Subintimal Layer Research Articles

Evaluation of the C-reactive protein serum levels in periodontitis patients with or without atherosclerosis – A clinical study

Background: Periodontitis is a bacterial infection that can originate in the gums and progress to damage the bone and other structures supporting the teeth. One indicator of systemic inflammation caused by various triggers such as injuries, infections, and hypoxia is the highly potent C-reactive protein (CRP). Atherosclerosis is a progressive arterial disease that is characterized by the thickening of subintimal layers in muscular and elastic arteries of intermediate-to-large size due to the accumulation of athermanous plaque. Therefore, our objective in this research is to evaluate CRP levels in individuals with periodontitis, with or without atherosclerosis, after diagnosis. It is important to determine if there is a correlation between periodontitis, a common inflammatory condition of the gums, and elevated CRP levels, which could potentially contribute to the development of atherosclerosis. Methodology: The Department of Periodontics and Implantology at Bhabha College of Dental Sciences conducted the study, which involved the random assignment of 60 participants of both sexes, aged between 30 and 60 years, using a cross-sectional design. The participants were selected from the Department of Periodontics and Implantology, and venous blood samples were taken before and after nonsurgical periodontal treatment to evaluate serum CRP and complete blood count levels. Results: Higher blood CRP levels were seen in individuals with chronic periodontitis, regardless of the presence or absence of atherosclerosis. When comparing the groups of people with and without atherosclerosis and chronic periodontitis, the rise in blood CRP levels was almost double in the group with atherosclerosis. To prevent the progression of atherosclerotic lesions, chronic periodontitis must be treated aggressively. Conclusion: Chronic periodontitis, with or without atherosclerosis, leads to high CRP levels in the blood. Those with both conditions have almost twice the CRP levels compared to those without atherosclerosis. Proper treatment is necessary due to chronic periodontitis’ association with atherosclerotic lesions.

From histology to sonography in synovitis: EURO-MUSCULUS/USPRM approach.

The aim of the present study was to propose a methodologically innovative sonographic approach for optimal evaluation of synovial tissues (starting from histopathology). Using high-frequency ultrasound probes and high-level ultrasound machines, we matched the histological microarchitecture of synovial tissues with multiple sonographic patterns in physiological and pathological conditions. Likewise, high-sensitive color/power Doppler assessments have also been performed to evaluate the microcirculation. Modern equipment allows for a macroscopic classification of synovial pathologies recognizing different morphological patterns; however, intimal and subintimal layers of the synovium cannot be distinguished from each other on ultrasound. High-sensitive Doppler imaging clearly defines the microvascular pattern, especially in patients with hypertrophic synovial pathologies. In clinical practice, using adequate technological equipment i.e. high-frequency B-mode and high-sensitive Doppler imaging, detailed sonographic assessment of synovial tissues can be performed - defining the main sono-histological patterns.

The inflammation influence on pathological remodeling of pulmonary arteries in monocrotaline-induced pulmonary hypertension in rats

Abstract Aim To investigate the inflammation role on pathological remodeling of pulmonary arteries (PA) in monocrotaline-induced pulmonary hypertension (mPAH) in rats with joint assessment serum and tissue inflammatory biomarkers and the morphological arteries changes. Methods The mPAH was induced by a subcutaneous monocrotaline injection (60 mg/kg) in male rats and control group received a single saline solution. Baseline and every 2, 4, 6, 8 weeks after the serum concentrations of interleukin-6 (IL-6), interleukin-10 (IL-10) were measured by enzyme-linked immunosorbent assay; matrix metalloproteinase-9 (MMP-9), interleukin-1β (IL-1β), collagen type 1 and 3, smooth muscle actin α (SMA-α) in lung tissue were investigated immunohistochemically and quantitative measurements of intima and media thickness were done by planimetry. The functional activity neutrophil changes measured by chemiluminescence and fluorescent methods. Results The IL-10 increased after 2 weeks of mPAH (5,9 vs 0,6 pg/ml, p<0,05) vs control and then it decreased to initial values by 8 weeks (0,06 vs 0,62 pg/ml, p>0,05). The increasing IL-6 (30,3 vs 0,01 pg/ml, p<0,05) and the maximum expression of IL-1β in the lung tissue (0,119 vs 0,099 index of expression (IE), p<0,05) we observed 4 weeks after mPAH. The SMA-α (29,4 vs 40,2 IE, p<0,05) and MMP-9 (1,6 vs 0,8 IE, p<0,05) expression level significantly raised 4 weeks later vs control and remained in a high level until 8 week. A significant increase of type 1 collagen expression was observed at all phases of the experiment, and high level type 3 collagen expression was observed from 4 to 8 weeks (7,0 vs 10,4 IE, p<0,05). The histological characteristics of remodeling these were a thickening of the media (30,6 vs 56,0 μm, p<0,05) and the subintimal layer (1,6 vs 10,9 μm, p<0,05) of the PA. 2 weeks after mPAH cell priming occurred which manifested by modification ROS generation systems, decrease NADPH oxidase activity, increase of myeloperoxidase secretion (MPO), enhance of unbound cytosolic calcium ions, mitochondrial potential reduction. From 4 to 8 weeks an increase NADPH oxidase activity and MPO secretion was revealed into the extracellular environment which leads to overproduction of hypochlorous acid. This functional activity reprogramming of circulating neutrophils indicate associated with time-development of mPAH. Conclusion The inflammation is the most important mediator of pathological remodeling processes in mPA. The monocrotaline launches a neutrophil reaction at an early stage of PAH with changes their functional activity which leads to immune cells recruitment into the lung tissue, producing inflammation and proliferation biomarkers. The hyperplasia of smooth muscle cells and reconstruction of the extracellular matrix are the result of this process and leads to increase intima and media thickness. The high MMP-9, SMA-α, IL-6 activity in 6–8 weeks reflects maintenance local inflammatory potential. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Basic and applied sciences - medicine, subprogram “Diagnostics and therapy of diseases” on the assignment “To establish the molecular-cellular mechanisms of the development of irreversible remodeling of pulmonary vessels in pulmonary arterial hypertension in an experiment in vivo.”

Development of PBPK model for intra-articular injection in human: methotrexate solution and rheumatoid arthritis case study

A physiologically based model describing the dissolution, diffusion, and transfer of drug from the intra-articular (IA) space to the plasma, was developed for GastroPlus® v9.8. The model is subdivided into compartments representing the synovial fluid, synovium, and cartilage. The synovium is broken up into two sublayers. The intimal layer acts as a diffusion barrier between the synovial fluid and the subintimal layer. The subintimal layer of the synovium has fenestrated capillaries that allow the free drug to be transported into systemic circulation. The articular cartilage is broken up into 10 diffusion sublayers as it is much thicker than the synovium. The cartilage acts as a depot tissue for the drug to diffuse into from synovial fluid. At later times, the drug will diffuse from the cartilage back into synovial fluid once a portion of the dose enters systemic circulation. In this study, a listing of all relevant details and equations for the model is presented. Methotrexate was chosen as a case study to show the application and utility of the model, based on the availability of intravenous (IV), oral (PO) and IA administration data in patients presenting rheumatoid arthritis (RA) symptoms. Systemic disposition of methotrexate in RA patients was described by compartmental pharmacokinetic (PK) model with PK parameters extracted using the PKPlus™ module in GastroPlus®. The systemic PK parameters were validated by simulating PO administration of methotrexate before being used for simulation of IA administration. For methotrexate, the concentrations of drug in the synovial fluid and plasma were well described after adjustments of physiological parameters to account for RA disease state, and with certain assumptions about binding and diffusion. The results indicate that the model can correctly describe PK profiles resulting from administration in the IA space, however, additional cases studies will be required to evaluate ability of the model to scale between species and/or doses.

Loss of Nfat5 promotes lipid accumulation in vascular smooth muscle cells.

The nuclear factor of activated T-cells 5 (NFAT5) is a transcriptional regulator of macrophage activation and T-cell development, which controls stabilizing responses of cells to hypertonic and biomechanical stress. In this study, we detected NFAT5 in the media layer of arteries adjacent to human arteriosclerotic plaques and analyzed its role in vascular smooth muscle cells (VSMCs) known to contribute to arteriosclerosis through the uptake of lipids and transformation into foam cells. Exposure of both human and mouse VSMCs to cholesterol stimulated the nuclear translocation of NFAT5 and increased the expression of the ATP-binding cassette transporter Abca1, required to regulate cholesterol efflux from cells. Loss of Nfat5 promoted cholesterol accumulation in these cells and inhibited the expression of genes involved in the management of oxidative stress or lipid handling, such as Sod1, Plin2, Fabp3, and Ppard. The functional relevance of these observations was subsequently investigated in mice fed a high-fat diet upon induction of a smooth muscle cell-specific genetic ablation of Nfat5 (Nfat5(SMC)-/- ). Under these conditions, Nfat5(SMC)-/- but not Nfat5fl/fl mice developed small, focal lipid-rich lesions in the aorta after 14 and 25weeks, which were formed by intracellular lipid droplets deposited in the sub-intimal VSMCs layer. While known for being activated by external stimuli, NFAT5 was found to mediate the expression of VSMC genes associated with the handling of lipids in response to a cholesterol-rich environment. Failure of this protective function may promote the formation of lipid-laden arterial VSMCs and pro-atherogenic vascular responses.

Evaluation of Villus Synovium From Unaffected Metacarpophalangeal Joints of Adult and Juvenile Horses

Horses are a widely accepted model for osteoarthritis (OA) research. Synovial tissue sampling is commonly used in studies to evaluate and grade the progress of OA or to assess treatment effects. Synovial explants play an important role in ex-vivo studies, increasingly replacing the use of living animals. To understand histomorphological changes in the process of joint-related diseases such as OA, detailed information about histomorphometric parameters of unaffected synovial villi is necessary.The objective of the present study was to evaluate the mean width of the intimal synovial lining and its cellularity as well as the vascularization of the subintimal layer in juvenile and adult horses not affected by a joint-related disease.One hundred synovial samples from both metacarpophalangeal joints from 25 horses (one day to 24 years old) were collected to evaluate the following parameters on digitalized hematoxylin-eosin stained samples: Width of intimal synovial lining measured by the distance from the inner joint surface to the subintimal layer; density of the cells making up the intimal synovial lining by counting cell nuclei; vascularization of the subintimal layer measured by the number and size of vessels in relation to the subintimal area.The median width of the intimal lining did not differ among juvenile (22.34 µm) and adult (23.34 µm) horses. The cellularity of the intimal lining was significantly lower in juvenile (one cell/143.8 µm2) than in adult (one cell /188.7µm2), (P < .001) horses as well as the density of blood vessels per mm2 within the subintimal layer (juveniles 1/mm2 vs. adults 0.05/mm2), (P < .001).This study provides morphometric data regarding synovial intimal width, intimal cellularity, and vascularization of equine synovial villi of unaffected horses. For future studies, age-related characteristics should be taken into consideration when synovial tissue samples are used for in-vivo and in-vitro studies.

Small caliber heparin loaded ultrafine microfiber woven graft achieved high patency rate in a preliminary study of canine carotid artery implantation

ObjectiveIn the past five decades, many small caliber vascular grafts have been developed as bypasses for infrapopliteal or coronary arteries. However, reliable grafts have not been obtained owing to poor patency, mainly caused by early thrombosis or neointimal hyperplasia in the intermediate period after implantation. We developed a novel small caliber heparin-loaded polyethylene terephthalate ultrafine microfiber (HL-PET) graft and evaluated the feasibility to overcome those main causes of graft failure in canine carotid artery implantation. MethodsThe HL-PET graft with a diameter of 3 mm and length of 30 mm was made with combination of three key technologies: (1) weaving with PET ultrafine microfiber with a high biological porosity allowing for cell ingrowth, (2) heparin loading on microfiber surfaces, and (3) an outer coating with a flexible bioabsorbable polymer for prevention of blood leakage and graft kinking. Kink resistance, water permeability, and loaded heparin were assessed. One HL-PET graft each was implanted into a carotid artery of six animals. Graft patency rate and healing were assessed 24 weeks after implantation. ResultsAmong the six grafts, five were deemed patent (patency rate of >83%), with one occluded 20 weeks after implantation. Histopathology of the patent grafts showed neointima formation with confluent endothelial cell lining (estimated mean endothelial cell coverage area, 89 ± 18%). Intimal hyperplasia at the anastomotic sites and severe chronic inflammatory responses were not observed. Immunohistochemistry with antibodies to endothelial nitric oxide synthase, alpha 2 smooth muscle actin and calponin 1 revealed luminal surface endothelial cell layer with expression of endothelial nitric oxide synthase and vascular smooth muscle cells with contractile phenotype in the subintimal layer. ConclusionsThe HL-PET graft showed no early postoperative thrombosis and was able to demonstrate a high patency rate with no severe biological response observed after 24 weeks. These results strongly suggest the potential of the HL-PET graft to be used for distal bypasses.

Dysphagia in a Patient With End-Stage Renal Disease

Question: A 65-year-old man with end-stage renal disease presented to the emergency department complaining of painful skin ulcers on his upper and lower extremities. The patient had been receiving peritoneal dialysis for the last 2 years, and he said that skin lesions developed 3 months prior. On further questioning, the patient described significant weight loss and 3 months of dysphagia to solids and liquids. On examination, the patient was cachectic and ill appearing with distal necrotic lesions of his upper and lower extremities with overlying eschars. His admission laboratory tests were significant for calcium of 6.8 mg/dL, phosphate of 12 mg/dL with a calcium-phosphorus product of 81.6 mg2/dL,2 and alkaline phosphatase of 326 μ/L. Esophagogastroduodenoscopy showed significant esophagitis with ulcerations throughout the length of the esophagus (Figure A, B). Biopsies taken during the esophagogastroduodenoscopy demonstrated prominent parakeratosis of the esophageal mucosa with calcium deposits in the subintimal layer (Figure C). What is the diagnosis? See the Gastroenterology web site (www.gastrojournal.org) for more information on submitting your favorite image to Clinical Challenges and Images in GI. The patient was given the diagnosis of calciphylaxis with associated esophageal calcinosis. Calciphylaxis is highly morbid disease process most commonly described in patients with end-stage renal disease. Although the pathogenesis is not well-understood, the literature demonstrates involvement of the mesentery, intestines, brain, eyes skeletal muscle, lung, and recently the esophagus.1Huber A.R. Sprung Jr., B.S. JM Findeis-Hosey J.J. Esophageal mucosal calcinosis: a rare site of gastrointestinal mucosal calcinosis.Am J Case Rep. 2018; 19: 406-409Crossref PubMed Scopus (3) Google Scholar Esophageal calcinosis is extremely rare, with only 4 previously reported cases. In those reports, patients had end-stage renal disease and concomitant calciphylaxis. These patients typically present with dysphagia and less commonly with gastrointestinal bleeding. Esophagogastroduodenoscopy windings in each case were consistent, demonstrating friable mucosa with areas of erosion and ulceration.2Garber A. Arora Z. Welch N. et al.Extraosseous calcification of the esophagus: clinicopathologic correlates of esophageal mucosal calcinosis.ACG Case Rep J. 2017; 4PubMed Google Scholar, 3Machavarapu A. Brown T.A. Nwakoby I.E. Rare case of hematemesis: calciphylaxis of the esophagus.Clin Gastroenterol Hepatol. 2018; 16: A35Abstract Full Text Full Text PDF PubMed Scopus (2) Google Scholar, 4Nigwekar S. Kroshinsky D. Nazarian R.M. et al.Calciphylaxis: risk factors, diagnosis, and treatment.Am J Kidney Dis. 2015; 66: 133-146Abstract Full Text Full Text PDF PubMed Scopus (255) Google Scholar Calciphlyaxis remains a highly morbid disease and treatment options are limited. Our patient was converted to hemodialysis, started on sodium thiosulfate thrice weekly, and started on proton pump inhibitors. The patient reported marked improvement in symptoms of dysphagia and improved oral intake, with eventual signs of weight gain after correcting his hyperphosphatemia, which corrected his calcium-phosphorus product to 35 mg2/dL.2

Coronary Artery Bypass Graft Surgery (CABG)

Coronary artery bypass grafting (CABG) is one of the procedure done worldwide with acceptable results and has the highest impact in the history of medicine. Atherosclerotic Plaque formation in the sub-intimal layer is the main pathophysiology which causes ischemia in cardiac muscle &amp; gives symptoms of coronary artery disease (CAD). There are many ways for revascularization but CABG is the mostly performed procedure &amp; still gold standard. Results of percutaneous coronary interventions (PCI) &amp; other novel approach to coronary revascularization is still compared with conventional CABG. Left internal mammary artery is the most durable conduits &amp; should be used for every patients unless contraindicated. In young non-diabetic patient as much possible arterial conduits should be used. In planned operations results are excellent with inhospital mortality &lt;1% with few morbidities like sternal wound infection &lt;3%, renal and neurological complications &lt;7% &amp; &lt;3%.KYAMC Journal Vol. 7, No.-2, Jan 2017, Page 795-803

Endothelial dysfunction mediated by interleukin-18 in patients with ischemic heart disease undergoing coronary artery bypass grafting surgery

When medication management or percutaneous coronary intervention is not successful in patients with advanced ischemic heart disease, surgical revascularisation—predominantly coronary artery bypass grafting (CABG)—is considered the gold standard. However, CABG surgery can lead to ischemia/reperfusion injury, which is characterized by a strong inflammatory response. Interleukin (IL)-18, is a strong inflammatory mediator, that is released from cardiomyocytes and can be found in the systemic circulation of patients during and immediately after CABG surgery. The existing damage of endothelial glycocalyx in patients with ischemic heart disease is further impaired concurrently during the surgery due to the anaesthesia-surgical technique used and intravascular fluid loading. This results in the increased incidence of adverse events, including myocardial infarction. IL-18 leads to the activation of lymphocyte cytotoxicity via cytotoxic mediators (Fas ligand, Tumour necrosis factor (TNF)-related apoptosis-inducing ligand, perforin, and granulysin). We hypothesize that IL-18 is released locally in the heart and the systemic circulation in patients undergoing CABG surgery and may be correlated with the level of activity of circulating lymphocytes. In turn, this may lead to lymphocyte-mediated cytotoxicity directed toward damaged and activated endothelial cells. Shear stress glycocalyx, as well as damaged and activated endothelial cells then become the main the source of pro-inflammatory cytokines, chemokines, and adhesion molecules. These attract activated lymphocytes to adhere to the endothelium or enter the subintimal layer, increasing existing or initiating the formation of new plaques, which leads to the development of myocardial infarction during or shortly after surgery. To evaluate our hypothesis, we will measure the local concentration of IL-18 in the sinus coronarius and systemic circulation. These values will then be correlated with immunological and biochemical parameters, predominantly with the concentration of degradation products of glycocalyx and cytotoxic mediators in activated lymphocytes. If our hypothesis is correct, measuring the IL-18 concentration that is responsible for glycocalyx deterioration, may become a useful tool for predicting myocardial infarction occurrence in patients undergoing CABG surgery.

Evaluation of sodium tetradecyl sulfate and polidocanol as sclerosants for leg telangiectasia based on histological evaluation with clinical correlation.

Introduction This study was designed to determine by histological evaluation and clinical correlation the most effective sclerosant concentration of Sotradecol® (sodium tetradecyl sulfate) and Asclera® (polidocanol) for the treatment of leg telangiectasia. Methods Histological studies were completed on 40 patients, all of whom were female with a mean age of 53. After sclerotherapy with varying concentrations of sclerosant solutions for the treatment of 0.8 mm and 1 mm leg telangiectasia, histological specimens were examined for the following criteria: luminal changes, subintimal changes, smooth muscle wall alterations, and vessel wall integrity. Six patients from this group were also treated with sodium tetradecyl sulfate foam 0.1%-0.2% or polidocanol foam 0.31% for the treatment of 2 mm reticular veins. In a second group of 20 patients, clinical results after treatment with sodium tetradecyl sulfate 0.15% and polidocanol 0.31% were evaluated. Results Histological findings in patients treated with sodium tetradecyl sulfate 0.05% and polidocanol 0.25% were minimal. Sclerosant concentrations of sodium tetradecyl sulfate ≥0.2% revealed intraluminal debris and greater smooth wall damage. Polidocanol 0.5% was equivalent in strength to sodium tetradecyl sulfate 0.2% on histological evaluation. Sodium tetradecyl sulfate 0.15% and polidocanol 0.31% had similar findings on histological examination with mild smooth muscle wall changes, endothelial cell lysis, and subintimal layer damage. Reticular veins treated with 0.1%, 0.15%, and 0.2% sodium tetradecyl sulfate foam and 0.31% polidocanol foam produced identical histological findings. Conclusion Sodium tetradecyl sulfate 0.15% and polidocanol 0.31% based on histological evaluations and clinical correlation are the best sclerosant concentrations for 0.8 mm to 1 mm leg telangiectasia. Sodium tetradecyl sulfate foam is comparable to polidocanol foam at these concentrations as well.

Distribution of Neuropeptides in the Synovium of Charcot Neuroarthropathy

Category: Basic Sciences/Biologics Introduction/Purpose: The pathology of Charcot neuroarthropathy (CNA) presents inflammation, and bone and cartilage destruction. It is still unclear how the neuropathic signals lead to joint inflammation and destruction. Neuropeptides are secreted by neurons and involve in the regulation of inflammation. Synovium is richly innervated and known to play a critical role in the pathology of inflammatory arthritis. This study is designed to investigate the distribution of several neuropeptides, such as vasoactive intestinal peptide (VIP), substance P (SP) and calcitonin gene-related peptide (CGRP), in CNA synovium, in comparison with non-CNA synovium, with immunohistochemistry. Methods: Synovial samples were collected from CNA patients (n=3) and patients with osteoarthritis and intraarticular fracture (non-CNA; n=3), during foot and ankle surgery (approved by IRB). The tissue samples were fixed with 4% paraformaldehyde, embedded in O.T.C media and sectioned with a cryostat. From each patient, 6-9 sections were randomly selected for immunohistochemistry. After blocking the tissue sections with normal serum, primary antibody of SP, VIP and CGRP was separately applied and incubated at 4ºC overnight. The biotinylated secondary antibody and ABC kit (Victor Laboratories) were applied subsequently. DAB (3,3’-diaminobenzidine) was used for colorimetric detection (brown) of immunohistochemical reactions. The cell nuclei were stained with hematoxylin. The staining was viewed under a microscope and images were taken with a digital camera. Results: In the non-CNA synovium, SP was stained around neuro-vascular bundles and intensely stained on the surface of the synovium, i.e. the intimal layer. In the CNA synovium, SP staining was increased and more diffuse into the subintimal layer of the synovium (Fig 1). In non-CNA synovium, VIP was detected along the surface area of the synovium but not limited in the intimal layer. In CNA synovium, VIP was a similar staining pattern with a reduced staining intensity. The CGRP staining was primarily limited in the intimal layer in the non-CNA synovium. In CNA synovium, CGRP was stained in the intimal layer with an increased intensity. Conclusion: SP, VIP and CGRP are well-studied neuropeptides and have broad biological and pathological functions. All of them were detected in the intimal layer, where fibroblast-like synoviocytes are located. Fibroblast-like synoviocytes are fundamental elements of joint inflammation and are capable of degradation of bone and cartilage directly and indirectly. The changed patterns of SP, VIP and CGRP distributions in CNA synovium revealed by immunohistochemistry indicate a possible pathological pathway in CNA development: the unbalanced neuropathic signals direct fibroblast-like synoviocytes to an inflammatory state, which trigger the cascade of bone and cartilage destruction in CNA.

Differential expression of interleukin-17 and interleukin-22 in inflamed and non-inflamed synovium from osteoarthritis patients

Synovitis associated with osteoarthritis (OA) is directly responsible for several clinical symptoms and reflects OA's structural progression. This study sought to analyze the expression of proinflammatory mediations, including Interleukin (IL)-17 and IL-22, which play key roles in regulating inflammatory processes, in inflamed and non-inflamed areas of osteoarthritic synovium. Synovium from knees of 32 OA patients were collected at surgery. Macroscopic evaluation of inflammation enabled inflamed and non-inflamed areas to be separated. Samples were incubated to obtain tissue-conditioned media. Quantitative mRNA expression of proinflammatory mediators was analyzed by RT-PCR and protein levels by ELISA and gelatin zymography. Immunohistochemistry and histology were performed. Inflamed synovium were characterized by increased leukocyte infiltration and a higher vessel-to-tissue area ratio than non-inflamed tissues. Macrophages, T and B lymphocytes, and some neutrophils were found only in the inflamed tissue, and only in the subintimal layer. Levels of proinflammatory cytokines and MMP-9 were significantly higher in tissue-conditioned media from inflamed than non-inflamed tissues. Inflamed areas were associated with higher expression of IL-17 and IL-22, both correlated with the combined release of IL-6, IL-23, and TGFβ1. Our results showed that inflammatory cytokines, including IL-17 and IL-22, are expressed at higher levels by inflamed OA synovium and suggest IL-22 involvement in OA pathophysiology. This study will help identify new therapeutic strategies for OA, especially the targeting of IL-22 to decrease inflammation.

Expression of interleukin-17 and interleukin-22 in non inflammatory and inflammatory synovial membranes from osteoarthritis patients

s / Osteoarthritis and Cartilage 22 (2014) S57–S489 S452 MMP1 and ADAMTS4 in FLSs at the gene expression level. Importantly, we also demonstrate that insulin blunts release of MMP1 and MMP13 protein into the culture media, counteracting the effect of TNFa and IL-1b. Thus, insulin appears to play a selective protective role in the diarthrodial joint by suppressing release of catabolic enzymes into the synovial fluid via its effects on FLSs. Further study is needed to determine if insulin resistance in obesity/diabetes impairs this critical role for insulin in protecting cartilage matrix in synovial joints. Although the mechanism of selective suppression of MMP1 and MMP13 by insulin is still unknown, insulin and insulinsensitizing agents could be new disease-modifying interventions in OA treatment. 822 EXPRESSION OF INTERLEUKIN-17 AND INTERLEUKIN-22 IN NON INFLAMMATORY AND INFLAMMATORY SYNOVIAL MEMBRANES FROM OSTEOARTHRITIS PATIENTS C. Deligne y,z, S. Casulli x,z, A. Pigenet y,z, C. Bougault y,z, L. Campillo-Gimenez x,z, F. Berenbaum k,y, C. Elbim x,z, X. Houard y,z. y INSERM UMRS938, Paris, France; zUPMC Univ Paris 06, Paris, France; x INSERM UMRS945, Paris, France; kDept. of Rheumatology, AP-HP SaintAntoine Hosp., Labex Transimmunom, Inflammation-ImmunopathologyBiotherapy Dept. (i2B), Paris, France Purpose: IL-17 and IL-22 are inflammatory cytokines classically involved in chronic inflammation in various diseases. Here, we investigated the expression and the release of IL-17 and IL-22 by OA synovial membranes, in relation to the inflammatory status of synovium. Methods: Synovial membranes from OA knee patients (n 1⁄4 32) were collected at surgery and inflammatory (Infl) and non-inflammatory (NI) areas were separated according to the macroscopic evaluation of inflammation that appeared as hypervascularized areas with hypertrophic and hyperaemic villi. A piece of each tissuewas frozen for mRNA extraction, fixed in paraformaldehyde for histology and incubated in serum free culture medium in order to obtain conditioned media containing soluble factors released by synovial tissues. IL-1b, IL-6, IL-8, IL18, IL-17, IL-22, IL-23, TNF-a, TGF-b1, myeloperoxidase (MPO) andMMP9were analyzed for mRNA expression by quantitative RT-PCR and/or for protein level by ELISA and gelatin zymography. Immunohistochemistry for endothelial cells (CD31) and leukocytes (CD45) among them macrophages (CD68), neutrophils (CD15) and Tand B-lymphocytes (CD3 and CD20, respectively) was performed. Results: Inflammatory areas of OA synovial membranes were characterized by increased CD45þ inflammatory cell infiltration and vessel area/tissue area as compared to non-inflammatory areas (p1⁄4 0.001 and p 1⁄4 0.009, respectively). Macrophages were present in the intimal layer of both NI and Infl OA synovial membranes with higher accumulation in Infl as compared to NI. Only the subintimal layer of Infl areas of OA synovial membranes contained macrophages, Tand B-lymphocytes and some neutrophils. Consistently, the inflammatory markers MMP-9 and MPO were released in significantly higher concentrations by Infl than by NI areas (p 1⁄4 0.026 and p 1⁄4 0.001, respectively). IL-17 and IL-22 were both expressed and released by OA synovial membranes. A stronger mRNA expression was found in Infl for both IL-17 and IL-22. Infl also released significantly higher levels of IL-22 than NI (p 1⁄4 0.046). Strong positive correlations were found between IL-17 and IL-22 at mRNA and protein levels. The expression of IL-17 and IL-22 is controlled by a subset of cytokines, including IL-1b, IL-6, IL-8, IL-18, IL-23, TGFb-1 and TNF-a. With the exception of TNF-a, all were released in significantly higher concentration by Infl as compared to NI areas. IL-12 was not detected in conditioned media of synovial tissues. Conclusions: Our results show an increased infiltration of lymphocytes as well as an increased release of inflammatory cytokines, including ILSynovial gene expression at day 1 after transection (as fold-change compared to healt

Utility of arthroscopic guided synovial biopsy in understanding synovial tissue pathology in health and disease states.

The synovium is the soft tissue lining diarthrodial joints, tendon sheaths and bursae and is composed of intimal and subintimal layers. The intimal layer is composed of type A cells (macrophages) and type B cells (fibroblasts); in health, the subintima has few inflammatory cells. The synovium performs several homeostatic functions and is the primary target in several inflammatory arthritides. Inflammatory states are characterised by thickening of the synovial lining, macrophage recruitment and fibroblast proliferation, and an influx of inflammatory cells including lymphocytes, monocytes and plasma cells. Of the various methods employed to perform synovial biopsies arthroscopic techniques are considered the "gold standard", and have an established safety record. Synovial biopsy has been of critical importance in understanding disease pathogenesis and has provided insight into mechanisms of action of targeted therapies by way of direct evidence about events in the synovial tissue in various arthritides. It has been very useful as a research tool for proof of concept studies to assess efficacy and mechanisms of new therapies, provide tissue for in vitro studies, proteomics and microarrays and allow evaluation for biomarkers that may help predict response to therapy and identify new targets for drug development. It also has diagnostic value in the evaluation of neoplastic or granulomatous disease or infection when synovial fluid analysis is non-contributory.

Expression of PRDM10 in arthritic synovial derived tissues

We previously reported the expression of PRDM10 in skeletal cartilage during mouse embryonic development. In this study, we examined PRDM10 expression in blood, synovial fluid and synovium from patients with rheumatoid arthritis (RA) or osteoarthritis (OA). PRDM10 mRNA was detected in all peripheral blood and RA synovial fluid, as well as in normal blood. We also performed immunohistochemistry with human synovium tissues. Compared to other PR/SET proteins including PRDM14, PRDM12, and SUV39H1, PRDM10 expression was conspicuous in cells of the synovial lining layer, subintimal layer, and perivascular layer in RA and OA synovium. Immunostaining using cultured fibroblast like synoviocytes detected PRDM10 mainly in the nucleus, with a small amount detected in the cytoplasm. Transforming growth factor-beta (TGF-β) treatment decreased PRDM10 expression in OA-derived FLS cells, whereas polyinosinic:polycytidylic acid or lipopolysaccharide treatment did not produce a distinct change of PRDM10 expression, PRDM10 expression was reversed by the smad inhibitor SB431542. Given that PRDM10 is highly expressed in human synovial tissues and is transcriptionally regulated by TGF-β, we suggest that PRDM10 might be involved in the pathogenesis of arthritis.

SAT0051 Vitamin D Levels and Inflammation in the Aortic Wall of Patients with Inflammatory Rheumatic Disease and Coronary Artery Disease

BackgroundVitamin D is involved in immune reactions, and vitamin D deficiency is associated with autoimmune diseases and with cardiovascular diseases (CVDs). In Feiring Heart Biopsy Study (FHBS), we previously demonstrated...

Isolated hypoglossal nerve palsy due to spontaneous carotid artery dissection: a neuroimaging study

Internal carotid artery dissection (ICAD) is a common cause of ischemic stroke in young patients. It usually presents with neck pain or headache, ipsilateral Horner Syndrome, followed by signs of cerebral ischemia. Palsy of XIIth nerve, alone or in combination with involvement of nerves IX, X or XI, may occur in about 5 % of patients with extracranial ICAD [1, 2]. Two mechanisms have been proposed to explain lower cranial nerves (LCN) involvement in the setting of ICAD, i.e., compression or stretching of nerves below the jugular foramen by an expanded or aneurysmatic ICA, or impairment (mechanical, embolic or hemodynamic) of the blood supply to the LCN [1]. Herein, we report a patient with ICAD presenting XIIth nerve palsy, in whom the MRI revealed compression of such nerve in parapharyngeal space. A previously healthy 37-year-old man presented to the emergency room with 6-day-history of left mandibular angle pain, followed by dysarthria 24 h after onset. No major neck trauma was reported. Neurological examination only revealed left deviation of protruded tongue (Fig. 1). Brain CT angiography scan revealed mild narrowing of the artery vessel with surrounding hematoma. MRI and time-of-flight magnetic resonance angiography (TOF-MRA) showed left ICAD with intramural hematoma surrounding the artery with a semilunar shape with mild narrowing of the arterial lumen; ICAD clearly expanded into parapharyngeal space (Fig. 2). MRI including the diffusion-weighted imaging (DWI) showed no brain or brainstem lesion. Patient was then given anticoagulant treatment with partial regression of symptoms at 1-month follow-up. We presented a young man with isolated XIIth nerve palsy. Isolated hypoglossal palsy may recognize different etiologies including base skull tumor, infections, injury, and carotid artery surgery, but often no etiology may be found [3]. In our patient, we considered that the isolated XIIth nerve palsy was due to the compression by ICAD in the parapharyngeal space. In the same patient, the supposed role of the reduction of blood supply to XII nerve [1] remains questionable. We also showed that both MRI and TOF-MRA are useful to visualize intramural hematoma with surrounding tissues, clarifying the pathogenesis of nerve palsy. The absence of brain ischemic lesion in our patient could probably be explained by the fact that hematoma was likely located in subadventitial layer, without significantly narrowing the lumen [3]. On the contrary, when hematoma locates in subintimal layer, with significant lumen narrowing or occlusion, embolism may easily lead to ischemic brain lesions [3]. Our report highlights the importance of looking for ICAD and to perform E. Ferlazzo S. Gasparini U. Aguglia Regional Epilepsy Centre and Department of Medical and Surgical Sciences, Magna Graecia University, Catanzaro, Italy

The Effect of Chlorpyrifos on Isolated Thoracic Aorta in Rats

This study investigated the effect of chlorpyrifos on thoracic aorta and on the level of NO in plasma and aorta. The effect of chlorpyrifos on thoracic aorta in organ bath was determined in 10 rats. Another 45 rats were assigned to 3 groups with 15 rats each: control group 1 received distilled water, control group 2 was given corn oil, and the last group was given 13.5 mg/kg chlorpyrifos dissolved in corn oil every other day for 8 weeks orally. Chlorpyrifos (10−10 M–10−5 M) showed no effect on isolated thoracic aorta. Plasma AChE activity was decreased, while LDH, ALT, GGT, and AST activities were increased in chlorpyrifos group compared to control groups. Plasma NO level was increased in chlorpyrifos group compared to control groups. iNOS expression was present in all groups in the cytoplasm of the endothelia and in the smooth muscle cells of aorta. According to semiquantitative histomorphological analysis, iNOS immunopositive reactions were seen in the decreasing order in chlorpyrifos, control 2, and control 1 groups. eNOS immunopositive reactions were observed in the endothelial cell cytoplasm, rarely in the subintimal layer, and the smooth muscle cells of aorta. There were no differences among the groups in terms of eNOS immunostaining. In conclusion, chlorpyrifos induced NO production in aorta following an increase in NOS expression.

H&amp;E 염색 이미지의 포토샵 분석을 이용한 골관절염과 류마티스 관절염 활막 세포의 정량 분석

활막조직은 관절부위에 존재하는 비연골성의 얇은 세포층으로 구성되어 있으며, 류마티스 관절염 등에서 활성화 되어진다. 우리는 골관절염(n=8)과 류마티스 관절염(n=5)에서 유래한 활막조직을 대상으로 활막조직내의 세포를 정량화하고 세포성분들을 비교 분석하고자 하였다. 활막조직을 H&E 염색한 후 광학현미경으로 관찰하였을 때 류마티스 관절염의 활막조직은 골관절염에 비해 형태적으로 두터워졌으며 비후된 양상이었다. 또한 CD68, CD90, PGP9.5 등과 마커들을 이용하여 IHC 분석을 수행한 결과 활막조직의 내막층과 내막하층에 존재하는 세포들을 특성을 분한 결과 내막층에는 대식세포가 집중적으로 분포하며, 내막하층에는 대식세포와 섬유아세포 유사 활막세포(FLS)가 존재한다는 사실을 알 수 있었다. H&E 이미지를 포토샵 프로그램을 이용하여 반전시켜 내막층과 내막하층 부위별로 세포계수 및 세포층계수를 수행하였다. 내막층 분석 결과 류마티스 관절염의 활막조직의 골괄절염보다 대식세포의 수와 층이 현저하게 증가된 것을 확인할수 있었다. 또한 류마티스 관절염의 활막조직의 내막하층분석결과 섬유아세포 유사 활막세포의 수적인 증가를 계수 할 수 있었다. 또한 류마티스 관절염의 경우 활막의 비후가 심하기 때문에 혈관의 위치가 내막층으로부터 상대적으로 멀리 위치하고 있음을 알 수 있었다. 그러므로 H&E 염색 이미지의 포토샵 분석을 이용한 골관절염과 류마티스 관절염 활막조직에 대한 정량 분석 방법은 류마티스 관절염의 발병과정에서 세포들이 활성화된다는 사실을 증명하는데 유용할 것으로 사료된다. Synovium is the soft tissue that lines the non-cartilaginous surfaces within joints. It has been reported that synovial cells are activated during the pathogenesis of rheumatoid arthritis. In this study, we quantitate and compare the cellular composition of synovia derived from individuals with non-inflammatory osteoarthritis (OA) and those with inflammatory rheumatoid arthritis (RA). Synovia from OA (n=8) and RA (n=5) patients were used for hematoxylin and eosin (H&E) staining. A light microscopic examination has shown that RA synovia were morphologically thickened and hypertrophied as compared to OA synovia. We also performed an immunohistochemistry (IHC) analysis to classify cell types in the synovia using CD68, CD90, or PGP9.5 markers. As a result, we obtained quantitative data regarding the cell populations, which are macrophages in the lining layer and FLSs in the subintimal layer of the synovium. Further Photoshop analyses of the H&E images could allow the counting of the number and layer of the cells in the synovium. The number and layers of the macrophage cells were increased in the lining layer of the RA synovia as compared to the OA synovia. FLS cells also were increased in the subintimal layer of RA synovia. Therefore, quantification of the H&E stained images via Photoshop is a possible analysis protocol for synovium study. This quantitation also supports the idea that the increases in cell number and cell activation are important processes for RA pathogenesis.