Nanotecnologia e Nanotecnologia e ambienteambiente

A.A. 2011-2012A.A. 2011-2012

The impact of new nanotechnology products with the environment are two-faced matter.

Several nanotech are environmentally friend and provide good means for soil remediation, water purification and energy storage.

On the other side, nanotechnology products can be considered a chemical hazard with unknown potential, poorly predictable from the knowledge about the corresponding bulk material.

The Janus-faced nanotechThe Janus-faced nanotech

Chemical hazard.Chemical hazard.

Nanomaterials represent a chemical hazard Nanomaterials represent a chemical hazard unpredictable from known properties of unpredictable from known properties of corresponding bulk materialcorresponding bulk materialOther variable tio be considered are:Other variable tio be considered are:1.1. Poorly controlled increment of productionPoorly controlled increment of production2.2. Poorly known dynamics in the environmental Poorly known dynamics in the environmental

matricesmatrices3.3. Poorly known environmental toxicityPoorly known environmental toxicity

Need for standardizationNeed for standardization

Standardization is recommended for:Standardization is recommended for:

• ProductionProduction• ManipulationManipulation• TransportTransport• Waste dispersionWaste dispersion• NanometrologyNanometrology• Toxicology studies in environmental matricesToxicology studies in environmental matrices

Agencies for nanotech monitoring: Agencies for nanotech monitoring: supernational.supernational.

Organization for Economic Co-operation Organization for Economic Co-operation and Development (OECD: WPN): and Development (OECD: WPN): http://www.oecd.org/http://www.oecd.org/

International Organization for International Organization for Standardization (ISO):Standardization (ISO):

http://www.iso.org/http://www.iso.org/

International Electrotechnical Commission International Electrotechnical Commission (IEC):(IEC):

http://www.iec.ch/http://www.iec.ch/

Environmental Protection Agency (EPA):Environmental Protection Agency (EPA):

http://www.epa.gov/http://www.epa.gov/

National Nanotechnology Initiative (NNI): National Nanotechnology Initiative (NNI): http://www.nano.gov/http://www.nano.gov/

Food and Drug Administration (FDA): Food and Drug Administration (FDA): http://www.fda.gov/http://www.fda.gov/

Agencies for nanotech monitoring: Agencies for nanotech monitoring: USAUSA

Community Research and Development Community Research and Development Information Service (CORDIS)Information Service (CORDIS): :

http://cordis.europa.eu/home_it.htmlhttp://cordis.europa.eu/home_it.html

Scientific Committee on Emerging and Scientific Committee on Emerging and Newly Identified Health Risks (Newly Identified Health Risks (SCENIHR): SCENIHR):

http://http://ec.europa.euec.europa.eu//healthhealth//scientific_committeesscientific_committees//emergingemerging/index_en.htm/index_en.htm

Agencies for nanotech monitoring: Agencies for nanotech monitoring: EUEU

The following list of needed methods for the

International Standardization for Nanotechnologies

Have been discussed by Dr Peter Hatto (Director of Research, IonBond Ltd, Chairman ISO TC 229 and UK NTI/1 Nanotechnologies Standardization committees) at the meeting organized at the Michigan State University (East Lansing, Michigan, USA, 7th February 2007).

Needs of standard methods for Needs of standard methods for NanoparticlesNanoparticles

1. Concentrations in air and water2. Stability, dissolution, aggregation and diffusion rates in the

environment3. Toxicological screening, physical and chemical hazard 4. Risk Assessments on exposure and use5. Environmental Toxicity Assessment for Nanomaterials6. Assessment of Product Degradation and Release of

Nanomaterials from Consumer Products7. Nanomaterial Product Labelling

Needs of standard methods for Needs of standard methods for NanotubesNanotubes

1.Eco-toxicology testing2.Exposure determination (environment and

food contamination)3. Inhalation testing4.Toxicology testing

An algorythm for NPs An algorythm for NPs environmental toxicity environmental toxicity

1.NPs characterization,

experimental plan

2.NPs interaction with

complex environments

3.Dissolution rate, end products

4.Solubility, diffusion

5.Evaluation of toxicity

If yes

Airborne NPs: models for Airborne NPs: models for toxicitytoxicity

SYSTEMSYSTEM TYPETYPE NPs studied NPs studied (exemples)(exemples)

Respiratory cellsRespiratory cells ““in vitro”in vitro” Metals, Metals, fullerenesfullerenes

Lung, rodentsLung, rodents ““in vivo”, experimentalin vivo”, experimental Fullerenes, Fullerenes, particulate particulate

matter (PM)matter (PM)

Brain, blood, Brain, blood, animalsanimals

““in vivo”, experimental in vivo”, experimental (diffusion from lung)(diffusion from lung)

fullerenesfullerenes

Human lungHuman lung ““in vivo”, professional or in vivo”, professional or accidentalaccidental

SWCNTsSWCNTs

Environmental pollution: airborneEnvironmental pollution: airborne

Nanometals permanently aggregate in air (right)….and enter the cells (left)

Nanoparticles in the lungNanoparticles in the lung

Carbon nanoparticles in the macrophages of the lung (mouse)

.

Cozzi E et al. Am J Physiol Heart Circ Physiol 2006;291:H894-H903

Mice lung (©2006 by American Physiological Society): A)normal aspectB) 24 h after exposure to ultrafine particulate matter (PM). Moderate neutrophil infiltration (arrows) and minimal edema, without increased alveolar macrophage and subpleural lymphocyte aggregates.

10 m 10 m

SWCNTs and the human lungSWCNTs and the human lung

Professional exposure to SWCNTs (Single Walled Carbon NanoTubes), Green regions inside the lung tissue show the production of reactive oxygen species.

Source: http://nanotech.dit.ie/nanotoxicology.html

Carbon nanospheres and cytotoxicityCarbon nanospheres and cytotoxicity

Carbon nanospheres CNs, functionalized with fluorescent dye at the surface. Vital staining, fluorescent dye.

Carbon nanospheres in the cells (B) and in the nucleus (C; time- dependent migration). Dissolution occurs, with delivery of the toxic functionalizing dye (a semiconductor or a QD). Dissolution is time-dependent and happens in the nucleus.

Selvi et al. Nano Letters 2008, 8(10): 3182–3188 DOI: 10.1021/nl801503m

““in vitro” models.in vitro” models.ORGANISMORGANISM EFFECTEFFECT TEST NAMETEST NAME

S. typhimurium, Escherichia coli

Genotoxicity, DNA oxidative damage

Ames Assay

Eukaryote, cellEukaryote, cell Genotoxicity, DNA oxidative damage

COMET assayCOMET assay

Eukaryote, cellEukaryote, cell Viability, apoptosisViability, apoptosis Trypan blue dye exclusion,Trypan blue dye exclusion,

MTT uptake test,MTT uptake test,

Apotosis genesApotosis genes

Eukaryote, cellEukaryote, cell Lipid peroxidationLipid peroxidation MDA determination assayMDA determination assay

Eukaryote, cellEukaryote, cell Substrate adhesionSubstrate adhesion Viability, light microscopyViability, light microscopy

Eukaryote, Eukaryote,

““in vitro” in vitro” developing developing

organsorgans

Gene expression, altered Gene expression, altered developmentdevelopment

Ex.: micro organ from Ex.: micro organ from cultured nasal epithelium, cultured nasal epithelium,

embryonic heartembryonic heart

Genotoxicity: Ames Assay in Salmonella typhimurium and Escherichia coli

Kumar et al., 2011. Chemosphere 83: 1124–1132

Toxicity in prokaryotes: eubacteria.Toxicity in prokaryotes: eubacteria.

Toxicity in prokaryotes: eubacteria.Toxicity in prokaryotes: eubacteria.

Damage to the membrane of E.coli exposed to TiO2 nanoparticles (10 mg/l). Cells stained red are damaged.

Toxicity in prokaryotes: cyanobacteriaToxicity in prokaryotes: cyanobacteria

Wang et al. 2011. Environ. Sci. Technol. 45: 6032–6040

““in vitro” cytotoxicity: nanometals. in vitro” cytotoxicity: nanometals.

Comparison of cytotoxic effects of CuO, micro (black) and nano-sized (gray) on A549 cells after 18 h exposure (40 μg/cm2)The percent of non-viable cells was evaluated by trypan blue staining. Means ± SD. ** < 0.05*** <0.01

The COMET assayThe COMET assay

Seriously (H2O2, 300 M) and moderately (nCo, 1 M) damagedDNA.

H2O2Co NPs

Hartung & Sabbioni. 2011. Nanomedicine and Nanobiotechnology, DOI: 10.1002/wnan.153

““in vitro” cell adhesion (Eukaryote)in vitro” cell adhesion (Eukaryote)

Quantification of human dermal fibroblast adhesion and viability on two different polymeric scaffolds (fibers diameter: 800 nm ca). Green: Viable cells Red: dead cells.

Grafahrend et al. 2011. Nature Materials, 10: 67–73. doi:10.1038/nmat2904

Models for aquatic toxicity (fresh Models for aquatic toxicity (fresh and and saltsalt water), invertebrates water), invertebrates

ORGANISMORGANISM Test approved by Test approved by ASTM/EPA/OECD/EU/ISOASTM/EPA/OECD/EU/ISO

Daphnids: D.magna, D. pulex, C.dubiaDaphnids: D.magna, D. pulex, C.dubia ++

Chydorus sphaericusChydorus sphaericus --

Thamnocephalus platyurusThamnocephalus platyurus --

Brachionus calyciflorusBrachionus calyciflorus ++

Hydra attenuataHydra attenuata ++

Elliptio complanataElliptio complanata --

Crustaceans: harpacticoida copepodsCrustaceans: harpacticoida copepods ++

Mytilus edulisMytilus edulis ++

Fluorescently labelled polymeric nanoparticles in the gut of Daphnia magna following ingestion after aqueous exposure.

(Image taken by Dag Altin, BioTrix)

Nanometals and fullerens intake by D.magna.

(Chen et al., 2008 Progress Report: Methodology Development for Manufactured Nanomaterial Bioaccumulation TestEPA Grant Number: R833327)

Models for toxicity in water: DafniaModels for toxicity in water: Dafnia

Mortality of D.pulex exposed to fullerenes and nTiO2 differs in relation with the chemical nature and concentration of the

chemical compound.Klaper et al., Environ. Poll. 2009. 157(4): 1152-1156

Models for aquatic toxicity Models for aquatic toxicity (sediments: fresh- and (sediments: fresh- and saltsalt water) water)

InvertebrateInvertebrate Test approved by international Test approved by international organismsorganisms

Hyalella atzecaHyalella atzeca ++

Lumbriculus variegatusLumbriculus variegatus ++

Leptocheirus plumulosusLeptocheirus plumulosus ++

Algae/PlantAlgae/Plant

Pseudokirchneriella Pseudokirchneriella subcapitatasubcapitata

ProkaryotesProkaryotes

Microcystis aeruginosaMicrocystis aeruginosa ++

Models for soil toxicity: Models for soil toxicity: invertebrates and plantsinvertebrates and plants

ORGANISMORGANISM Test approved by Test approved by ASTM/EPA/OECD/EU/ISOASTM/EPA/OECD/EU/ISO

Eisenia sp.Eisenia sp. ++

Enchytraeus crypticusEnchytraeus crypticus ++

Lumbricus terrestrisLumbricus terrestris --

Porcellio scaberPorcellio scaber --

PLANTSAllium cepa, Hordeum vulgare, Lolium perenne, Phaseolus radiatus,

Solanum lycopersicum, Spinacia oleracea, Triticum aestivum

Effect of nanoZnO on the primary root tip of L.perenne:

A)ControlB)ZnO NPs, 20 nm,

1000 mg/l

Lin & Xing 2008. Environ. Sci. Technol., 42(15): 5580–5585

Models for terrestrial toxicity: plantsModels for terrestrial toxicity: plantsChromosomal aberrations observed in Allium cepa meristematic cells exposed to ZnO NPs.

Sticky chromosomes (A1, B1); binucleated cells (c1); chromosomal break (a2); nuclei with pronucleus (c2).

Metaphase Anaphase Prophase

Anaphase Multipolar Anaphase Interphase

Kumari et al. 2011. Journal of Hazardous Materials. 190: 613–621

X1000’X1000’

X1000’ X1000’

X1000’ X1000’ X1000’

Light microscopy images from Apostain stained tissues (945× magnification) of Lombriculus terrestris exposed to 0 (plates a and c) or 100 mg l− 1 of TiO2 nanoresidues (plates b and d) in water for 7 days distinguishing cuticule (a and b) and intestinal epithelium (c and d). Arrows indicate negative (N) and positive (P) apoptotic responses in stained cells.

(Lapied et al., 2011, Environ. Int. 37(6): 1105–1110)

Models for aquatic toxicity: Models for aquatic toxicity: vertebratevertebrate

The fishes O. mykiss, O.latipes and D.rerio have been studied during exposure to NPs, in natural or experimental conditions.

The developmental effects on the model organism X.laevis, a clawed frog, have been also studied in laboratory conditions.

Nanometals: toxicity in zebrafishNanometals: toxicity in zebrafish

Fully developed zebrafish embryo with accumulated particles of nAg in different areas of the body:i) Retinaii) brain (mesencephalon

cavity)iii) Heartiv) gill archesv) Tail.

(Lee et al., 2007. ACSnano, 1(2):133)

nAg: embryotoxicity in zebrafishnAg: embryotoxicity in zebrafish

(Lee et al., 2007. ACSnano, 1(2):133)

And now, the good news.

Nanotechnologies for soil and Nanotechnologies for soil and groundwater remediationgroundwater remediation

NPsNPs EnvironmentEnvironment Ref.Ref.

amphiphilic amphiphilic polyurethane (APU)polyurethane (APU)

Groundwater and soils Groundwater and soils from polynuclear from polynuclear

aromatic aromatic hydrocarbons.PAHshydrocarbons.PAHs

DOI: 10.1021/es0348997

Nanoscale or emulsified zero-valent

iron (NZVI, EZVI) (NZVI, EZVI)

EPA 542-F-08-009, 2008

http://www.epa.gov/tio/download/remed/542-f-08-009.pdf

bi-metallic nanoscale particles (BNPs)

Au-polymeric nanofoam

Soils from oil, volatile Soils from oil, volatile haromatic haromatic

DOI: 10.1002/cssc.201000410

Nanotechnology for wastewater Nanotechnology for wastewater purificationpurification

NPsNPs PollutantsPollutants

DendrimersDendrimers toxic metal ions, radionuclide and inorganic

anions, delivery vehicles for antimicrobial

Metal nanoparticlesMetal nanoparticles Antimicrobial action, remove As (ZnO NPs)

Zeolite Zeolite nanoparticlesnanoparticles

Remove Cr(III), Ni(II), Zn(II), Cu(II)

and Cd(II). Reverse osmosis for desalination.

Carbon-based nanomaterials

Wide-range applications

Boron nanotubes Desalination

Tiwari et al., 2008 World Applied Sciences Journal 3 (3): 417-433,

Nanotechnology for solar power.Nanotechnology for solar power.Photoactive materials: Semiconductors with appropriate band gap

and high absorption coefficient.1. Inorganic Semiconductors: Si, Ge, CdS, GaAs, CdTe, TiO2,

etc.2. Organic Semiconductors: A. Small molecules: phthalocyanines, porphyrins, etc. B. Conducting polymers: Polythiophenes, polypyrrole,

polyaniline, etc.

Solid-state polymers have lesser problems than liquid ones.Coupling solid-state polymers with C60 improves the performance.

Teketel Y. , Solid-State Photoelectrochemical Solar Energy Conversion Based On Conducting Polymers. ICPC NanoNet 3rd Annual Workshop, May 2011, St.Petersburg, Russia.

Nanotechnology for solar power.Nanotechnology for solar power.

Photoactive materials: Semiconductors with appropriate band gap and high absorption coefficient.

1. Inorganic Semiconductors: Si, Ge, CdS, GaAs, CdTe, TiO2, etc.1. Organic Semiconductors: A. Small molecules: phthalocyanines, porphyrins, etc. B. Conducting polymers: Polythiophenes, polypyrrole, polyaniline, etc.