Bi-layered silane-TiO2/collagen coating to control biodegradation and biointegration of Mg alloys

International audienceMagnesium alloys have shown high potential as biodegradable implants for bone repair applications. However, their fast degradation in physiological media demands tuning their corrosion rate to accompany the natural tissue healing processes. Here, a new bi-layered silane-TiO2/collagen coating efficient in stabilizing and biofunctionalizing the surface of AZ31 and ZE41 Mg alloys is presented. Corrosion tests performed in cell culture medium over 7 weeks showed that the bi-layered coating promotes the formation of a stable layer of Mg(OH)2/MgCO3/CaCO3 that provides effective protection to the alloys at advanced immersion stages. The intrinsic reactivity of each alloy plus formation of transitory calcium phosphate phases, resulted in distinct corrosion behavior in the short term. Cell experiments showed that the bi-layered coating improved osteoblasts and fibroblasts proliferation compared to bare and silane-TiO2-coated alloys. Different responses in terms of cell adhesion could be related to the intrinsic corrosion rate of each alloy and some toxicity from the alloying elements. The results evidenced the complex interplay between alloy nature, coating-alloy combination and cell type. The silane-TiO2/collagen coating showed to be a promising strategy to improve cell response and viability and to control degradation rate of Mg alloys in the long term

255 In vivo detection of non-occlusive thrombi in drug-eluting stents by scintigraphy and radio-labelled annexin V in a rabbit model

IntroductionThrombi in contact with non re-endothelialized stent struts associated with drug-eluting stent (DES) thrombosis. Hence, detection of thrombi in DES could help to evaluate the risk of DES thrombosis. Annexin V radio-labelled with 99mTechnetium (99mTc) is a radio-tracer with a high affinity for activated platelets.ObjectivesOur objectives were: 1) to develop an animal model of non-occlusive thrombosis of stents, 2) to evaluate the ability of annexin V 99mTc for the detection of in-stent thrombi using scintigraphy.MethodsRight carotid arteries of NZW rabbits (n=14) fed a high cholesterol diet were implanted with overlapping DES (n=7) or bare-metal stents (BMS; n=7). Four weeks after stent implantation, rabbits underwent a first scintigraphy 3 hours after injection of 200 MBq of radio-labelled annexin V 99mTc. At the end of the first scintigraphy, a suture was placed surgically proximal to the stented carotid arteries in order to induce a thrombus-prone flow limiting stenosis. Four days later, a second scintigraphy was performed. After the second scintigraphy, stents were excised, imaged ex vivo and then fixed for histological examination and scanning electron microscopy (SEM).ResultsActivities measured in vivo in the stented carotid arteries after injection of annexin V 99mTc were higher on the second scintigraphy after creation of a surgical stenosis as compared to the first scintigraphy (0.24 vs. 0.15 counts/pixel/MBq, respectively; p<0.05). On the second scintigraphy, activities were higher in DES vs. BMS (0.26 vs. 0.19 counts/pixel/MBq, respectively; p < 0.005). High activities measured in stents in vivo were associated with the detection of thrombi on corresponding histological sections and SEM.ConclusionsIn this work, we developed a rabbit model of non-occlusive thrombosis of stents in carotid arteries. In this model, in-stent thrombi could be detected using annexin V 99mTc scintigraphy

In vitro studies and preliminary in vivo evaluation of silicified concentrated collagen hydrogels

Hybrid and nanocomposite silicacollagen materials derived from concentrated collagen hydrogels were evaluated in vitro and in vivo to establish their potentialities for biological dressings. Silicification significantly improved the mechanical and thermal stability of the collagen network within the hybrid systems. Nanocomposites were found to favor the metabolic activity of immobilized human dermal fibroblastswhile decreasing the hydrogel contraction. Cell adhesion experiments suggested that in vitro cell behavior was dictated by mechanical properties and surface structure of the scaffold. First-to-date in vivo implantation of bulk hydrogels in subcutaneous sites of rats was performed over the vascular inflammatory period. These materials were colonized and vascularized without inducing strong inflammatory response. These data raise reasonable hope for the future application of silicacollagen biomaterials as biological dressings.Fil: Desimone, Martín Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; ArgentinaFil: Hélary, Christophe. Université Pierre et Marie Curie; FranciaFil: Quignard, Sandrine. Université Pierre et Marie Curie; FranciaFil: Rietveld, Ivo B. Universite de Paris; FranciaFil: Bataille, Clement. Université de Versailles Saint-quentin-en-yvelines.; FranciaFil: Copello, Guillermo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; ArgentinaFil: Mosser, Gervaise. Université Pierre et Marie Curie; FranciaFil: Giraud Guille, Marie-Madeleine. Université Pierre et Marie Curie; FranciaFil: Livage, Jacques. Université Pierre et Marie Curie; FranciaFil: Meddahi Pellé, Anne. Université de Versailles Saint-quentin-en-yvelines.; FranciaFil: Coradin, Thibaud. Université Pierre et Marie Curie; Franci

Recent advances in biomaterials for tissue engineering and controlled drug delivery

In this review, recent advances in biomaterials developed to favor tissue repair are presented. The focus is particularly on devices used to promote bone repair, skin wound healing and nerve regeneration. In each case, the specifications for an ideal substitute and the recent advances in the field of these biomaterials are presented. Alternatively, drug delivery systems associated with biomaterials have been developed over the recent decades to stimulate wound healing without any side effects. For this purpose, the overview presents recent advances in medicated dressings for controlled release of antibiotic to prevent infections, growth factors to promote tissue regeneration and gene delivery to modulate cell phenotype.Fil: Hélary, Christophe. Universite de Paris Vi; Francia. Centre National de la Recherche Scientifique; FranciaFil: Desimone, Martín Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Metabolismo del Fármaco. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Metabolismo del Fármaco; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentin

Hyperglycemia acts in synergy with hypoxia to maintain the pro-inflammatory phenotype of macrophages

International audienceDiabetic foot ulcers (DFUs) are characterized by a chronic inflammation state which prevents cutaneous wound healing, and DFUs eventually lead to infection and leg amputation. Macrophages located in DFUs are locked in an pro-inflammatory phenotype. In this study, the effect of hyperglycemia and hypoxia on the macrophage phenotype was analyzed. For this purpose, a microarray was performed to study the gene expression profile of macrophages cultivated in a high glucose concentration. Hyperglycemia upregulated the expression of pro-inflammatory cytokines such as TNF-α, IL-1, IL-6, chemokines and down-regulated the expression of two receptors involved in phagocytosis (CD 36 and Class B scavenger type I receptors). In addition, eleven anti-apoptotic factors were upregulated whereas three pro-apoptotic genes were downregulated. Subsequently, the contribution of hypoxia and hyperglycemia to chronic inflammation and their potential synergistic effect was evaluated on activated THP-1 derived macrophages. A long term post activation effect (17 hours) was only observed on the upregulation of pro-inflammatory cytokines when hyp-oxia was combined with a high glucose concentration. In contrast, hyperglycemia and hyp-oxia did not have any effect on wound healing molecules such as TGF-β1. Taken together, the results show that hyperglycemia acts in synergy with hypoxia to maintain a chronic inflammation state in macrophages

3D models of dilated cardiomyopathy: Shaping the chemical, physical and topographical properties of biomaterials to mimic the cardiac extracellular matrix

International audienceThe pathophysiology of dilated cardiomyopathy (DCM), one major cause of heart failure, is characterized by the dilation of the heart but remains poorly understood because of the lack of adequate in vitro models. Current 2D models do not allow for the 3D organotypic organization of cardiomyocytes and do not reproduce the ECM perturbations. In this review, the different strategies to mimic the chemical, physical and topographical properties of the cardiac tissue affected by DCM are presented. The advantages and drawbacks of techniques generating anisotropy required for the cardiomyocytes alignment are discussed. In addition, the different methods creating macroporosity and favoring organotypic organization are compared. Besides, the advances in the induced pluripotent stem cells technology to generate cardiac cells from healthy or DCM patients will be described. Thanks to the biomaterial design, some features of the DCM extracellular matrix such as stiffness, porosity, topography or chemical changes can impact the cardiomyocytes function in vitro and increase their maturation. By mimicking the affected heart, both at the cellular and at the tissue level, 3D models will enable a better understanding of the pathology and favor the discovery of novel therapies

Generation of an Adequate Perfusion Network within Dense Collagen Hydrogels Using Thermoplastic Polymers as Sacrificial Matrix to Promote Cell Viability

International audienceTDense collagen hydrogels are promising biomaterials for several tissue-engineering applications.They exhibit high mechanical properties, similar to physiological extracellular matrices,and do not shrink under cellular activity. However, they suffer from several drawbacks, such asweak nutrient and O2 diffusion, impacting cell survival. Here, we report a novel strategy to createa perfusion system within dense and thick collagen hydrogels to promote cell viability. The 3Dprinting of a thermoplastic filament (high-impact polystyrene, HIPS) with a three-wave shape is usedto produce an appropriate sacrificial matrix. The HIPS thermoplastic polymer allows for good shapefidelity of the filament and does not collapse under the mechanical load of the collagen solution. Afterthe collagen gels around the filament and dissolves, a channel is generated, allowing for adequateand rapid hydrogel perfusion. The dissolution process does not alter the collagen hydrogel’s physicalor chemical properties, and the perfusion is associated with an increased fibroblast survival. Here,we report the novel utilization of thermoplastics to generate a perfusion network within biomimeticcollagen hydrogels

MnO2-gated Nanoplatforms with Targeted Controlled Drug Release and Contrast-Enhanced MRI Properties: from 2D Cell Culture to 3D Biomimetic Hydrogels

International audienceMultifunctional nanomaterials combining diagnosis and therapeutic properties have attracted a considerable attention in cancer research. Yet some important challenges are still to be faced, including an optimal coupling between these two types of properties that would be effective within complex biological tissues. To address these points, we have prepared novel nanoplatforms associating controlled drug delivery of doxorubicin and Magnetic Resonance Imaging (MRI) contrast-enhancement that exhibit high specificity towards cancer cells compared to normal cells and evaluated them both in 2D cultures and within 3D tissue-like biomimetic matrices. Methods: Nanoplatforms were prepared from hollow silica nanoparticles coated with MnO2 nanosheets and conjugated with the AS1411 aptamer as a targeting agent. They were fully characterized from a chemical and structural point of view as well as for drug release and MRI signal enhancement. Standard two-dimensional monolayer cultures were performed using HeLa and Normal Human Dermal Fibroblasts (NHDF) cells to testify targeting and cytotoxicity. Cellularized type I collagen-based hydrogels were also used to study nanoparticles behavior in 3D biomimetic environments. Results: The as-established nanoplatforms can enter HeLa cells, leading to the dissociation of the MnO2 nanosheets into Mn 2+ that enhanced T1 magnetic resonance signals and concomitantly release doxorubicin, both effects being markedly more significant than in the presence of NHDFs. Moreover, particles functionality and specificity were preserved when the cells were immobilized within type I collagen-based fibrillar hydrogels. Conclusion: The use of MnO2 nanosheets as glutathione-sensitive coatings of drug loaded nanoparticles together with surface conjugation with a targeting aptamer offers an effective strategy to obtain efficient and specific nanotheranostic systems for cancer research, both in 2D and 3D. The here-described tissue-like models should be easy to implement and could constitute an interesting intermediate validation step for newly-developed theranostic nanoparticles before in vivo evaluation

Surface reactivity of hydroxyapatite nanocoatings deposited on iron oxide magnetic spheres toward toxic metals

International audienceHypothesis : Hydroxyapatite and magnetite are two environmentally-friendly mineral phases that have fruitful properties for remediation process. The formation of magnetic core@sorbent shell nanostructures should provide efficient materials for toxic metal removal from aqueous media. However the nanoscale confinement of hydroxyapatite may influence its reactivity.Experiments : Fe3O4@Hydroxyapatite nanocomposites were prepared by surface-controlled precipitation of hydroxyapatite layers from 10 nm to 150 nm in thickness on iron oxide spheres. The surface reactivity of the core–shell particles toward selected inorganic ions of environmental relevance (Pb(II), Y(III), Eu(III), Sb(III)) was studied by batch sorption experiments, X-ray diffraction and electron microscopy.Findings : The reactivity of the hydroxyapatite coating varied from partial cation exchange to dissolution/transformation of the shell. The nature and extent of the reactions depended significantly on the hydroxyapatite layer structure but was not significantly influenced by the magnetic core. These novel nanocomposites should be useful for environmental applications

A specific beta 3-adrenoceptor agonist induces increased pancreatic islet blood flow and insulin secretion in rats

International audienc