Treatment Of Joint Diseases Research Articles

Local Cellular Responses to Metallic and Ceramic Nanoparticles from Orthopedic Joint Arthroplasty Implants.

Over the last decades, joint arthroplasty has become a successful treatment for joint disease. Nowadays, with a growing demand and increasingly younger and active patients accepting these approaches, orthopedic surgeons are seeking implants with improved mechanical behavior and longer life span. However, aseptic loosening as a result of wear debris from implants is considered to be the main cause of long-term implant failure. Previous studies have neatly illustrated the role of micrometric wear particles in the pathological mechanisms underlying aseptic loosening. Recent osteoimmunologic insights into aseptic loosening highlight the important and heretofore underrepresented contribution of nanometric orthopedic wear particles. The present review updates the characteristics of metallic and ceramic nanoparticles generated after prosthesis implantation and summarizes the current understanding of their hazardous effects on peri-prosthetic cells.

An immortalized human adipose-derived stem cell line with highly enhanced chondrogenic properties

Human adipose-derived stem cells (ASCs) are a commonly used cell type for cartilage tissue engineering. However, donor-to-donor variability, cell heterogeneity, inconsistent chondrogenic potential, and limited expansion potential can hinder the use of these cells for modeling chondrogenesis, in vitro screening of drugs and treatments for joint diseases, or translational applications for tissue engineered cartilage repair. The goal of this study was to create an immortalized ASC line that showed enhanced and consistent chondrogenic potential for applications in cartilage tissue engineering as well as to provide a platform for investigation of biological and mechanobiological pathways involved in cartilage homeostasis and disease. Starting with the ASC52telo cell line, a hTERT-immortalized ASC line, we used lentivirus to overexpress SOX9, a master regulator of chondrogenesis, and screened several clonal populations of SOX9 overexpressing cells to form a new stable cell line with high chondrogenic potential. One clonal line, named ASC52telo-SOX9, displayed increased GAG and type II collagen synthesis and was found to be responsive to both mechanical and inflammatory stimuli in a manner similar to native chondrocytes. The development of a clonal line such as ASC52telo-SOX9 has the potential to be a powerful tool for studying cartilage homeostasis and disease mechanisms in vitro, and potentially as a platform for in vitro drug screening for diseases that affect articular cartilage. Our findings provide an approach for the development of other immortalized cell lines with improved chondrogenic capabilities in ASCs or other adult stem cells.

Novel Framework for Knee Arthroscopic Image Enhancement

An arthroscope is a tool for allowing an endoscope to be inserted directly into the inside of a joint to observe its structure, in contrast to X-rays, computed tomography, and magnetic resonance imaging, which directly capture pictures of a joint. Therefore, it can effectively treat joint diseases by identifying causes of pain that are not found by, e.g., computed tomography and magnetic resonance imaging. However, joint endoscopy has a very high cost, is very burdensome for patients, and has problems in regards to infection when being re-used. Thus, we developed disposable joint endoscopic camera modules for preventing re-use and infection, and researched approaches to reducing patient waiting times and cost burdens. In that regard, it is necessary to improve the brightness and color of the images, as they are used for compacting and disposal of the camera modules. In addition, we studied methods for improving automatic images, as image colors may vary (owing to blood or other foreign substances) when observed using the arthroscope. The proposed framework is divided into two sequences. First, we perform a histogram modification algorithm as an image enhancement technique. This results in a brightness optimization effect on the arthroscopic image. Second, we conduct a high saturation color mapping before proceeding to the next step. In particular, one of the reference points for diagnosing a disease is color information; thus, the improvement of color saturation is considered first in the color mapping. The proposed method provides better brightness values while preserving color information.

Mass spectrometric analysis of the in vitro secretome from equine bone marrow-derived mesenchymal stromal cells to assess the effect of chondrogenic differentiation on response to interleukin-1\u03b2 treatment

BackgroundSimilar to humans, the horse is a long-lived, athletic species. The use of mesenchymal stromal cells (MSCs) is a relatively new frontier, but has been used with promising results in treating joint diseases, e.g., osteoarthritis. It is believed that MSCs exert their main therapeutic effects through secreted trophic biomolecules. Therefore, it has been increasingly important to characterize the MSC secretome. It has been shown that the effect of the MSCs is strongly influenced by the environment in the host compartment, and it is a crucial issue when considering MSC therapy. The aim of this study was to investigate differences in the in vitro secreted protein profile between naïve and chondrogenic differentiating bone marrow-derived (BM)-MSCs when exposed to an inflammatory environment.MethodsEquine BM-MSCs were divided into a naïve group and a chondrogenic group. Cells were treated with normal expansion media or chondrogenic media. Cells were treated with IL-1β for a period of 5 days (stimulation), followed by 5 days without IL-1β (recovery). Media were collected after 48 h and 10 days. The secretomes were digested and analyzed by nanoLC-MS/MS to unravel the orchestration of proteins.ResultsThe inflammatory proteins IL6, CXCL1, CXCL6, CCL7, SEMA7A, SAA, and haptoglobin were identified in the secretome after 48 h from all cells stimulated with IL-1β. CXCL8, OSM, TGF-β1, the angiogenic proteins VCAM1, ICAM1, VEGFA, and VEGFC, the proteases MMP1 and MMP3, and the protease inhibitor TIMP3 were among the proteins only identified in the secretome after 48 h from cells cultured in normal expansion media. After 10-day incubation, the proteins CXCL1, CXCL6, and CCL7 were still identified in the secretome from BM-MSCs stimulated with IL-1β, but the essential inducer of inflammation, IL6, was only identified in the secretome from cells cultured in normal expansion media.ConclusionThe findings in this study indicate that naïve BM-MSCs have a more extensive inflammatory response at 48 h to stimulation with IL-1β compared to BM-MSCs undergoing chondrogenic differentiation. This extensive inflammatory response decreased after 5 days without IL-1β (day 10), but a difference in composition of the secretome between naïve and chondrogenic BM-MSCs was still evident.

Synthesis and Characterization of Coated Iron Oxide Nanoparticles Produced for Drug Delivery in Viscoelastic Solution

Drug delivery systems enable transportation of drugs in the body, controlling the time, rate or place of the release rendering them ideal for local treatments. When treating joint diseases such as osteoarthritis of the knee the therapeutically substances are given intra-particularly. Magnetic nanoparticles are used so that the solution containing the treatment can be easily directed from the outside with the help of magnets allowing the drug to reach the maximum concentration at the area of interest. Functionalization of nanoparticles is necessary to obtain viscoelastic solutions with optimal physico-chemical and medical properties. This paper presents a superior method of delivering intraarticular hyaluronic acid using iron oxide magnetic nanoparticles (Fe2O3) which were embedded in tetramethylammonium hydroxide (TMOH) and coated using a layer-by-layer technique with hyaluronic acid and inulin. TMOH was chosen for a better dispersion of nanoparticles in the viscoelastic solution, eliminating the risk of agglomerations, hyaluronic acid and inulin being used for medical purposes.

Efficiency of Glucosamine Sulphate and Chondroitin Sulphate Сombination in Osteoarthritis Treatment from Evidence-Based Medicine Positions

Osteoarthritis remains by far the most common form of joint damage and the leading cause of disability worldwide. Unfortunately, today there are no radical methods that can stop the «epidemic» of OA and completely cure patients with this disease. In recent decades, the attention of scientists and clinicians has been shrouded in the possibility of using symptomatic or diseasemodifying slow-acting drugs that are potentially not only able to influence the symptoms of OA (primarily pain), but also to have a lasting structural-modifying effect on the joint. The results of clinical studies, as well as a deeper understanding of the differences in the pharmacokinetics and properties of molecules of different chondroprotectors have aroused scientific and practical interest in approaches to the treatment of joint diseases using a combination of chondroitin sulfate and glucosamine sulfate.

Deciphering the potential pharmaceutical mechanism of Guzhi Zengsheng Zhitongwan on rat bone and kidney based on the \u201ckidney governing bone\u201d theory

BackgroundGuzhi Zengsheng Zhitongwan (GZZSZTW) is an effective Chinese medicinal formulation for the treatment of osteoarthritis (OA) designed according to the “kidney governing bone” theory, which has been widely used as a golden guide for treating bone and cartilage diseases in traditional Chinese medicine. The aim of this study was to explore the molecular mechanism underlying its effects on the bone and kidney.MethodsPreparation and quality control were performed as previously described. Since GZZSZTW is orally administered in the form of pills prepared in boiled water, the Chinese materia medica (CMM) mixture of this formula was extracted with distilled water by a reflux method and was then filtered through a 0.45-μm Hollow Fiber Cartridge (GE Healthcare, USA). The filtrate was freeze-dried by a Heto PowerDry LL3000 Freeze Dryer (Thermo, USA) and stored at − 80 °C. The effects of GZZSZTW on gene expression and regulation of both kidney and bone tissues were investigated using a state-of-the-art RNA-seq technology.ResultsWe demonstrated that GZZSZTW could enhance kidney function and suppress bone formation and resorption by modulating the activities of osteoblast and osteoclast, and might subsequently contribute to the inhibition of osteophyte formation during the process of OA. These effects might be achieved by the synergistic interactions of various herbs and their active components in GZZSZTW, which increased the expression levels of functional genes participating in kidney function, regulation, and repair, and then decreased the expression levels of genes involved in bone formation and resorption. Thus, our findings were consistent with the “kidney governing bone” theory, which has been widely used as a guide in clinical practice for thousands of years.ConclusionsThis study has deepened the current knowledge about the molecular effects of GZZSZTW on bone and kidney regulation. Furthermore, this study might be able to provide possible strategies to further prevent and treat joint diseases by using traditional Chinese medicinal formulations following the “kidney governing bone” theory.

Cannabidiol: A Brief Review of Its Therapeutic and Pharmacologic Efficacy in the Management of Joint Disease

Cannabis use in the management of musculoskeletal diseases has gained advocacy since several states have legalized its recreational use. Cannabidiol (CBD), a commercially available, non-neurotropic marijuana constituent, has shown promise in arthritic animal models by attenuating pro-inflammatory immune responses. Additional research has demonstrated the benefit of CBD in decreasing the endogenous pain response in mice subjected to acute arthritic conditions, and further studies have highlighted improved fracture healing following CBD use in murine mid-femoral fractures. However, there is a lack of high-quality, novel research investigating the use of CBD in human musculoskeletal diseases aside from anecdotal accounts and retrospective reviews, perhaps due to legal ramifications limiting the enrollment of patients. The purpose of this review article is to highlight the extent of current research on CBD and its biochemical and pharmacologic efficacy in the treatment of joint disease, as well as the evidence for use of CBD and cannabis in patients undergoing joint arthroplasty. Based on available literature relying on retrospective data and case reports, it is challenging to propose a recommendation for CBD use in perioperative pain management. Additionally, a number of CBD products currently available as supplements with different methods of administration, and it is important to remember that these products are non-pharmaceuticals. However, given the increased social relevance of CBD and cannabis-based medicines, future, prospective controlled studies evaluating their efficacy are needed.

Identification and Quantification of Transient Receptor Potential Vanilloid 1 (TRPV1) in Equine Articular Tissue.

Simple summaryOsteoarthritis is affecting several species including the horse. Transient receptor potential vanilloid 1 (TRPV1)—also known as the ‘chili receptor’—is currently being investigated as a target for treating osteoarthritis in humans. To evaluate whether it could be a potential target in treating osteoarthritis in horses, we collected synovial membrane samples from healthy horse joints and horse joints with joint disease to investigate the expression of the TRPV1. Laboratory analysis showed that TRPV1 is present in horse joints, and that the levels might be elevated in diseased joints, even at a chronic state of disease. This means that TRPV1 could be used as a potential target for osteoarthritis treatment in horses. Future studies in this area will not only be beneficial to horses and their owners, but more knowledge about TRPV1, the mode of action, and possible side effects will also benefit translation into human osteoarthritis research.Joint pain and osteoarthritis (OA) are some of the most common causes of lameness in horses, and most of the available treatments focus on symptomatic relief without a disease-modifying effect. TRPV1 is a potential target for treating joint diseases, including OA, and the present study aims to investigate if the TRPV1 receptor is present in equine articular tissue and determine whether the number of receptors is upregulated in joint inflammation. Metacarpo/metatarsophalangeal (MCP/MTP) joints from 15 horses euthanised for reasons unrelated to this study were included. Based on synovial fluid analysis, macroscopic evaluation, and magnetic resonance imaging (MRI), joints were divided into two groups: healthy joints and joints with pathology. ELISA analysis was performed on synovial tissue harvested from all joints. TPRV1 was found in all joints. The mean concentration of TRPV1 compared to total protein in healthy joints (8.4 × 10−7 ng/mL) and joints with pathology (12.9 × 10−7 ng/mL) differed significantly (p = 0.01, t-test with Welch correction). Quantitative real-time reverse transcriptase PCR analysis was performed on RNA isolates from synovial tissue from all joints. TRPV1 mRNA expression ratio normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in healthy joints (0.16 (SD: 0.19)) and joints with pathology (0.24 (SD: 0.14)) did not differ significantly (p = 0.43, t-test with Welch correction). mRNA expression of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) was very low for both groups. In conclusion, TRPV1 was detected both on mRNA and the protein level, with a higher expression of TRPV1 in samples from joints with pathology. Future studies will determine the clinical potential of equine TRPV1 as a target in the management of joint pain and inflammation.

Dexamethasone causes calcium deposition and degeneration in human anterior cruciate ligament cells through endoplasmic reticulum stress

BackgroundDexamethasone is widely used in the treatment of joint diseases due to its anti-inflammatory properties. However, it can cause serious adverse effects. The anterior cruciate ligament (ACL) is an important stabilizer of the knee joint. However, the effect of dexamethasone treatment on the ACL is unclear. ObjectiveThis study aims to explore the effects of dexamethasone on ACL tissues and cells through in vitro and in vivo experiments. ResultsIn vitro, we found that after treatment with dexamethasone, human ACL cell apoptosis was increased, type I collagen (COL1A1) content was decreased, mineralization related genes (ENPP1 and ANKH) and calcified nodules were increased, and endoplasmic reticulum stress (ERS) was enhanced. However, ERS inhibitors could significantly inhibit the increase in calcification and the decrease in COL1A1 induced by dexamethasone. In vivo, Wistar rats received the infra-articular injection with dexamethasone (0.5 mg/kg) for 8 weeks. We found that dexamethasone treatment decreased the COL1A1 content and increased the COL2A1 content in the ACL tissues of rats and that chondroid differentiation and mineralization occurred. Meanwhile, the expression of ERS-related proteins was increased. ConclusionDexamethasone increased the calcification of ACL cells and caused ACL degeneration through ERS, suggesting that long-term treatment with dexamethasone may cause adverse effects on ACL tissue and increase the risk of long-term rupture.

Catalpol promotes mitochondrial biogenesis in chondrocytes

The chondrocyte mitochondrial dysfunction has been considered to be associated with the pathogenesis of joint diseases. Catalpol is an active traditional Chinese medicine ingredient named Di-Huang, which is used widely to treat different diseases. In this study, we found the addition of catalpol in chondrocytes induced the expression of crucial mitochondrial regulators, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), and mitochondrial transcription factor A (TFAM). Catalpol promoted mitochondrial biogenesis, as revealed by the induction on the mitochondrial DNA/nuclear DNA (mtDNA/nDNA) and the expression of several mitochondrial genes including translocase of outer mitochondrial membrane 22 (Tomm22), translocase of outer mitochondrial membrane 70 (Tomm70), mitochondrial import inner membrane translocase subunit 50 (Timm50), NADH dehydrogenase [ubiquinone] iron-sulphur protein 3 (NDUFS3), adenosine triphosphate (ATP) synthase subunit D (ATP5d), and cytochrome B. Consequently, catalpol increased cytochrome c oxidase activity, the mitochondrial respiratory rate, and the extracellular ATP production, indicating that catalpol boosted mitochondrial function. Mechanistically, catalpol increased the activation of the cAMP-responsive element-binding protein (CREB), and the inhibition of CREB abolished catalpol-mediated promotion on mitochondrial biogenesis. In summary, this study demonstrated that catalpol has the potential to be used in the treatment of joint diseases.

The role of rehabilitation and apparatus physiotherapy in rheumatic disease treatment strategies

Rheumatic diseases are a serious medical and social problem. The variability of mechanisms for the development of this group of diseases requires different approaches: strategies of drug and non-medicament therapy in modern rheumatology are designed to complement each other. EULAR recommendations pay insufficient attention to the use of non-medical methods of joint disease treatment. According to modern research, multi-disciplinary rehabilitation strategies, including, in addition to drug therapy, educational programmes, physical training of various intensities, as well as the use of apparatus rehabilitation methods, have proven to be the most effective. Hardware physiotherapy seems to be optimal for the treatment of rheumatic patients due to its higher accessibility and lower cost in comparison with classical balneo- and peloidotherapy. With the use of magnetotherapy there is also greater safety and fewer contraindications to the procedures. The method of using pulsed magnetic fields is the most justified in the treatment of joint diseases, since the sensitivity of biological tissues to them is the highest. The clinical effectiveness of pulsed magnetotherapy has been demonstrated in several randomized placebo-controlled multi-centre studies. Against the background of carried out magnetic field therapy in patients were statistically significant reduction of pain and stiffness, improvement of joint function, which indicates the expediency of including the physical factor in the strategy of treatment of rheumatic diseases in combination with drug therapy.

АКТИВНІСТЬ АНТИОКСИДАНТНИХ ФЕРМЕНТІВ У СИРОВАТЦІ КРОВІ ЩУРІВ ЗА УМОВ КАРРАГІНАН-ІНДУКОВАНОГО ЗАПАЛЕННЯ СУГЛОБУ ТА ПРОФІЛАКТИЧНОГО ВВЕДЕННЯ ХОНДРОЇТИН СУЛЬФАТУ

Among the diseases of the musculoskeletal system, the leading place is occupied by joint diseases. It is important to conduct effective prevention of these diseases in order to stop or slow down the development of pathological changes in the body. In this regard, an important issue is the search for means to restore joints. Promising in the prevention and treatment of joint diseases is the use of chondroitin sulfate, which is a natural component of the intercellular substance of cartilage. The aim of this work was to investigate the prophylactic effect of chondroitin sulfate on the activity of superoxide dismutase and catalase in rat blood serum under conditions of carrageenan-induced joint inflammation. The studies were conducted on white non-linear, sexually mature male rats weighing 180–240 g, in compliance with the general ethical principles of experiments on animals. All animals were divided into four experimental groups. The first group – control: animals sub-planar injected 0.1 ml of 0,9 % NaCl solution into the posterior right limb. The second group – animals received a therapeutic dose of 3 mg x kg-1 chondroitin sulfate daily for 28 days daily. The third group – animals were infused intramuscularly with 0,1 ml of 0,9 % NaCl solution in the posterior right limb for 28 days and for 29 days inflammatory edema of the limb was stimulated (animals were sub-planar injected with 0,1 ml of 1% carrageenan solution to the posterior right limb ) The fourth group – for 28 days rats were daily intramuscularly injected with a therapeutic dose of 3 mg x kg-1 chondroitin sulfate, after which on 29th day, inflammatory edema of the limb was stimulated. The total number of animals involved in experimental studies was 40 individuals. Superoxide dismutase activity was assessed by the ability of the enzyme to compete with nitro blue tetrazolium for superoxide radicals. Catalase activity was measured by the amount of intact hydrogen peroxide in the sample. Protein content was measured by the Lowry method. It was revealed that under conditions of carrageenan-induced joint inflammation in the blood serum, the antioxidant defense system is disrupted: the activity of superoxide dismutase decreases by 1,5 times, while the catalase activity increases by 2,1 times compared to the control. Prophylactic administration of chondroitin sulfate to animals with carrageenan-induced joint inflammation contributes to the restoration of the antioxidant system.

ЭФФЕКТИВНОСТЬ ФОТОМАГНИТОТЕРАПИИ ПРИ ЭКСПЕРИМЕНТАЛЬНОМ АРТРИТЕ

Background. The choice of physiotherapeutic method in the treatment of joint diseases remains an urgent problem in physiotherapy. The aim: а comparative study of the efficiency of a low-frequency pulsed magnetic field (LPMF) and its combination with infrared light (photomagnetic therapy) in experimental arthritis. Material and methods. Studies were performed on 50 male rats. A zymosan model of arthritis was used. The animals were divided into 4 groups, the 1st group – intact animals, the 2nd – animals with arthritis without treatment, in the 3rd group they underwent course exposure to LPMF, in the 4th group LPMF in combination with infrared light. Clinical, laboratory, morphological and histochemical studies were performed. Results. It was revealed that photomagnetic therapy to a greater extent than a LPMF reduces the activity of the inflammatory process, and also leads to a partial normalization of the structure of cartilage tissue. Conclusions. The data obtained can be used to develop differentiated methods of physiotherapy in rheumatology.

ЭФФЕКТИВНОСТЬ ПРИМЕНЕНИЯ ФОТОМАГНИТОТЕРАПИИ ПРИ РЕВМАТОИДНОМ АРТРИТЕ

Background. The choice of physiotherapeutic method in the treatment of joint diseases remains an urgent problem in physiotherapy. Aim: to evaluate the efficiency of photomagnetic therapy in the treatment of patients with rheumatoid arthritis. Material and methods. The studies were performed on 72 patients with rheumatoid arthritis. We conducted a clinical and laboratory study, assessment of functional ability using HAQ questionnaire. Results. It was revealed that photomagnetic therapy helps to reduce pain, to reduce the duration of morning stiffness, to increase the strength of hand compression and to improve the functional ability of patients. Photomagnetic therapy has an anti-inflammatory effect and leads to a decrease in C-reactive protein level. Conclusions. Photomagnetic therapy is effective in patients with rheumatoid arthritis.

Virtual Screening Technique Used to Estimate the Mechanism of Adhatoda vasica Nees for the Treatment of Rheumatoid Arthritis Based on Network Pharmacology and Molecular Docking.

Adhatoda vasica Nees (AVN) is commonly used to treat joint diseases such as rheumatoid arthritis (RA) in ethnic minority areas of China, especially in Tibetan and Dai areas, and its molecular mechanisms on RA still remain unclear. Network pharmacology, a novel strategy, utilizes bioinformatics to predict and evaluate drug targets and interactions in disease. Here, network pharmacology was used to investigate the mechanism by which AVN acts in RA. The chemical compositions and functional targets of AVN were retrieved using the systematic pharmacological analysis platform PharmMapper. The targets of RA were queried through the DrugBank database. The protein-protein interaction network (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of key targets were constructed in the STRING database, and the network visualization analysis was performed in Cytoscape. Maestro 11.1, a type of professional software, was used for verifying prediction and analysis based on network pharmacology. By comparing the predicted target information with the targets of RA-related drugs, 25 potential targets may be related to the treatment of RA, among which MAPK1, TNF, DHODH, IL2, PTGS2, and JAK2 may be the main potential targets for the treatment of RA. Finally, the chemical components and potential target proteins were scored by molecular docking, and compared with the ligands of the protein, the prediction results of network pharmacology were preliminarily verified. The active ingredients and mechanism of AVN against RA were firstly investigated using network pharmacology. Additionally, this research provided a solid foundation for further experimental studies.

EPCR Deficiency Prevents Development of Hemophilic Arthropathy

Recent studies from our laboratory showed that clotting factor VIIa (FVIIa), whose primary function is to initiate the coagulation cascade following its binding to procoagulant cofactor tissue factor (TF), also binds anticoagulant cofactor, endothelial cell protein C receptor (EPCR). EPCR plays a key role in the activated protein C (APC)-mediated anticoagulant pathway by promoting the activation of protein C bound to it. Pharmacological concentrations of rFVIIa could compete with protein C for the EPCR and downregulate APC generation by preventing protein C binding to the EPCR. Our recent studies showed that FVIIa binding to EPCR also induces barrier-protective and anti-inflammatory effects both in vitro and in vivo model systems. Bio-distribution of rFVIIa in mice revealed that a small fraction of rFVIIa administed to mice i.v. accumulates in knee joints and retained there for three to seven days. The present study was conducted to investigate the relevance of FVIIa binding to EPCR in pharmacological FVIIa treatment of joint disease in hemophilia. For this, we first generated hemophilia A mice lacking EPCR or overexpressing EPCR. To generate EPCR deficient FVIII-/- mice, Procrflox/flox and Procr+/floxMeox2+/cre mice were first backcrossed with FVIII-/- mice to generate Procrflox/flox/FVIII-/- and Meox2+/cre/FVIII-/- mice. EPCR deficient FVIII-/- mice (Procr-/-/FVIII-/-) were generated by breeding female Procrflox/flox/FVIII-/- with male Procr+/floxMeox2+/cre /FVIII-/- mice. EPCR- overexpressing FVIII-/- were generated by crossing Tie2-EPCR mice with FVIII-/- mice. We induced joint bleeding in FVIII-/-, EPCR deficient FVIII-/-, and EPCR-overexpressing FVIII-/- mice by needle puncture injury. Right knee joint intra articular space was pierced with 30G needle to induce joint bleeding; left knee joint served as an uninjured control. Injured mice were treated with a single dose of rFVIIa (1 mg/kg) at 20 min following the injury or three doses of rFVIIa (1 mg/Kg) at 20 min, 24 h and 72 h following the injury. In controls, saline was administered in place of rFVIIa. The knee injury was evaluated by measuring knee joint diameter and visual bleeding score. At the end of 14 days post injury, mice were euthanized, and the knee joints were excised and fixed for immunohistochemical analysis. Hemophilic arthropathy was evaluated by scoring a set of pathological parameters (e.g., synovial hyperplasia, neoangiogenesis, presence of blood, iron score, villus formation, and cartilage degeneration). A subset of mice was used to collect synovial fluid at 7 days post injury. Assessment of joint bleeding at 5 h following the needle injury, by extracting and measuring hemoglobin levels in joint tissues and monitoring hematocrit, showed an equal amount of bleeding in all three genotypes of mice (FVIII-/-, EPCR deficient FVIII-/-, and EPCR-overexpressing FVIII-/- mice). Administration of three doses of FVIIa was effective in preventing hemophilic arthropathy in FVIII-/- and EPCR-overexpressing FVIII-/- mice, whereas a single dose of rFVIIa had a minimal effect in reducing hemophilic arthropathy. Interestingly, a single dose of FVIIa fully corrected the needle injury-induced hemophilic arthropathy in EPCR-deficient FVIII-/- mice. More importantly, EPCR-deficient FVIII-/- mice failed to develop full-blown hemophiic arthropathy even in the absence of any treatment. Measurement of inflammatory cytokines in the synovial fluid showed a robust increase in IL-6 levels in FVIII-/- mice and a single dose of FVIIa substantially reduced IL-6 levels. Interestingly, IL-6 levels in the synovial fluid of injured EPCR-overexpressing FVIII-/- mice were markedly higher compared to injured FVIII-/- mice, three doses of rFVIIa was required to attain a significant reduction in IL-6 levels in these mice. In contrast to these data, we found very little IL-6 levels in the synovial fluids of injured EPCR-deficient FVIII-/- mice. Overall, these data indicate that EPCR levels profoundly influence hemophilic arthorpathy. EPCR-deficiency protects from development hemophilic arthropathy because down-regulation of APC generation in EPCR deficiency could allow sufficient thrombin generation in hemophilia to prevent joint bleeding and bleeding-associated inflammation. Disclosures Rao: Takeda: Research Funding.

Anatomical and ultrasound navigation of intra joint injections

Optimization of joint syndrome treatment methods, including those based on the intra- and periarticular drug administration and invasive diagnostic techniques, remains high on the agenda of modern clinical rheumatology. The implementation and quite widely spread use of ultrasonographic visualization has been an impetus to the development of this type of treatment for joint diseases. Without any doubt, the quality of intraarticular injection performance mainly depends on the professional level of the specialist and his/hers procedural skills. However, here comes a predictable question: are these conditions sufficient to enable maximal precision, safety, and efficacy of intraarticular interventions? From this perspective, it is interesting to study the possibilities to improve the results of local treatments for the joint syndrome by means of the ultrasound navigation technique. Based on data presented in the literature review, we compared a “blind” invasive treatment method to the ultrasound navigation-guided intra- and periarticular interventions in patients with skeletomuscular and connective tissue disorders. The authors of the studies published point to higher safety, efficacy, procedure precision, and diagnostic quality of the information obtained by the ultrasound navigation. Its important advantages include wider possibilities and availability of this method in outpatient settings, due to its rather low costs and patients' safety. The information from the current literature review reflects an initial stage of studies on the evaluation of the role, significance, determination of potential of the ultrasound navigation to enhance the quality of diagnosis and invasive treatment in patients with joint syndromes of various origins and to minimize adverse effects.

Efficiency of Glucosamine Sulphate and Chondroitin Sulphate Сombination in Osteoarthritis Treatment from Evidence-Based Medicine Positions

Osteoarthritis remains by far the most common form of joint damage and the leading cause of disability worldwide. Unfortunately, today there are no radical methods that can stop the «epidemic» of OA and completely cure patients with this disease. In recent decades, the attention of scientists and clinicians has been shrouded in the possibility of using symptomatic or disease-modifying slow-acting drugs that are potentially not only able to influence the symptoms of OA (primarily pain), but also to have a lasting structural-modifying effect on the joint. The results of clinical studies, as well as a deeper understanding of the differences in the pharmacokinetics and properties of molecules of different chondroprotectors have aroused scientific and practical interest in approaches to the treatment of joint diseases using a combination of chondroitin sulfate and glucosamine sulfate.

Medical imaging and diagnosis of subpatellar vertebrae based on improved Laplacian image enhancement algorithm

ObjectiveNowadays, arthroscopy is widely applied to the treatment of joint diseases. The experimental trials were designed to determine whether the infrapatellar plica was symptomatic or not, and to appraise the medical effects of these patients who underwent arthroscopic treatment. An improved Laplacian image enhancement algorithm is added to the experiment. The medical image of the Subpatellar vertebral body under arthroscope is processed by the algorithm. The processed image is compared with the original image, and the advantages and disadvantages of the improved Laplacian image enhancement algorithm are analyzed. MethodsRetrospective Medical trial design was executed in our study. In addition, X-ray film and magnetic resonance imaging (MRI) were included in the study. Visual Analogue Scale (VAS) and Lysholm Score were carried out. Arthroscopy results, MRI findings, and Medical features were researched and analyzed carefully. Then we use the improved Laplacian image enhancement algorithm to process the image, which makes the image more convenient for analysis and improves the diagnostic accuracy. ResultsSome of the experimental protomedical images are not clear enough, and the details and textures are difficult to judge, which hinders the diagnosis. After the improved Laplacian algorithm processing, the image effect has been significantly improved. From the image we get the result, although the wound healed after surgery, some patients have existence of transient swelling in recovery process but no effusion. The pain of all patients knee was sharply relieved and the function was improved. All patients’ conditions were most satisfactory. ConclusionThe findings in this study demonstrate a significant reduction in knee pain and improvement in function by releasing and removal of the symptomatic infrapatellar plica under arthroscopic surgery. The image processed by the improved Laplacian image enhancement algorithm can effectively retain the image details, which is conducive to diagnosis and improve the diagnostic accuracy.