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![Page 1: Laser Generated Pure Nanoparticulate Reference Material for Risk Assessment Studies S. Barcikowski, J. Jakobi, A. Hahn, J. Walter, S. Petersen NanoMed.](https://reader035.fdocuments.net/reader035/viewer/2022062423/56649e535503460f94b49d80/html5/thumbnails/1.jpg)
Laser GeneratedPure Nanoparticulate Reference Material
for Risk Assessment Studies
S. Barcikowski, J. Jakobi, A. Hahn, J. Walter, S. Petersen
NanoMed 2009, March 6th, Berlin
JRG Nanoparticles
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"Healthy" organic nanoparticles… …from Hannover brewery
0 100 200 300 400 500 600 700 800 900 10000
1
2
3
4
5
6Lindener Spezial Bier
Particle Diameterd
90 = 699.50 nm
d50
= 337.00 nmNumber of Particles: 452
Log-Nomal Fit of Hydrodyn. Diam. Distrib.
= 0.55733xc = 338.801 ±64.378w = 0.688 ±0.199
Rel
ativ
e P
artic
le N
umbe
r F
req
uenc
y P
N (
%)
Hydrodynamic Diameter dh (nm)
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Example of a Risk Assessment Approach: Airborne Nanoparticles in the Human Respiratory Tract
Animal Model Human Respiratory Tract
In vitro
Inhalation
Instillation
Aerosol
Colloid
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Biological response is correlated to nanoparticle SURFACE
ZrO2: Y
Ref.: Stoeger et al.,GSF Annul. Rep. (2004) 43-48
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"Gold Colloid" (Material Data Sheet)
NaN3
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Uptake Mediated by Cell Penetrating Peptides
Yang et al., Transferrin-Mediated Gold Nanoparticle Cellular Uptake. Bioconjugate Chem., Vol. 16, No. 3, 2005 495
Transferrin-Mediated Cellular Uptake by Endocytosis. 5-h treatment, green labeling, exc. 488 nm, emiss. 515 nm (A) Control cells (human nasopharyngeal carcinoma cells)(B) cells treated with Au nanoparticles(C) cells incubated with Au-Transferrin nanoparticles;(D) cells treated with Au-albumin nanoparticles; (E and F) cells co-treated with different proportions of Au-TF versus holo-TF (1:2 and 1:5, respectively).
Transferrin Ligand:
not toxic but prolongs
nanoparticle penetration
into cytoplasma
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Requirements to Nanoparticulate Reference Material for Risk Assessment Studies
1. Same material composition of aerosol and colloid
- aerosol: inhalation (in vivo)
- colloid: instillation (in vivo / in vitro)
2. High purity
minimized cross effects
3. Size control
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Nanoparticle Generation Processes
Form-in-Place
Wet Chemistry
Mechanical Synthesis
Gas Phase Synthesis
• Lithography • Chemical Vapour Deposition • Physical Vapour Deposition
• Sol-Gel-Process• Precipitation of Salts
• Ball Mills, Planet Mills• Kryo-Milling• Homogenisation (organics)
• Flame Hydrolysis• Flame Pyrolysis • Cemical Vapour Synthesis• CO2 Laser Pyrolysis
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Start
Ref.: Fonds der Chem. Industrie, Nanobox (2006)
Gas Phase Synthesis / Wet Chemistry
Sol-Gel / Precipitation
- scalability, monodisperse colloids
- chemical precursors and additives
Gasphasensynthese Gasphasenreaktor
Nukleation Wachstum Aggregation
Vorläufer-Dampf
Oxid-Dampf Kügelchen Verklumpung
Reaktions-bereich/Flammzone
Keimbildung
Temperatur / Zeit
Gasphasensynthese Gasphasenreaktor
Nukleation Wachstum Aggregation
Vorläufer-Dampf
Oxid-Dampf Kügelchen Verklumpung
Reaktions-bereich/Flammzone
Keimbildung
Temperatur / Zeit
Gas phase Synthesis- multi-ton scale, gaseous precursors- powders / agglomerates
Precursor Nucleation Aggregation
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Nanoparticle Generation Processes
• Availability of precursors Limited Nanomaterials• Agglomeration (powder) Re-Dispersion • Additives and chemicals Purification
Limitations in Risk Assessment Studies
Form-in-Place
Wet Chemistry
Mechanical Synthesis
Gas Phase Synthesis
• Lithography • Chemical Vapour Deposition • Physical Vapour Deposition
• Sol-Gel-Process• Precipitation of Salts
• Ball Mills, Planet Mills• Kryo-Milling• Homogenisation (organics)
• Flame Hydrolysis• Flame Pyrolysis • Cemical Vapour Synthesis• CO2 Laser Pyrolysis
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Start
Laser generated
Aerosols
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Kammer
Strahlteiler
P
Linse
SampleHolder P
Laser
SampleV
CCD
ELPI
Lens
Beam Splitter
Chamber
Continuous Generation of Nanoparticles in Gasesby Laser Ablation from Solid Targets
Sample Holder
SampleWindow
Inlet
Exit
Experimental Setup Process Chamber
• continuous ablation of any solid target material(e.g. titanium, silver, alloys, …)
• no chemical precursors
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Particle Size Distribution During fs-Laser Ablation of Graphite
0%
20%
40%
60%
80%
100%
0.00
70.
030.
06 0.1
0.16
0.25
0.39
0.63
0.98
1.59
2.43
3.93
Aerodyn. Diameter [µm]
Nu
mb
er
Fre
qu
en
cy
Pulse Energy: 50 µJ
Pulse Energy: 300 µJ
S. Barcikowski,et al., ICALEO 2005
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Influence of the Process Gas on the Particle Size during fs-laser ablation of Titanium
0%
20%
40%
60%
80%
100%
Press. Air Helium Nitrogen
3.93 µm
2.43 µm
1.59 µm
0.98 µm
0.63 µm
0.39 µm
0.25 µm
0.16 µm
0.10 µm
0.06 µm
0.03 µm
0.007 µm
Nano
Mic
ron a
nd S
ubm
icro
n
S. Barcikowski, A. Hahn, B. Chichkov. J. Laser Appl., Vol. 19, No. 2, May 2007
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Particle size distribution during laser ablation of zirconia using different types of laser
0,01 0,1 10
10
20
30
40
50
60
70
80
rel.
fre
qu
en
cy o
f pa
rtic
le n
um
be
r [%
]
aerodynamic particle diameter [µm]
Nd:YAG-Laser: 1 kJ/m CO2-Laser: 20 kJ/m Ti:Sa-Laser: 3 kJ/m
Zirconia
S. Barcikowski, A. Hahn, B. Chichkov, J. Laser Appl., Vol. 19, No.2, pp. 65-73 (2007))
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Composition of the nanoparticulate matter
Zr
YO
Zr
Y
Y2O3-doped ZrO2
Laboratory of Biomaterials Dept. of Neurosciences, University of Modena and Reggio Emilia, Italy
S. Barcikowski, J. Walter, A. Hahn, J. Koch, H. Haloui, T. Herrmann, A. Gatti. Proc. LPM 2008. Subm. to Journal of Laser Micro/Nanoengineering (2009)
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Composition of the nanoparticles:Energy Electron Loss Spectroscopy
Laboratory of Biomaterials Dept. of Neurosciences, University of Modena and Reggio Emilia, Italy
ZrO2: Y
S. Barcikowski, J. Walter, A. Hahn, J. Koch, H. Haloui, T. Herrmann, A. Gatti. Proc. LPM 2008. Subm. to Journal of Laser Micro/Nanoengineering (2009)
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Cr
Fe
Ni
FeCrNi alloy (stainless steel)
Composition of the nanoparticulate matter sampled at the workplace
Laboratory of Biomaterials Dept. of Neurosciences, University of Modena and Reggio Emilia, Italy
S. Barcikowski, J. Walter, A. Hahn, J. Koch, H. Haloui, T. Herrmann, A. Gatti. Proc. LPM 2008. Subm. to Journal of Laser Micro/Nanoengineering (2009)
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Start
Laser generated
Colloids
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100% pure
fully dispersed and stable
Laser Ablation in Liquids
Pulsed Laser Beam
Liquid
Solid
Pulsed Laser BeamPulsed Laser Beam
Liquid
Solid
safe
unlimited materials and liquids
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Laser Generated Colloidal Nanoparticles
300 nm500 nm
50µm
Platinum Silver
500 nm
Copper
Gold ConjugatesNiFe
100 nm
SilicaPVP
FeOx
100 nm
SilicaPVP
FeOx
Iron Oxide
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Limitations in fabricating Colloidal nanoparticle alloy from Sm2Co17
Problem: Disproportionation!
Enrichment of
- Co in small
- Sm in big
Nanoparticles.
Possible reason:
Difference (130%) Heat of Evap.
- Co: 377 kJ/mol
- Sm: 166 kJ/mol
cause segregation in
fast (Sm) and slow (Co)
component in laser plume.
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PtIr Alloy Nanoparticle Colloid
Pt-Ir Alloy Similar (18%) Heat of Evap.
- Pt: 510 kJ/mol
- Ir: 604 kJ/mol
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Electro-Deposition at Neural Electrodes / Implant Surface
50µm 500nm50µm
Uncoated NiTi-Microstructure… …coated with NiTi-Nanoparticles
Menendez et al, JLMN 2009 // Barcikowski et al.,Biomaterialien 2007
PtIr
NiTi
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Barcikowski, Hahn, Guggenheim, Reimers, Vogt; unveröffentlicht
Human adipose-tissue derived mesenchymal stem cells grown on NiTi-Nanoparticles
(FE-ESEM at 99.7% humidity)
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Start
Stabilisation
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Ex-situ Stablisation with Albumin
Albumin ist contained in...
- Blood, Human Serum
- Cell culture media (DMEM, RPMI)
V. Amendola, M. Meneghetti. J. Mater. Chem., 2007, 17, 4705–4710
No Albumin
Stability in 0.07 M KCl
Albumin
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U=0 V Brownian
Motion
v
vi
Gold Electrodes
Laser Scattering
Nanoparticle MotionSet-Up
Kinetics
Motion and kinetics of laser-generated Nanoparticles
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Vector field of electro-mobility
at U=20 V
(-)
(+)
v
vi
Gold Electrodes
Laser Scattering
Nanoparticle MotionSet-Up
Motion and kinetics of laser-generated Nanoparticles
Kinetics
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In-situ Functionalisation
AuS
S
S
S
S S
S
S
S
fs laser beam
variable target (e.g. Au)
variable
ligand
SH COOHNH2
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Laser generated NiTi-alloy Nanoparticles
-60
-50
-40
-30
-20
-10
0
10
0.01 0.1 1 10 100
Concentration [mmol/l]
Zet
a P
ote
nti
al [
mV
]
Cysteine
Citrate
In-situ conjugation with citrate or cysteine
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Stability in Saline Solutions
Stability of conjugates is evidenced in saline solutions
0.0 0.5 1.0 1.5 2.00.0
0.2
0.4
0.6
conjugated particles pure particles
A (
80
0 n
m/ 3
80
nm
)
[NaCl] [M]
physiological
salinity
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400 600 8000.0
0.2
0.4
0.2
0.4 Au NPA
bso
rba
nce
without salt +0.15M NaCl (=physiological) +2M NaCl transfection buffer
Conjugates
Wavelength
0
-20
-40
surf
ace
po
ten
tial [
mV
]
Au NP Conjugates
Stability in Saline Solutions or Buffers
S. Petersen, S. Barcikowski, Advanced Funct. Materials, in press (2009)
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Gold Nanoparticles
Nano-Bioconjugates
0 50 100
AUZ
200 nm
200 nm
10 nm
Laser generated Nano-Bio-Conjugates are monodisperse
TEM Micrographs Size distribution
Frequency
Diameter
S. Petersen, S. Barcikowski, Advanced Funct. Materials, in press (2009)
10 100
DLS
0 25 50
TEM
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Conclusion
Laser ablation in gases and liquids:
Contribution to systematic studies
on adverse health effects
of engineered nanoparticles?
Nanoparticle reference material:
- purity,
- size,
- composition
matters!
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Contributions at LZH-Nanomaterials Group:
Niko BärschAnne HahnJurij Jakobi Ana MenendezChristin MennekingSvea Petersen Laszlo SajtiRamin SattariAndreas SchwenkePhilipp WagenerJohanna WalterJürgen Walter
Funding:European Commission: integrated project LAUNCH-MICRO (NMP2-CT-2005-011795)
German Research Foundation (DFG):Junior Research Group 'Nanoparticles'within Excellence Cluster REBIRTH
Contribution from Univ. Bologna:
Antionetta Gatti
Acknowledgement
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Acknowledgement
Contributions at LZH-Nanomaterials Group:Stephan BarcikowskiNiko BärschAnne HahnJurij Jakobi Ana MenendezChristin MennekingSvea Petersen Laszlo SajtiRamin SattariAndreas SchwenkePhilipp WagenerJohanna WalterJürgen Walter
Contribution from Univ. Bologna:
Antionetta Gatti
Thank you !
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2 min
1 min
30 s0 s
Duraion of Irradiatin
Laser fragmentation of gold nanoparticles in water
Abs
orpt
ion
[-]
Wavelenghth [nm]