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International audience; BackgroundNeuromuscular blocking agents (NMBAs) are among the leading cause of perioperative anaphylaxis, and most of these reactions are IgE mediated. Allergic sensitisation induced by environmental exposure to other quaternary ammonium-containing compounds, such as pholcodine, has been suggested. The aim of this study was to assess the relationship between pholcodine exposure and NMBA-related anaphylaxis.MethodsALPHO was a multicentre case-control study, comparing pholcodine exposure within a year before anaesthesia between patients with NMBA-related perioperative anaphylaxis (cases) and control patients with uneventful anaesthesia in France. Each case was matched to two controls by age, sex, type of NMBA, geographic area, and season. Pholcodine exposure was assessed by a self-administered questionnaire and pharmaceutical history retrieved from pharmacy records. The diagnostic values of anti-pholcodine and anti-quaternary ammonium specific IgE (sIgE) were also evaluated.ResultsOverall, 167 cases were matched with 334 controls. NMBA-related anaphylaxis was significantly associated with pholcodine consumption (odds ratio 4.2; 95% confidence interval 2.3–7.0) and occupational exposure to quaternary ammonium compounds (odds ratio 6.1; 95% confidence interval 2.7–13.6), suggesting that apart from pholcodine, other environmental factors can also lead to sensitisation to NMBAs. Pholcodine and quaternary ammonium sIgEs had a high negative predictive value (99.9%) but a very low positive predictive value (

International audience; Urothelium is a highly specialized epithelium covering the entire urinary excretory system. Tissue engineering of this urinary tract may allow to consider its reconstruction to perform in vitro studies or in vivo replacement. Therefore, the question of specific reconstruction of the urothelium arises in order to guarantee the neotissue’s ability to act as a barrier against highly cytotoxic urine. This literature review describes the different cell types and strategies available for this reconstruction. The non-reconstruction of urothelium relies on the colonization of a biomaterial by the adjacent healthy tissue but allows only incomplete reconstruction and fibrosis. The use of autologous urothelial cells requires preliminary surgery and has not been successful enough in humans. Research has therefore focused on the use of stem cells. Adipose Derived Stem Cells (ADSCs) and Bone Marrow Derived Stem Cells (BMSCs) allow the reconstruction of the smooth muscle layer, but have little effect on urothelium reconstruction. Urine Derived Stem Cells (UDSCs) and Bladder mesenchymal Stem Cells (BSCs) are very promising because they allow the achievement of a differentiated urothelium. Induced Pluripotent Stem Cells (IPSCs) and Embryonic Stem Cells (ESCs) can be differentiated towards urothelial phenotype but their use is restricted by ethics.

Worldwide, the consumption of dietary supplements for the enhancement of sexual performance is common. Consumers are generally fond of these products because they often want to avoid drugs, preferring "natural" than "chemical" solutions. This is challenging, as many of these supplements labelled "herbal" or "natural" are actually adulterated with drugs, mainly phosphodiesterase-5 inhibitors. This phenomenon is facilitated by fewer demanding regulations for marketing supplements. Thus, consumers may be widely exposed to serious adverse events, such as acute liver injury, kidney failure, pulmonary embolism, stroke or even death. We aim to warn physicians about this issue. This multidisciplinary review simultaneously deals with clinical consequences of this phenomenon, analytical toxicology, and regulation. Indeed, after outlining this worldwide issue and highlighting that a drug-adulterated dietary supplement is actually a falsified drug, we discuss its main contributing factors. Then, we describe some examples of adverse events of which a case of sildenafil-tadalafil-induced ischaemic stroke that benefited medical care in our hospital. Furthermore, we present some means to avoid adulteration and discuss their limitations that may be explained by the heterogeneity of the regulation of dietary supplements between countries. Doing so, we point out the requirement of a global harmonization of this regulation for an efficient eradication of this public health threat. Meanwhile, dietary supplements should be considered adulterated until proven otherwise. Thus, we encourage physicians to investigate these products in the drug histories of their patients, especially when clinical conditions cannot be explained by classical aetiologies.

Hyaline cartilage is deficient in self-healing properties. The early treatment of focal cartilage lesions is a public health challenge to prevent long-term degradation and the occurrence of osteoarthritis. Cartilage tissue engineering represents a promising alternative to the current insufficient surgical solutions. 3D printing is a thriving technology and offers new possibilities for personalized regenerative medicine. Extrusion-based processes permit the deposition of cell-seeded bioinks, in a layer-by-layer manner, allowing mimicry of the native zonal organization of hyaline cartilage. Mesenchymal stem cells (MSCs) are a promising cell source for cartilage tissue engineering. Originally isolated from bone marrow, they can now be derived from many different cell sources (e.g., synovium, dental pulp, Wharton’s jelly). Their proliferation and differentiation potential are well characterized, and they possess good chondrogenic potential, making them appropriate candidates for cartilage reconstruction. This review summarizes the different sources, origins, and densities of MSCs used in extrusion-based bioprinting (EBB) processes, as alternatives to chondrocytes. The different bioink constituents and their advantages for producing substitutes mimicking healthy hyaline cartilage is also discussed.

International audience

International audience

Background: Liver toxicity during immune checkpoint inhibitor treatment is mostly due to immune mediated hepatitis. Viral hepatitis, as well as auto-immune or metabolic hepatitis, are considered as exclusion criteria for ICI induced immune hepatitis diagnosis. However, considering the high prevalence of viral hepatitis B infection and the increasing prescription of immune checkpoint inhibitors, their use in patients with HBV chronic viral infection may be common, even more if patients are treated for hepatocellular carcinoma. Few clinical studies directly deal with the risk of HBV reactivation during ICI therapy and real-life data is currently based on five reported cases of HBV reactivation, one with fatal outcome. In this review, we summarize the current available clinical information about HBV reactivation risk during ICI treatment, its hypothetic mechanism, and propose practical recommendations about verifying and monitoring HBV status throughout the treatment.

Resume Objectif L’objectif de cette etude etait de realiser un etat des lieux des usages, mesusages et des cas d’abus/dependance avec les formes non injectables de fentanyl (forme de fentanyl transdermique et formes de fentanyl transmuqueuses) en France. Methode La consommation problematique de la forme de fentanyl transdermique et des formes de fentanyl transmuqueuses a ete evaluee en utilisant une approche combinant plusieurs sources d’information : (i) notifications spontanees (NotS) recensees sur 6 ans (2010–2015) pour la forme transdermique et sur 3 ans (2013–2015) pour les formes transmuqueuses rapportees aux reseaux de pharmacosurveillance et (ii) donnees des enquetes specifiques du reseau francais d’addictovigilance recensees sur 6 ans (2010–2015). Resultats Au total, 147 NotS ont ete recensees pour la forme de fentanyl transdermique et 109 NotS pour les formes de fentanyl transmuqueuses. Les profils mis en evidence dans les cas d’addictovigilance different en fonction de la forme galenique. Pour la forme de fentanyl transdermique, 2 profils : 1/recherche d’effets antalgiques (74 %) : femmes (61 %), âge median 47 ans, antecedents d’abus et/ou psychiatriques (46 %), traitees pour des douleurs chroniques d’origine non cancereuse (93 %), 2/recherche d’effets psychiques positifs autres qu’antalgiques (26 %) : hommes (82 %), âge median 32 ans, antecedents addictifs et/ou psychiatriques (87 %), obtention illegale (60 %). Pour les formes de fentanyl transmuqueuses, 1 seul profil : femmes (52 %), âge median 48 ans, antecedents d’abus (24 %) et psychiatriques (28 %), indications hors autorisation de mise sur le marche (AMM) [72 %] (indication hors cancer et/ou patients sans traitement de fond). Le mesusage des formes de fentanyl transmuqueuses comporte un risque eleve d’effets indesirables : ceux deja connus des opioides, parfois letaux, (syndrome de sevrage, depression respiratoire et du systeme nerveux central,…) mais aussi des reactions graves au niveau du site d’administration (buccal ou nasal). Pour la forme de fentanyl transdermique, 27 cas d’intoxications, dont 25 graves, lors d’une consommation problematique ont ete signales. A noter, 19 cas de deces impliquant les deux formes de fentanyl ont ete mis en evidence (2 pour les formes de fentanyl transmuqueuses et 17 concernant la forme de fentanyl transdermique). Conclusion Nos resultats rapportent un mesusage important et preoccupant des formes de fentanyl transmuqueuses avec une large utilisation hors AMM et egalement une dependance primaire au fentanyl, quelle que soit la forme galenique, chez des patients traites pour des douleurs chroniques d’origine non cancereuse. Devant les risques importants du fentanyl, il est necessaire de poursuivre la surveillance du mesusage, en particulier, grâce aux activites du reseau francais d’addictovigilance permettant une analyse multisource des informations concernant les cas d’abus, de mesusage et de dependance.

International audience; Introduction : L’impression 3D s’avère un outil révolutionnaire en médecine régénératricepersonnalisée pour façonner des tissus et des organes bio-imprimés [1] à partir de cellules du patient.Les cellules souches mésenchymateuses (CSMs) se présentent comme une alternative prometteuse pourl’ingénierie tissulaire du cartilage [2] par ses capacités de prolifération et de différentiation. Notreobjectif a été d’évaluer la chondrogenèse des CSMs de moelle osseuse au sein de substituts produits parimpression 3D.Méthodes : Les CSMs issues de moelle osseuse humaine ont été ensemencées dans une bio-encreà base d’alginate à une densité cellulaire de 1 million de cellules/mL. La bio-encre a été imprimée couchepar couche grâce à un système d’impression 3D par bio-extrusion. Les substituts produits ont été divisésen deux groupes, un premier groupe contrôle (ITS) et le second cultivé avec un facteur de croissancechondro-inducteur, le TGF-β1 (10 ng/mL). La chondrogenèse a été évaluée après 28 jours (J28) et 56jours (J56) par analyses des expressions géniques (RT-PCRq) et de la synthèse matricielle (histologie etimmunohistochimie) au sein des substituts produits.Résultats : L’analyse des marqueurs chondrogéniques (COL2A1, COLIIB, ACAN, SOX9 etCOMP) par RT-PCRq a montré qu’à J28 leurs expressions étaient significativement plus importantessous l’effet de l’ajout de TGF-β1. En revanche, aucune différence a été constatée à J56 avec uneexpression proche du niveau d’expression basale pour l’ensemble des gènes. Les analyses histologiqueset immunohistologiques ont montré que le TGF-ß1 a induit la synthèse de protéoglycannes et decollagène de type II dense et homogène à J56.Conclusion : Notre procédé d’impression 3D associé à des CSMs humaines et à un facteurchondro-inducteur a permis d’obtenir un substitut cartilagineux de bonne qualité, avec une matriceextracellulaire dense en collagène de type II et en protéoglycannes.[1] Mandrycky C, Wang Z, Kim K, Kim DH. 3D bioprinting for engineering complex tissues. Biotechnol Adv2015, S0734-9750(15)30066-5.[2] Baghaban Eslaminejad M, Malakooty Poor E. Mesenchymal stem cells as a potent cell source for articularcartilage regeneration. World J Stem Cells 2014, 6(3):344-54.

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International audience

The flesh of the Pinctada radiata pearl oyster from coastal Tunisia is considered as a high source of n-3 and n-6 and its shell nacre layer is a promising osteogenic biomaterial. Fatty acid (FA) analysis showed that the major components found in total FA (TFA) were 14:0, 16:0, and 18:0 saturated FA (SFA); 16:1, 18:1, and 20:1 monoenoic FA; 20:4n-6 (ARA), 22:5n-3 (DPA). Characteristically high levels of 20:5n-3 (EPA) and 22:6n-3 (DHA) (6.53-89.75 mg/100 g TFA) polyunsaturated FA (PUFA) were found, respectively, in the TFA of nacre and flesh. Evaluated the effects in vitro of lipids extracted from nacre (Ln) and from flesh (Lc) of P. radiata on growth and the differentiation of osteoblasts. Cytotoxicity tests (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide [MTT] and lactic acid dehydrogenase c [LDH]) demonstrated that both extracts are nontoxic. Alizarin Red staining was used in an osteoblast differentiation model using the osteoblast MC3T3-E1 cell line. It showed that the FA of both extracts induced osteoblast differentiation leading to mineralization. Reverse transcription-polymerase chain reaction (RT-PCR) showed a significantly higher expression of osteocalcin (Bglap) and runt-related transcription (Runx2) in MC3T3-E1 cells in the presence of Ln. No difference of osteopontin (Spp1) and Collagen type I (Col1a1) genes compared to the control was observed. In conclusion, these results supported, obtained from our in vitro experimental model used, the interest/potential of lipids extracted from nacre and P. radiata flesh to stimulate bone formation.

Amiodarone treatment is contraindicated during breastfeeding. As the regional pharmacovigilance centre, we were contacted for information relative to the possibility of breastfeeding after single intravenous administration of 450 mg amiodarone to a breastfeeding woman. A monitoring of amiodarone concentration in plasma and milk was performed in the mother. At day 4, milk concentration of amiodarone reached a peak (233 μg/L) and milk to plasma ratio was determined to 3.5. Milk concentration was still detectable at day 10 (132 μg/L). The maximal relative infant dose was estimated to be 0.6% of the maternal weight-adjusted dosage, corresponding to 0.18% of the usual posology used in children by parenteral route. The review of the literature retrieved one publication suggesting that a single intravenous administration of 150 mg of amiodarone to a mother represents a negligible infant risk based on low breast milk concentration. The French National Pharmacovigilance database query did not disclose any case of side effects during breastfeeding after a single dose of amiodarone. A very limited exposition of breastfed newborns to amiodarone, as well as a low risk of side effects, is expected after a single administration of amiodarone to their mothers.

Angiogenesis is a critical parameter to consider for the development of tissue-engineered bone substitutes. The challenge is to promote sufficient vascularization in the bone substitute to prevent cell death and to allow its efficient integration. The capacity of nacre extract to restore the osteogenic activity of osteoarthritis osteoblasts has already been demonstrated. However, their angiogenic potential on endothelial progenitor cells (EPCs) was not yet explored. Therefore, the current study aimed at investigating if nacreous molecules affect EPC behavior. The gene and protein expression levels of endothelial cell (EC)-specific markers were determined in EPCs cultivated in presence of a nacre extract (ethanol soluble matrix [ESM] at two concentrations: 100 μg/mL and 200 μg/mL (respectively abbreviated ESM100 and ESM200)). Cell functionality was explored by proangiogenic factors production and in vitro tube formation assay. ESM200 increased the expression of some EC-specific genes. The in vitro tube formation assay demonstrated that ESM200 stimulated tubulogenesis affecting angiogenic parameters. We demonstrated that a stimulation with 200 μg/mL of ESM increased angiogenesis key elements. This in vitro study strongly highlights the proangiogenic effect of ESM. Due to its osteogenic properties, previously demonstrated, ESM could constitute the key element to develop an ideal prevascularized bone substitute. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2019.

International audience

International audience

International audience; BACKGROUND: Mesenchymal stem cells (MSCs) are found in synovial fluid (SF) and can easily be harvested during arthrocentesis or arthroscopy. However, SF-MSC characterization and chondrogenicity in collagen sponges have been poorly documented as well as their hypothetical in vivo chondroprotective properties with intra-articular injections during experimental osteoarthritis (OA).METHODS: SF-MSCs were isolated from human SF aspirates in patients suffering from advanced OA undergoing total knee joint replacements. SF-MSCs at passage 2 (P2) were characterized by flow cytometry for epitope profiling. SF-MSCs at P2 were subsequently cultured in vitro to assess their multilineage potentials. To assess their chondrogenicity, SF-MSCs at P4 were seeded in collagen sponges for 4 weeks under various oxygen tensions and growth factors combinations to estimate their gene profile and matrix production. Also, SF-MSCs were injected into the joints in a nude rat anterior cruciate ligament transection (ACLT) to macroscopically and histologically assess their possible chondroprotective properties.RESULTS: We characterized the stemness (CD73+, CD90+, CD105+, CD34-, CD45-) and demonstrated the multilineage potency of SF-MSCs in vitro. Furthermore, the chondrogenic induction (TGF-ß1 ± BMP-2) of these SF-MSCs in collagen sponges demonstrated a good capacity of chondrogenic gene induction and extracellular matrix synthesis. Surprisingly, hypoxia did not enhance matrix synthesis, although it boosted chondrogenic gene expression (ACAN, SOX9, COL2A1). Besides, intra-articular injections of xenogenic SF-MSCs did exert neither chondroprotection nor inflammation in ACLT-induced OA in the rat knee.CONCLUSIONS: Advanced OA SF-MSCs seem better candidates for cell-based constructs conceived for cartilage defects rather than intra-articular injections for diffuse OA.

International audience

Cartilage tissue engineering is making progress, but the competing available strategies still leave room for improvement and consensual overviews regarding the best combinations of scaffolds and cell sources are limited by the capacity to compare them directly. In addition, because most strategies involve autologous cell transfer, once these are optimized, the resulting implants require individual quality control prior to grafting in order to emphasize patient-to-patient differential responsiveness to engineering processes. Here, cartilage substitutes prepared from human mesenchymal stem cells undergoing chondrogenic differentiation within distinct scaffolds were used as pilot samples to investigate the pertinence of a novel method with the aim of characterizing the implants. The limits and advantages of analysing, by label-free liquid chromatography-coupled matrix-assisted laser desorption and ionization (LC-MALDI) mass spectrometry, the secreted proteome released into culture medium by engineered cartilage tissues were investigated and compared with more classically used methods for biomaterial characterization. This method did not require sacrificing the biomaterials and robustly evidenced their chondrogenic statuses. In more detail, the method highlighted differences between batches prepared from distinct donors. It was adapted to distinct scaffolds and allowed a comparison of the influence of individual engineering steps, such as growth factor combinations and oxygen tension. Finally, it evidenced subtle changes between replicate substitutes within a series, thereby distinguishing the least and most accomplished ones. We conclude that relative quantification of secreted proteins through label-free LC-MALDI will be useful, not only to orientate engineering methodologies, but also to ultimately provide non-invasive quality control of engineered tissue substitutes for the repair of cartilage and possibly other connective tissues.

Introduction Les inhibiteurs du point de controle immunitaire (ICI) representent une avancee considerable dans la prise en charge et la survie de cancers comme le melanome ou le carcinome bronchique non a petites cellules. Cependant ils induisent des effets indesirables inhabituels, tels que des hypophysites, peu decrites par ailleurs. Notre etude retrospective decrit les caracteristiques des hypophysites declarees au reseau des centres regionaux de pharmacovigilance. Materiel et methodes Requete sur l’ensemble des cas d’endocrinopathies enregistres dans la Base nationale de pharmacovigilance avant le 30 avril 2017, avec selection des cas d’hypophysite imputables au nivolumab, a l’ipilimumab ou au pembrolizumab et revues par un endocrinologue et un pharmacologue. Resultats Environ 61 hypophysites ont ete retenues, concernant 31 femmes et 30 hommes. L’ipilimumab etait la molecule la plus representee (59 %). La plupart des cas (51 %) correspondait a une atteinte de grade 3 et la majorite (88 %) a un deficit corticotrope. Les cas avec atteinte thyreotrope et/ou gonadotrope etaient respectivement de 20 et 2 %. Cinq patients (8 %) ont presente un panhypopituitarisme. L’IRM hypophysaire etait en faveur d’une hypophysite dans 50 % lorsqu’elle etait realisee. Aucun patient n’a recupere sa fonction hormonale anterieure. Le delai moyen de survenue etait significativement plus court avec l’ipilimumab (93 jours vs. 213 p = 0,005). Conclusion Les hypophysites sont des effets indesirables caracteristiques des ICI et generent des deficits qui ne recuperent pas a distance de l’evenement, contrairement a ce qui est observe avec les thyroidites. Les patients doivent alors beneficier d’une prise en charge coordonnee et adaptee aux deficits du patient.

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Introduction De nombreux tests de biologie comportent des reactifs biotinyles. La prise de biotine a haute dose (BHD 300 mg/jour ; QIZENDAY®) dans le cadre de la sclerose en plaques peut induire des interferences et fausser leurs resultats. Nous rapportons 2 cas de patientes traitees par BHD illustrant cette interaction. Cas cliniques Madame XX1, 48 ans, traitee par BHD, presente une thyroidite multinodulaire a l’echographie, tout en etant asymptomatique avec un examen clinique normal. A la biologie, TSH a 0,005 mUI/L (VN 0,27–4,2), T4L a 59,9 ng/L (VN 7,8–19,4), anticorps anti-TPO positifs a > 600 UI/L (seuil > 34) et anti-TG a 591 UI/L (seuil > 115). La biologie faite 5 jours apres l’arret de la BHD montre une normalisation (TSH a 0,58 mUI/L, T4L a 9,5 ng/L). Madame XX2, 42 ans, presente une biologie avec TSH a 0,08 mUI/L, T4L a 41 ng/L et anti-TPO positifs a 364 UI/L avec des signes cliniques etonnamment moderes de thyrotoxicose (perte de 2 kg, irritabilite, thermophobie, absence de tachycardie). Le doute sur une interaction avec la BHD ainsi que son inefficacite sur la SEP conduisent a son arret. Deux semaines plus tard, la biologie est normale (TSH a 1,82 mUI/L et T4L a 9,6 ng/L) avec negativation des anti-TPO. Conclusion La BHD peut simuler de veritables thyroidites biologiques avec anticorps faussement positifs. De nombreux parametres biologiques peuvent ainsi etre perturbes. Un contact avec le biologiste est indispensable face a un tableau clinicobiologique douteux ou discordant.

Purpose The aim of this work is to compare the energy response of a scintillating fiber to that of standard detectors used for dose monitoring in Computed Tomography (CT) and confirm the validity of Plastic Scintillating Detectors in CT dosimetry. We also seek to model the scintillating fiber response in realistic irradiation conditions for future applications on a complete dose measurement system. For this, we have modeled an X-ray generator emission and validated the model using its measured X-rays spectrum. Materials and methods Irradiations were performed in an X-ray generator at voltage varying from 80 to 140 kV. The Plastic Scintillating Detector (PSD) was compared with a pencil ion chamber and Thermo-Luminescent Detectors (TLD). Monte Carlo simulations of each detector response were compared to measurements in order to validate the simulation in a simple case. Results The Monte Carlo simulations reproduced accurately the response of each detector with less than 3% and 5% of difference for the ion chamber and the TLDs respectively. When comparing the PSD with the two standard detectors, we observed that while the plastic scintillator detector and the ion chamber responses presented less than 5% discrepancy, they both were very different than TLDs response on the whole investigated energy range. The Monte Carlo simulation of the PSD reproduced accurately the response of the plastic scintillating detector with less than 3% of difference. Conclusion The X-ray spectrum has been accurately measured which allowed to successfully reproduce the energy response of the PSD using Monte Carlo simulations. Although this study proved that the scintillating fibers energy response is appropriate for CT dose measurement, further experiments must be performed in clinical conditions.

International audience; Opioid antagonists such as naltrexone and nalmefene are used in drug therapy for alcoholism. Nalmefene, approved in Europe in February 2013 for the reduction of alcohol consumption, is used in patients with alcohol dependence. We report 11 cases of opioid withdrawal syndrome after a single dose of nalmefene in patients usually treated with methadone, buprenorphine, but also with fentanyl or loperamide. Nalmefene is both a partial agonist and an antagonist of opioid receptors. Regarding to its opioid antagonist activity, nalmefene is contraindicated in patients with an opioid treatment. Therefore, when prescribing or delivering nalmefene, healthcare professionals need to be vigilant about any type of opioid exposure, even masked or hidden, to avoid these potential life-threatening syndromes.

Resume Introduction Le paracetamol est la substance active la plus impliquee dans les intoxications medicamenteuses volontaires et involontaires. Neanmoins, sa mise a disposition hors officine, via la grande distribution, est actuellement debattue en France. Methode Nous avons realise une evaluation prospective de la connaissance, de l’usage et du risque de surdosage en paracetamol a l’aide d’un questionnaire autoadministre dans la patientele de cabinets de medecine generale de la zone urbaine de Metz Metropole, entre mai 2015 et fevrier 2016. La prevalence des patients en risque potentiel de surdosage involontaire a ete estimee par methode de capture-recapture. Resultats Parmi les 819 patients ayant repondu, 17,9 % avaient une connaissance satisfaisante et 20,3 % etaient a risque de surdosage potentiel, majoritairement des patients âges de plus de 55 ans ou appartenant aux categories socioprofessionnelles des ouvriers et des inactifs. Un bon score de connaissance etait un facteur protecteur du risque de surdosage ( p Conclusion La dispensation du paracetamol hors des pharmacies d’officine ne saurait etre recommandee en l’etat. Des campagnes d’information sont necessaires pour limiter le risque de surdosage involontaire et de ses consequences hepatotoxiques.

Resume Introduction Le tocilizumab (TCZ) est un anticorps monoclonal qui inhibe la voie de signalisation de l’interleukine 6 (IL-6). C’est un traitement tres efficace de la polyarthrite rhumatoide (PR), qui peut cependant s’accompagner d’infections graves avec des tableaux cliniques trompeurs. Nous rapportons un cas de dermo-hypodermite bacterienne atypique chez une femme traitee par TCZ. Observation Une femme de 80 ans, traitee par methotrexate (MTX) et TCZ pour une PR, presentait une dermo-hypodermite de jambe gauche 8 jours apres une 13e perfusion de TCZ. Il n’y avait ni fievre ni syndrome inflammatoire biologique. Une antibiotherapie orale par amoxicilline (3 g/j) etait debutee en ambulatoire. Deux semaines plus tard, la patiente restait apyretique et le bilan biologique normal mais les signes inflammatoires locaux persistaient. La perfusion de TCZ etait reportee et un traitement par amoxicilline intraveineuse (4 g/j) etait instaure pendant 2 jours avec un relai per os, permettant une guerison rapide. Les cures suivantes de TCZ se sont deroulees sans incident particulier. Discussion Cette patiente a presente au cours du traitement par TCZ une dermo-hypodermite bacterienne (DHB) circonscrite a type d’erysipele, particuliere par son caractere apyretique et l’absence de syndrome inflammatoire biologique. Des DHB severes ont ete rapportees lors de traitements par TCZ, mais pas d’erysipele a notre connaissance. L’attenuation des symptomes locaux et systemiques d’inflammation est connue, directement liee a l’activite anti-IL-6 du TCZ. Les patients atteints de PR sont particulierement sensibles aux infections opportunistes ou severes, du fait de la maladie elle-meme et de ses traitements ; une grande vigilance est necessaire vis-a-vis d’infections adulterees par le TCZ et potentiellement plus severes.

Introduction Differentes sources de cellules souches mesenchymateuses (CSMs) sont etudiees pour une utilisation en ingenierie cellulaire et tissulaire du cartilage : la moelle osseuse, le tissu adipeux, la gelee de Wharton, la membrane synoviale et le liquide synovial. Les CSMs issues du liquide synovial (CSMs LS), representent un interet majeur de par leur localisation articulaire et leur facilite de prelevement par ponction. L’objectif de cette etude etait d’optimiser les conditions de cultures pour isoler les cellules issues du liquide synovial et de les caracteriser en comparaison avec des CSMs issues de la moelle osseuse (CSMs MO). Materiels et methodes A partir des prelevements de liquide synovial, differentes methodes ont ete testees pour isoler les cellules : centrifugation, digestion a la collagenase B, digestion a la hyaluronidase et dilution du liquide synovial dans du milieu de culture. Apres optimisation des conditions d’ensemencement, nous avons etudie l’expression des marqueurs de surface par cytometrie en flux, le taux d’expression des genes d’interet du cartilage (marqueurs chondrogenique, osteogenique et fibrotique) par PCR en temps reel, ainsi que la capacite de differenciation des cellules vers les voies chondrogenique, osteogenique et adipogenique a la fin du deuxieme passage. Dans un second temps, les CSMs de LS ont ete ensemencees dans un biomateriau a base de collagene a raison de 500000 cellules par biomateriau et cultivees pendant 28 jours dans un milieu temoin (1 % ITS) ou enrichi en facteur de croissance (TGF-1) dans un environnement normoxique (20 % O2). Au sein des implants obtenus, nous avons analyse l’activite mitochondriale, le taux d’expression relative des genes d’interet du cartilage et la qualite de la matrice synthetisee. Resultats Concernant l’isolement des cellules, la dilution au 1/6 dans du milieu de culture represente la methode la plus efficace pour obtenir un isolement correct avec une expansion rapide des cellules. L’analyse de l’expression des marqueurs de surface par cytometrie en flux a permis de mettre en evidence la presence des marqueurs caracteristiques des CSMs (CD90+, CD105+, CD73+, CD34− et CD45−). Les expressions des genes d’interet du cartilage (COL2A1, SOX9, ACAN) sont similaires entre les deux types cellulaires. Cependant, les expressions de certains genes hypertrophiques et/ou osteogeniques sont plus faibles pour les CSMs LS (BGLAP, COL1A1, COL10A1, RUNX2, VCAN). Les CSMs LS possedent des capacites de differenciation similaires vers les voies chondrogenique, osteogenique et adipogenique aux CSMs de moelle osseuse. L’activite mitochondriale montre une adaptation satisfaisante de nos cellules au sein de l’implant. L’analyse de l’expression des genes met en evidence une augmentation de l’expression des genes d’interet du cartilage (col2a1, Sox9, acan) en condition TGF-s1De plus, la matrice extracellulaire synthetisee au sein des implants est riche en proteoglycanes et en collagene. Conclusion Les CSMs du liquide synovial representent une source cellulaire prometteuse en ingenierie cellulaire et tissulaire du cartilage. Les expressions plus faibles des marqueurs hypertrophiques et osteogeniques suggerent une differenciation chondrogenique moins propice a une differenciation terminale osseuse observee avec les CSMs de la moelle osseuse. Travaux de recherche subventionnes par la Societe francaise de rhumatologie.

International audience

International audience

Selected paper from the 6th China–France International Symposium “Stem Cells and Regenerative Medicine” and the first meeting of the France–China International CNRS Network (GDRI) CeSMeR, Nancy, 10–13 July 2016; International audience; Mesenchymal stem cells (MSCs) are a common tool in regenerative medicine. The nanoscale extracellular vesicles (nEVs) secreted by these cells were recently brought up to light thanks to their therapeutic potential. In this study, we assessed the in vitro behaviour of human umbilical vein endothelial cells (HUVECs) exposed to nEVs derived from human umbilical cord mesenchymal stem cells (hUC-MSCs). Nanoscale extracellular vesicles were isolated and characterized by NanoSight® and flow cytometry. HUVECs were stimulated with various concentrations of nEVs. To assess nEV interactions with HUVECs, confocal microscopy and angiogenesis assay were performed. The use of nEVs derived from hUC-MSCs was able to produce positive outcomes on HUVECs by acting on their angiogenic potential.

International audience

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International audience

International audience

Nacre is able to induce bone-forming cells mineralization, and gains widely interest in bone regeneration. While, the osteoinductive compounds are not yet identified. ESM (Ethanol Soluble Matrix), a nacre extract from powder of Pinctada margaritifera pearl oyster shell, has been firstly proven having the capacity to induce mineralization and to restore mineralization defect in vitro. It is suitable to treat ESM as a source of osteoinductive compounds. Herein, we develop a new method for separating and purifying nacre extracts by an ionic approach. At first, cationic ESM (ESMc) and anionic ESM (ESMa) were achieved with ion-exchange resin. Then, ESM was separated and collected on cation exchange HPLC. Scanning Electron Microscopy coupled with Energy Dispersive X-ray Spectrometry (EDS) was used to reveal the concentrated elements in ESM fractions. A coupled cell models were used to test the ESM fractions. Alizarin Red staining was performed and quantified to evaluate the mineralization level. ESMc and 2 HPLC fractions stimulated the mineralization in both cells. EDS demonstrated the abundant presence of calcium and chloride in the osteogenic fractions. To validate, pure CaCl2 was tested and proven having an osteogenic effect in both cells, but less stable than ESM. The mineralization nodules induced by ESM fractions and CaCl2 differed in both cells. In conclusion, a new method was developed for separating and purifying nacre extracts by an ionic approach. By which, the osteoinductive compounds in ESM were proven cationic, and calcium in ESM was demonstrated to play a role in inducing the cell mineralization.

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National audience

Introduction Nalmefene (NMF – SELINCRO®), a drug for alcohol withdrawal management (AWM), is both a partial agonist and an antagonist of opioids receptors. Inherent to the latter activity, NMF is contra-indicated in patients with opioid treatment. However, despite this contra-indication, cases of opioid withdrawal syndrome (OWS) in patients treated with opioid maintenance therapy (OMT) are reported [1] , [2] . We aim to report 2 additional cases. Cases description A 46 and a 34-year-old men, with a history of alcoholism and opioid dependence, had OMT with methadone since a few years and were prescribed NMF. Thirty minutes after their single first dose of NMF, they experienced nausea, diarrhoea, vomiting and psychomotor excitement. First patient also had clonus, abundant sweating and anxiety. Both were transferred to the emergency department where OWS was diagnosed. They both received intravenous morphine, plus oxazepam (first patient) or a mix of benzodiazepines and neuroleptics (second patient). On the following day, first patient still presented digestive symptoms leading to hypokalaemia. A symptomatic treatment was initiated, morphine was stopped and methadone was reintroduced. He totally recovered on the 3rd day and was discharged. The second patient was transferred into intensive care unit (ICU) as he required intubation and sedation. Two days later, morphine was stopped and methadone was reintroduced. The patient had totally recovered and was discharged two days later. Discussion–conclusion Our two cases must be added to the nine other recently reported cases of OWS soon after a single first NMF intake in opioid dependent patients [1] , [2] . This condition always required hospitalization and sometimes ICU stay. No patient died. The summary of product characteristics of Selincro® has been updated in July 2015 with a clearer additional assertion to emphasize the need for healthcare professionals to be aware of their patients’ opioid consumption, especially OMT, before prescribing or delivering NMF. As co-existence of alcoholism and OMT is not rare, healthcare professionals really need to be vigilant about the contra-indication of NMF in patients under OMT.

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International audience; Osteoarthritis (OA) is the most common cause of joint chronic pain and involves the entire joints. Subchondral osteoarthritic osteoblasts present a mineralization defect and, to date, only a few molecules (Vitamin D3 and Bone Morphogenetic Protein2) could improve the mineralization potential of this cell type. In this context, we have tested for the first time the effect of nacre extract on the mineralization capacity of osteoblasts from OA patients. Nacre extract is known to contain osteogenic molecules which have demonstrated their activities notably on the MC3T3 pre-osteoblastic cell line. For this goal, molecules were extracted from nacre (ESM, Ethanol Soluble Matrix) and tested on osteoblasts of the subchondral bone from OA patients undergoing total knee replacement and on MC3T3 cells for comparison. We chose to investigate the mineralization with Alizarin Red staining and with the study of extracellular matrix (ECM) structure and composition. In a complementary way the structure of the ECM secreted during the mineralization phase was investigated using second harmonic generation (SHG). Nacre extract was able to induce the early presence (after 7 days) of precipitated calcium in cells. Raman spectroscopy and electron microscopy showed the presence of nanograins of an early crystalline form of calcium phosphate in OA osteoblasts ECM and hydroxyapatite in MC3T3 ECM. SHG collagen fibers signal was present in both cell types but lower for OA osteoblasts. In conclusion, nacre extract was able to rapidly restore the mineralization capacity of osteoarthritis osteoblasts, therefore confirming the potential of nacre as a source of osteogenic compounds. (C) 2015 Elsevier Inc. All rights reserved.

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Introduction La reaction anaphylactique aux curares est une complication qui peut concerner et preoccuper l’ensemble des anesthesistes car elle peut etre severe voire fatale. Le but de cette etude etait d’evaluer son taux de mortalite en France, d’identifier les facteurs de risque de mortalite et de decrire la prise en charge therapeutique des cas mortels. Materiel et methodes La base de donnees du systeme national de pharmacovigilance etait analysee afin de selectionner les cas de reactions d’hypersensibilites aux curares survenus entre janvier 2000 et decembre 2011. Un questionnaire supplementaire etait envoye aux centres regionaux afin d’ameliorer l’information sur la gestion therapeutique des cas a issue fatale. Les cas mortels etaient separes en deux groupes : – « deces precoces » : issue fatale immediate (arret cardiovasculaire non recupere) ou apres mise en place d’une assistance cardiocirculatoire ; – « deces tardifs » liees a une defaillance multiviscerale secondaire a l’arret cardiovasculaire recupere. Une analyse multivariee etait realisee afin de determiner les facteurs de risque independants de mortalite. Resultats Deux mille vingt-deux cas de reaction d’hypersensibilite a un curare etaient analyses dont 84 cas mortels (taux de mortalite moyen 4,1%). A partir des 1247 cas de reactions severes (grade 3 et 4) ont pu etre mis en evidence les facteurs de risques independants de mortalite reportes dans le Tableau 1 . Le curare le plus souvent incrimine etait le suxamethonium ( n = 815, 65,4 %) suivi de l’atracrium ( n = 181, 14,5 %), rocuronium ( n = 105, 8,4 %), cisatracrium ( n = 94, 7,5 %) et vecuronium ( n = 23, 1,8 %). Aucun d’entre eux n’etait identifier comme facteur de risque de surmortalite. Les details de la prise en charge des reactions mortelles etaient obtenus pour 31 patients. Le delai moyen de survenu du premier symptome etait de 2,1 minutes, le delai moyen d’injection du premier bolus d’adrenaline etait de 3,4 minutes et la dose totale d’adrenaline injectee en bolus etait de 15 mg. Il n’y avait aucune difference significative dans la prise en charge entre les groupes deces tardif et precoce. Aucun des patients places sous assistance cardiocirculatoire ne survivait. Discussion Le sexe masculin, l’obesite, un antecedent de maladie cardiovasculaire, un traitement chronique par betabloquant et le contexte d’urgence sont des facteurs de risques independants de mortalite a une reaction d’hypersensibilite aux curares. Le taux de mortalite est de 4,1 %. La prise en charge des cas mortels etait conforme aux recommandations internationales, avec un delai court de mise en œuvre des therapeutiques specifiques. On peut donc supposer qu’il existe des cas de resistance a l’adrenaline. Il est donc necessaire de developper de nouvelles approches therapeutiques.

Résumé; International audience

Joint Bone Spine - In Press.Proof corrected by the author Available online since vendredi 21 juin 2013

International audience; In Magnetic Resonance Imaging (MRI), multi-channel coil arrays are increasingly being used to improve Signal-to-Noise Ratio (SNR) in order to increase spatial and/or temporal image resolution. Recent decoupling technique allows conception of two-channel surface transceiver coil arrays; this technique based on common conductor does not require additional preamplifier for the decoupling between elements. In this case the coil array loops are directly connected to the independent transmit/receive switches and preamplifiers of the MR system. Using this common conductor decoupling technique, a topology of a four-channel coil array was developed and described in this paper. A four-channel surface receiver coil array was designed to perform the simultaneous acquisition of both rat knee joints at 7T. Without the use of additional preamplifiers, a good decoupling between channels was obtained and very high spatial resolution 3D images with a voxel size of 49 × 49 × 98 µm 3 were achieved in 1h22min scan time. Acquisitions allowed the quantification of cartilage morphological parameters such as thickness and volume

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The extracellular matrix (ECM) has long been viewed primarily as an organized network of solid-phase ligands for integrin receptors. During degenerative processes, such as osteoarthritis, the ECM undergoes deterioration, resulting in its remodeling and in the release of some of its components. Matrilin-3 (MATN3) is an almost cartilage specific, pericellular protein acting in the assembly of the ECM of chondrocytes. In the past, MATN3 was found required for cartilage homeostasis, but also involved in osteoarthritis-related pro-catabolic functions. Here, to better understand the pathological and physiological functions of MATN3, its concentration as a circulating protein in articular fluids of human osteoarthritic patients was determined and its functions as a recombinant protein produced in human cells were investigated with particular emphasis on the physical state under which it is presented to chondrocytes. MATN3 down-regulated cartilage extracellular matrix (ECM) synthesis and up-regulated catabolism when administered as a soluble protein. When artificially immobilized, however, MATN3 induced chondrocyte adhesion via a α5β1 integrin-dependent mechanism, AKT activation and favored survival and ECM synthesis. Furthermore, MATN3 bound directly to isolated α5β1 integrin in vitro. TGFβ1 stimulation of chondrocytes allowed integration of exogenous MATN3 into their ECM and ECM-integrated MATN3 induced AKT phosphorylation and improved ECM synthesis and accumulation. In conclusion, the integration of MATN3 to the pericellular matrix of chondrocytes critically determines the direction toward which MATN3 regulates cartilage metabolism. These data explain how MATN3 plays either beneficial or detrimental functions in cartilage and highlight the important role played by the physical state of ECM molecules.

International audience; Introduction: We have taken advantage of the large screening capacity of a multiplex immunoassay to better define the respective contribution of articular versus systemic cytokines in experimental arthritis.Methods: We performed a follow up (from 7 hours to 14 days) multiplex analysis of 24 cytokines in synovial fluid and sera of rats developing Antigen-Induced Arthritis (AIA) and confronted their protein level changes with molecular, biochemical, histological and clinical events occurring in the course of the disease.Results: The time-scheduled findings in arthritic joints correlated with time-dependent changes of cytokine amounts in joint effusions but not with their blood levels. From seven hours after sensitization, high levels of chemokines (MCP-1, MIP1α, GRO/KC, RANTES, eotaxin) were found in synovial fluid of arthritic knees whereas perivascular infiltration occurred in the synovium; local release of inflammatory cytokines (IFNγ, IL-1β, IL-6) preceded the spreading of inflammation and resulted in progressive degradation of cartilage and bone. Finally a local overexpression of several cytokines/adipocytokines poorly described in arthritis (IL-13, IL-18, leptin) was observed.Conclusions: Distinct panels of cytokines were found in arthritic fluid during AIA, and the expected effect of mediators correlated well with changes occurring in joint tissues. Moreover, multiplex analysis could be helpful to identify new pathogenic mediators and to elucidate the mechanisms supporting the efficacy of putative targeted therapies.