References of "De Pauw-Gillet, Marie-Claire"
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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 detailPotential diagnostic biomarkers of Ulcerative colitis-associated colorectal dysplasia
Merli, Angela-Maria ULiege; MASSOT, Charlotte ULiege; BLETARD, Noëlla ULiege et al

in Acta Gastro-Enterologica Belgica (2018)

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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 detailProteomic differential distribution of 53BP1 in serrated and conventional adenomas validated by histological characterisation
QUESADA-CALVO, Florence ULiege; Merli, Angela-Maria ULiege; MASSOT, Charlotte ULiege et al

Poster (2017, February 10)

INTRODUCTION: Sessile serrated adenoma/polyp (SSA/p) is a precancerous lesion, mostly located in the right side of the colon (cecum, ascending and transverse colon). The difficulty is to visualize this ... [more ▼]

INTRODUCTION: Sessile serrated adenoma/polyp (SSA/p) is a precancerous lesion, mostly located in the right side of the colon (cecum, ascending and transverse colon). The difficulty is to visualize this lesion during colonoscopy because of its subtle appearance. MATERIAL AND METHOD: We compared proteomes of serrated polyps (SSA/p) and conventional adenomas using residual human formalin fixed paraffin embedded (FFPE) samples. FFPE-FASP method was applied on samples before label free proteomic analysis. Immunohistochemistry (IHC) characterisation of one candidate marker was performed for tissue validation on an independent set of samples including: conventional adenomas (low and high-grade dysplasia), serrated polyps (hyperplastic polyps, SSA/p and traditional serrated adenoma) and finally normal colon (taken at the margin of colorectal cancer (CRC) or of diverticular disease). RESULTS: Proteomics provided 765 proteins (out of 5992 proteins identified) significantly discriminating conventional adenomas from serrated lesions. We selected 53BP1 (Tumor suppressor p53-binding protein 1) among these for IHC validation, because of its tumor suppressor gene function and role as a mediator of DNA damage checkpoint. 53BP1 appeared significantly up-regulated in proteomes of low and high grade adenomas compared to these of normal tissue and SSA/p. 53BP1 IHC signal was located in the nucleus and the percentage of positive nucleus decreased in serrated polyps, especially in crypts and in the border epithelium, confirming part of the proteomic results. CONCLUSION: This study highlights potential marker proteins, including 53BP1 from which IHC signal was strongly decreased in some serrated polyps. The loss of 53BP1 has been associated with tumour progression and poor prognosis, while little is currently known about its involvement in precancerous CRC lesions. 53BP1 decrease of expression in the nucleus and therefore possible loss of function in some epithelial cells could reflect important changes occurring during dysplasia to neoplasia progression in serrated lesions. [less ▲]

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See detailIdentification of proteins discriminating inflammation induced dysplasia from simple inflammation in ulcerative colitis by laser capture microdissection and label free proteomics – a pilot study
Merli, Angela-Maria ULiege; QUESADA-CALVO, Florence ULiege; MASSOT, Charlotte ULiege et al

Conference (2017, February 09)

Chronic colonic inflammation in ulcerative colitis (UC) may induce dysplasia, which can itself progress and transform into neoplasia. Diagnosis of dysplasia in UC remains difficult particularly when ... [more ▼]

Chronic colonic inflammation in ulcerative colitis (UC) may induce dysplasia, which can itself progress and transform into neoplasia. Diagnosis of dysplasia in UC remains difficult particularly when tissue inflammation is present. The aim of this retrospective pilot study was to highlight proteins specifically associated with inflammation induced dysplasia in UC. We performed a pilot experiment on 15 Formalin-Fixed, Paraffin-Embedded (FFPE) samples isolated from 5 cases of UC patients with a Polypoïd Pedunculated dysplasia (UC-PP). We compared the proteomes of the UC-PP, the inflammatory (UC-I) and the normal (UC-NL) tissues of each patient. We performed Laser Capture Microdissection (LCM) in order to collect only epithelial cells, avoiding inflammatory infiltrating ones. Label free proteomic analysis using a 2D-nanoUPLC coupled with a hybrid Quadrupole-Orbitrap was applied, as well as differential analysis on the paired samples. Immunohistochemistry (IHC) characterisation of one of the selected proteins of interest was used for validation. Out of 985 quantified proteins, 7 were found significantly more abundant in UC-PP compared to UC-I tissues, with 6 being only detected in UC-PP using proteomics. One of these is Solute Carrier Family 12 member 2 (SLC12A2), also known as Na-K-2Cl co-transporter 1 (NKCC1), a protein involved in ionic balance, in T-cell migration promotion and in some features involved in cancer development like proliferation, migration or invasion. IHC results obtained were in correlation with proteomic results and showed that SLC12A2 was more abundant in UC-PP tissue than in UC-I and UC-NL tissues, with a signal clearly delimiting the dysplastic region from the surrounding inflammatory tissue. This pilot experiment shows a different proteomic profile in inflammation-associated dysplasia and simple inflammation. This should be replicated using other types of dysplasia in IBD. SLC12A2 could be a potential biomarker of inflammation-associated dysplasia. [less ▲]

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See detailIdentification of proteins discriminating inflammation induced dysplasia from simple inflammation in ulcerative colitis by laser capture microdissection and label free proteomics – a pilot study
Merli, Angela-Maria ULiege; QUESADA-CALVO, Florence ULiege; MASSOT, Charlotte ULiege et al

Poster (2017, February 01)

Chronic colonic inflammation in ulcerative colitis (UC) may induce dysplasia, which can itself progress and transform into neoplasia. Diagnosis of dysplasia in UC remains difficult particularly when ... [more ▼]

Chronic colonic inflammation in ulcerative colitis (UC) may induce dysplasia, which can itself progress and transform into neoplasia. Diagnosis of dysplasia in UC remains difficult particularly when tissue inflammation is present. The aim of this retrospective pilot study was to highlight proteins specifically associated with inflammation induced dysplasia in UC. We performed a pilot experiment on 15 Formalin-Fixed, Paraffin-Embedded (FFPE) samples isolated from 5 cases of UC patients with a Polypoïd Pedunculated dysplasia (UC-PP). We compared the proteomes of the UC-PP, the inflammatory (UC-I) and the normal (UC-NL) tissues of each patient. We performed Laser Capture Microdissection (LCM) in order to collect only epithelial cells, avoiding inflammatory infiltrating ones. Label free proteomic analysis using a 2D-nanoUPLC coupled with a hybrid Quadrupole-Orbitrap was applied, as well as differential analysis on the paired samples. Immunohistochemistry (IHC) characterisation of one of the selected proteins of interest was used for validation. Out of 985 quantified proteins, 7 were found significantly more abundant in UC-PP compared to UC-I tissues, with 6 being only detected in UC-PP using proteomics. One of these is Solute Carrier Family 12 member 2 (SLC12A2), also known as Na-K-2Cl co-transporter 1 (NKCC1), a protein involved in ionic balance, in T-cell migration promotion and in some features involved in cancer development like proliferation, migration or invasion. IHC results obtained were in correlation with proteomic results and showed that SLC12A2 was more abundant in UC-PP tissue than in UC-I and UC-NL tissues, with a signal clearly delimiting the dysplastic region from the surrounding inflammatory tissue. This pilot experiment shows a different proteomic profile in inflammation-associated dysplasia and simple inflammation. This should be replicated using other types of dysplasia in IBD. SLC12A2 could be a potential biomarker of inflammation-associated dysplasia. [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 detailBiocompatibility of polymer-infiltrated-ceramic-network (PICN) materials with Human Gingival Fibroblasts (HGFs).
GRENADE, Charlotte ULiege; De Pauw-Gillet, Marie-Claire ULiege; Gailly, Patrick ULiege et al

in Dental Materials (2016), 32(9), 1152-1164

OBJECTIVES: Polymer-infiltrated-ceramic-network (PICN) materials constitute an innovative class of CAD-CAM materials offering promising perspectives in prosthodontics, but no data are available in the ... [more ▼]

OBJECTIVES: Polymer-infiltrated-ceramic-network (PICN) materials constitute an innovative class of CAD-CAM materials offering promising perspectives in prosthodontics, but no data are available in the literature regarding their biological properties. The objective of the present study was to evaluate the in vitro biocompatibility of PICNs with human gingival fibroblasts (HGFs) in comparison with materials typically used for implant prostheses and abutments. METHODS: HGF attachment, proliferation and spreading on discs made of PICN, grade V titanium (Ti), yttrium zirconia (Zi), lithium disilicate glass-ceramic (eM) and polytetrafluoroethylene (negative control), were evaluated using a specific insert-based culture system (IBS-R). Sample surface properties were characterized by XPS, contact angle measurement, profilometry and SEM. RESULTS: Ti and Zi gave the best results regarding HGF viability, morphology, number and coverage increase with time in comparison with the negative control, while PICN and eM gave intermediate results, cell spreading being comparable for PICN, Ti, Zi and eM. Despite the presence of polymers and their related hydrophobicity, PICN exhibited comparable results to glass-ceramic materials, which could be explained by the mode of polymerization of the monomers. SIGNIFICANCE: The results of the present study confirm that the currently employed materials, i.e. Ti and Zi, can be considered to be the gold standard of materials in terms of HGF behavior, while PICN gave intermediate results comparable to eM. The impact of the present in vitro results needs to be further investigated clinically, particularly in the view of the utilization of PICNs for prostheses on bone-level implants. [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 detailA new method using insert-based systems (IBS) to improve cell behavior study on flexible and rigid biomaterials.
GRENADE, Charlotte ULiege; Moniotte, Nicolas; Rompen, Eric ULiege et al

in Cytotechnology (2016)

In vitro studies about biomaterials biological properties are essential screening tests. Yet cell cultures encounter difficulties related to cell retention on material surface or to the observation of ... [more ▼]

In vitro studies about biomaterials biological properties are essential screening tests. Yet cell cultures encounter difficulties related to cell retention on material surface or to the observation of both faces of permeable materials. The objective of the present study was to develop a reliable in vitro method to study cell behavior on rigid and flexible/permeable biomaterials elaborating two specific insert-based systems (IBS-R and IBS-F respectively). IBS-R was designed as a specific cylindrical polytetrafluoroethylene (PTFE) system to evaluate attachment, proliferation and morphology of human gingival fibroblasts (HGFs) on grade V titanium and lithium disilicate glass-ceramic discs characteristics of dental prostheses. The number of cells, their covering on discs and their morphology were determined from MTS assays and microscopic fluorescent images after 24, 48 and 72 h. IBS-F was developed as a two components system to study HGFs behavior on guided bone regeneration polyester membranes. The viability and the membrane barrier effect were evaluated by metabolic MTS assays and by scanning electron microscopy. IBS-R and IBS-F were shown to promote (1) easy and rapid handling; (2) cell retention on biomaterial surface; (3) accurate evaluation of the cellular proliferation, spreading and viability; (4) use of non-toxic material. Moreover IBS-F allowed the study of the cell migration through degradable membranes, with an access to both faces of the biomaterial and to the bottom of culture wells for medium changing. [less ▲]

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See detailEffects of Methylmercury on Harbour Seal Peripheral Blood Leucocytes In Vitro Studied by Electron Microscopy
Dupont, Aurélie; De Pauw-Gillet, Marie-Claire ULiege; Schnitzler, Joseph ULiege et al

in Archives of Environmental Contamination and Toxicology (2016), 70(1), 133-142

Methylmercury MeHg is highly immunotoxic and can alter the health status of the harbour seal, Phoca vitulina, from the North Sea. To investigate the mechanism of MeHg-induced toxicity in harbour seal ... [more ▼]

Methylmercury MeHg is highly immunotoxic and can alter the health status of the harbour seal, Phoca vitulina, from the North Sea. To investigate the mechanism of MeHg-induced toxicity in harbour seal lymphocytes, ConA-stimulated peripheral blood leucocytes were exposed in vitro to sub-lethal concentrations of MeHgCl (0.2, 1 and 2 µM) for 72h, and then analysed for their viability and ultrastructure. After 72h of incubation, cells were counted with a propidium iodide staining technique, a metabolic MTS assay was performed, and cells exposed to 1 µM MeHgCl were observed by transmission electron microscopy (TEM). Alive cell numbers decreased with increased MeHgCl concentrations. In presence of ConA and 1µM of MeHgCl, TEM images revealed a higher frequency of apoptotic cells. Exposed cells displayed condensation of the chromatin at the nuclear membrane and mitochondrial damages. The results suggest that in vitro MeHgCl induced apoptosis in harbour seal lymphocytes through a mitochondrial pathway. [less ▲]

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See detailAbsence of selenium protection against methylmercury toxicity in harbour seal leucocytes in vitro
Das, Krishna ULiege; Dupont, Aurélie; De Pauw-Gillet, Marie-Claire ULiege et al

in Marine Pollution Bulletin (2016), 108

Previous studies described high concentrations of mercury (Hg) and selenium (Se) in the blood of harbour seals, Phoca vitulina from the North Sea. In the present study, we evaluated the in vitro potential ... [more ▼]

Previous studies described high concentrations of mercury (Hg) and selenium (Se) in the blood of harbour seals, Phoca vitulina from the North Sea. In the present study, we evaluated the in vitro potential protective effects of sodium selenite (Na2SeO3) and selenomethionine (SeMet) on cell proliferation of harbour seal lymphocytes exposed to MeHgCl 0.75 µM. In vitro exposure of ConA-stimulated T lymphocytes resulted in severe inhibition of DNA synthesis, likely linked to severe loss of mitochondrial membrane potential at 0.75 µM. Neither selenite nor SeMet showed a protective effect against MeHg toxicity expressed at the T lymphocyte proliferation level for harbour seals. Selenite and SeMet did not show negative effects regarding lymphocyte proliferation and mitochondrial membrane potential. To conclude, our results clearly demonstrated that MeHg affected in vitro immune cells exposure with no protective effects of selenium at a molar ratio Hg:Se of 1:10 in harbour seals from the North Sea. [less ▲]

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See detailEffet inflammatoire des extraits de Cupressus sempervirens planté dans l’Est de l’Algérie : du modèle murin à la stimulation des cellules BEAS-2B par l’allergène majeur Cup s 1
Bouguenoun, Imene; De Pauw-Gillet, Marie-Claire ULiege; Bensakhri, Z et al

in Revue Française d'Allergologie (2016), 56

Cupressus sempervirens est l’une des espèces les plus répandues en Algérie ayant une capacité allergénique très élevée. Nous avons étudié,d’abord, comment l’instillation nasale des différentes fractions ... [more ▼]

Cupressus sempervirens est l’une des espèces les plus répandues en Algérie ayant une capacité allergénique très élevée. Nous avons étudié,d’abord, comment l’instillation nasale des différentes fractions de l’extrait pollinique de cette espèce peut moduler la réponse immunitaire chez les souris, et, ensuite, la stimulation des cellules épithéliales bronchiques humaines (BEAS-2B) par l’allergène majeur, identifié par spectrométrie de masse et purifié par chromatographie échangeuse d’ions. La sensibilisation des souris a révélé, d’une part, une diminution du nombre de neutrophiles et une augmentation des éosinophiles, et, d’autre part, une augmentation significative du total cellulaire dans le liquide du lavage bronchoalvéolaire.L’étude histologique des poumons a montré une présence d’infiltrat inflammatoire et d’oedème péri-bronchique. La spectrométrie de masse nous a permis de mettre en évidence la présence de l’allergène majeur Cup s 1 dans toutes les fractions avec une présence spécifique du Cup s 3 et Cupa 1 dans la fraction 4. Cet allergène avait le potentiel pour stimuler la libération de l’IL-8 et l’IL-6 par la ligné cellulaire, BEAS-2B, in vitro d’une manière dose-dépendante [less ▲]

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