage I (the IBs haven't emerged) to stage IV (all of the IBs ossified in the tail with the mature morphology), the expression profiles revealed that bmp16 was the most abundant transcript while bmp4 had the lowest abundance. The mRNA levels of bmp3 , bmp4 , bmp5 and bmp8a increased significantly at stage II, suggesting their roles in stimulating IB formation. The expression of bmp7b reached the highest level at stage III (the rapid period of IB development), suggesting potential involvement of bmp7b in promoting osteoblast differentiation. With the exception of bmp7b and bmp16 , most bmp genes appeared a significant increase at IB maturation phase (stage IV), which means that they may play important roles in maintenance of IB morphogenesis. Spatial tissue distribution of bmp genes showed that most bmp genes were observed at the highest level in developing IBs at one year old fish. Spatiotemporal expression patterns suggest the potential key roles of these bmp genes in IBs formation and maintenance in fish, being as possible promoters or inhibitors. [ABSTRACT FROM AUTHOR]

Xenopus laevis (an anuran amphibian) shows limb regeneration ability between that of urodele amphibians and that of amniotes. Xenopus frogs can initiate limb regeneration but fail to form patterned limbs. Regenerated limbs mainly consist of cone-shaped cartilage without any joints or branches. These pattern defects are thought to be caused by loss of proper expressions of patterning-related genes. This study shows that hyperinnervation surgery resulted in the induction of a branching regenerate. The hyperinnervated blastema allows the identification and functional analysis of the molecules controlling this patterning of limb regeneration. This paper focuses on the nerve affects to improve Xenopus limb patterning ability during regeneration. The nerve molecules, which regulate limb patterning, were also investigated. Blastemas grown in a hyperinnervated forelimb upregulate limb patterning-related genes ( shh, lmx1b, and hoxa13 ). Nerves projecting their axons to limbs express some growth factors ( bmp7, fgf2, fgf8, and shh ). Inputs of these factors to a blastema upregulated some limb patterning-related genes and resulted in changes in the cartilage patterns in the regenerates. These results indicate that additional nerve factors enhance Xenopus limb patterning-related gene expressions and limb regeneration ability, and that bmp, fgf, and shh are candidate nerve substitute factors. [ABSTRACT FROM AUTHOR]

Background: Patients presenting with cam deformity of the femoral head and neck sustain repeated trauma to the articular cartilage of the superior acetabulum, with chondral delamination injuries found during hip arthroscopy. Two previous studies reveal conflicting chondrocyte viability data in these traumatic cartilage injuries. The full-thickness nature of flaps may suggest that chondrocytes residing in the cartilage flap matrix in the joint environment would remain viable despite shear trauma. Hypothesis/Purpose: The purpose of this study is to determine the in vivo tissue viability of acetabular chondral flaps in patients with femoroacetabular impingement (FAI) when samples are analyzed immediately after biopsy. We hypothesize that the majority of the tissue in acetabular chondral flaps is viable in the joint microenvironment. Study Design: Descriptive laboratory study. Methods: Partially detached cartilage flaps from 10 patients undergoing arthroscopic hip surgery for FAI were biopsied in a minimally traumatic manner before chondroplasty and microfracture. Samples were placed in cold Hank’s Balanced Salt Solution without phenol red solution and immediately transported on ice to our laboratory. The edge of the samples was trimmed and further cut into 3 separate, 1-mm-thick sections. Sections were stained using a live/dead staining kit. Images were obtained with confocal microscopy, and the percentage of live cells was quantified. Results: Patients averaged 36 ± 11 years (range, 18-48 years), and 2 patients were female. The mean body mass index was 28.9 ± 5.6 kg/m2. The total proportion of live cells from all sections analyzed was 85.8%. The proportion of live cells per patient was 87% ± 10%. Conclusion: We determined that acetabular chondral flaps are approximately 87% live cells when analyzed immediately after biopsy, with 6 of 10 patients having greater than 90% live cells. These data point to the importance of laboratory techniques in making viability judgments in biologic systems. Clinical Relevance: Full-thickness cartilage loss is a difficult problem for all active people but particularly in the young population in whom joint preservation is key. We describe the viability of chondrocytes present in full-thickness acetabular-based chondral flaps encountered during hip arthroscopy. Identification of greater than 85% chondrocyte viability supports a foundation for evaluation and creation of novel clinical innovations for repair and replacement techniques using the flap as donor tissue, as alternatives to chondroplasty and microfracture. [ABSTRACT FROM AUTHOR]

Background: Osteochondral lesions of the talar dome (OLTs) involve the articular cartilage and subchondral bone. The subchondral bone plate (SBP) plays a crucial role in cartilage metabolism, and computed tomography (CT) can provide more precise information on subchondral bone. CT image prediction of the histological findings on OLT will be useful to determine the most appropriate therapeutic strategy. Purpose: To evaluate the unique features of the CT findings that relate to the condition of the articular cartilage in OLT. Study Design: Cohort study; Level of evidence, 3. Methods: Thirty ankles in 29 patients who had OLT with an osteochondral fragment were retrospectively reviewed. At surgery, the osteochondral fragment of 19 ankles could be preserved (preservation group) by fixation or drilling, and in the remaining 11 ankles, the osteochondral fragment was removed (excision group). Preoperative CT findings were compared between the 2 groups. Biopsies of the osteochondral fragment from 13 ankles were performed. The relationship between the CT and histological findings were evaluated. Results: The area of lesion in the preservation group was significantly larger than that in the excision group. The CT images of the lesion showed the rate of absorption of the SBP in the preservation group to be lower than that in the excision group. As for the lesion bed absorption, it was higher in the preservation group than in the excision group. All cases in the excision group showed bed sclerosis, compared with 42.1% in the preservation group. The specimens with disruption of the SBP exhibited cartilage degeneration and abundant chondrocyte cloning. OLT with absorption of the SBP on CT showed severe cartilage degeneration, while the remaining SBP on CT showed low-grade cartilage degeneration. Conclusion: This study shows the features of preoperative CT and histological findings in OLT with osteochondral fragments. The condition of the SBP affects the cartilage degeneration. CT findings provide important information for the determination of surgical treatment. [ABSTRACT FROM AUTHOR]

Background: Osteoarthritis of the medial tibiofemoral joint (MTFJ) is prevalent among patients undergoing anterior cruciate ligament reconstruction (ACLR). Magnetic resonance T1ρ and T2 relaxation times provide noninvasive methods to quantify early cartilage degeneration. Altered sagittal-plane gait biomechanics have been observed after ACLR, but their associations with longitudinal changes in MTFJ cartilage T1ρ and T2 remain unclear. Hypothesis/Purpose: To examine whether the peak knee flexion moment (KFM), knee flexion angle (KFA), and vertical ground-reaction force (vGRF) during gait are associated with prospective changes in medial tibiofemoral cartilage T1ρ and T2 in ACL-reconstructed knees and to compare these gait characteristics between patients undergoing ACLR and healthy control participants. We hypothesized that a higher KFM, KFA, and vGRF would be associated with greater increases in cartilage relaxation times and that patients undergoing ACLR would demonstrate altered gait characteristics compared with healthy controls. Study Design: Controlled laboratory study. Methods: Thirty-three patients undergoing ACLR underwent gait analysis before and 6 months and 1 year after ACLR and knee magnetic resonance imaging (MRI) before and 6 months, 1 year, and 2 years after ACLR. Twelve healthy controls underwent knee MRI and gait analysis at baseline and 1 year. Cartilage T1ρ and T2 were calculated for the medial tibia and medial femoral condyle. Linear regressions were used to evaluate associations between gait characteristics and changes in cartilage relaxation times from before ACLR to follow-up time points. Independent t tests were used to compare differences in gait between patients undergoing ACLR and control participants. Results: A higher KFM and KFA before ACLR were related to greater increases in medial femoral condyle T1ρ and T2 at 6 months after ACLR. Similarly, a higher KFM, KFA, and vGRF at 6 months were associated with greater increases in medial tibia and medial femoral condyle T1ρ and T2 at 1 and 2 years after ACLR. Gait characteristics at 1 year were not associated with changes in cartilage relaxation times at 2 years after ACLR. Compared with healthy controls, patients undergoing ACLR demonstrated a lower KFM at 6 months after ACLR. Conclusion/Clinical Relevance: The findings of this study revealed that a higher KFM, KFA, and vGRF during gait, especially at 6 months after ACLR, were associated with greater deterioration of MTFJ cartilage health at later time points. [ABSTRACT FROM AUTHOR]

The mandibular condylar cartilage (MCC) shoulders force for the subchondral bone during mastication. The cartilage matrix contains various large molecules, such as type I, II, and X collagens and proteoglycans (PGs), which jointly play essential roles in maintaining cartilage characteristics. PGs play key roles in maintaining the elasticity of cartilage and providing a cushion against mastication forces. In addition to the well-known PGs, DMP1-PG, which is the PG form of dentin matrix protein 1 (DMP1), is a newly identified PG. DMP1 is proteolytically processed in vivo, and the N-terminus is glycosylated into its PG form-that is, DMP1-PG, which is highly expressed not only in tooth and bone but also in the matrix of the MCC. However, the specific functions of DMP1-PG in the MCC remain unclear. In human temporomandibular joint osteoarthritis and hyperocclusion model rat specimens, PGs are significantly downregulated, and DMP1-PG is the most prominently affected PG. To further investigate the role of DMP1-PG in condylar chondrogenesis, a glycosylation site mutant (S89-G89) mouse model was established with knock-in methods. In the MCC of the S89G-DMP1 mice, the glycosylation level of DMP1 was significantly downregulated, and a series of abnormal developmental and pathologic changes could be observed. The morphologic changes included thinner cartilage layers, deformations of the MCC, and disordered arrangements of the chondrocytes, and an earlier onset of temporomandibular joint osteoarthritis-like changes was observed. In addition, markers of chondrogenesis were downregulated, and the matrix of the MCC displayed OA phenotypes in the S89G-DMP1 mice. Further investigations showed that the transforming growth factor β signaling molecules were affected in the MCC after the loss of DMP1-PG. In addition, the loss of DMP1-PG significantly accelerated the progression of cartilage injuries in the hyperocclusion models. Given these findings, we investigated the significant role of DMP1-PG in the chondrogenesis and maintenance of MCC. [ABSTRACT FROM AUTHOR]

Fatigue failure of bones of the metacarpo-phalangeal (fetlock, MCP) joint is common in thoroughbred racehorses. Stresses within the fetlock joint cartilages are affected by the morphology of the third metacarpal bone (MC3) and proximal phalangeal bone, and the steepness of the median sagittal ridge of MC3 is believed to be associated with fracture. This study investigated the influence of the steepness of the sagittal ridge on cartilage stress distribution using a finite element model of the joint. Changes to the steepness of the sagittal ridge were made by applying a parabolic function to the mesh, creating four different models with sagittal ridge angles ranging from 95° to 105°. In the fetlock joint of Thoroughbred racehorses, sagittal ridge angles of >100° were associated with higher Von Mises stresses in cartilage at the palmar aspect of the condylar groove than such stresses in joints with sagittal ridge angles of <100°. Stresses were high in the specific region where fractures are known to originate in MC3. This aspect of morphology of the fetlock joint thus appears to play an important role in the magnitude and distribution of cartilage stresses, which, when acting on the underlying hard tissues of the articular calcified cartilage and subchondral bone may play a role in the initiation of fatigue fracture in the third metacarpal bone. [ABSTRACT FROM AUTHOR]

Cartilage fatigue may be a factor in the precocious development of degenerative changes in the temporomandibular joint (TMJ). This cross-sectional study estimated potential for cartilage fatigue via TMJ energy densities (ED) and jaw muscle duty factors (DF), which were combined to calculate mechanobehavioral scores (MBS) in women with (+) and without (-) bilateral TMJ disc displacement (DD). All subjects gave informed consent to participate and were examined using Diagnostic Criteria (DC) for Temporomandibular Disorders (TMD) and magnetic resonance (MR) and computed tomography (CT) images. Forty-seven subjects were categorized into +DD (n = 29) and -DD (n = 18) groups. Dynamic stereometry (MR images combined with jaw-tracking data) characterized individual-specific data of TMJ stress-field mechanics to determine ED (ED = W/Q mJ/mm³, where W = work done, Q = volume of cartilage) during 10 symmetrical jaw-closing cycles with a 20-N mandibular right canine load. Subjects were trained to record masseter and temporalis electromyography over 3 days and 3 nights. Root mean square electromyography/bite-force calibrations determined subject-specific masseter and temporalis muscle activities per 20-N bite-force (T20 N, μV), which defined thresholds. Muscle DF (DF = % duration of muscle activity/total recording time) were determined for a range of thresholds, and MBS (ED² × DF) were calculated. Intergroup differences in ED, DF, and MBS were assessed via analyses of variance with Bonferroni and Tukey honest significant difference post hoc tests. Average ED for contralateral TMJs was significantly larger (P = 0.012) by 1.4-fold in +DD compared to -DD subjects. Average DF were significantly larger (all P < 0.01) for +DD compared to -DD subjects by 1.7-, 2.5-, and 1.9-fold for day, night, and overall, respectively. Daytime MBS were significantly larger (all P < 0.04) by up to 8.5-fold in +DD compared to -DD subjects. Significantly larger ED, DF, and MBS were shown in women with compared to women without bilateral TMJ DD. [ABSTRACT FROM AUTHOR]

There is evidence that low-grade inflammation may be responsible for pain and development of degenerative changes in temporomandibular joint internal derangement. This article reviews the current knowledge of the molecular mechanisms behind TMJ internal derangements. A non-systematic search was carried out in PubMed, Embase and the Cochrane library for studies regarding pathophysiological mechanisms behind internal derangements focusing on pain-mediating inflammatory and cartilage-degrading molecules. Recent data suggest that release of cytokines may be the key event for pain and cartilage destruction in TMJ internal derangements. Cytokines promote the release of matrix metalloproteinases ( MMPs), and due to hypoxia, vascular endothelial growth factor ( VEGF) is released. This activates chondrocytes to produce MMPs and reduce their tissue inhibitors ( TIMPs) as well as the recruitment of osteoclasts, ultimately leading to cartilage and bone resorption. Also, proteoglycans have an important role in this process. Several cytokines, MMPs, TIMPs and VEGF have been identified in higher concentrations in the TMJ synovial fluid of patients with painful internal derangements and shown to be associated with the degree of degeneration. Other molecules that show elevated levels include hyaluronic acid synthase, disintegrin and metalloproteinase with thrombospondin motifs ( ADAMTs), aggrecan, fibromodulin, biglycan and lumican. Taken together, more or less pronounced inflammation of TMJ structures with release of cytokines, MMPs and other molecular markers that interact in a complex manner may be responsible for tissue degeneration in internal derangements. As internal derangements may be symptom-free, the degree of inflammation, but also other mechanisms, may be important for pain development. [ABSTRACT FROM AUTHOR]

Objective Proinflammatory cytokines such as interleukin-1 (IL-1) are found in elevated levels in diseased or injured tissues and promote rapid tissue degradation while preventing stem cell differentiation. This study was undertaken to engineer inflammation-resistant murine induced pluripotent stem cells (iPSCs) through deletion of the IL-1 signaling pathway and to demonstrate the utility of these cells for engineering replacements for diseased or damaged tissues. Methods Targeted deletion of the IL-1 receptor type I (IL-1RI) gene in murine iPSCs was achieved using the RNA-guided, site-specific clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 genome engineering system. Clonal cell populations with homozygous and heterozygous deletions were isolated, and loss of receptor expression and cytokine signaling was confirmed by flow cytometry and transcriptional reporter assays, respectively. Cartilage was engineered from edited iPSCs and tested for its ability to resist IL-1-mediated degradation in gene expression, histologic, and biomechanical assays after a 3-day treatment with 1 ng/ml of IL-1α. Results Three of 41 clones isolated possessed the IL-1RI+/− genotype. Four clones possessed the IL-1RI−/− genotype, and flow cytometry confirmed loss of IL-1RI on the surface of these cells, which led to an absence of NF-κB transcription activation after IL-1α treatment. Cartilage engineered from homozygous null clones was resistant to cytokine-mediated tissue degradation. In contrast, cartilage derived from wild-type and heterozygous clones exhibited significant degradative responses, highlighting the need for complete IL-1 blockade. Conclusion This work demonstrates proof-of-concept of the ability to engineer custom-designed stem cells that are immune to proinflammatory cytokines (i.e., IL-1) as a potential cell source for cartilage tissue engineering. [ABSTRACT FROM AUTHOR]

Several immune cell populations are involved in cartilage damage, bone erosion, and resorption processes during osteoarthritis. The purpose of this study was to investigate the role of NK cells in the pathogenesis of experimental osteoarthritis and whether and how neutrophils can regulate their synovial localization in the disease. Experimental osteoarthritis was elicited by intra-articular injection of collagenase in wild type and Cxcr3-/- 8-wk old mice. To follow osteoarthritis progression, cartilage damage, synovial thickening, and osteophyte formation were measured histologically. To characterize the inflammatory cells involved in osteoarthritis, synovial fluid was collected early after disease induction, and the cellular and cytokine content were quantified by flow cytometry and ELISA, respectively. We found that NK cells and neutrophils are among the first cells that accumulate in the synovium during osteoarthritis, both exerting a pathogenic role. Moreover, we uncovered a crucial role of the CXCL10/CXCR3 axis, with CXCL10 increasing in synovial fluids after injury and Cxcr3-/- mice being protected from disease development. Finally, in vivo depletion experiments showed that neutrophils are involved in an NK cell increase in the synovium, possibly by expressing CXCL10 in inflamed joints. Thus, neutrophils and NK cells act as important disease-promoting immune cells in experimental osteoarthritis and their functional interaction is promoted by the CXCL10/CXCR3 axis. [ABSTRACT FROM AUTHOR]

Magnetic resonance imaging (MRI) is widely used in the clinical setting as well as for research applications. Since its inception, technical development has broadly progressed as a response to challenges in both the clinical and research settings. Higher magnetic field strength and advances in hardware and software have revolutionized the diagnostic potential of MRI and moved well beyond diagnosis to characterization of tissue metabolism, biochemistry, disease pathogenesis, and material property, to name a few. This article focuses on state-of-the-art clinical and cutting-edge novel pulse sequences applied to knee MRI. [ABSTRACT FROM AUTHOR]

Purpose: Mesenchymal stem cells (MSCs) have been introduced for cell therapy strategies in osteoarthritis (OA). Despite of their capacity for differentiation into chondrocyte, there are some evidences about their life-threatening problem after transplantation. So, some researchers shifted on the application of stem cells conditioned medium. The goal of this study is to evaluate whether Wharton's jelly derived stem cell conditioned medium (WJSCs-CM) can enhance the gene expression profile by chondrocytes in monolayer and mass culture systems. Methods: Conditioned medium was obtained from WJSCs at fourth passage. Isolated chondrocytes were plated at density of 1×106 for both monolayer and high density culture. Then cells in both groups were divided into control (received medium) and experiment group treated with WJ-CM for 3 and 6 days. Samples were prepared to evaluate gene expression profile of collagen II, aggrecan, cartilage oligomeric matrix protein (COMP) and sox-9 using real-time RT-PCR. Results: After 3 days, Chondrocytes treated with WJSCs-CM expressed significantly higher level of genes compared to the control group in both culture systems. After 6 days, the expression of genes in monolayer cultivated chondrocytes was decreased but that of the mass culture were up-regulated significantly. Conclusion: WJ-SCs-CM can increase the expression of cartilage-specific genes and can be introduced as a promoting factor for cartilage regeneration. [ABSTRACT FROM AUTHOR]

Objective: In previous experiments, the crosshatching incision has been shown to be an effective method for the correction of cartilaginous deviations. Although the settings of the experiments were different from that of septoplasty, the crosshatching incision has been considered a useful method for septoplasty. Therefore, we attempted to determine the efficacy of the crosshatching incision technique under actual septoplasty surgical settings.Methods: Commercial pig ear cartilages were used for the following experiments: firstly, the crosshatching incision was performed with the cartilage in a partially fixed state (in order to approximate caudal and dorsal fixation of septal cartilage); secondly, for the purpose of approximating L-strut preservation in septoplasty, the crosshatching incision was performed while excluding a marginal area of 1cm on any two contiguous borders. After the experiments, the change of curvature was assessed.Results: Under fixation of two contiguous borders, the curvature of the cartilage did not straighten after using the crosshatching incision. Incisions preserving the L-strut were not effective either. Furthermore, unpredicted deviation and splitting of the cartilage developed after the crosshatching incision.Conclusions: Under actual surgical settings, the crosshatching incision was ineffective for the correction of septal deviation. Therefore, usefulness of the crosshatching incision needs to be re-evaluated. [ABSTRACT FROM AUTHOR]

Although the pharyngeal wall is well known to have high elasticity, the distribution of submucosal elastic fibers has not been described. Observations of histological sections of the mid and lower pharyngeal walls from 15 elderly donated cadavers were made. We found two distinct submucosal tissue layers with a high content of elastic fibers (tentatively termed the "submucosal elastic laminae"). The inferolateral elastic lamina was restricted to the level from the upper part of the arytenoid to the lower end of the inferior cornu of the thyroid cartilage. It originated from the pharyngeal submucosa, extended laterally along the inner aspect of the thyropharyngeal muscle, and inserted into the posterior margin of the thyroid cartilage including the cornu. The posteromedial lamina extended along the supero-inferior axis from a level above the greater horn of the hyoid bone to reach the muscularis mucosae of the cervical esophagus. The inferolateral and posteromedial laminae were connected at levels below the cricoarytenoid joint. Individual variations were evident in their thicknesses (ranging from almost absent to 0.3 mm) as well as the extent of connection between them. In association with striated muscle function, the inferolateral lamina seemed to suspend the lower pharyngeal mucosa, while the posteromedial lamina seemed to provide mucosal fold forcing smoothly peristaltic conveyance of a bolus during swallowing. [ABSTRACT FROM AUTHOR]

Purpose: Knowledge of the effect of hip pathologies on hip biomechanics is important to the understanding of the development of osteoarthritis, and the contribution of the labrum to hip joint stability has had limited study. The purpose of this study was to evaluate the effect of labral injury to stability of the femoral head in the acetabular socket.Methods: Ten cadaver hip specimens were tested using a robotic system under four different loading conditions: axial loading (80 N) along the femoral axis and axial loading (80 N) combined with either anterior, posterior or lateral loading (60 N). The hip states were examined were intact, with a 1.5 cm capsulotomy and with a 1 cm resection of the anterosuperior labrum.Results: At 30° of flexion, under axial load, the displacement of the hip with capsulotomy and labral resection (9.6 ± 2.5 mm) was significantly larger then the hip with capsulotomy alone (5.6 ± 4.1 mm, p = 0.005) and the intact hip (5.2 ± 3.8 mm, p = 0.005). Also, at 30° of flexion, the displacement under combined axial and anterior/posterior load was increased with capsulotomy and labral resection.Conclusion: The acetabular labrum provides stability to the hip joint in response to a distraction force and combined distraction and translation forces. One centimetre of labral resection caused significant displacement ("wobbling" effect) of the femoral head within the acetabulum with normal range of motion. Successful labral repair could be crucial for restoration of the hip biomechanics and prevention of coxarthrosis. [ABSTRACT FROM AUTHOR]

Our previous study identified that endoplasmic reticulum stress (ERS) plays a critical role in chondrocyte apoptosis and mandibular cartilage thinning in response to compressive mechanical force, although the underlying mechanisms remain elusive. Because the endoplasmic reticulum (ER) is a primary site of intracellular Ca(2+) storage, we hypothesized that Ca(2+)-dependent ERS might be involved in mechanical stress-mediated mandibular cartilage thinning. In this study, we used in vitro and in vivo models to determine Ca(2+) concentrations, histological changes, subcellular changes, apoptosis, and the expression of ERS markers in mandibular cartilage and chondrocytes. The results showed that in chondrocytes, cytosolic Ca(2+) ([Ca(2+)]i) was dramatically increased by compressive mechanical force. Interestingly, the inhibition of Ca(2+) channels by ryanodine and 2-aminoethoxydiphenyl borate, inhibitors of ryanodine receptors and inositol trisphosphate receptors, respectively, partially rescued mechanical force-mediated mandibular cartilage thinning. Furthermore, chondrocyte apoptosis was also compromised by inhibiting the increase in [Ca(2+)]i that occurred in response to compressive mechanical force. Mechanistically, the ERS induced by compressive mechanical force was also repressed by [Ca(2+)]i inhibition, as demonstrated by a decrease in the expression of the ER stress markers 78 kDa glucose-regulated protein (GRP78) and 94 kDa glucose-regulated protein (GRP94) at both the mRNA and protein levels. Collectively, these data identified [Ca(2+)]i as a critical mediator of the pathological changes that occur in mandibular cartilage under compressive mechanical force and shed light on the treatment of mechanical stress-mediated cartilage degradation. [ABSTRACT FROM AUTHOR]

Tendons are transition tissues that transfer the contractile forces generated by the muscles to the bones, allowing movement. The region where the tendon attaches to the bone is called bone-tendon junction or enthesis and may be classified as fibrous or fibrocartilaginous. This study aims to analyze the collagen fibers and the cells present in the bone-tendon junction using light microscopy and ultrastructural techniques as scanning electron microscopy and transmission electron microscopy. Forty male Wistar rats were used in the experiment, being 20 adult rats at 4 months-old and 20 elderly rats at 20 months-old. The hind limbs of the rats were removed, dissected and prepared to light microscopy, transmission electron microscopy and scanning electron microscopy. The aging process showed changes in the collagen fibrils, with a predominance of type III fibers in the elderly group, in addition to a decrease in the amount of the fibrocartilage cells, fewer and shorter cytoplasmic processes and a decreased synthetic capacity due to degradation of the organelles involved in synthesis. [ABSTRACT FROM AUTHOR]

Background: Medial femoral condyle (MFC) chondral defects cause knee pain. Clinical studies have shown worse functional outcomes and cartilage defect fill rates after microfracture in obese patients (BMI ≥30) and for defects with size ≥2 cm2. Purpose: To determine the effect of obesity, defect size, and cartilage thickness on the force sustained at the base of full-thickness MFC cartilage defects during weightbearing. Study Design: Controlled laboratory study. Methods: Eight human cadaveric knees were loaded in 15° of flexion. A sensor measured force across the medial compartment. The area at the base of the defect protected from load, termed the “area of containment,” was quantified, and loads simulating weightbearing for BMIs of 20, 30, and 40 were applied. A full-thickness cartilage defect was created on the MFC. Cycles of loads were applied for defect sizes with diameters of 6, 8, 10, 12, 14, 16, 18, and 20 mm. A second sensor recorded force at the base of the defect for defects with diameters of 14, 16, 18, and 20 mm. Results: Loads simulating BMI ≥30 led to a decrease in the area of containment for all defects ≥14 mm in diameter (P ≤ .038). Base of defect force increased for defects ≥16 mm in diameter (area, ≥2 cm2) between loaded and unloaded states (P ≤ .042) and for loads simulating BMI ≥30 (P ≤ .045). Cartilage rim thickness <2 mm showed higher base of defect force than did thickness ≥2 mm, for all BMI groups (P ≤ .025). Conclusion: Increased force at the base of MFC cartilage defects was observed for weightbearing loads simulating BMI ≥30, for defect size ≥2 cm2, and for rim thickness <2 mm. This may lead to a biomechanically unfavorable environment after microfracture in these patient subsets. Clinical Relevance: These biomechanical findings corroborate clinical studies that have noted worse outcomes after microfracture in patients with BMI ≥30 and cartilage defects of size ≥2 cm2. Further clinical studies are needed to compare microfracture with other cartilage restoration procedures in these patient subsets. [ABSTRACT FROM AUTHOR]

Despite its pervasive use, the clinical efficacy of platelet-rich plasma (PRP) therapy and the different mechanisms of action have yet to be established. This overview of the literature is focused on the role of PRP in bone, tendon, cartilage, and ligament tissue regeneration considering basic science literature deriving from in vitro and in vivo studies. Although this work provides evidence that numerous preclinical studies published within the last 10 years showed promising results concerning the application of PRP, many key questions remain unanswered and controversial results have arisen. Additional preclinical studies are needed to define the dosing, timing, and frequency of PRP injections, different techniques for delivery and location of delivery, optimal physiologic conditions for injections, and the concomitant use of recombinant proteins, cytokines, additional growth factors, biological scaffolds, and stems cells to develop optimal treatment protocols that can effectively treat various musculoskeletal conditions. [ABSTRACT FROM AUTHOR]

Objective Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into chondrocytes. Articular cartilage contains MSC-like chondroprogenitor cells, which suggests their involvement in the maintenance of cartilage homeostasis by a self-repair mechanism. Interleukin-6 (IL-6) is a cytokine with a wide range of physiologic functions, which are produced by MSCs in a steady manner and in large quantities. The purpose of this study was to investigate the involvement of IL-6 signaling in MSC differentiation into chondrocytes. Methods Human bone marrow-derived MSCs were cultured using a pellet culture system in medium containing transforming growth factor β3. Chondrogenic differentiation was detected by cartilage matrix accumulation and chondrogenic marker gene expression. Results IL-6 was detected at a high concentration in culture supernatants during chondrogenic differentiation. The expression of the IL-6 receptor (IL-6R) was significantly increased, accompanied by markedly increased phosphorylation and expression of STAT-3. Addition of IL-6 and soluble IL-6R (sIL-6R) to the chondrogenic culture resulted in concentration-dependent increases in cartilage matrix accumulation and cartilage marker gene expression (type II collagen/aggrecan/type X collagen). Phosphorylation of the master transcription factor SOX9 was enhanced upon addition of IL-6 and sIL-6R. STAT-3 knockdown suppressed chondrogenic differentiation. IL-6 and the MSC markers CD166 and nestin were colocalized in macroscopically normal human cartilage taken from the lateral femoral compartment of knees with medial tibiofemoral osteoarthritis. Conclusion During differentiation of human MSCs into chondrocytes, the activation of IL-6/STAT-3 signaling positively regulated chondrogenic differentiation. The presence of IL-6 around MSC-like cells in the cartilage tissue was identified, suggesting that IL-6 contributes to homeostasis and cartilage self-repair by promoting chondrogenic differentiation. [ABSTRACT FROM AUTHOR]

It has been considered that concentrations of certain amino acids in the egg are not sufficient to fully support embryonic development of modern broilers. In this study we evaluated embryo growth and development with particular emphasis on one of the major components of connective tissue, collagen. Experiments were performed on Ross 308 chicken embryos from 160 fertilised eggs. Experimental solutions of silver nanoparticles (Ag), hydroxyproline solution (Hyp) and a complex of silver nanoparticles with hydroxyproline (AgHyp) were injected into albumen, and embryos were incubated until day 20. An assessment of the mass of embryo and selected organs was carried out followed by measurements of the expression of the key signalling factors’ fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor-A (VEGF-A). Finally, an evaluation of collagen microstructure using scanning electron microscopy was performed. Our results clearly indicate that Hyp, Ag and AgHyp administeredin ovoto chicken embryos did not harm embryos. Comparing to the control group, Hyp, Ag and the AgHyp complex significantly upregulated expression of the FGF-2 at the mRNA and protein levels. Moreover, Hyp, Ag and, in particular, the complex of AgHyp significantly increased blood vessel size, cartilage collagen fibre lattice size and bundle thickness. The general conclusion from this study is that AgHyp treatment may help to build a stronger and longer lasting form of collagen fibres. [ABSTRACT FROM AUTHOR]

Articular cartilage lacks an intrinsic capacity for self-repair, and once damaged, it never heals. This creates an opportunity for surgical intervention, whether to stimulate a healing response that results in the formation of a lower-quality fibrocartilaginous scar or formal cartilage repair in the form of cartilage transplants. This article will review the nature of cartilage injury and discuss indications and techniques for repair. [ABSTRACT FROM AUTHOR]

Chondromodulin-I (ChM-I) is a 20–25 kDa anti-angiogenic glycoprotein in cartilage matrix. In the present study, we identified a novel 14-kDa species of ChM-I by immunoblotting, and purified it by immunoprecipitation with a newly raised monoclonal antibody against ChM-I. The N-terminal amino acid sequencing indicated that it was an N-terminal truncated form of ChM-I generated by the proteolytic cleavage at Asp37-Asp38. This 14-kDa ChM-I was shown by the modified Boyden chamber assay to have very little inhibitory activity on the VEGF-A-induced migration of vascular endothelial cells in contrast to the intact 20–25 kDa form of ChM-I (ID50 = 8 nM). Immunohistochemistry suggested that 20–25 kDa ChM-I was exclusively localized in the avascular zones, i.e. the resting, proliferating, and prehypertrophic zones, of the cartilaginous molds of developing long bone, whereas the 14-kDa form of ChM-I was found in hypertrophic and calcified zones. Immunoblotting demonstrated that mature growth-plate chondrocytes isolated from rat costal cartilage actively secrete ChM-I almost exclusively as the intact 20–25 kDa form into the medium in primary culture. Taken together, our results suggest that intact 20–25 kDa ChM-I is stored as a component of extracellular matrix in the avascular cartilage zones, but it is inactivated by a single N-terminal proteolytic cleavage in the hypertrophic zone of growth-plate cartilage. [ABSTRACT FROM AUTHOR]

Objective To investigate the role of the newly discovered epigenetic mark 5-hydroxymethylcytosine (5hmC) and its regulators in altered gene expression in osteoarthritis (OA). Methods Cartilage was obtained from OA patients undergoing total knee arthroplasty and from control patients undergoing anterior cruciate ligament reconstruction. Global levels of 5hmC and 5-methylcytosine (5mC) were investigated using immunoblotting, enzyme-linked immunosorbent assays, and cellular staining. Gene expression changes were monitored by quantitative polymerase chain reaction (PCR) analysis. Levels of locus-specific 5hmC and 5mC at CpG sites in the matrix metalloproteinase 1 (MMP-1), MMP-3, ADAMTS-5, and hypoxanthine guanine phosphoribosyltransferase 1 (HPRT-1) promoters were quantified using a glucosylation and enzyme digestion-based method followed by quantitative PCR analysis. Global and locus-specific 5hmC levels and gene expression changes were monitored in normal chondrocytes stimulated with inflammatory cytokines to identify the effect of joint inflammation. Results A global 5-6-fold increase in 5hmC concomitant with a loss of TET1 was observed in human OA chondrocytes compared to normal chondrocytes. Enrichment of 5hmC was observed in promoters of enzymes critical to OA pathology, MMP-1 and MMP-3. Short-term treatment of normal chondrocytes with inflammatory cytokines induced a rapid decrease in TET1 expression but no global or locus-specific 5hmC enrichment. Conclusion This study provides the first evidence of an epigenetic imbalance of the 5hmC homeostasis in OA leading to TET1 down-regulation and 5hmC accumulation. Our experiments identify 5hmC and its regulators as potential diagnostic and therapeutic targets in OA. [ABSTRACT FROM AUTHOR]

The present study was intended to examine the stroboscopic parameters with regard to the extent and type of deviancies or variations from normalcy in perceptually phononormic individuals. Fifty perceptually phononormic individuals who served as the participants for the study were made to undergo Videostroboscopic examinations by an experienced Otolaryngologist. Later, all the recordings were examined, and under glottal activity majority of the parameters inclined towards normalcy but for certain deviations noticed for glottis closure and amplitude. Under supraglottal activity variations were noticed mainly under the structure and functioning of epiglottis, ventricular folds and arytenoids. These findings of the current study advocate that the presence of slight anatomically deviant parameters (probable normal variations) alone without any related presenting complaints do not serve as diagnostic hallmarks, mainly in cases of Functional or Non organic dysphonia and serves to caution in exercising just the theoretical framework in diagnosis of primarily functional dysphonia. [ABSTRACT FROM AUTHOR]

Objective: Application of distraction is an alternative to bone graft or flap operations for repairing bone defects. Bone can be extended in width and length without using a new donor area. Operation time is shortened and failures are reduced without any concern about graft or flap loss. However, there are no distraction methods known for generating new cartilage as an alternative to graft or flap applications in repairs of cartilage defects This experimental study was planned to determine whether or not chondroneogenesis can be obtained via application of distraction. Methods: Fourteen New Zealand rabbits were used in this study. Distractions at a rate of 0.50 mm/day, 0.25 mm/day and 0.25 mm on alternate days were applied on 3 rabbits consecutively and the ideal distraction rate was determined as 0.25 mm once every two days. On alternate days 0.25 mm of distraction was applied for two months aiming to generate 7.5 mm of new cartilage. Results: In the rabbits in which 0.25 mm of distraction on alternate days had been applied, regular new cartilage formation was observed. Conclusions: According to these findings, the slow metabolic rate of avascular cartilage tissue affects distraction rates and rhythms. However, by applying distraction and taking advantage of the regenerative capacity of the perichondrium, it is possible to generate new and regular chondrocytes and extend the cartilage. [ABSTRACT FROM AUTHOR]

Distal radio-ulnar joint (DRUJ) instability is increasingly recognised and assessment can be subjective and difficult. Previous research has used cadaveric models or in-vivo with CT, with variable results. A test device was designed to establish normal values of in-vivo DRUJ dorso-palmar translation. Twenty volunteers were recruited. Those with previous wrist/forearm injuries were excluded. The device held the elbow at 90° flexion and neutral forearm rotation, with the distal ulna secured. A dorso-palmar shear force was applied to the distal radius and displacement measured three times on each wrist alternately by the same operator. The mean translation of the DRUJ is 5.5 mm. Same-sided mean measurements for two subjects taken days apart varied by 1 mm. The intra-class correlation coefficient was 0.93- The device is reliable, reproducible and appears to be a simple valid test. Contralateral sides were comparable. It will primarily be a research device to guide clinical practice in DRUJ instability. [ABSTRACT FROM AUTHOR]

Objective We and others previously demonstrated that sirtuin 1 (SIRT-1) regulates apoptosis and cartilage-specific gene expression in human chondrocytes and mouse models. This study was undertaken to determine if SIRT-1 enzymatic activity plays a protective role in cartilage homeostasis in vivo, by investigating mice with SIRT-1 mutations to characterize their cartilage. Methods Articular cartilage was harvested from the paws and knees of 5- and 6-month-old wild-type (WT) mice and mice homozygous for SIRT-1tm2.1Mcby (SIRT-1y/y), an allele carrying a point mutation that encodes a SIRT-1 protein with no enzymatic activity (y/y mice). Mice ages 2 days old and 6-7 days old were also examined. Mouse joint cartilage was processed for histologic examination or biochemical analyses of chondrocyte cultures. Results We found that articular cartilage tissue sections from y/y mice of up to 6 months of age contained reduced levels of type II collagen, aggrecan, and glycosaminoglycan compared to sections from WT mice. In contrast, protein levels of matrix metalloproteinase 8 (MMP-8), MMP-9, and MMP-13 were elevated in the cartilage of y/y mice. In addition, chondrocyte apoptosis was elevated in SIRT-1 mutant mice as compared to their WT littermates. Consistent with these observations, protein tyrosine phosphatase 1b was elevated in the y/y mice. Conclusion Our in vivo findings in this animal model demonstrate that mice with defective SIRT-1 also have defective cartilage, with elevated rates of cartilage degradation with age. Hence, normal cartilage homeostasis requires enzymatically active SIRT-1 protein. [ABSTRACT FROM AUTHOR]

The author discusses fibroblast growth factors (FGFs), which has been regarded as important homeostatic factors within joint tissues and the controversy regarding them. She comments on a study by Yan and colleagues, which demonstrated that FGF receptor (FGFR) expression in articular chondrocytes is dynamic, thus explaining the fibroblast growth factor paradox in osteoarthritis (OA). She also highlights the clinical significance of a study by Weng and colleagues in the current issue.

Objective To investigate aggrecan degradation in juvenile idiopathic arthritis (JIA). Methods The pattern and abundance of aggrecan fragments in synovial fluid (SF) aspirates from JIA patients were analyzed and compared with aggrecan fragments in SF from patients with other arthritides, children with knee injury, and a knee-healthy reference group. Concentrations of sulfated glycosaminoglycan (sGAG) in SF were measured by Alcian blue precipitation assay. Aggrecan fragments were purified by dissociative CsCl density-gradient centrifugation, deglycosylated, and analyzed by Western blot using antibodies specific for either aggrecanase-derived ARGS, SELE, and KEEE neoepitopes or the aggrecan G3 domain. Results The concentration of sGAG in SF from patients with JIA was significantly lower compared with that in SF from patients with osteoarthritis (OA) ( P < 0.001), patients with juvenile knee injury ( P = 0.006), and knee-healthy controls ( P = 0.022). Western blot analysis revealed KEEE, SELE, and G3 fragments generated by aggrecanase cleavage in the chondroitin sulfate-rich region of aggrecan in patients with JIA. The pattern of aggrecan fragments in JIA patients was not identical to that in pooled OA SF, although there were notable similarities. Surprisingly, aggrecanase-derived ARGS fragments were barely detectable in JIA SF, in marked contrast to levels in OA SF. Conclusion Aggrecanases appear to cleave minimally in the interglobular domain of aggrecan in JIA patients despite robust levels of cleavage in the chondroitin sulfate-rich region. These results suggest that in JIA, unlike other arthritides, aggrecanase cleavage in the aggrecan interglobular domain might not be a major pathogenic event. [ABSTRACT FROM AUTHOR]

Objective Fibroblast growth factor (FGF) family members are involved in the regulation of articular cartilage homeostasis. The aim of this study was to investigate the function of FGF receptor 1 (FGFR-1) in the development of osteoarthritis (OA) and its underlying mechanisms. Methods FGFR-1 was deleted from the articular chondrocytes of adult mice in a cartilage-specific and tamoxifen-inducible manner. Two OA models (aging-associated spontaneous OA, and destabilization-induced OA), as well as an antigen-induced arthritis (AIA) model, were established and tested in Fgfr1-deficient and wild-type (WT) mice. Alterations in cartilage structure and the loss of proteoglycan were assessed in the knee joints of mice of either genotype, using these 3 arthritis models. Primary chondrocytes were isolated and the expression of key regulatory molecules was assessed quantitatively. In addition, the effect of an FGFR-1 inhibitor on human articular chondrocytes was examined. Results The gross morphologic features of Fgfr1-deficient mice were comparable with those of WT mice at both the postnatal and adult stages. The articular cartilage of 12-month-old Fgfr1-deficient mice displayed greater aggrecan staining compared to 12-month-old WT mice. Fgfr1 deficiency conferred resistance to the proteoglycan loss induced by AIA and attenuated the development of cartilage destruction after surgically induced destabilization of the knee joint. The chondroprotective effect of FGFR-1 inhibition was largely associated with decreased expression of matrix metalloproteinase 13 (MMP-13) and up-regulation of FGFR-3 in mouse and human articular chondrocytes. Conclusion Disruption of FGFR-1 in adult mouse articular chondrocytes inhibits the progression of cartilage degeneration. Down-regulation of MMP-13 expression and up-regulation of FGFR-3 levels may contribute to the phenotypic changes observed in Fgfr1-deficient mice. [ABSTRACT FROM AUTHOR]

Magnetic resonance imaging (MRI) allows one to analyze cartilage physiology in vivo. Cartilage deforms during loading, but little is known about its recovery after deformation. Here we study 'nocturnal' changes in knee cartilage thickness and whether postexercise deformation differs between morning and evening. Axial magnetic resonance (MR) images were acquired in the right knees of 17 healthy volunteers (age 23.5 ± 3.0 years) after a normal day, and then after 30 deep knee bends. Coronal images were additionally acquired in 8 of these volunteers after a normal day and then after 2 min of static loading of the leg with 150% body weight. The volunteers then remained unloaded overnight and the same protocol was repeated in the morning. A significant increase (p < 0.01) in cartilage thickness was observed between evening (preexercise) and morning (preexercise): +2.4% in the patella, +8.4% in the medial tibia and +6.2% in the lateral tibia. Deformation in the morning (-6.8/ -4.6/-5.1%) was generally greater than that in the evening (-5.4/-3.2/-3.7%), but this difference did not reach statistical significance. No significant difference in the nocturnal thickness increase (or postexercise deformation) was observed between men and women. We conclude that knee cartilage (thickness) recovers overnight by approximately 2-8%, independent of sex. Given the lack of 'predeformation' after nocturnal periods of unloading, morning postexercise deformation of the cartilage may have a greater magnitude than evening postexercise deformation. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Objective Human mesenchymal stem cells (hMSCs) represent one of the most promising stem cell therapies for traumatic injury and age-related degenerative diseases involving cartilage. However, few genetic factors regulating chondrogenesis of MSCs have been identified. One study showed that zinc-finger protein 145 (ZNF145), a transcription factor, was up-regulated during 3-lineage differentiation of hMSCs. The present study was undertaken to validate whether this novel transcription factor is useful for the repair and regeneration of cartilage. Methods Human MSCs were transfected with lentiviral short hairpin RNA (for small interfering RNA knockdown of ZNF145) and a lentiviral vector for overexpression of ZNF145, and the effects of ZNF145 on chondrogenesis were studied using quantitative polymerase chain reaction and immunostaining. Microarray and transient expression analyses were used to determine whether ZNF145 is a factor operating upstream of SOX9. Allogeneic transplantation of hMSCs into osteochondral defects in rats was performed to determine the effects of ZNF145 on repair of cartilage in vivo. Results Small interfering RNA-mediated gene silencing of ZNF145 slowed down chondrogenesis, whereas overexpression of ZNF145 enhanced chondrogenesis. Global gene expression profiling showed up-regulated gene expression in ZNF145-overexpressing MSCs, and transient overexpression of ZNF145 enhanced the expression of SOX9, suggesting that ZNF145 acts as a factor upstream of SOX9, the master regulator of chondrogenesis. Moreover, allogeneic transplantation of hMSCs into osteochondral defects of rat knees showed that ZNF145-overexpressing MSCs repaired cartilage defects better and earlier than empty control MSCs. Conclusion These findings suggest that ZNF145 gene therapy may be a very useful strategy for improving the quality of cartilage regeneration and repair. [ABSTRACT FROM AUTHOR]

Objective Although both cartilage and synovium are affected in osteoarthritis (OA), no in vitro coculture models of human OA tissue have been described. The aim of this study was to develop an in vitro model that includes both the synovium and cartilage of patients with knee OA. Methods Explants of human OA cartilage and synovium were cultured alone or in coculture for 21 days. Histologic evaluation and analyses of lactate dehydrogenase release, matrix metalloproteinase (MMP) activity, content, release, and synthesis of glycosaminoglycan (GAG), and cytokine production were used to evaluate synovial tissue functionality and its effect on cartilage metabolism. To assess the possibility of intervention in the model system, the effect of triamcinolone was studied. Results Throughout the entire culture period, OA synovial tissue remained viable and produced cytokines. Monocultures of synovial and cartilage explants produced different cytokine subsets, with the subsets found in coculture being most similar to those previously described in OA synovial fluid. MMP activity was detectable only in the synovial explant monoculture and in coculture. Cocultures showed a reduction in final GAG content ( P < 0.02), attributable to an inhibition of GAG production ( P < 0.001) rather than an increase in GAG release. Addition of triamcinolone inhibited cytokine production and MMP activity in coculture and synovial tissue monoculture and counteracted the inhibition of GAG production induced by coculture. In cartilage monoculture, however, triamcinolone reduced GAG production. Conclusion OA synovium affects cartilage metabolism by reducting GAG production. Triamcinolone can relieve this effect of synovial tissue, while being inhibitory when added to cartilage monoculture. These results clearly indicate the importance of tissue coculture as a promising tool for studying OA pathophysiology and for development of possible interventions. [ABSTRACT FROM AUTHOR]

Objectives. To investigate whether maintaining the chondrocyte’s native pericellular matrix prevents collagen-induced up-regulation of collagenase-3 (MMP-13) and whether integrin α1 (ITGα1) and/or discoidin domain receptor 2 (DDR2) modulate MMP-13 expression and which signalling pathway plays a role in collagen-stimulated MMP-13 expression.Methods. Goat articular chondrocytes and chondrons were cultured on collagen coatings. Small interfering RNA (siRNA) oligonucleotides targeted against ITGα1 and DDR2 were transfected into primary chondrocytes. Chemical inhibitors for mitogen-activated protein kinase kinase (MEK1) (PD98059), focal adhesion kinase (FAK) (FAK inhibitor 14), mitogen-activated protein kinase 8 (JNK) (SP600125) and protein kinase C (PKC) (PKC412), and a calcium chelator (BAPTA-AM) were used in cell cultures. Real-time PCR was performed to examine gene expression levels of MMP-13, ITGα1 and DDR2 and collagenolytic activity was determined by measuring the amount of hydroxyproline released in the culture medium.Results. Maintaining the chondrocyte’s native pericellular matrix prevented MMP-13 up-regulation and collagenolytic activity when the cells were cultured on a collagen coating. Silencing of ITGα1 and DDR2 reduced MMP-13 gene expression and collagenolytic activity by primary chondrocytes cultured on collagen. Incubation with the PKC inhibitor strongly reduced MMP-13 gene expression levels. Gene expression levels of MMP-13 were also decreased by chondrocytes incubated with the MEK, FAK or JNK inhibitor.Conclusion. Maintaining the native pericellular matrix of chondrocytes prevents collagen-induced up-regulation of MMP-13. Both ITGα1 and DDR2 modulate MMP-13 expression after direct contact between chondrocytes and collagen. PKC, FAK, MEK and JNK are involved in collagen-stimulated expression of MMP-13. [ABSTRACT FROM PUBLISHER]

Injuries to the knee meniscus, particularly those in the avascular region, pose a complex problem and a possible solution is tissue engineering of a replacement tissue. Tissue engineering of the meniscus involves scaffold selection, addition of cells, and stimulation of the construct to synthesize, maintain, or enhance matrix production. An acellular collagen implant is currently in clinical trials and there are promising results with other scaffolds, composed of both polymeric and natural materials. The addition of cells to these constructs may promote good matrix production in vitro, but has been studied in a limited manner in animal studies. Cell sources ranging from fibroblasts to stem cells could be used to overcome challenges in cell procurement, expansion, and synthetic capacity currently encountered in studies with fibrochondrocytes. Manipulation of construct culture with exogenous growth factors and mechanical stimulation will also likely play a role in these strategies. [ABSTRACT FROM AUTHOR]

1. The 5-HT receptor subtype that mediates bronchocontraction and the involvement of neuronal and non-neuronal acetylcholine was assessed in murine isolated tracheae. 2. Atropine (1-10 nM) caused a rightward shift of the methacholine concentration-effect curves (pA(2)=9.0) but reduced the maximum response to 5-HT, suggesting that 5-HT acts as an indirect agonist. The potency of 5-HT receptor agonists (alpha-methyl-5-HT approximately 5-HT>5-carboxamidotryptamine), together with the competitive antagonism of 5-HT by ketanserin (pA(2)=9.4), suggests the involvement of the 5-HT(2A) receptor. 3. While cholinergic twitch responses to electrical field stimulation were abolished by the fast sodium channel inhibitor tetrodotoxin (300 nM), as well as by combined blockade of N-, P- and Q-type voltage-operated calcium channels by omega-conotoxin GVIA (30 nM) and agatoxin IVA (100 nM), responses to 5-HT were unaffected. Similarly, botulinum toxin A (50 nM) inhibited EFS twitch responses, but not contractions to 5-HT. 4. Choline acetyltransferase immunoreactivity was localised to ganglia and nerve fibres as well as approximately half the epithelial cells in the preparation. Removal of the epithelial layer markedly attenuated the contractile response to 5-HT, but had no effect on contractions to either methacholine or EFS. 5. These findings suggest that 5-HT, acting at 5-HT(2A) receptors on mouse tracheal epithelial cells, stimulates these cells to release acetylcholine, which then causes contraction of airway smooth muscle. This phenomenon should be borne in mind in when interpreting studies of murine models of airway disease. [ABSTRACT FROM AUTHOR]

Release of mitochondrial calcium has been shown to occur concomitant with mineral ion loading of matrix vesicles at the onset of mineralization in epiphyseal growth plate cartilage. Matrix vesicles contain amorphous calcium phosphate (ACP), a mineral form that usually results from rapid precipitation at high initial levels of Ca2+ and/or inorganic P (Pi). Since the cytosol of growth plate chondrocytes has been found to contain high levels of Pi, rapid release of mitochondrial Ca2+ into the cytosol may cause local precipitation of calcium phosphate and thus be coupled with matrix vesicle formation. Studies were carried out to determine the kinetics and nature of mineral formation that occur when small amounts of Ca2+ are added under various conditions to a Pi buffer composed of electrolytes matched in concentrations and pH to that of the cytosol of epiphyseal chondrocytes. Depending on the manner in which Ca2+ was added, ACP, dicalcium phosphate dihydrate (DCPD), or apatite (HA) first formed. In the presence of ATP, ACP was the only solid phase detected, being stable for at least 24 h. However, in its absence, ACP rapidly transformed into DCPD. Increasing the pH of the reaction buffer from 6.9 to 7.5 increased the amount of ACP initially formed, but DCPD was consistently found upon ACP transformation. Yet at pH 8.0, ACP persisted for at least 24 h. The amount of precipitate formed was proportional to the level of added Ca2+; precipitates formed when as little as 1.0 mmole was added per liter of buffer. Our findings support the possibility that rapid release of mitochondrial Ca2+ may cause localized intracellular precipitation of ACP. Since nascent ACP is known to stimulate membrane fusion and blebbing of vesicles, these findings may explain the presence of ACP in matrix vesicles. The rapid conversion of ACP to DCPD in the absence of ATP under these conditions may also explain the reported occurrence of DCPD in samples of early mineralizing tissue. [ABSTRACT FROM AUTHOR]

The author discusses cell clustering in osteoarthritis (OA) pathology, and focuses on the vertical tissue sections observed in the organization of chondrocytes. He refers to a mathematical algorithm introduced in a study by Rolauffs and colleagues, and highlights that this novel spatial measurement demonstrates the differences between species on the basis of rdered and unordered cellular spatial placements between 2 neighboring chondrocytes.

The regeneration-competent adult animals have ability to regenerate their lost complex appendages with a near-perfect replica, owing to the positional identity acquired by the progenitor cells in the blastema, i.e. the blastemal cells. CD59, a CD59/Ly6 family member, has been identified as a regulator of positional identity in the tail blastemal cells of Gekko japonicus. To determine whether this function of CD59 is unique to the regenerative amniote(s) and how CD59 mediates PD axis patterning during tail regeneration, we examined its protective role on the complement-mediated cell lysis and intervened CD59 expression in the tail blastemal cells using an in vivo model of adenovirus transfection. Our data revealed that gecko CD59 was able to inhibit complement-mediated cell lysis. Meanwhile, CD59 functioned on positional identity through expression in cartilage precursor cells. Intervening positional identity by overexpression or siRNA knockdown of CD59 resulted in abnormal cartilaginous cone patterning due to the decreased differentiation of blastemal cells to cartilage precursor cells. The cartilage formation-related genes were found to be under the regulation of CD59. These results indicate that CD59, an evolutionarily transitional molecule linking immune and regenerative regulation, affects tail regeneration by mediating cartilage patterning. [ABSTRACT FROM AUTHOR]