VITO

Name of the infrastructure: VITO-TA

Location (town, country): Mol, BE

Web site address: www.vito.be; www.cardam.eu

Legal name of organisation operating the infrastructure: Vlaamse Instelling voor Technologisch Onderzoek(VITO NV)

Location of organisation (town, country): Mol, Belgium

 

Description of the Infrastructure

MATERIALS AND EXPERTISE: The key component of the VITO offer is its platform of alternative test methodsfor assessment of nanotoxicity, called the Centre for Advanced R&D on Alternative Methods (CARDAM;www.cardam.eu). CARDAM has class III facilities for tissue culture (primary cells and cell lines) and nanoparticleexposure, and test performance is according to GLP standards and/or Good Cell Culture Practice (GCCP)principles. Technical and scientific expertise of a variety of core facilities has been adapted for application tonanoparticles, such as –omics facilities and (eco)toxicity test facilities. In addition, VITO has state-of-the-artanalytical facilities and dedicated laboratories and workshops, distributed over two closely located sites. Theextensive collection of instruments and accumulation of expertise through engineers, physicists, materialscientists, chemists, pharmacists, cell and molecular biologists, all specialising in different aspects ofnanomaterials technology, characterization and risk assessment, is unique and rare in Europe by its size.VITO has an internationally recognised research base of more than 320 full time researchers and academicemployees, supported by some 200 curatorial and technical staff. Of the latter, 63 technical staff members willprovide key expertise to support Users when accessing VITO’s state-of-the-art analytical facilities. VITO hostssome 30 PhD students and post-doctoral fellows, and stimulates an outstanding research environment throughclose collaboration and exchange of expertise with universities and (inter)national research institutes. All Userswill be provided with individually-tailored training in the use of specific analytical facilities as a minor componentof their visit, ensuring Health and Safety regulations are adhered to.

 

SUPPORTING FACILITIES AND SERVICES:

VITO will be offering access to two components listed in theproposal, as follows:

 

Particle characterization:

Mass spectrometry and chemical analysis: a mass spectrometry facility covering the latest evolutions in ionoptics, ionisation sources and chromatographic hyphenation techniques is available. Of particular interest is agas chromatograph with a high-resolution mass spectrometer which allows for the detection and quantificationof GC amenable compounds at very low concentrations based on their accurate masses. For elemental andspeciation analysis a high-resolution ICP-MS, a quadrupole ICP-MS and 3 ICP-AES are available in VITO'slaboratory which can be coupled to HPLC. For direct element determination of solid material (e.g. fine dust) aWD-XRF, ED-XRF and μ-XRF are available.

 

Materials characterization and imaging facilities: encompasses a vast range of analytical equipment forthermogravimetric analysis, BET surface area and XPS surface analysis, size and zeta potential determination,depth sensing indentation, determination of thin coating adhesion, X-ray powder diffraction, and electron imagingvia FEG-SEM and electron probe microanalysis. Highlight is a high-resolution ‘in-lens’ Schottky field emissiongun electron microprobe analysis (FEG-EPMA) system with a cathodoluminescence detection system (expected2010). For preparation, processing and shaping of ceramic, metallic or other materials, it has extensivesupporting facilities at its disposal, including sieves, mixers (Z-blade, US, high shear,..), mills (ball mill, attritionmill, 3-roll-mill,..), semi-industrial spray drying equipment, furnaces (for calcining), (reaction-)sintering, either withoxidising, reducing or inert atmosphere up to temperatures of 2200°C), presses (uniaxial and CIP), surface andprofile grinding machines. A workshop dedicated for machining ceramic materials is available.

 

Particle Exposure assessment:

Proteomics and Transcriptomics facilities: A nano-UPLC-high resolution linear ion trap – orbitrap MS platformcombined with field-assisted ion mobility spectrometry allows for spearheading proteomics research, includingproteotyping, biomarker discovery and post-translational modification discovery. VITO has an up-to-datesample preparation facility for characterization of nanoparticle filters, speciation of nanoparticles and proteinidentification of nanoparticle surfaces. Automated and robotic sample handling is available and high-throughputcharacterization is at hand in a BELAC-certified lab environment. A fully equipped microarray and real-time PCRplatform, including Agilent Hybridization oven, Tecan HS4800Pro (4x44K chambers), Agilent DNA MicroarrayScanner, Tecan Microarray Scanner LS200 and 4 real-time qPCR cyclers is supported by excellent technicalsTAf and 2 biostatisticians. Dedicated RNase-free laboratories for sample handling and quality control arepresent.

 

In vitro toxicity testing facility: a GLP facility for handling and characterisation of (potentially) harmfulnanomaterials (CARDAM; www.cardam.eu) is equipped with glove boxes, biosafety cabinets class I, II and III,and fume cupboards to ensure safe handling of nanomaterials. Several cell culture laboratories, acclimatizedrooms and cultivation rooms are in place. Additional supporting equipment include a scanning fluorescencemicroscope with automated slide feeder and image analysis for genotoxicity assays, inverted fluorescencemicroscope with time lapse and multiplate readers for cellular assays, a flow cytometer BD FACSCalibur, anda Luminex system. Each suite of tests is supported by a dedicated technical support and is ensured by theavailability of at least one back-up technical staff member.

 

Occupational exposure assessment facilities: Multiple up-to-date laboratories are equipped with different gassupplies and filtering systems. Multiple fume cupboards are available for particle generation during synthesis,calibration and validation experiments, as well as a testing chamber (pressurized and HEPA-filtered) fornano-aerosol experiments.

 

Research supported by the infrastructure:

VITO performs research with 3 specific foci: 1. industrial innovation, 2. energy and 3. quality of the environment.Technology development and research activities take place in accordance with international quality standards(incl. ISO9001-2000, GLP). VITO research is regularly subjected to a critical evaluation by internationalspecialists (incl. for co-financing) and scientific advice councils (strategic advice council or SAC).More specifically, VITO’s research on nanomaterials focuses on technology development, characterization andrisk assessment as part of focus 1 and 3:

 

Particle characterization:

Advanced (functionalising) surface treatments of (nano)particles by laser and plasma technologies areinvestigated to improve the dispersion of these particles in coatings and bulk materials. Another focus is oneconomic powder processing methods based on aqueous solution synthesis to produce ceramic componentsof high quality. Porous materials with pore sizes from 1 nm to a few mm are produced, both for ceramicmembranes and on ceramic and metal foams. Several manufacturing routes are developed and optimized inorder to cover this wide range of material properties, which is inevitably accompanied with full characterisationof the obtained components, including TGA, DLS, BET, XRD, SDC, zeta potential, EPMA, FEG-SEM, AFM andXPS.

 

Particle Exposure assessment:

Research areas for the -omics platforms include in vivo and in vitro health risk assessment of nanoparticleexposure expressed at the protein and gene level. Use of nanoparticles in pharmaceutical applications (therapeutic protein drug administration, antibodies, proteotoxins) is a new area of investigation and the proteinsurface analysis of these compounds are key to understanding their effectiveness. The health effect of personalexposure to nanoparticles and the intake of xenobiotics is assessed at the protein level by means of studyingdifferential proteomics and protein adduct formation. Microarray and real-time PCR analyses are being usedto reveal gene expression patterns that discriminate between controls and exposure scenarios with the aim todiscover novel biomarkers for the impact of nanoparticles on functionally relevant (e.g. immune-related) geneprofiles, which are further developed and applied in alternative toxicity testing and in human biomonitoring.Investigations are being performed in vitro or on different tissues after in vivo exposure.

Alternative methods are applied to assess possible (eco)toxicological effects of (engineered) nanoparticles,including MW-CNT, according to OECD guidelines.

 

Engineered airborne nanoparticles are monitored in occupational settings using a wide range of onlineinstrumentation, sampling and characterization techniques resulting in identification of possible suitable metricsand measurement methods for the exposure assessment of nanoparticles. The nano-aerosol instrumentation(number concentration, surface area, size distribution) is specifically designed for the nanosafety area.

 

Services currently on offer and scientific highlights

VITO offers a dynamic interdisciplinary research environment, the particular strength of which is the vast rangeof facilities and expertise from a broad area of disciplines, as well as a close collaboration with universities andthe availability of full technical support when accessing the core facilities and technical equipment.