Personne : Chevallier, Pascale
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Chevallier
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Pascale
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Université Laval. Département de génie des mines, de la métallurgie et des matériaux
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ncf11892659
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- PublicationAccès libreSingle or mixed tethered peptides to promote hMSC differentiation toward osteoblastic lineage(American Chemical Society, 2018-11-27) Padiolleau, Laurence; Chanseau, Christel; Laroche, Gaétan; Durrieu, Stephanie; Chevallier, Pascale; Durrieu, Marie-ChristineThe commitment and differentiation of human mesenchymal stem cells (hMSCs) are guided by bioactive molecules within the extracellular matrix. Among the various approaches to design biomaterials, the functionalization of biomaterial surfaces with peptides from the sequence of proteins from the extracellular matrix is quite common. The purpose of this functionalization is to recruit hMSCs and promote their differentiation into the appropriate lineage. The aim of this work was to investigate the influence of RGD and FHRRIKA peptides and peptide sequences taken from bone morphogenic protein (BMP-2) and histone H4 (osteogenic growth peptide; OGP) either tethered alone or as a mixture on the surface of a model material and to also examine the level of hMSC osteogenic commitment without using a differentiation medium. Grafting of the different peptides was assessed by X-ray photoelectron spectroscopy (XPS), while their surface density was quantified by fluorescence microscopy, and their surface properties were assessed by atomic force microscopy (AFM) and contact angle (CA). The osteogenic commitment of hMSCs cultured on the different surfaces was characterized by immunohistochemistry using Runx-2 as an earlier osteogenic marker and OPN, a late osteogenic marker, and by RT-qPCR through the expression of ColI-a1, Runx-2, and ALP. Biological results show that the osteogenic commitment of the hMSCs was increased on surfaces tethered with a mixture of peptides. Results indicate that tethered peptides in the range of pmol mm–2 were indeed effective in inducing a cellular response after 2 weeks of cell culture without using an osteogenic media. These findings contribute to the research efforts to design biomimetic materials able to induce a response in human stem cells through tethered bioactive molecules for bone tissue engineering.
- 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 libreMilkweed scaffold : a new candidate for bone cell growth(Taylor & Francis Online, 2019-06-17) Bilem, Ibrahim; Laroche, Gaétan; Naudé, Nicolas; Elkoun, S.; Chevallier, Pascale; Robert, Mathieu; Soulié, SimonThis study aims to evaluate the potential of milkweed as potential candidate to construct biodegradable scaffold for bone regeneration. A mat made of milkweed, polyethylene, and polypropylene was treated with an atmospheric pressure plasma to functionalize the surface of the polymer assembly with carboxylic acid groups, which enable to conjugate bioactive molecules, while accelerating the degradation of milkweed. Degradation tests demonstrated substantial decrease of the weight of the treated polymer mat as compared to untreated one. Biological assays revealed that the polymer assembly promoted preosteoblast MC3T3 cells recruitment with a significant enhancement observed on the RGD-grafted polymer mat.
- PublicationAccès libreEvaluating poly(Acrylamide-co-Acrylic Acid) hydrogels stress relaxation to direct the osteogenic differentiation of mesenchymal stem cells(Wiley, 2021-04-19) Prouvé, Émilie; Drouin, Bernard; Laroche, Gaétan; Rémy-Zolghadri, Murielle; Chevallier, Pascale; Durrieu, Marie-ChristineThe aim of this study is to investigate polyacrylamide-based hydrogels stress relaxation and the subsequent impact on the osteogenic differentiation of human mesenchymal stem cells (hMSCs). Different hydrogels are synthesized by varying the amount of cross-linker and the ratio between the monomers (acrylamide and acrylic acid), and characterized by compression tests. It has been found that hydrogels containing 18% of acrylic acid exhibit an average relaxation of 70%, while pure polyacrylamide gels show an average relaxation of 15%. Subsequently, hMSCs are cultured on two different hydrogels functionalized with a mimetic peptide of the bone morphogenetic protein-2 to enable cell adhesion and favor their osteogenic differentiation. Phalloidin staining shows that for a constant stiffness of 55 kPa, a hydrogel with a low relaxation (15%) leads to star-shaped cells, which is typical of osteocytes, while a hydrogel with a high relaxation (70%) presents cells with a polygonal shape characteristic of osteoblasts. Immunofluorescence labeling of E11, strongly expressed in early osteocytes, also shows a dramatically higher expression for cells cultured on the hydrogel with low relaxation (15%). These results clearly demonstrate that, by fine-tuning hydrogels stress relaxation, hMSCs differentiation can be directed toward osteoblasts, and even osteocytes, which is particularly rare in vitro.
- 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 libreHigh-sensitivity permeation analysis of ultra-small nanoparticles across the skin by positron emission tomography (PET)(American Chemical Society, 2021-03-10) Fortin, Marc-André; Omar, Mahmoud; Chevallier, Pascale; Tuduri, Ludovic; Laprise-Pelletier, MyriamUltrasmall nanoparticles (US-NPs; <20 nm in hydrodynamic size) are now included in a variety of pharmacological and cosmetic products, and new technologies are needed to detect at high sensitivity the passage of small doses of these products across biological barriers such as the skin. In this work, a diffusion cell adapted to positron emission tomography (PET), a highly sensitive imaging technology, was developed to measure the passage of gold NPs (AuNPs) in skin samples in continuous mode. US-AuNPs (3.2 nm diam.; TEM) were functionalized with deferoxamine (DFO) and radiolabeled with 89Zr(IV) (half-life: 3.3 days, matching the timeline of diffusion tests). The physicochemical properties of the functionalized US-AuNPs (US-AuNPs-PEG-DFO) were characterized by FTIR (DFO grafting; hydroxamate peaks: 1629.0 cm–1, 1569.0 cm–1), XPS (presence of the O═C–N C 1s peak of DFO at 287.49 eV), and TGA (organic mass fraction). The passage of US-AuNPs-PEG-DFO-89Zr(IV) in skin samples was measured by PET, and the diffusion parameters were extracted thereby. The signals of radioactive US-AuNPs-PEG-DFO-89Zr(IV) leaving the donor compartment, passing through the skin, and entering the acceptor compartment were detected in continuous at concentrations as low as 2.2 nM of Au. The high-sensitivity acquisitions performed in continuous allowed for the first time to extract the lag time to the start of permeation, the lag time to start of the steady state, the diffusion coefficients, and the influx data for AuNPs permeating into the skin. PET could represent a highly valuable tool for the development of nanoparticle-containing topical formulations of drugs and cosmetics.
- 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 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 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.
- PublicationAccès libreFibronectin-modified surfaces for evaluating the influence of cell adhesion on sensitivity of leukemic cells to siRNA nanoparticles(London Future Medicine, 2016-04-13) Valencia-Serna, Juliana; Laroche, Gaétan; Chevallier, Pascale; Bahadur K.C., Remant; Uludağ, HasanAim: This study aimed to create fibronectin (FN)-grafted polymeric surfaces to investigate the influence of leukemic cell adhesion on siRNA treatment. Materials & methods: FN was grafted on plasma-treated PTFE surfaces using chemical crosslinkers. Adhesion and growth of chronic myeloid leukemia K562 cells on modified surfaces were investigated. The silencing effect of siRNA/lipid-polymers nanoparticles on cells grown on FN-grafted surfaces was evaluated. Results: Crosslinker-mediated immobilization showed significant FN grafting on surfaces, which provided K562 cell adhesion and growth advantage. siRNA nanoparticle silencing was similarly effective on FN-adhered and suspension-growing K562 cells. Conclusion: This study provided initial data to develop a cell-adhesive system to investigate therapeutic effects on leukemic cells. The response of chronic myeloid leukemia cells to siRNA nanoparticles was independent on cell attachment.