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See detailDevelopment of bimetallic nanoparticles for Surface-Assisted Laser Desorption/Ionisation Mass Spectrometry imaging of small molecules
Verdin, Alexandre ULiege; Malherbe, Cédric ULiege; Bertrand, Virginie ULiege et al

Poster (2019, June 04)

Mass Spectrometry Imaging (MSI) allows the simultaneous visualisation of hundreds to thousands of molecules in biological tissue slides enabling the comprehension of complex biochemical processes ... [more ▼]

Mass Spectrometry Imaging (MSI) allows the simultaneous visualisation of hundreds to thousands of molecules in biological tissue slides enabling the comprehension of complex biochemical processes. Additionally, the analysis of untargeted MSI data with advanced statistical tools has the potential to reveal pertinent features of the sample such as the progression or outcome of a disease or the response to a treatment. Recently, metallic nanoparticles (NPs) have received increasing attention as desorption/ionization substrate for Surface-Assisted Laser Desorption/Ionization-MS (SALDI-MS). Metallic NPs provide several advantages over organic MALDI matrices for the analysis of small molecules: high absorption coefficients of the ionization laser (in the UV-Visible range), the ability to load small molecules on their surface by physical or chemical adsorption, little interferences in the low m/z range and a simple sample preparation (no crystallization required). SALDI-MSI is adapted to study the variations of the molecular composition associated with various diseases and to visualize the localization of pertinent small molecules in complex biological tissues. Innovative Au@Ag (core@shell) bimetallic NPs are interesting for SALDI applications, especially because controlling the thickness of the Ag shell around the Au core allows the tuning of the absorption in near-UV. These Au@Ag NPs permits to maximize the energy absorption for an efficient ionization and desorption of analytes adsorbed to the NPs. Moreover Au@Ag NPs can also be used for Surface-Enhanced Raman Spectroscopy (SERS) imaging which is a highly complementary technique. We report here a study of ovarian cancer tissue using Au@Ag NPs to characterize the overexpression of membrane receptor in the tumor by SERS imaging and to gain molecular insights by SALDI-MSI. [less ▲]

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See detailDevelopment of bimetallic nanoparticles for Surface-Assisted Laser Desorption/Ionisation Mass Spectrometry imaging of small molecules
Verdin, Alexandre ULiege; Malherbe, Cédric ULiege; Bertrand, Virginie ULiege et al

Poster (2019)

Metallic nanoparticles (NPs) receive increasing attention as desorption/ionization substrate for Surface-Assisted Laser Desorption Ionization-MS (SALDI-MS) [1]. Metallic NPs offer high extinction ... [more ▼]

Metallic nanoparticles (NPs) receive increasing attention as desorption/ionization substrate for Surface-Assisted Laser Desorption Ionization-MS (SALDI-MS) [1]. Metallic NPs offer high extinction coefficients of the ionization laser (in the UV-Visible range of wavelengths) and are able to ionize small molecules with little interferences in the low m/z range with a simple sample preparation [2]. SALDI-MSI is adapted to study the variations of the molecular composition associated with various diseases and to visualize the localization of pertinent small molecules in complex biological tissues. Innovative Au@Ag (core@shell) bimetallic NPs are interesting for SALDI applications, especially because controlling the thickness of the Ag shell around the Au core allows the tuning of the absorption in near-UV. The Au@Ag NPs permits to vary the energy transfer for an efficient ionization and desorption of analytes adsorbed to the NPs. Moreover Au@Ag NPs can also also be used for Surface-Enhanced Raman Spectroscopy (SERS) imaging. We report here a study of ovarian cancer tissue using Au@Ag NPs to localize the tumor by SERS imaging and to gain molecular insight of tumors by SALDI-MSI. [less ▲]

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See detailMetallic nanoparticles: an interesting option for mass spectrometry imaging of small molecules in tissues
Verdin, Alexandre ULiege; Müller, Wendy ULiege; Bertrand, Virginie ULiege et al

Poster (2018, July 05)

MALDI mass spectrometry imaging is a central technique for the visualisation and identification of a wide variety of molecules in biological samples, especially in tissues slides. However, organic ... [more ▼]

MALDI mass spectrometry imaging is a central technique for the visualisation and identification of a wide variety of molecules in biological samples, especially in tissues slides. However, organic matrices used for the desorption and ionisation of molecules suffers from major limitations, particularly for the analysis of small molecules due to interferences from the matrix in the low m/z range. In this preliminary study we compared the performances of 9-aminoacridine, a well known organic matrix used for small molecules anaylsis, and metallic nanoparticles to evaluate the information obtained from both method. Ionisation by nanoparticles is known as Surface-Assisted Laser Desorption Ionisation (SALDI). Metallic nanoparticles seem to be a promising substrate for MS analysis of small compounds in tissues and reveals information that are complementary to those obtained by MALDI. [less ▲]

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See detailSmall molecule analysis on FFPE-TMA samples using MALDI-mass spectrometry imaging
Massonnet, Philippe ULiege; Kriegsmann, Mark; Smargiasso, Nicolas ULiege et al

Poster (2018, March 29)

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See detailDevelopment of highly sensitive SERS nanoprobes for cellular imaging of Folate Receptor alpha
Verdin, Alexandre ULiege; Malherbe, Cédric ULiege; Cambroisier, Florie ULiege et al

Poster (2018, March)

During the last decade, spectroscopic cellular imaging has increasingly become a powerful characterization method for the study of all kind of cellular-related disease. In this context, Surface-Enhanced ... [more ▼]

During the last decade, spectroscopic cellular imaging has increasingly become a powerful characterization method for the study of all kind of cellular-related disease. In this context, Surface-Enhanced Raman Spectroscopy (SERS) is a powerful technique with many advantages over other spectroscopic techniques such as fluorescence or infrared spectroscopy. The SERS effect is capable of providing huge enhancement factors, highly increasing the sensitivity of Raman spectroscopy. One interesting approach is to use so called “SERS nanoprobes” to selectively target and detect a particular receptor, protein, DNA sequence, etc. In the area of SERS detection of cancerous cells, the target of these nanoprobes is usually a surface receptor that is characteristic of the cancerous state of the cell, or that is overexpressed in a cancerous cell in comparison to a healthy situation. In this study, we investigated the targeting of the Folate Receptor alpha (FRα) which is overexpressed in several cancers (ovarian and lung adenocarcinoma, among others) and can therefore be used for the imaging and detection of cancerous cells. We focused on the development of highly sensitive SERS nanoprobes, combining bimetallic nanoparticles and resonant Raman-active molecules. We successfully imaged Folate Receptor α at the surface of two kinds of cancerous cells (KB and PC-3) thanks to the high confocality of the Raman micro-spectrometer. Moreover, we were able to distinguish these two kinds of cells by measuring the SERS intensity coming from each cell population, since both kinds of cells have a different expression level of the FRα receptor and will therefore accumulate different amounts of nanoprobes. Our approach provides new perspectives toward the discrimination of cancerous and healthy cells in real samples. [less ▲]

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See detailTD-GCxGC-HRTOFMS in biological applications 
Focant, Jean-François ULiege; Pesesse, Romain ULiege; Dubois, Lena ULiege et al

Conference (2017, September)

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See detailOLFM4, KNG1 and Sec24C identified by proteomics and immunohistochemistry as potential markers of early colorectal cancer stages
QUESADA-CALVO, Florence ULiege; MASSOT, Charlotte ULiege; Bertrand, Virginie ULiege et al

in Clinical Proteomics (2017), 24(9),

Abstract Background: Despite recent advances in colorectal cancer (CRC) diagnosis and population screening programs, the identification of patients with preneoplastic lesions or with early CRC stages ... [more ▼]

Abstract Background: Despite recent advances in colorectal cancer (CRC) diagnosis and population screening programs, the identification of patients with preneoplastic lesions or with early CRC stages remains challenging and is important for reducing CRC incidence and increasing patient’s survival. Methods: We analysed 76 colorectal tissue samples originated from early CRC stages, normal or inflamed mucosa by label-free proteomics. The characterisation of three selected biomarker candidates was performed by immunohisto‑ chemistry on an independent set of precancerous and cancerous lesions harbouring increasing CRC stages. Results: Out of 5258 proteins identified, we obtained 561 proteins with a significant differential distribution among groups of patients and controls. KNG1, OLFM4 and Sec24C distributions were validated in tissues and showed differ‑ ent expression levels especially in the two early CRC stages compared to normal and preneoplastic tissues. Conclusion: We highlighted three proteins that require further investigations to better characterise their role in early CRC carcinogenesis and their potential as early CRC markers. [less ▲]

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See detailBiocompatibility of polymer-infiltrated-ceramicnetwork (PICN) materials with Human Gingival Keratinocytes (HGKs)
GRENADE, Charlotte ULiege; De Pauw-Gillet, Marie-Claire ULiege; PIRARD, Catherine ULiege et al

in Dental Materials (2017), 33

Objective. Biocompatibility of polymer-infiltrated-ceramic-network (PICN) materials, a new class of CAD–CAM composites, is poorly explored in the literature, in particular, no data are available regarding ... [more ▼]

Objective. Biocompatibility of polymer-infiltrated-ceramic-network (PICN) materials, a new class of CAD–CAM composites, is poorly explored in the literature, in particular, no data are available regarding Human Gingival Keratinocytes (HGK). The first objective of this study was to evaluate the in vitro biocompatibility of PICNs with HGKs in comparison with other materials typically used for implant prostheses. The second objective was to correlate results with PICN monomer release and indirect cytotoxicity. Methods. HGK attachment, proliferation and spreading on PICN, grade V titanium (Ti), yttrium zirconia (Zi), lithium disilicate glass-ceramic (eM) and polytetrafluoroethylene (negative control) discs were evaluated using a specific insert-based culture system. For PICN and eM samples, monomer release in the culture medium was quantified by high performance liquid chromatography and indirect cytotoxicity tests were performed. Results. Ti and Zi exhibited the best results regarding HGK viability, number and coverage. eM showed inferior results while PICN showed statistically similar results to eM but also to Ti regarding cell number and to Ti and Zi regarding cell viability. No monomer release from PICN discs was found, nor indirect cytotoxicity, as for eM. Significance. The results confirmed the excellent behavior of Ti and Zi with gingival cells. Even if polymer based, PICN materials exhibited intermediate results between Ti–Zi and eM. These promising results could notably be explained by PICN high temperature–high pressure (HT–HP) innovative polymerization mode, as confirmed by the absence of monomer release and indirect cytotoxicity [less ▲]

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See detailPhotoreversibility and biocompatibility of polydimethylsiloxane-coumarin as adjustable intraocular lens material
Jellali, Rachid; Bertrand, Virginie ULiege; Alexandre, Michaël et al

in Macromolecular Bioscience (2017)

Polydimethylsiloxane (PDMS) constitutes an interesting material for a variety of biomed- ical applications, especially as intraocular lenses (IOLs), for its excellent transparency. In this work, a ... [more ▼]

Polydimethylsiloxane (PDMS) constitutes an interesting material for a variety of biomed- ical applications, especially as intraocular lenses (IOLs), for its excellent transparency. In this work, a photoreversible PDMS-coumarin network, whose shape and properties can be adjusted postoperatively in a noninvasive manner, is developed. The synthesis of PDMS-cou- marin is achieved by amidation of a coumarin acid chloride derivative with amine-function- alized PDMSs. Under exposure of λ > 300 nm, these polymers can be cured by dimerization of coumarin. The cured polymers can be uncrosslinked via photocleavage of cyclobutane dimers upon illumination at λ < 290 nm. The diffusion of linear PDMSs in a crosslinked network and the controlled shape modification are studied, which demonstrate that these polymers are good candidates for adjustable IOL application. IOL disks prepared from these materials show high hydrophobicity and good transparency. In vitro cytotoxicity, lens epithelial cell adhesion assays, and rabbit host reaction against implanted disks demonstrate the biocompatibility of the polymer. [less ▲]

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See detailGC×GC-(HR)TOFMS in Cancer Research
Pesesse, Romain ULiege; Stefanuto, Pierre-Hugues ULiege; Bertrand, Virginie ULiege et al

Conference (2016, May 30)

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See detailExploring the volatome of different cancer cell lines
Stefanuto, Pierre-Hugues ULiege; Pesesse, Romain ULiege; Bertrand, Virginie ULiege et al

in European Respiratory Journal (2015), 46(suppl 59), 3975

As reported by the American Cancer Society, lung cancers have the highest death rate, compared to other types of cancer. The main reason is the lack of early stage non-invasive screening methods. Indeed ... [more ▼]

As reported by the American Cancer Society, lung cancers have the highest death rate, compared to other types of cancer. The main reason is the lack of early stage non-invasive screening methods. Indeed, the death rate of the other most abundant cancers is decreasing due to the implementation of large-scale population screening techniques. It is believed that the death rate of lung cancer could be lowered by developing and implementing more efficient diagnostic methods1. A large number of diseases appear to have a volatile signature present in exhaled air. In practice, analytical breath profiling offers solutions for early detection of different kind of lung infections2. For example, cystic fibrosis patients show a different volatile organic compound (VOC) profile because of the bacterial colonization that localizes in their lungs. A similar behavior is observed for patient suffering from lung cancer. However, the list of recognized volatile biomarkers of lung cancer is still scarce and could be improved. In spite of this, the biological variability of exhaled air profile makes the biomarkers identification challenging. In the hope of contributing to a better understanding, we used comprehensive two-dimensional gas chromatography coupled to time of light mass spectrometry (GC×GC-TOFMS) to investigate the headspace of cancer cells. A procedure has been developed to sample cell culture headspace. After proper data treatment, univariate and multivariate statistics were compared for isolation of potential cancer biomarkers. Further tentative identification was also carried out by using high-accuracy MS analyzers as detection techniques. 1 Siegel, R. et al. Cancer statistics 2013 2 Di Francesco, F. et al. Microchemical Journal 2005 . [less ▲]

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See detailComparison of early stages of colorectal cancer by label free proteomics
QUESADA CALVO, Florence ULiege; MEUWIS, Marie-Alice ULiege; Bertrand, Virginie ULiege et al

in Acta Gastroenterologica (2015, February 27)

Introduction and objectives: Colorectal cancer (CRC) is the second most frequent cancer in women and the third in men. Identification of the mechanisms of progression in these early CRC stages is ... [more ▼]

Introduction and objectives: Colorectal cancer (CRC) is the second most frequent cancer in women and the third in men. Identification of the mechanisms of progression in these early CRC stages is important to develop new diagnostic and therapeutic tools. Formalin-Fixed Paraffin-Embedded (FFPE) specimens are materials that enable proteomic clinical research. Hence our aim was to address the comparison of FFPE samples from early CRC stages patients using shotgun proteomic analysis. Methods: We performed a retrospective study on 36 CRC tissue samples (pT1N0M0, n=16 and pT2N0M0, n=20) compared together and with 40 control tissue samples (20 patients with diverticulitis, using paired inflamed (DI) and healthy tissue (DH)). Each tissue slice was macrodissected to enrich in epithelial cells. We used FFPE-FASP kit (Expedeon) for sample preparation and protein digests were analyzed using 2D-nanoAquity UPLC separation online with Q-Tof Synapt HDMSTM G2 using ion mobility as additional separation. We performed protein identification and differential analysis using Progenesis QI for proteomics (Nonlinear Dynamics). Results and discussion: We selected 149 proteins differentially distributed between T1 and T2 CRC stages which were not significantly different between CRC and DH or DI. Only 30 proteins were significantly more abundant in T1 versus T2 and 119 were distributed inversely, with a minimum fold ratio of 2. Among those, ATP synthase subunit beta, Aspartate-tRNA ligase, Haptoglobin and Kininogen were identified. . Moreover, we validated Kininogen and 3 others proteins with a significant differential distribution between pT1N0M0 and pT2N0M0 stages by immunohistochemistry. Conclusion: This FFPE retrospective study comparing T1 and T2 CRC highlighted proteins already previously identified as potential CRC biomarkers. These proteins may reflect important early changes in cancer development and may help understanding early tumor progression. [less ▲]

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See detailRGD surface functionalization of the hydrophilic acrylic intraocular lens material to control posterior capsular opacification
Huang, Yi-Shiang ULiege; Bertrand, Virginie ULiege; Bozukova, Dimitriya et al

in PLoS ONE (2014), 9(12), 32

Posterior Capsular Opacification (PCO) is the capsule fibrosis developed on implanted IntraOcular Lens (IOL) by the de-differentiation of Lens Epithelial Cells (LECs) undergoing Epithelial Mesenchymal ... [more ▼]

Posterior Capsular Opacification (PCO) is the capsule fibrosis developed on implanted IntraOcular Lens (IOL) by the de-differentiation of Lens Epithelial Cells (LECs) undergoing Epithelial Mesenchymal Transition (EMT). Literature has shown that the incidence of PCO is multifactorial including the patient’s age or disease, surgical technique, and IOL design and material. Reports comparing hydrophilic and hydrophobic acrylic IOLs have shown that the former has more severe PCO. On the other hand, we have previously demonstrated that the adhesion of LECs is favored on hydrophobic compared to hydrophilic materials. By combining these two facts and contemporary knowledge in PCO development via the EMT pathway, we propose a biomimetically inspired strategy to promote LEC adhesion without de-differentiation to reduce the risk of PCO development. By surface grafting of a cell adhesion molecule (RGD peptide) onto the conventional hydrophilic acrylic IOL material, the surface-functionalized IOL can be used to reconstitute a capsule-LEC-IOL sandwich structure, which has been considered to prevent PCO formation in literature. Our results show that the innovative biomaterial improves LEC adhesion, while also exhibiting similar optical (light transmittance, optical bench) and mechanical (haptic compression force, IOL injection force) properties compared to the starting material. In addition, compared to the hydrophobic IOL material, our bioactive biomaterial exhibits similar abilities in LEC adhesion, morphology maintenance, and EMT biomarker expression, which is the crucial pathway to induce PCO. The in vitro assays suggest that this biomaterial has the potential to reduce the risk factor of PCO development. [less ▲]

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See detailEtude multiparamétrique de polymères acryliques, modèles de lentilles intraoculaires : recherche d'indicateurs de risque de développement de la cataracte secondaire.
Bertrand, Virginie ULiege

Doctoral thesis (2014)

In this study we compared 3 biomaterials (supplied by the firm PhysIOL , Science park, Liège, Belgium). We have at first to estimate certain physico-chemical properties such as the surface hydrophilicity ... [more ▼]

In this study we compared 3 biomaterials (supplied by the firm PhysIOL , Science park, Liège, Belgium). We have at first to estimate certain physico-chemical properties such as the surface hydrophilicity, the adhesion force (Atomic Force Microscopy). We were then interested in the bioadhesive character of these biomaterials by estimating in vitro, the adsorption of BSA and the cellular adhesion (lens epithelial cells, LECs), the ex vivo capsular adhesion and the in vivo tissular reaction (subcutaneous implant of polymers for 1 month (rabbit)). We then realized a proteomic analysis using mass spectrometry : LECs used during our in vitro tests, proteins adsorbed on biomaterials after incubation in a complex medium, the fibrous capsule surrounding the biomaterial after its subcutaneous implantation and the lens (MALDI imaging). [less ▲]

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See detailEvaluation of a class of polyurethane materials for intraocular lens manufacturing.
Bozukova, Dimitriya; Bertrand, Virginie ULiege; Pagnoulle, Christophe et al

in Journal of Biomedical Materials Research. Part B, Applied Biomaterials (2014)

Ophthalmic lenses are medical devices with considerable requirements in terms of optical, biomechanical and biological performance. There is limited number of materials used for their manufacturing ... [more ▼]

Ophthalmic lenses are medical devices with considerable requirements in terms of optical, biomechanical and biological performance. There is limited number of materials used for their manufacturing, comprising mainly silicones and poly(meth)acrylates. This series of publications aims at investigating the applicability of thermoplastic polyurethane elastomers (TPU) for the manufacturing of ophthalmic lenses and examining the properties of the respective devices. This study is related to the synthesis of TPUs with chemical compositions that comprise chemically grafted filters for the hazardous-light. GC-MS, attenuated total reflectance Fourier transform infrared spectroscopy, and UV-vis spectroscopies confirmed the reaction completion and the beneficial effect of the filters on the light transmittance, respectively. Relatively high refractive index of the material was measured and allows for the manufacturing of thinner lenses. The contrast sensitivity determined for a model intraocular lens (IOL) was satisfactory. Few optical defects were, however, present on the model lens prepared by thermoplastic injection molding. The elasticity of the materials was evaluated in view to their potential applicability as foldable IOLs by determining their glass transition temperature and their Young modulus and measuring their shore A. The TPU materials demonstrated more bioadhesive character compared with a benchmark hydrophilic acrylic reference material, which is already used for IOL manufacturing. (c) 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2014. [less ▲]

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See detailPolymer based intraocular lens adsorbome: a bottom up proteomic study
Bertrand, Virginie ULiege; Yi-Shiang, Huang; Mazzucchelli, Gabriel ULiege et al

Poster (2014, May 26)

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