Personne : Laroche, Gaétan
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Université Laval. Département de génie des mines, de la métallurgie et des matériaux
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- PublicationAccès libreRapid nucleation of iron oxide nanoclusters in aqueous solution by plasma electrochemistry(ACS Publications, 2015-06-18) Turgeon, Stéphane; Fortin, Marc-André; Laroche, Gaétan; Sarra-Bournet, Christian; Lagueux, Jean; Létourneau, Mathieu; Chevallier, Pascale; Laprise-Pelletier, Myriam; Bouchard, MathieuProgresses in cold atmospheric plasma technologies have made possible the synthesis of nanoparticles in aqueous solutions using plasma electrochemistry principles. In this contribution, a reactor based on microhollow cathodes and operating at atmospheric pressure was developed to synthesize iron-based nanoclusters (nanoparticles). Argon plasma discharges are generated at the tip of the microhollow cathodes, which are placed near the surface of an aqueous solution containing iron salts (FeCl₂ and FeCl₃) and surfactants (biocompatible dextran). Upon reaction at the plasma−liquid interface, reduction processes occur and lead to the nucleation of ultrasmall iron-based nanoclusters (IONCs). The purified IONCs were investigated by XPS and FTIR, which confirmed that the nucleated clusters contain a highly hydrated form of iron oxide, close to the stoichiometric constituents of α-FeOOH (goethite) or Fe₅O₃(OH)₉ (ferrihydrite). Relaxivity values of r₁ = 0.40 mM−¹ s−¹ and r₂/r₁ = 1.35 were measured (at 1.41 T); these are intermediate values between the relaxometric properties of superparamagnetic iron oxide nanoparticles used in medicine (USPIO) and those of ferritin, an endogenous contrast agent. Plasma-synthesized IONCs were injected into the mouse model and provided positive vascular signal enhancement in T₁-w. MRI for a period of 10−20 min. Indications of rapid and strong elimination through the urinary and gastrointestinal tracts were also found. This study is the first to report on the development of a compact reactor suitable for the synthesis of MRI iron-based contrast media solutions, on site and upon demand.
- PublicationAccès libreAtmospheric pressure cold plasma versus wet-chemical surface treatments for carboxyl functionalization of polylactic acid : a first step toward the immobilization of bioactive molecules(Elsevier, 2020-02-08) Laroche, Gaétan; Rodríguez Durán, Iván; Vanslambrouck, Stéphanie; Chevallier, PascaleThe use of polylactic acid (PLA) has attracted growing interest, particularly in recent years, for biomedical applications because of its mechanical properties, biocompatibility, and biodegradability. Despite this, features such as surface hydrophobicity and the absence of suitable functional groups for covalent immobilization of bioactive molecules, make it challenging to endow PLA-based medical devices with additional features and thus broaden their range of applicability. In the present study, we demonstrate the suitability of atmospheric pressure dielectric barrier discharges operating in the Townsend regime as a promising alternative to other surface treatments, such as diazonium and alkali hydrolytic treatments, for carboxyl functionalization of PLA. Chemical changes in PLA surfaces are evaluated by contact angle measurements and by X-ray photoelectron spectroscopy while physical changes are investigated by scanning electron microscopy and atomic force microscopy. The amount of carboxyl groups generated on PLA surfaces is assessed by toluidine blue O assay and substantiated by grafting, through carboxyl groups, a fluorescent probe containing amino functionalities. All of the surface treatments have proven to be very effective in generating carboxylic groups on the PLA surface. Nevertheless, plasma treatment is shown to not degrade the PLA surface, in sharp contrast with diazonium and alkali hydrolytic treatments.
- PublicationAccès libreSynthesis, characterization, and functionalization of ZnO nanoparticles by N-(trimethoxysilylpropyl) ethylenediamine triacetic acid (TMSEDTA) : investigation of the interactions between Phloroglucinol and ZnO@TMSEDTA(King Saud University, 2016-05-14) Barrak, Haythem; Laroche, Gaétan; Saied, Taieb; Chevallier, Pascale; M’nif, Adel; Hamzaoui, Ahmed HichemThe use of semiconductor oxides, such as chemical or biological sensors, requires their functionalization with appropriate molecules displaying specific interaction with the substance to be detected. Generally, the support materials used are TiO₂ or SiO₂. In the present work, zinc oxide nanoparticles (ZnO NPs), known for its reactivity and high specific area, were used. The synthesis of nanoscale ZnO was advantageously performed by precipitation at low temperature (60 °C). To our knowledge, it was the first time that this material was synthesized at such a low temperature, therefore lowering production cost. Moreover, the surface functionalization of ZnO was performed with N-(trimethoxysilylpropyl) ethylenediamine triacetic acid (TMSEDTA) in ethanol. This allowed shortening the functionalization reaction duration as compared to previously published literaturein the field. The samples obtained were analyzed by XRD, TEM, DLS, FTIR, TGA and XPS, which all concur with the successful synthesis of ZnO nanoparticles as well as the efficiency of TMSEDTA grafting on ZnO. Then, the interactions of this functionalized material, ZnO@TMSEDTA, with the Phloroglucinol (drug) were evaluated by using cyclic voltammetry measurements in solution. The cyclic voltammograms showed an intense cathodic peak which was correlated to the initial concentration of free Phloroglucinol. This cathodic peak was degraded upon addition of ZnO@TMSEDTA particles due to the drug interactions with free available carboxylic groups on the functionalized NPs. Based on a calibration curve, the drug concentration uptake can be therefore quantified. Thus, these results establish a big step to develop a Phloroglucinol sensor.
- PublicationAccès libreEffect of linking arm hydrophilic/hydrophobic nature, length and end-group on the conformation and the RGD accessibility of surface-immobilized fibronectin(Elsevier, 2019-10-30) Laroche, Gaétan; Vanslambrouck, Stéphanie; Guay-Bégin, Andrée-Anne; Chevallier, PascaleIn order to stimulate the cellular response to implant materials, extracellular matrix (ECM) proteins, such as collagen and fibronectin (FN), are immobilized on the implant surface. Amongst all ECM proteins used for biomimetic materials for medical applications, FN is one of the most investigated proteins thanks to its ability to promote cell adhesion and its contribution to important physiological processes. However, its conformation and hence its bioactivity strongly depend on the hydrophilic/hydrophobic nature of the surface as well as on immobilization strategies. This work investigates the effect of these two parameters, as well as the effect of the crosslinker length. FN was grafted onto silicon wafers using eights different linking arms presenting different lengths, hydrophilic/hydrophobic characters and binding sites. The protein was linked through either its amino groups (lysine amino acids) or sulfhydryl functionalities (cysteine amino acids). The grafting of each crosslinker and subsequent FN conjugation onto the surfaces was evidenced by X-ray photoelectron spectroscopy, while the surface hydrophilicity was determined by contact angle measurements. Moreover, atomic force microscopy images revealed that the conformation of surface conjugated FN only depends on the hydrophilicity of the linking arm. The FN conformation was also probed by enzyme-linked immunosorbent assays (ELISA). ELISA data demonstrated that all of the three investigated parameters linking arm parameter (length, hydrophobic/hydrophilic character, and terminal end-group) somewhat influence the RGD accessibility.
- PublicationAccès libreInterplay of geometric cues and RGD/BMP-2 crosstalk in directing stem cell fate(American Chemical Society, 2017-08-21) Bilem, Ibrahim; Laroche, Gaétan; Plawinski, Laurent; Chevallier, Pascale; Sone, E. (Eli); Durrieu, Marie-ChristineWithin the native microenvironment, extracellular matrix (ECM) components are thought to display a complex and heterogeneous distribution, spanning several length scales. Herein, the objective is to mimic, in vitro, the hierarchical organization of proteins and growth factors as well as their crosstalk. Photolithography technique was used to adjacently pattern geometrically defined regions of RGD and BMP-2 mimetic peptides onto glass substrates. These ECM-derived ligands are known to jointly regulate mesenchymal stem cells (MSCs) osteogenic differentiation. By manipulating the spatial distribution of dually grafted peptides, the extent of human MSCs osteogenic differentiation was significantly affected, depending on the shape of peptide micropatterns. Our data highlight the existence of a strong interplay between geometric cues and biochemical signals. Such in vitro systems provide a valuable tool to investigate mechanisms by which multiple ECM cues overlap to regulate stem cell fate, thereby contributing to the design of bioinspired biomaterials for bone tissue engineering applications.
- PublicationAccès libreHuman saphenous vein endothelial cell adhesion and expansion on micropatterned polytetrafluoroethylene(Wiley, 2012-08-31) Boivin, Marie-Claude; Laroche, Gaétan; Hoesli, Corinne A.; Lagueux, Jean; Bareille, Reine; Rémy-Zolghadri, Murielle; Chevallier, Pascale; Bordenave, Laurence; Durrieu, Marie-ChristineIntimal hyperplasia and thrombosis are responsible for the poor patency rates of small-diameter vascular grafts. These complications could be avoided by a rapid and strong adhesion of endothelial cells to the prosthetic surfaces, which typically consist of expanded polytetrafluoroethylene (PTFE) for small-diameter vessels. We have previously described two peptide micropatterning strategies that increase the endothelialization rates of PTFE. The micropatterns were generated either by inkjet printing 300 μm squares or by spraying 10.1 ± 0.1 μm diameter droplets of the CGRGDS cell adhesion peptide, while the remaining surface was functionalized using the CWQPPRARI cell migration peptide. We now directly compare these two micropatterning strategies and examine the effect of hydrodynamic stress on human saphenous vein endothelial cells grown on the patterned surfaces. No significant differences in cell adhesion were observed between the two micropatterning methods. When compared to unpatterned surfaces treated with a uniform mixture of the two peptides, the cell expansion was significantly higher on sprayed or printed surfaces after 9 days of static cell culture. In addition, after 6 h of exposure to hydrodynamic stress, the cell retention and cell cytoskeleton reorganization on the patterned surfaces was improved when compared to untreated or random treated surfaces. These results indicate that micropatterned surfaces lead to improved rates of PTFE endothelialization with higher resistance to hydrodynamic stress.
- PublicationAccès libreA fluorophore-tagged RGD peptide to control endothelial cell adhesion to micropatterned surfaces(ScienceDirect, 2013-10-31) Hoesli, Corinne A.; Duchesne, Carl; Juneau, Pierre-Marc; Laroche, Gaétan; Chevallier, PascaleThe long-term patency rates of vascular grafts and stents are limited by the lack of surface endothelialisation of the implanted materials. We have previously reported that GRGDS and WQPPRARI peptide micropatterns increase the endothelialisation of prosthetic materials in vitro. To investigate the mechanisms by which the peptide micropatterns affect endothelial cell adhesion and proliferation, a TAMRA fluorophore-tagged RGD peptide was designed. Live cell imaging revealed that the micropatterned surfaces led to directional cell spreading dependent on the location of the RGD-TAMRA spots. Focal adhesions formed within 3 h on the micropatterned surfaces near RGD-TAMRA spot edges, as expected for cell regions experiencing high tension. Similar levels of focal adhesion kinase phosphorylation were observed after 3 h on the micropatterned surfaces and on surfaces treated with RGD-TAMRA alone, suggesting that partial RGD surface coverage is sufficient to elicit integrin signaling. Lastly, endothelial cell expansion was achieved in serum-free conditions on gelatin-coated, RGD-TAMRA treated or micropatterned surfaces. These results show that these peptide micropatterns mainly impacted cell adhesion kinetics rather than cell proliferation. This insight will be useful for the optimization of micropatterning strategies to improve vascular biomaterials.
- PublicationAccès libreFibronectin grafting to enhance skin sealing around transcutaneous titanium implant(John Wiley & Sons, 2021-04-30) Bilem, Ibrahim; Ghadhab, Souhaila; Ruel, Jean; Laroche, Gaétan; Auger, François A.; Guay-Bégin, Andrée-Anne; Pauthe, Emmanuel; Chevallier, PascaleIntraosseous transcutaneous amputation prosthesis is a new approach in orthopedic implants that overcomes socket prosthesis problems. Its long-term performance requires a tight skin-implant seal to prevent infections. In this study, fibronectin (Fn), a widely used adhesion protein, was adsorbed or grafted onto titanium alloy. Fn grafting was performed using two different linking arms, dopamine/glutaric anhydride or phosphonate. The characterization of Fn-modified surfaces showed that Fn grating via phosphonate has led to the highest amount of Fn cell-binding site (RGD, arginine, glycine, and aspartate) available on the surface. Interestingly, cell culture studies revealed a strong correlation between the amount of available RGD ligands and cellular behavior, since enhanced proliferation and spreading of fibroblasts were noticed on Fn-grafted surfaces via phosphonate. In addition, an original in vitro mechanical test, inspired from the real situation, to better predict clinical outcomes after implant insertion, has been developed. Tensile test data showed that the adhesion strength of a bio-engineered dermal tissue was significantly higher around Fn-grafted surfaces via phosphonate, as compared to untreated surfaces. This study sheds light on the importance of an appropriate selection of the linking arm to tightly control the spatial conformation of biomolecules on the material surface, and consequently cell interactions at the interface tissue/implant.
- PublicationAccès libreRGD and BMP-2 mimetic peptides crosstalk enhances osteogenic commitment of human bone marrow stem cells(Elsevier, 2016-03-18) Bilem, Ibrahim; Laroche, Gaétan; Plawinski, Laurent; Chevallier, Pascale; Stone, E.; Durrieu, Marie-ChristineHuman bone marrow mesenchymal stem cells (hBMSCs) commitment and differentiation are dictated by bioactive molecules sequestered within their Extra Cellular Matrix (ECM). One common approach to mimic the physiological environment is to functionalize biomaterial surfaces with ECM-derived peptides able to recruit stem cells and trigger their linage-specific differentiation. The objective of this work was to investigate combinatorial effects of RGD and BMP-2 mimetic peptides on the osteogenic commitment of hBMSCs, without supplementing the media with pro-osteogenic factors. The RGD peptide promotes cell adhesion via cell transmembrane integrin receptors, while the BMP-2 peptide, corresponding to residues 73-92 of Bone Morphogenetic Protein-2, was shown to induce hBMSCs osteoblast differentiation. The immobilization of peptides on aminated glass was ascertained by X-ray Photoelectron Spectroscopy (XPS), the density of grafted peptides was quantified by fluorescence microscopy and the surface roughness was evaluated using Atomic Force Microscopy (AFM). The osteogenic commitment of hBMSCs cultured on RGD and/or BMP-2 surfaces was characterized by immunohistochemistry using STRO-1 as specific stem cells marker and Runx-2 as an earlier osteogenic marker. Biological results showed that the osteogenic commitment of hBMSCs was enhanced on bifunctionalized surfaces as compared to surfaces containing BMP-2, while on RGD surfaces cells mainly preserved their stemness character. These results demonstrated that RGD and BMP-2 mimetic peptides act synergistically to enhance hBMSCs osteogenesis without supplementing the media with osteogenic factors. These findings contribute to the development of biomimetic materials, allowing a deeper understanding of signaling pathways that govern the transition of stem cells towards the osteoblastic lineage.
- PublicationAccès libreGrafting of a model protein on lactide and caprolactone based biodegradable films for biomedical applications(Taylor & Francis, 2014-01-23) Larrañaga, Aitor; Laroche, Gaétan; Guay-Bégin, Andrée-Anne; Chevallier, Pascale; Sabbatier, Gad; Fernández, Jorge; Sarasua, José-RamónThermoplastic biodegradable polymers displaying elastomeric behavior and mechanical consistency are greatly appreciated for the regeneration of soft tissues and for various medical devices. However, while the selection of a suitable base material is determined by mechanical and biodegradation considerations, it is the surface properties of the biomaterial that are responsible for the biological response. In order to improve the interaction with cells and modulate their behavior, biologically active molecules can be incorporated onto the surface of the material. With this aim, the surface of a lactide and caprolactone based biodegradable elastomeric terpolymer was modified in two stages. First, the biodegradable polymer surface was aminated by atmospheric pressure plasma treatment and second a crosslinker was grafted in order to covalently bind the biomolecule. In this study, albumin was used as a model protein. According to X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM), albumin was efficiently immobilized on the surface of the terpolymer, the degree of albumin surface coverage (ΓBSA) reached ~35%. Moreover, gel permeation chromatography (GPC) studies showed that the hydrolytic degradation kinetic of the synthesized polymer was slightly delayed when albumin was grafted. However, the degradation process in the bulk of the material was unaffected, as demonstrated by Fourier transform infrared (FTIR) analyses. Furthermore, XPS analyses showed that the protein was still present on the surface after 28 days of degradation, meaning that the surface modification was stable, and that there had been enough time for the biological environment to interact with the modified material.