Osteoarthritic Cartilage Lesions Research Articles

MiR-708–5p deficiency involves the degeneration of mandibular condylar chondrocytes via the TLR4/NF-κB pathway

ObjectiveAgeing and aberrant biomechanical stimulation are two major risk factors for osteoarthritis (OA). One of the main characteristics of aged cartilage is cellular senescence. One of the main characteristics of osteoarthritic joints is cartilage degeneration. The cells in the temporomandibular joint (TMJ) cartilage are zonally arranged. The deep zone cells are differentiated from the superficial zone cells (SZCs). The purpose of the present study was to investigate whether degenerative shear stress (SS) stimulates the senescence programme in TMJ SZCs, and to determine which miRNA is involved in this process. MethodSZCs were isolated from the TMJ condyles of 3-week-old rats and treated with continuous passaging or SS. RNA sequencing was conducted to identify miRNA(s) that overlap with those involved in the replication senescence process and the SS-induced degeneration programme. Unilateral anterior crossbite (UAC), which is TMJ-OA inducible, was applied to 2-month-old and 12-month-old mice for 3 weeks. The effect of TMJ local injection of agomiR-708-5p was evaluated histologically. ResultsBoth replication and SS treatment induced SZC senescence. miR-708-5p was identified. Knocking down miR-708-5p in SS treated SZCs led to more severe senescence by alleviating the inhibitory impact of miR-708-5p on the TLR4/NF-κB pathway. miR-708-5p expression in mouse TMJ cartilage decreased with age. UAC induced more severe osteoarthritic cartilage lesions in 12-month-old mice than in 2-month-old mice. Injection of agomiR-708-5p suppressed UAC-induced osteoarthritic cartilage lesions. ConclusionsAge-related miR-708-5p deficiency is involved in the mechanically stimulated OA process. Intra-articular administration of agomiR-708-5p is a promising new strategy for OA treatment. Data availabilityThe small RNA and mRNA sequencing data included in this study are available in the NCBI Gene Expression Omnibus (GEO) database (accession numbers GSE236980, GSE236977, and GSE 236978).

Covalent Conjugation of Small Molecule Inhibitors and Growth Factors to a Silk Fibroin-Derived Bioink to Develop Phenotypically Stable 3D Bioprinted Cartilage.

Implantation of a phenotypically stable cartilage graft could represent a viable approach for repairing osteoarthritic (OA) cartilage lesions. In the present study, we investigated the effects of modulating the bone morphogenetic protein (BMP), transforming growth factor beta (TGFβ), and interleukin-1 (IL-1) signaling cascades in human bone marrow stromal cell (hBMSC)-encapsulated silk fibroin gelatin (SF-G) bioink. The selected small molecules LDN193189, TGFβ3, and IL1 receptor antagonist (IL1Ra) are covalently conjugated to SF-G biomaterial to ensure sustained release, increased bioavailability, and printability, confirmed by ATR-FTIR, release kinetics, and rheological analyses. The 3D bioprinted constructs with chondrogenically differentiated hBMSCs were incubated in an OA-inducing medium for 14 days and assessed through a detailed qPCR, immunofluorescence, and biochemical analyses. Despite substantial heterogeneity in the observations among the donors, the IL1Ra molecule illustrated the maximum efficiency in enhancing the expression of articular cartilage components, reducing the expression of hypertrophic markers (re-validated by the GeneMANIA tool), as well as reducing the production of inflammatory molecules by the hBMSCs. Therefore, this study demonstrated a novel strategy to develop a chemically decorated, printable and biomimetic SF-G bioink to produce hyaline cartilage grafts resistant to acquiring OA traits that can be used for the treatment of degenerated cartilage lesions.

Arthroscopic Implantation of Adipose-Derived Stromal Vascular Fraction Improves Cartilage Regeneration and Pain Relief in Patients With Knee Osteoarthritis

PurposeTo compare the pain relief and cartilage repair status of patients with knee osteoarthritis who received arthroscopic treatment with or without stromal vascular fraction (SVF) implantation.MethodsWe retrospectively evaluated the patients who were examined with 12-month follow-up magnetic resonance imaging (MRI) after arthroscopic treatment for knee osteoarthritis from September 2019 to April 2021. Patients were included in this study if they had grade 3 or 4 knee osteoarthritis according to the Outerbridge classification in MRI. The visual analog scale (VAS) was used for pain assessment over the follow-up period (baseline and at 1-, 3-, 6-, and 12-month follow-ups). Cartilage repair was evaluated using follow-up MRIs based on Outerbridge grades and the Magnetic Resonance Observation of Cartilage Repair Tissue scoring system.ResultsAmong 97 patients who received arthroscopic treatment, 54 patients received arthroscopic treatment alone (conventional group) and 43 received arthroscopic treatment along with SVF implantation (SVF group). In the conventional group, the mean VAS score decreased significantly at 1-month post-treatment compared with baseline (P < .05), and gradually increased from 3 to 12 months’ post-treatment (all P < .05). In the SVF group, the mean VAS score decreased until 12 months post-treatment compared with baseline (all P < .05 except P = .780 in 1-month vs 3-month follow-ups). Significantly greater pain relief was reported in the SVF group than in the conventional group at 6 and 12 months’ post-treatment (all P < .05). Overall, Outerbridge grades were significantly greater in the SVF group than in the conventional group (P < .001). Similarly, mean Magnetic Resonance Observation of Cartilage Repair Tissue scores were significantly greater (P < .001) in the SVF group (70.5 ± 11.1) than in the conventional group (39.7 ± 8.2).ConclusionsThe results regarding pain improvement and cartilage regeneration and the significant correlation between pain and MRI outcomes at 12-months follow-up indicate that the arthroscopic SVF implantation technique may be useful for repairing cartilage lesions in knee osteoarthritis.Level of EvidenceLevel III, retrospective comparative study. To compare the pain relief and cartilage repair status of patients with knee osteoarthritis who received arthroscopic treatment with or without stromal vascular fraction (SVF) implantation. We retrospectively evaluated the patients who were examined with 12-month follow-up magnetic resonance imaging (MRI) after arthroscopic treatment for knee osteoarthritis from September 2019 to April 2021. Patients were included in this study if they had grade 3 or 4 knee osteoarthritis according to the Outerbridge classification in MRI. The visual analog scale (VAS) was used for pain assessment over the follow-up period (baseline and at 1-, 3-, 6-, and 12-month follow-ups). Cartilage repair was evaluated using follow-up MRIs based on Outerbridge grades and the Magnetic Resonance Observation of Cartilage Repair Tissue scoring system. Among 97 patients who received arthroscopic treatment, 54 patients received arthroscopic treatment alone (conventional group) and 43 received arthroscopic treatment along with SVF implantation (SVF group). In the conventional group, the mean VAS score decreased significantly at 1-month post-treatment compared with baseline (P < .05), and gradually increased from 3 to 12 months’ post-treatment (all P < .05). In the SVF group, the mean VAS score decreased until 12 months post-treatment compared with baseline (all P < .05 except P = .780 in 1-month vs 3-month follow-ups). Significantly greater pain relief was reported in the SVF group than in the conventional group at 6 and 12 months’ post-treatment (all P < .05). Overall, Outerbridge grades were significantly greater in the SVF group than in the conventional group (P < .001). Similarly, mean Magnetic Resonance Observation of Cartilage Repair Tissue scores were significantly greater (P < .001) in the SVF group (70.5 ± 11.1) than in the conventional group (39.7 ± 8.2). The results regarding pain improvement and cartilage regeneration and the significant correlation between pain and MRI outcomes at 12-months follow-up indicate that the arthroscopic SVF implantation technique may be useful for repairing cartilage lesions in knee osteoarthritis.

Intra-Articular Injection of Adipose-Derived Stem Cells Ameliorates Pain and Cartilage Anabolism/Catabolism in Osteoarthritis: Preclinical and Clinical Evidences.

Background: Osteoarthritis (OA) is the most common joint disorder, lacking disease-modifying treatments. Adipose-derived mesenchymal stem cells (ADSCs) are adult multipotent stromal cells obtained from fat tissue, which holds great potential in treating OA. This study aimed to evaluate the anti-OA efficacy of ADSCs from preclinical and clinical facets and explore the underlying mechanism of action. Methods: In vivo, a single dose of 5 × 105 ADSCs was injected into the knee joints of monoiodoacetate-induced OA rat model. The levels of metabolic and hypertrophic molecules (MMP13, Collagen II, Collagen X) of chondrocytes were measured by immunohistochemistry. In vitro, cell viability assay was conducted to detect the proliferation ability of chondrocytes treated with ADSCs conditioned medium (ADSCs-CM). Quantitative real-time polymerase chain reaction and Western blot assays were applied to explore the mechanism of action of ADSCs. Moreover, a retrospective analysis was conducted to determine the clinical efficacy and safety of ADSCs on OA patients. Results: The animal study showed that ADSCs significantly alleviated OA cartilage lesions in rats, as was confirmed by downregulation of the MMP13 and Collagen X and upregulation of the Collagen II. In vitro data showed that ADSCs-CM promoted the proliferation of chondrocytes, and significantly restored the IL-1β-induced abnormal expressions of molecular markers IL-6, Aggrecan, MMP3, MMP13, Collagen II, Collagen X, ADAMTS5, ADAMTS9, SOX6, and SOX9 in chondrocytes. Such regulatory effects of ADSCs-CM on the proliferation and these anabolic, catabolic, and hypertrophic markers of chondrocytes suggested a paracrine-based mode of action of ADSCs. Furthermore, the clinical data showed that ADSCs reduced pain and repaired cartilage damage in OA patients, with no adverse events. Conclusion: This study demonstrated the anti-OA efficacy, safety, and a paracrine-based mechanism of ADSCs, providing a promising cell-based therapeutic option for OA treatment.

Human adipose-derived stromal/stem cells expressing doublecortin improve cartilage repair in rabbits and monkeys

Localized cartilage lesions in early osteoarthritis and acute joint injuries are usually treated surgically to restore function and relieve pain. However, a persistent clinical challenge remains in how to repair the cartilage lesions. We expressed doublecortin (DCX) in human adipose-derived stromal/stem cells (hASCs) and engineered hASCs into cartilage tissues using an in vitro 96-well pellet culture system. The cartilage tissue constructs with and without DCX expression were implanted in the knee cartilage defects of rabbits (n = 42) and monkeys (n = 12). Cohorts of animals were euthanized at 6, 12, and 24 months after surgery to evaluate the cartilage repair outcomes. We found that DCX expression in hASCs increased expression of growth differentiation factor 5 (GDF5) and matrilin 2 in the engineered cartilage tissues. The cartilage tissues with DCX expression significantly enhanced cartilage repair as assessed macroscopically and histologically at 6, 12, and 24 months after implantation in the rabbits and 24 months after implantation in the monkeys, compared to the cartilage tissues without DCX expression. These findings suggest that hASCs expressing DCX may be engineered into cartilage tissues that can be used to treat localized cartilage lesions.

General Caspase Inhibition in Primary Chondrogenic Cultures Impacts Their Transcription Profile Including Osteoarthritis-Related Factors.

The knowledge about functions of caspases, usually associated with cell death and inflammation, keeps expanding also regarding cartilage. Active caspases are present in the growth plate, and caspase inhibition in limb-derived chondroblasts altered the expression of osteogenesis-related genes. Caspase inhibitors were reported to reduce the severity of cartilage lesions in osteoarthritis (OA), and caspase-3 might represent a promising biomarker for OA prognosis. The objective of this investigation was to decipher the transcriptomic regulation of caspase inhibition in chondrogenic cells. Limb-derived chondroblasts were cultured in the presence of 2 different inhibitors: Z-VAD-FMK (FMK) and Q-VD-OPH (OPH). A whole transcriptome RNA sequencing was performed as the key analysis. The analysis revealed a statistically significant increase in the expression of 252 genes in the FMK samples and 163 genes in the OPH samples compared with controls. Conversely, there was a significant decrease in the expression of 290 genes in the FMK group and 188 in the OPH group. Among the top up- and downregulated genes (more than 10 times changed), almost half of them were associated with OA. Both inhibitors displayed the highest upregulation of the inflammatory chemokine Ccl5, the most downregulated gene was the one for mannose receptors Mrc1. The obtained datasets pointed to a significant impact of caspase inhibition on the expression of several chondro-/osteogenesis-related markers in an in vitro model of endochondral ossification. Notably, the list of these genes included some encoding for factors associated with cartilage/bone pathologies such as OA.

Back Propagation Neural Network-Based Ultrasound Image for Diagnosis of Cartilage Lesions in Knee Osteoarthritis.

Objective To explore the application value of ultrasound image based on back propagation (BP) neural network algorithm in knee osteoarthritis (KOA) and evaluate the application effect and value of ultrasound image technology based on the BP neural network in the diagnosis of knee osteoarthritis cartilage lesions, 98 patients who were admitted to our hospital were diagnosed with KOA and had undergone arthroscopic soft tissue examinations were randomly selected. According to whether image processing was performed, the ultrasound images of all patients were divided into two groups. The control group was image before processing, and the experimental group was image after processing optimization. The consistency of the inspection results of the ultrasound images before and after the processing with the arthroscopy results was compared. The results showed that the staging accuracy of the control group was 68.3% and that of the experimental group was 76.9%. The accuracy of staging cartilage degeneration of the experimental group was higher than that of the control group, and the difference was not remarkable (P > 0.05). The kappa coefficient of the experimental group was 0.61, and that of the control group was 0.40. The kappa coefficient of the experimental group was higher than that of the control group, and the difference was significant (P < 0.05). Conclusion The inspection effect of the ultrasound image processed by the BP neural network was superior to that of the conventional ultrasound image. It reflected the good adoption prospect of neural networks in image processing.

Genetic deletion of nuclear protein-1 reduces the severity of cartilage lesions in a mouse model of post-traumatic osteoarthritis

Purpose: We recently found that Nuclear protein 1 (Nupr1), a stress-inducible transcription factor, is highly expressed in aged and osteoarthritic cartilage and in the knee joints of mice fed on high-fat diet. However, the role of Nupr1 in the development of OA is not clear. This study aimed to determine if genetic deletion of Nupr1 reduces the severity of cartilage lesions in surgically-induced osteoarthritis (OA) in mice

Bert model fine-tuning for text classification in knee OA radiology reports

Purpose: Traditional Natural Language Processing (NLP) techniques do a good job of understanding relationships between adjacent or nearby words. However, clinical text data such as patient notes or radiology reports require capturing interactions between distant words. BERT (Bidirectional Encoder Representations from Transformers), a new form of deep neural network recently introduced by Google, overcomes this challenge given its ability to comprehend long range word interactions. While BERT has dramatically improved outcomes in NLP tasks in the general domain such as optimizing search results, its performance in domain specific tasks such as analyzing biomedical data is still being explored. We report the findings of using a pre-trained BERT model further trained on knee radiology reports with the aim of testing its ability to identify cartilage lesions in osteoarthritis (OA) patients, the first such effort in musculoskeletal imaging. Methods: 1521 MR scans of patients with osteoarthritis and corresponding radiology reports were assessed by the clinician for all 6 cartilage compartments (Patella, Trochlea, Medial Femur, Medial Tibia, Lateral Femur and Lateral Tibia) according to the Whole-Organ Magnetic Resonance Imaging Score (WORMS) system for knee OA (Figure 1). These gradings were further simplified into binary (Normal and Abnormal) classes (WORMS target). The clinician also assessed (blinded) 89 radiology reports and provided binary targets (NLP target). Radiology reports contains Technique, Findings and Impression sections, due to reports vary in length, with median 329 and max 731 tokens for our dataset, and BERT has 512 tokens limitation, Findings section was selected as dataset for both classification tasks, Logistic Regression and BERT fine-tuning. Both models have 80/20 train test data split, trains on all 6 compartments and use the binary WORMS target. For baseline Logistic Regression model, texts were cleaned and tokenized with Spacy then applied Term Frequency Inverse Document Frequency (TF-IDF) transforming texts into numeric vectors as input and trained with class weighted logistic regression. After the training process, 5-fold cross validation was performed to evaluate performance. To fine-tune BERT text classification task, reports were preprocessed with lower cased text, added special tokens to mark beginning and end sentences, padded if reports are shorter than 512 tokens, then generated vector representations with bert-base-uncased pre-trained model into training features along with input mask, and segment id. In evaluation step, the fine-tuned BERT models were then tested on each compartment for both WORMS target and NLP target. Results: For both training and testing datasets, the ratio of normal to abnormal cases varies. Patella has the most abnormal making it more balanced while MT has the least (Figure 2A). In the more balanced compartments such as Patella and Trochlea, LFC, BERT significantly improved the classification results compares to the baseline model. However, for the imbalanced dataset, even though BERT shows higher accuracy, it overfits and predicted Normal for all. This can be solved by hyper-parameter tuning and adjusting class weight in loss function for each model. Each fine-tuned BERT model is tested with NLP Target (labels on 89 radiology reports) and compared with the earlier WORMS target (gradings from images) (Figure 2B). Accuracy significantly improves on the NLP target with more balanced dataset (Figure 3). However, the overfit problem persists on MFC, MT, LT compartments. This implies the discrepancies between MR images and the content in the radiology reports, using NLP target which provides a more accurate annotations on reports will further improve the model’s prediction abilities. Conclusions: Our results in cartilage abnormality prediction esp. in compartments with balanced data holds promise and calls for further investigating use of BERT for domain specific tasks. The ability to develop a model which can accurately decipher text sentiment from radiology reports avoiding the need of time-consuming image annotation will accelerate and augment image based deep learning approaches for anomaly detection and improve clinical workflow.View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Effect of high molecular weight hyaluronic acid in treatment of osteoarthritic temporomandibular joints of rats

ObjectiveThe aim of this study was to determine the histological and biochemical effects of high molecular weight hyaluronic acid on experimentally induced osteoarthritic changes in male rat temporomandibular joint (TMJ). DesignIn this in-vivo study, 36 male albino rats were divided into 3 groups; control non-injected, disease and treatment group. Osteoarthritis was induced using a single intra-articular injection of Complete Freund’s adjuvant (CFA) (50 μl). High molecular weight hyaluronic acid (HMWHA) was injected intra-articularly once a week, for a total of 3 injections. TMJ tissue samples were dissected from control, disease and treatment groups for biochemical analysis using ELISA test to detect the levels of matrix metalloproteinase-3 (MMP-3) on day 28. Histological examination was done using Hematoxylin &eosin, Mallory’s trichrome and Alcian blue stains. Data was analyzed using Kruskal-Wallis test followed by Dunn-Sidek method, with a 5 % significance level. ResultsTreatment group showed regaining of the normal histological features of the TMJ and decreased levels of MMP-3 when compared to disease group. ConclusionsThese findings may suggest that HMWHA plays a role in the management of CFA-induced osteoarthritic cartilage lesions.

Effects of rAAV-Mediated sox9 Overexpression on the Biological Activities of Human Osteoarthritic Articular Chondrocytes in Their Intrinsic Three-Dimensional Environment.

Gene therapy for osteoarthritis offers powerful, long-lasting tools that are well adapted to treat such a slow, progressive disorder, especially those therapies based on the clinically adapted recombinant adeno-associated viral (rAAV) vectors. Here, we examined the ability of an rAAV construct carrying a therapeutic sequence for the cartilage-specific SOX9 transcription factor to modulate the phenotype of human osteoarthritic articular chondrocytes compared with normal chondrocytes in a three-dimensional environment where the cells are embedded in their extracellular matrix. Successful sox9 overexpression via rAAV was noted for at least 21 days, leading to the significant production of major matrix components (proteoglycans, type-II collagen) without affecting the proliferation of the cells, while the cells contained premature hypertrophic processes relative to control conditions (reporter rAAV-lacZ application, absence of vector treatment). These findings show the value of using rAAV to adjust the osteoarthritic phenotype when the chondrocytes are confined in their inherently altered environment and the possibility of impacting key cellular processes via gene therapy to remodel human osteoarthritic cartilage lesions.

Biplanar ultrasonografik incelemenin femoral artiküler kıkırdaktaki dejeneratif değişiklikleri saptamadaki tanı değeri

Aim: The aim of our study was to assess the diagnostic value of biplanar ultrasonography in detecting degenerative changes of femoral articular cartilage using magnetic resonance imaging as the reference method.Material and Methods: Femoral articular cartilage examination was performed with ultrasonography and magnetic resonance imaging in 40 patients with knee pain. Magnetic resonance imaging of the knees were carried out on a 1.5 T scanner with proton density weighted, fat suppressed sequence in three orthogonal planes. Ultrasonographic evaluations of the knees were performed in both longitudinal and transverse planes. Taking the findings of magnetic resonance imaging as a reference, the diagnostic effectiveness of biplanar US in detecting medial condylar, lateral condylar and intercondylar notch cartilage lesions were calculated.Results: The sensitivity of biplanar ultrasonography in detecting medial condylar, lateral condylar, and intercondylar notch cartilage lesions were 93.55%, 61.11%, and 53.85%, respectively. The specificity of the technique in detecting the lesions in these locations were 88.89%, 90.91%, and 100.00%, respectively. Conclusion: Ultrasonography is a relatively inexpensive, easily accessible and a non-invasive imaging tool. We found that biplanar ultrasonography is a reliable method in detecting medial condylar cartilage lesions in knee osteoarthritis, and we recommend using it for the purposes of screening and follow-up evaluation of medial type knee osteoarthritis where MRI is not accessible.

3D convolutional neural networks for detection and severity staging of meniscus and PFJ cartilage morphological degenerative changes in osteoarthritis and anterior cruciate ligament subjects.

Semiquantitative assessment of MRI plays a central role in musculoskeletal research; however, in the clinical setting MRI reports often tend to be subjective and qualitative. Grading schemes utilized in research are not used because they are extraordinarily time-consuming and unfeasible in clinical practice. To evaluate the ability of deep-learning models to detect and stage severity of meniscus and patellofemoral cartilage lesions in osteoarthritis and anterior cruciate ligament (ACL) subjects. Retrospective study aimed to evaluate a technical development. In all, 1478 MRI studies, including subjects at various stages of osteoarthritis and after ACL injury and reconstruction. 3T MRI, 3D FSE CUBE. Automatic segmentation of cartilage and meniscus using 2D U-Net, automatic detection, and severity staging of meniscus and cartilage lesion with a 3D convolutional neural network (3D-CNN). Receiver operating characteristic (ROC) curve, specificity and sensitivity, and class accuracy. Sensitivity of 89.81% and specificity of 81.98% for meniscus lesion detection and sensitivity of 80.0% and specificity of 80.27% for cartilage were achieved. The best performances for staging lesion severity were obtained by including demographics factors, achieving accuracies of 80.74%, 78.02%, and 75.00% for normal, small, and complex large lesions, respectively. In this study we provide a proof of concept of a fully automated deep-learning pipeline that can identify the presence of meniscal and patellar cartilage lesions. This pipeline has also shown potential in making more in-depth examinations of lesion subjects for multiclass prediction and severity staging. 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;49:400-410.

Cartilage repair in degenerative osteoarthritis mediated by squid type II collagen via immunomodulating activation of M2 macrophages, inhibiting apoptosis and hypertrophy of chondrocytes

Cartilage lesions in degenerative osteoarthritis (OA) are involved with pathological microenvironmental alterations induced by inflammatory macrophages, and apoptotic and/or hypertrophic chondrocytes. However, current non-operative therapies for cartilage repair in OA can rarely achieve long-term and satisfactory outcomes. This study aims to evaluate a newly developed squid type II collagen (SCII) for repairing OA-induced cartilage lesions. Our in vitro data show that SCII induces M2 polarization of macrophages, and activates macrophages to express pro-chondrogenic genes (TGF-β and IGF), which greatly improves the microenvironment around chondrocytes to produce type II collagen and glycosaminoglycan. In addition, glycine in SCII activates glycine receptors on inflammatory chondrocytes to decrease intracellular calcium concentration, leading to effective inhibition of chondrocyte apoptosis and hypertrophy. The in vitro effects of SCII are further confirmed in vivo. In a rat model of OA, SCII increases the ratio of M2 macrophages, elevates the levels of pro-chondrogenic cytokines (TGF-β1 and TGF-β3) in synovial fluid, and inhibits chondrocyte apoptosis and MMP13 production. Our findings show that SCII immunomodulates M2 activation of macrophages to skew the local OA microenvironment towards a pro-chondrogenic atmosphere, and promotes cartilage repair under inflammatory condition. It shows great potential for SCII to be a novel biomaterial for cartilage repair in OA.

A Review of Commercially Available Point-of-Care Devices to Concentrate Bone Marrow for the Treatment of Osteoarthritis and Focal Cartilage Lesions

Mesenchymal stem cells (MSCs) are a promising cell-based therapy treatment option for several orthopedic indications. Because culture expansion of MSC is time and cost intensive, a bedside concentration of bone marrow (BM) aspirate is used as an alternative. Many commercial systems are available but the available literature and knowledge regarding these systems is limited. We compared different point-of-care devices that concentrate BM (BMC) by focusing on technical features and quality parameters to help surgeons make informed decisions while selecting the appropriate device. We compared published data on the BMC devices of Arteriocyte, Arthrex, Celling Biosciences, EmCyte, Exactech, ISTO Tech, Harvest Tech/Terumo BCT, and Zimmer/BIOMET regarding technical features (centrifugation speed/time, input/output volume, kit components, type of aspiration syringes, filter usage) and quality parameters of their final BMC product (hematocrit, concentration of platelets and total nucleated cells, concentration of MSC and connective tissue progenitor cells). The systems differ significantly in their technical features and centrifugation parameters. Only the fully automated systems use universal kits, which allow processing different volumes of BM. Only the Arthrex system allows selection of final hematocrit. There was no standardized reporting method to describe biologic potency. Based on the data obtained in this review, recommending a single device is not possible because the reported data could not be compared between devices. A standardized reporting method is needed for valid comparisons. Furthermore, clinical outcomes are required to establish the true efficacy of these systems. We are conducting additional studies for more careful comparison among the devices.

Pentosan Polysulfate Sodium Restores the Phenotype of Dedifferentiated Monolayer Canine Articular Chondrocytes Cultured in Alginate Beads

Autologous chondrocyte transplantation is a promising option for the repair of isolated osteoarthritic cartilage lesions that requires isolation and expansion of chondrocytes from a small cartilage biopsy prior to implantation. However, when cultured in vitro, chondrocytes lose their stable phenotype and dedifferentiate to fibroblastic-like cells. The study investigated the potential of pentosan polysulfate (PPS) sodium to restore the phenotype of dedifferentiated monolayer articular chondrocytes. Canine articular chondrocytes isolated from four cartilage samples were culture expanded to establish primary culture. First passage chondrocytes were cultured as alginate beads for 18 days under normoxia in PPS concentrations of 0, 1, 5, 15 and 40 μg/mL in 20% DMEM. Effect of PPS on type I, II and X collagen, aggrecan and Runx2 gene expression were evaluated by real-time PCR. Runx2, HIF-1α and HIF-2α protein expression were evaluated by Western blot and proteoglycan deposition was determined by Alcian blue stain. Dedifferentiated chondrocytes fully retained their phenotype as evidenced by increased synthesis of cartilage-specific genes, type II collagen and aggrecan mRNA with complete suppression of type I and X collagen at PPS concentrations of 15 and 40 μg/mL. Compared to the control, type II collagen and aggrecan mRNA were significantly upregulated (P<0.05) at 5, 15 and 40 μg/mL and 5 and 15 μg/mL PPS, respectively. PPS significantly enhanced proteoglycan with peak deposition at 5 μg/mL compared to control. HIF-1α and HIF-2α proteins were detectable at protein level for the first time under normoxia condition in alginate culture. The study demonstrates for the first time the restoration of dedifferentiated canine articular chondrocytes phenotype by combining alginate encapsulation with culture in PPS without the addition of known chondrocytic growth factors. The study confirms PPS as novel chondroinductive factor with potential to offer a solution to the major challenges that exist in cartilage tissue engineering.

Effect of Kaempferia parviflora extract on knee osteoarthritis.

Knee osteoarthritis (OA) is becoming more prevalent worldwide due to increases in the numbers of elderly and obese patients. Currently, pharmaceutical medicines used for the treatment of OA are for symptomatic therapy and therefore new therapeutic agents are needed. Kaempferia parviflora (KP) is a plant growing naturally in Southeast Asia and has various pharmacological effects including an anti-inflammatory effect, but no effect on OA has yet been reported. We therefore conducted a search for the effects KP and the active components of KP extract (KPE) exert on OA as well as its mechanism of action. Results from a study of KPE using the monoiodoacetic acid rat OA model revealed that KPE reduced the pain threshold and severity of osteoarthritic cartilage lesions. The mechanism of action and active components were then investigated using IL-1β-treated human knee-derived chondrocytes. KPE, as well as 5,7-dimethoxyflavone and 5,7,4'-trimethoxyflavone, which are key constituents of KPE and highly absorbable into the body, reduced the expression of matrix metalloproteinases (MMPs), which are the main extracellular matrix enzymes that degrade collagen within cartilage. As mentioned above, KPE acted to suppress OA and 5,7-dimethoxyflavone and 5,7,4'-trimethoxyflavone were shown to be involved as part of KPE's mechanism that inhibits MMPs.

Elevated adiabatic T1ρ and T2ρ in articular cartilage are associated with cartilage and bone lesions in early osteoarthritis: A preliminary study.

To evaluate adiabatic T1ρ and T2ρ of articular cartilage in symptomatic osteoarthritis (OA) patients and asymptomatic volunteers, and to determine their association with magnetic resonance imaging (MRI)-based structural abnormalities in cartilage and bone. A total of 24 subjects (age range: 50-68 years; 12 female) were enrolled, including 12 early OA patients and 12 volunteers with normal joint function. Patients and volunteers underwent 3T MRI. T2 , adiabatic T1ρ , and T2ρ relaxation times of knee articular cartilage were measured. Proton density (PD)- and T1 -weighted MR image series were also obtained and separately evaluated for morphological changes using the MRI OA Knee Scoring (MOAKS) system. Comparisons using the Mann-Whitney nonparametric test were performed after dividing the study participants according to physical symptoms as determined by Western Ontario and McMaster Universities (WOMAC) score or presence of cartilage lesions, bone marrow lesions, or osteophytes. Elevated adiabatic T1ρ and T2ρ relaxation times of articular cartilage were associated with cartilage loss (P = 0.024-0.047), physical symptoms (0.0068-0.035), and osteophytes (0.0039-0.027). Elevated adiabatic T1ρ was also associated with bone marrow lesions (0.033). Preliminary data suggest that elevated adiabatic T1ρ and T2ρ of cartilage are associated with morphological abnormalities of cartilage and bone, and thus may be applicable for in vivo OA research and diagnostics. 2 Technical Efficacy: Stage 2 J. MAGN. RESON. IMAGING 2017;46:678-689.

PDK2 promotes chondrogenic differentiation of mesenchymal stem cells by upregulation of Sox6 and activation of JNK/MAPK/ERK pathway.

This study was undertaken to clarify the role and mechanism of pyruvate dehydrogenase kinase isoform 2 (PDK2) in chondrogenic differentiation of mesenchymal stem cells (MSCs). MSCs were isolated from femurs and tibias of Sprague-Dawley rats, weighing 300-400 g (5 females and 5 males). Overexpression and knockdown of PDK2 were transfected into MSCs and then cell viability, adhesion and migration were assessed. Additionally, the roles of aberrant PDK2 in chondrogenesis markers SRY-related high mobility group-box 6 (Sox6), type ΙΙ procollagen gene (COL2A1), cartilage oligomeric matrix protein (COMP), aggrecan (AGC1), type ΙX procollagen gene (COL9A2) and collagen type 1 alpha 1 (COL1A1) were measured by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The expressions of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK) and extracellular regulated protein kinase (ERK) were measured. Overexpressing PDK2 promoted cell viability, adhesion and inhibited cell migration in MSCs (all P<0.05). qRT-PCR assay showed a potent increase in the mRNA expressions of all chondrogenesis markers in response to overexpressing PDK2 (P<0.01 or P<0.05). PDK2 overexpression also induced a significant accumulation in mRNA and protein expressions of JNK, p38MAPK and ERK in MSCs compared to the control (P<0.01 or P<0.05). Meanwhile, silencing PDK2 exerted the opposite effects on MSCs. This study shows a preliminary positive role and potential mechanisms of PDK2 in chondrogenic differentiation of MSCs. It lays the theoretical groundwork for uncovering the functions of PDK2 and provides a promising basis for repairing cartilage lesions in osteoarthritis.

Systemic inhibition of IL-6/Stat3 signalling protects against experimental osteoarthritis

ObjectiveTo investigate the impact of systemic inhibition of interleukin 6 (IL-6) or signal transducer and activator of transcription (Stat3) in an experimental model of osteoarthritis (OA).MethodsExpression of major catabolic and...