Development and biological evaluation of Iinkjet printed drug coatings on intravascular stent

Inkjet–printing technology was used to apply biodegradable and biocompatible polymeric coatings of poly(D, L lactide) with the antiproliferative drugs simvastatin (SMV) and paclitaxel (PCX) on coronary metal stents. A piezoelectric dispenser applied coating patterns of very fine droplets (300 xL) and ink jetting was optimized to develop uniform, accurate and reproducible coatings of high yields on the stent strut. The drug loaded polymeric coatings were assed by scanning electron microscopy (SEM), atomic force microscopy (AFM) and transition thermal microscopy (TTM) where a phase separation was observed for SMV/PLA layers while PCX showed a uniform distribution within the polymer layers. Cytocompatibility studies of PLA coatings showed excellent cell adhesion with no decrease of cell viability and proliferation. In vivo stent implantation studies showed significant intra stent restenosis (ISR) for PXC/PLA and PLA plain coatings similar to marketed Presillion (bare metal) and Cypher (drug eluting) stents. The investigation of several cytokine levels after seven days of stent deployment showed no inflammatory response and hence no in vivo cytotoxicity related to PLA coatings. Inkjet printing can be employed as a robust coating technology for the development of drug eluting stents compared to the current conventional approaches

Marine-inspired enzymatic mineralization of dairy-derived whey protein isolate (WPI) hydrogels for bone tissue regeneration

Whey protein isolate (WPI) is a by-product from the production of cheese and Greek yoghurt comprising β-lactoglobulin (β-lg) (75%). Hydrogels can be produced from WPI solutions through heating; hydrogels can be sterilized by autoclaving. WPI hydrogels have shown cytocompatibility and ability to enhance proliferation and osteogenic differentiation of bone-forming cells. Hence, they have promise in the area of bone tissue regeneration. In contrast to commonly used ceramic minerals for bone regeneration, a major advantage of hydrogels is the ease of their modification by incorporating biologically active substances such as enzymes. Calcium carbonate (CaCO3) is the main inorganic component of the exoskeletons of marine invertebrates. Two polymorphs of CaCO3, calcite and aragonite, have shown the ability to promote bone regeneration. Other authors have reported that the addition of magnesium to inorganic phases has a beneficial effect on bone-forming cell growth. In this study, we employed a biomimetic, marine-inspired approach to mineralize WPI hydrogels with an inorganic phase consisting of CaCO3 (mainly calcite) and CaCO3 enriched with magnesium using the calcifying enzyme urease. The novelty of this study lies in both the enzymatic mineralization of WPI hydrogels and enrichment of the mineral with magnesium. Calcium was incorporated into the mineral formed to a greater extent than magnesium. Increasing the concentration of magnesium in the mineralization medium led to a reduction in the amount and crystallinity of the mineral formed. Biological studies revealed that mineralized and unmineralized hydrogels were not cytotoxic and promoted cell viability to comparable extents (approximately 74% of standard tissue culture polystyrene). The presence of magnesium in the mineral formed had no adverse effect on cell viability. In short, WPI hydrogels, both unmineralized and mineralized with CaCO3 and magnesium-enriched CaCO3, show potential as biomaterials for bone regeneration

A New Hemodynamic Ex Vivo Model for Medical Devices Assessment

Introduction: In stent restenosis (ISR) remains a major public health concern with an increased morbidity, mortality and health-related costs. Drug-eluting stents (DES) have reduced ISR, but are associated with healing-related issues or hypersensitivity reactions, leading to an increased risk of late acute stent thrombosis. Evaluations of new DES are based on animal models or in vitro release systems which show several limitations. The role of flow and shear stress on endothelial cell and ISR has also been emphasized. The aim of this work was to design and first evaluate an original bioreactor, reproducing ex vivo hemodynamic and biological conditions similar to human conditions, to further evaluate new DES. Methods & Results: This bioreactor was designed to study up to 6 stented arteries connected in bypass, immersed in a culture box, in which circulated a physiological systolo-diastolic resistive flow. Two centrifugal pumps drove the flow. The principal pump generated pulsating flows by modulation of rotation velocity, and the second pump worked at constant rotation velocity, ensuring the counter pressure levels and backflows. The flow rate, the velocity profile, the arterial pressure and the resistance of the flow were adjustable. The bioreactor was placed in an incubator to reproduce a biological environment. A first experience of feasibility was realized over a period of 24 days. Three rat aortic thoracic arteries were placed into the bioreactor, immersed in cell culture medium change every 3 days, and with a circulating systole diastolic flux circulating among the entire experimentation. There was no infection, no leak. At the end of experimentation, a morphometric analysis was performed confirming the viability of the arteries. Conclusion: We design and patent an original hemodynamic ex vivo model to further study new DES and ISR. We will next validate this ex vivo model of ISR reproducing this experimentation with stented arteries. Once validated, this bioreactor will allow characterization of the velocity field and drug transfers within a stented artery with new functionalized DES, with experimental means not available in vivo. Another main point will be the reduction of animal experimentation, and the availability of first results of new DES in human tissues (human infra popliteal or coronary arteries collected during human donation)

Phenolic plant extract enrichment of enzymatically mineralized hydrogels

Hydrogel mineralization with calcium phosphate (CaP) and antibacterial activity are desirable for applications in bone regeneration. Mineralization with CaP can be induced using the enzyme alkaline phosphatase (ALP), responsible for CaP formation in bone tissue. Incorporation of polyphenols, plant-derived bactericidal molecules, was hypothesized to provide antibacterial activity and enhance ALP-induced mineralization. Three phenolic rich plant extracts from: (i) green tea, rich in epigallocatechin gallate (EGCG) (herafter referred to as EGCG-rich extract); (ii) pine bark and (iii) rosemary were added to gellan gum (GG) hydrogels and subsequently mineralized using ALP. The phenolic composition of the three extracts used were analyzed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MSn). EGCG-rich extract showed the highest content of phenolic compounds and promoted the highest CaP formation as corroborated by dry mass percentage meassurements and ICP-OES de-termination of mass of elemental Ca and P. All three extracts alone exhibited antibacterial activity in the following order EGCG-rich > PI > RO, respectively. However, extract-loaded and mineralized GG hydro-gels did not exhibit appreciable antibacterial activity by diffusion test. In conclusion, only the EGCG-rich extract promotes ALP-mediated mineralization

Mineralization of gellan gum hydrogels with calcium and magnesium carbonates by alternate soaking for bone regeneration applications

Mineralization of hydrogels is desirable prior to applications in bone regeneration. CaCO3 is a widely used bone regeneration material and Mg, when used as a component of calcium phosphate biomaterials, has promoted bone‐forming cell adhesion and proliferation and bone regeneration. In this study, gellan gum (GG) hydrogels were mineralized with carbonates containing different amounts of calcium (Ca) and magnesium (Mg) by alternate soaking in, firstly, a calcium and/or magnesium ion solution and, secondly, a carbonate ion solution. This alternate soaking cycle was repeated five times. Five different calcium and/or magnesium ion solutions, containing different molar ratios of Ca to Mg ranging from Mg‐free to Ca‐free were compared. Carbonate mineral formed in all sample groups subjected to the Ca:Mg elemental ratio in the carbonate mineral formed was higher than in the respective mineralizing solution. Mineral formed in the absence of Mg was predominantly CaCO3 in the form of a mixture of calcite and vaterite. Increasing the Mg content in the mineral formed led to the formation of magnesian calcite, decreased the total amount of the mineral formed and its crystallinity. Hydrogel mineralization and increasing Mg content in mineral formed did not obviously improve proliferation of MC3T3‐E1 osteoblast‐like cells or differentiation after 7 days

Mise en évidence et caractérisation de l'excrétion pre-partum d'oocystes de Cryptosporidium chez la chèvre adulte

L objectif de cette étude était de caractériser l excrétion pre-partum d oocystes de Cryptosporidium chez la chèvre adulte et d identifier les espèces excrétées. 618 chèvres issues de 31 élevages ont été prélevées dans les 15 jours précédant leur mise-bas. Après concentration au chlorure de césium, les échantillons ont été analysés par immunofluorescence. Les échantillons positifs en immunofluorescence ont été soumis à une PCR 18S afin d identifier les espèces de Cryptosporidium. 48 chèvres positives ont été détectées sur les 618 chèvres prélevées, soit une prévalence d excrétion de 7,8%. Parmi ces positives, la médiane d excrétion est de 13 opg [6-2140 opg] et seules 3 chèvres excrètent plus de 300 opg. Aucun échantillon positif en immunofluorescence n a pu être amplifié par PCR 18S du fait des faibles excrétions. Le rôle des chèvres adultes dans la contamination des chevreaux est probablement mineur du fait des faibles prévalences et niveaux d excrétion. Cependant l impossibilité d identification des espèces ne permet pas d exclure leur rôle de réservoir.TOULOUSE-EN Vétérinaire (315552301) / SudocNANTES-Ecole Nat.Vétérinaire (441092302) / SudocSudocFranceF

Imperméabilisation, traitement antiseptique et teinture d'une prothèse vasculaire fonctionnalisée par la cyclodextrine

Les infections post-opératoires surviennent dans 1 à 6% des cas d'implantation de prothèses vasculaires. Afin de résoudre ce problème, nous avons envisagé de fonctionnaliser la prothèse vasculaire en polyester par une méthode développée dans notre laboratoire et permettant l'enrobage des fibres par un polymère de cyclodextrines, ce dernier permettant l'encapsulation de nombreux principes actifs, dont des agents antimicrobiens. Les résultats de DSC, de MEB et d'ATG ont montré que le polymère de cyclodextrines était bien fixé physiquement sur la prothèse et qu'il ne modifiait aucunement ses propriétés. Des essais de dégradation ont montré que le revêtement de cyclodextrine se résorbait sur environ 10 semaines dans le plasma sanguin et que les produits de dégradation n'étaient pas toxiques. Enfin, les essais histopathologiques in vivo et d'hémocompatibilité in vitro ont confirmé que cette prothèse était parfaitement biocompatible. Ce revêtement nous a permis de fixer de façon efficace le bleu de méthylène sur la prothèse, connu dans le domaine médical pour ses propriétés antiseptiques. Les essais bactériologiques ont démontré une efficacité prolongée de la prothèse contre les bactéries à gram (+) et gram (-) dans des conditions in vitro. D'autre part, ce colorant nous a également permis de résoudre un problème rencontré en coelioscopie, en évitant le phénomène d'interférences par réduction de la réverbération de la lumière blanche sur les parois de la prothèse, ceci favorisant les conditions de l'intervention chirurgicale. Un deuxième volet de notre travail a consisté à substituer les hydrogels d'origine animale, encore couramment utilisés pour imperméabiliser la prothèse textile, par des polymères synthétiques. Ces recherches nous ont permis de répondre au cahier des charges en trouvant le meilleur compromis entre souplesse et imperméabilité du greffon synthétique tout en exprimant une parfaite innocuité, supérieure à celle du témoin traité au collagène vis-à-vis des cellules endothéliales et du sang à travers les tests de cytocompatibilité et d'hémocompatibilité. Pour conclure, nos travaux ont permis de mettre au point une prothèse vasculaire qui apporte trois qualité supplémentaires et donc trois avantages par rapports aux produits actuellement utilisés, c'est à dire une activité antiseptique, une intervention chirurgicale facilitée, et une réduction totale des risques de transmission d'agents pathogènes (virus, prions ...) issus des produits d'origine animale.Post operative infections occur in 1 to 6% of patients implanted with a vascular prosthesis. To solve this problem, we functionalised the polyester prostheses by a method developed in our laboratory. This method can be used to coat the fibre by a cyclodexrin polymer, which allows the complexation of drugs like antiseptics. DSC, SEM and TGA results showed that cyclodextrin polymer was physically fixed onto the prosthesis and that no properties of the prosthesis were modified. Degradation tests revealed that cyclodextrin coating was eliminated after about 10 weeks in human blood plasma and that degradation products were non-toxic. Finally, in vivo histopathological tests and in vitro hemocompatibility tests confirmed that this new prosthesis was perfectly biocompatible. This coating allows us to impregnate methylen-blue known in the medical field to have antiseptic properties. Bacteriological in vitro tests have shown a prolonged efficiency of the functionalised prosthesis against gram(+) and gram(-) bacteria compared to the virgin prosthesis. Moreover, this dye allows to solve the problem met in coelioscopie surgery avoiding the interference phenomena by reducing white light reflection onto the prosthesis wall. The operating conditions can subsequently be improved. A second component of our work was to blood proof the prosthesis by replacing the still widely used hydrogels of animal origin by synthetic polymers. These investigations have allowed us to fulfil the specifications requires by elaborating the best compromise between flexibility and blood proofing of the synthetic graft. Cytocompatibility and hemocompatibility results also ensure a higher safety against cells and blood of the functionalised prosthesis compared to the collagen-treated prosthesis used as the control. ln conclusion, our work has allowed us to develop a vascular prosthesis, which provides three additional qualities an therefore three advantages with respect to the currently used products, i.e. an antiseptic activity, improved surgical conditions, and a total reduction of risks induced by pathogenic agents (viruses, prions...) from products of animal originLILLE1-Bib. Electronique (590099901) / SudocSudocFranceF

Implants pour la délivrance de principes actifs

Les implants permettant de contrôler la libération d’un principe actif peuvent grandement améliorer l’efficacité d’un traitement médical tout en réduisant le risque d’apparition d’effets secondaires toxiques. Dans cet article, quatre stratégies différentes sont développées à titre d’exemple : des substituts osseux hybrides alliant de l’hydroxyapatite et des hydrogels de chitosan, des stents vasculaires recouverts d’un polymère bio-inspiré, des implants cochléaires pour la délivrance locale de dexaméthasone et des implants se formant in situ pour le traitement des parodontites. Mais cela ne représente qu’une petite sélection et de nombreuses autres approches et applications basées sur des implants libérant un principe actif (ou une combinaison d’actifs) existent. Comparés aux implants ou formes galéniques conventionnelles, ils peuvent offrir des avantages potentiellement décisifs

Sulfonated and sulfated chitosan derivatives for biomedical applications: A review

International audienceFrom 20th century, chitosan, a natural polysaccharide, has received much attention for use in biomedical applications thanks to its remarkable properties, such as biodegradability, biocompatibility, hemostasis and antibacterial activity. Over the last decades, many researchers have attempted to generate new chitosan derivatives-based biomaterials though chemical modifications, especially through sulfonation or sulfation reactions in order to tailor the physicochemical and biochemical properties. Due to the presence of residual amino groups, the generated polyampholytic derivatives are characterized by convenient biological properties, such as anti-oxidation, antiviral activity, anticoagulation and bone regeneration, expanding their application scope. This paper provides an overview of the strategies used to chemically modify chitosan by introduction of sulfonate groups on chitosan backbone, focusing on various sulfonating or sulfating agents used and substitution regioselectivity, and highlights their applications in biomedical field

Influence of a Double-Lumen Extension Tube on Drug Delivery: Examples of Isosorbide Dinitrate and Diazepam

International audiencePURPOSE: Plastic materials such as polyurethane (PUR), polyethylene (PE), polypropylene (PP) and polyvinyl chloride (PVC) are widely used in double-lumen extension tubing. The purposes of our study were to 1) compare in vitro drug delivery through the double extension tubes available on the market 2) assess the plastic properties of PUR in infusion devices and their impact on drug delivery.METHODS: The study compared eight double-lumen extension tubes in PUR, co-extruded (PE/PVC) plastic and plasticised PVC from different manufacturers. Isosorbide dinitrate and diazepam were used as model compounds to evaluate their sorption on the internal surface of the infusion device. Control experiments were performed using norepinephrine known not to absorb to plastics. Drug concentrations delivered at the egress of extension tubes were determined over time by an analytical spectrophotometric UV-Vis method. The main characteristics of plastics were also determined.RESULTS: Significant differences in the sorption phenomenon were observed among the eight double-lumen extension tubes and between pairs of extension tubes. Mean concentrations of isosorbide dinitrate delivered at the egress of double-lumen extension tubes after a 150-minute infusion (mean values ± standard deviation in percentage of the initial concentrations in the prepared syringes) ranged between 80.53 ± 1.66 (one of the PUR tubes) and 92.84 ± 2.73 (PE/PVC tube). The same parameters measured during diazepam infusion ranged between 48.58 ± 2.88 (one of the PUR tubes) and 85.06 ± 3.94 (PE/PVC tube). The double-lumen extension tubes in PUR were either thermosetting (resin) or thermoplastic according to reference.CONCLUSIONS: Clinicians must be aware of potential drug interactions with extension tube materials and so must consider their nature as well as the sterilisation method used before selecting an infusion device