NMs and Safety Experiences€¦ · NMs and Safety Experiences Fernando Pastrana MD, MSc, PhD....
Transcript of NMs and Safety Experiences€¦ · NMs and Safety Experiences Fernando Pastrana MD, MSc, PhD....
NMs and Safety Experiences
Fernando Pastrana MD, MSc, PhD.
Departamento de Ingeniería Eléctrica y Electrónica
Alba Avila PhD
Profesor Asociado, Departamento de Ingeniería Eléctrica y Electrónica
Bogotá D.C., February 14 and 15, 2017 1
Nanomaterials in the market
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Beilstein J. Nanotechnol. 2015, 6, 1769–1780.
Outline
• Nanomaterials in products• From the bench to the market
• Toxic effects of NMs
• NMs characterization• Physic-chemical to biological identity
• Labeled free cytotoxicity assessment
• Implementing a safety culture
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• Oxidative Stress• ROS• NOS
• Inflammation• Interleukins• Complement activation• Neutrophils
• Mechanical Toxicity• Cytoskeleton disruption
Cells toxicity mechanism
Elsaesser, C.V. Howard, Toxicology of nanoparticles., Adv. Drug Deliv. Rev. 64 (2012) 129–37.
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Carbon-based nanomaterials
• Mesothelioma
SiO2nanoparticles
• Platelet aggregation,• Infarcts
• Fibrosis• Hepatic steatosis
Dendrimers
Nanomaterial Organ Toxic Effect Reference
Potential Organ toxicity of NMs
Part. Fibre Toxicol. 6 (2009) 2.
Int. J. Nanomedicine. 7 (2012) 631–9
Arch. Toxicol. 88 (2014) 1939–64
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How a nanomedical product can be in the market?
Concept
Pre-clinical
•In-silicon
•In-vitro
•In-vivo
Clinical trials
• Three phases
Approval
1 Year
3 Years
5 – 7 Years
Aprox 10 Y
Bogotá D.C., February 14 and 15, 2017
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Clinical trials with nano
Source: clinicaltrials.gov acceded on Feb 15 2017
Outline
• Nanomaterials in products• From the bench to the market
• Toxic effects of NMs
• NMs characterization• Physic-chemical to biological identity
• Labeled free cytotoxicity assessment
• Implementing a safety culture
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Physic-chemical properties
Size Biodistribution
Translocation
Biological barriers
Specific surface area
Surface area/volume ratio
Shape and aspect ratio
Fiber like
Planar
Amorphous
Spherical
Surface reactivity
Surface chemistry
Photocatalytic activity
ROS production
Composition Oxides
Polymers
Carbon
Crystalline structure
Defects
Aggregation & Agglomeration
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Biological Characterization
Sterility Endotoxin detection
Microbial contamination
Bacterial Contamination
Mycoplasma contamination
Hemocompatibility Hemolytic properties
Platelet aggregation by cell counting
Platelet aggregation by light transmission
Complement activation In-vitro immunology
Leukocyte proliferation
Chemotaxis
Phagocytosis
Cytokines, Chemokines and Interferons
Toxicity Oxidative stress
Necrosis
Apoptosis , viability
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Toxicity
Biological Characterization
Oxidative stress Viability Apoptosis
DHE
CellROX
MTT
Calcein
Anexin V
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Safety assessment or nanomaterials interference
Biological Characterization
Agglomeration effectsInterference with assay components
- Increase reagent biotransformation- Bioactive protein sites sequestration
Assays dependence on optical - NMs blocking the light path- NMs reflecting the light path- NMs fluorescence at the same
wavelength than assay dyes
NR: neutral redMTT: tetrazolium to formazan96AQ: MTS to formazanaB: rezasurin to red resorufinCTB: CellTiter-BlueCTO: CytoTox OneToxicology and Applied Pharmacology 234 (2009) 222–235
SWCNT
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Confusing principle
Bogotá D.C., February 14 and 15, 2017
Outline
• Nanomaterials in products• From the bench to the market
• Toxic effects of NMs
• NMs characterization• Physic-chemical to biological identity
• Mechanical indicators-Labeled free cytotoxicity assessment
• Implementing a safety culture
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New labeled free approaches
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Impedance (electrical current)
high sensitivity from no label
Real time recording
Microfluidics (improve current biochemical label assays)
Improve of ENM dispersion, better correlation dose toxicity, physiological environment
Biological Physiology
Multi (Proteo and metabo) nomics
specific effects, cellular pathways
Identification of the individual pathways involves on cytotoxicity
Atomic force microscopy (mechanical response to toxicity
Corroboration of the observed results by immunofluorescence techniques
No label required, in-vivo and environmental assessment
Nanomaterials 2015, 5(3), 1181-1199
Mechanical cellular response
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Pastrana et, al
Cell viabilityAssay (MTT)
Basic Logic Gate diagram of Cytotoxicity assessment of NIH3T3 Exposed to 0.05wt% CBNs after 24h
No toxicity, cells viableCell viability
(Trypan Blue)
Cell viabilityAssay (Live/dead) No toxicity
orToxic NMsCellular stress
(CellROX®)
Cell viabilityAssay (MTT)
Cell viability(Live/dead)
Cellular stress (CellROX®)
Mechanical Toxicity(AFM)
Cytotoxic nanomaterial
Pastrana et al, thesis 2016
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1 terminal
2 terminal
3 terminal
Number of terminals:
SGM SMM
CAFM SSRMPFM
SCMEFM-SKFM
Electrical Measurements
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Fundamental concepts on microscopy
DIDACTIC MODELS FOR KIDS
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Clubes de ciencias600 studentes
Ingenieros sin fronteras
Teusaca
Outline
• Nanomaterials in products• From the bench to the market
• Toxic effects of NMs
• NMs characterization• Physic-chemical to biological identity
• Mechanical indicators-Labeled free cytotoxicity assessment
• Implementing a safety culture
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Nano activities distribution at UniandesDistribución de actividades nano en Uniandes
Nano-Risk ChallengeRetos con el Riesgo-Nano¿How to manage the safety of a laboratory where nano-
materials are handled?¿Cómo gestionar la seguridad de laboratorios que sintetizan o manipulan nano-materiales?
¿How to turn research & development processes intoacademic oportunities to construct safety cultutre?
¿Cómo convertir los procesos de investigación y desarrollo en oportunidades académicas para construir cultura de seguridad?
¿How to include the emerging risks related to the handlingof nano-materials or nano-composites in the HSE
management?¿Cómo incluir los riesgos emergentes asociados al uso de nano-materiales o nano-
compuestos en la gestión HSE?
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Ti NPZn NPAg NP
Carbon Based Nanomaterials
Potential exposed groups
Potential nano
materials hazards
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ISI and Scopus publication trackSeguimiento de publicaciones en ISI y Scopus
Purchasing department requestSolicitud al departamento de compras
Pre-training surveyEncuesta pre-entrenamiento
• Physics• Chemist• Engineering Dep.
Scenaries and context definitionDefinición de contexto y escenarios
• Engineering barriers• PPE Barriers
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Frequently used nanomaterialNanomateriales mas usados
Risks identificationIdentificación de riesgos
High
Low to Medium
Medium
Medium
High
Low to High
Hazard*
Peligrosidad
*Musee N: Oct 4 2010
MWCNT
Ag Nanoparticles
Corrective controlsControles correctivos
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Safe handling of carbon based nanomaterials’ protocols
Public access to the protocols by intranet
Identify the different CNTs
Explain the potential health risk
Assess the risk according with the CNT & Activities
• Administrative
• Engineering
• Personal protective equipment
• Waste disposal
• Accident management
Suggest the appropriate exposure barriers
web
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Self-assessment
Nanomaterial hazard identification
Exposure probability
• Protective equipment
• Engineered barriers
Safety culture
nanoseguridad.uniandes.edu.co
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NanoRisk App geography
Source google analytics
0
50
100
150
200
250
300
350
400
450
500
ago-15 sep-15 oct-15 nov-15 dic-15 ene-16 feb-16 mar-16 abr-16 may-16 jun-16 jul-16 ago-16 sep-16 oct-16
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Some Strategic Programmes
Thanks
Acknowledgements
F. Muñoz . Profesor Ingeniería Química. Universidad de los AndesAlexis Vignes, INERISCandace Tsai, Colorado State UniversityA. Raman, Purdue UniversityAlexander Cartagena, NIH
Fluorescence image of NIH3T3 cell exposed to graphene
3D figure of AFM measurements on NIH3T3 cell
Force map of NIH3T3 cell by AFM
Topographic map of NIH3T3 cell by AFM
ROS RO
S
Protein media adhesion
ROS
F-actin
depolimerization
ROS production
ROS
Surface chemistry• X-ray photoelectron spectroscopy (XPS)
http://www.nuance.northwestern.edu
Sensitivity 0,1%Chemical state infoThickness up to 8nm
Photoelectric effect
KE = hν - (EB+ϕ)
Photoemission
h𝑣
Detector
Probe https://www.helmholtz-berlin.de http://biointerface.org/surface/xps/
Composition• Atomic absorbance spectrometry (AAS)
Lab-training.com
Skidmore's Analytical Interdisciplinary Laboratory (SAIL)
Pastrana et, al
Composition
• Mass spectroscopy
Pastrana et, al
0,00E+00
2,00E+00
4,00E+00
6,00E+00
8,00E+00
1,00E+01
1,20E+01
1,40E+01
1,60E+01
MWCNT [A]
0,00E+00
5,00E-07
1,00E-06
1,50E-06
2,00E-06
2,50E-06
Graphene [A]
Series2DR. DEEPAK BHANOT
Main objectivesObjetivos principales
Provide safe environment to develop nanotechnology activities at Universidad de Los Andes
Protect human health
Protect environment
Protect facilities
Protect institution goodwill
Promote sustainable research and innovation development
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Proteger la salud humana
Proteger en medio ambiente
Proteger la infraestructura
Proteger la reputación de la institución
Promover el desarrollo sostenible de investigación e innovación
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