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H. Hofman Powder Technology Laboratory
Overview of Nanoparticle Technology and Characterisation
www.framingnano.eu
Nanotechnology products sorted by application area
Materialshttp://www.nanotechproject.org/inventories/consumer/.
www.framingnano.eu
Examples
Superparamagnetic Iron oxide nanoparticle
Si O
O
NH2
Si O
Si
O OH
O
NH2
NH2
Si
OSi
Si
OSi
OSiH
O
SiO
Si
OSi
O
OH
OH
O
NH2
NH2
NH2
NH
OH
O
NH2
NH2
NH2
OH
O
NH
N
N+
O-
O
NH
R
Fe2O3
= Avidin
= Biotin
T T T T
T T T T
+ +
SiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSi
O OOOOOOOOOOOOOOOOOOOHHHHHHHHHH
O
NNNNNNNHHHHHHHHHHHHHHHHHHHHHHHHHH222222
NNNNHH
TT TT TT
TTTT TTTT
Quantum Dots
4
Materialshttp://www.nanotechproject.org/inventories/consumer/.
www.framingnano.eu
Nanomaterials (2009!)
Nan
opar
ticle
s 6
Nam et al. Advanced Drug Delivery Reviews 65 (2013) 622–648
Synthesis of nanoparticles 7
Liquid phase synthesis • Coprecipitation • Sol-Gel • Microemulsion • Hydrothermal • Solvothermal • Microwave • Sonochemistry • Template synthesis • Biomimetic synthesis
Gas/vapor-phase synthesis
Methods using solid precursors • Inert gas condensation • Pulsed Laser Ablation • Spark Discharge Generation • Ion sputtering Methods using liquid or vapor precursors • CVD • Spray pyrolysis • Thermal Plasma synthesis • Flame synthesis • Flame sparay synthesis
Challenges • Means to achieve mono-
dispersity • Size and shape control • Reproducibility • Scale up • Building complex
nanoastructures
8
Mamaeva et al. Advanced Drug Delivery Reviews 65 (2013) 689–702
Nanoparticles for medical applications Important parameters
9 9 9
Structure of inorganic nanoparticles for medical applications Core is superparamagnetic iron
oxide
1. Shell: silica
2. Shell: Polymer with functional groups
Antibodies for target coupling to cells
Peptides for cell up-take
Stabilisiers
APS-SPIONs (~25-35 nm; + 33 mV)
PEG - APS-SPIONs 1 hr recirculation
(~70 nm & -20 mV)
FL-protein-PEG - APS-SPIONs
cysteine-protein-FL-PEG - APS-SPIONs (~80 nm & -13 mV)
APS: aminopropyltriethoxysilane
Si O
O
NH2
Si O
Si
O OH
O
NH2
NH2
Si
OSi
Si
OSi
OSiH
O
SiO
Si
OSi
O
OH
OH
O
NH2
NH2
NH2
NH
OH
O
NH2
NH2
NH2
OH
O
NH
N
N+
O-
O
NH
R
Fe2O3
= Avidin
= Biotin
T T T T
T T T T
+
+SiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSiSi
O OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOHHHHHHHHHHHHHHHHHH
O
NNNNNNNNNNNNNNNNNNNNHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH222222222222222222222
AAAAAAAPPPPPPSSS::: aaammmmiNNNNN
HH
TTT TTT TT
TTTT TTTTT
10 10 10
Key: Reproducible manufacturing of functionalized nanoparticles
CSEM Reactor inegrated into the EPFL set-up
• Standard operation protocols for synthesis, coating characterization
• Preparation for GMP • High potential for tarnsfer to industrial
production
Core and Surface properties 11
Nanoeffects Ferro- to superparamagnetism Band gab change (fluorescence) Surface plasmon
Scaling efffects Increased reactivity per mass Solubility Colloidal stability Penetration through barriers
Mechanistic injury pathways 12
Nanomaterial Toxicity Testing in the 21st Century Nel et al.
Band gap energy of Oxides 13
Zhang,†, et al. VOL. 6 ’ NO. 5 ’ 4349–4368 ’ 2012 ACSano
Influence of pH and size on Ec , Ev level and Band gap
12 2 3
2 2 20 0 0 0 0 0
1 1 1.8 0.124 1 12 4 (4 )
0.5 0.059( )
0.5 0.059( )
bulkgap gap
e h e h
C gap
V gap
e eE Eer m m m m r m m m m
E E PZC pHE E PZC pH
ππεε πεε
χ
χ
−
= + + − − +
= − + + −
= − − + −
Brus, L. E. J. Phys. Chem. 1986, 90, 2555
+ e 00 000000000000000000000000000000000
0mmmmm mmmm0mmmmmmmmmmmmmmmmm mmmmmmmmmmmmmmmmm +0000000000000000m m0hhmmmm mmmmm
TiO2 (anatase)
-9.000
-8.000
-7.000
-6.000
-5.000
-4.000
-3.000
-2.000
-1.000
0.000
0.0 5.0 10.0 15.0 20.0 25.0
Ban
d en
ergy
(eV)
Particle radius (nm)
pH 1 pH4 Ph7.4 pH 8
pH 1 pH4 Ph7.4 pH 8
1 CuO2
2 Alpha Fe2O3
3 Gamma Fe2O3
4 Fe3O4
5 WO3
6 CoO
7 Co3O4
8 Mn2O3
9 Ni2O3
10 TiO2 Anatase
11 TiO2Rutile
12 SnO2 Rutile
13 CeO2
14 Sb2O3
15 NiO
Physico-chemical reactivity of Nanoparticles
-6
-5.5
-5
-4.5
-4
-3.5
-3
-2.5
-2
0 1 2 3 4 5
Ener
gy le
vel o
f con
duct
ance
ban
d (e
V)
Band gap (eV)
Datenreihe1
5 nm
25 nm
Electron exitation by visible light
2 1 4 3
5
11 10
9
8 7 6
15
14 13
12
Particle diameter
11111111111111 1111110000
9999999999
88888888777 666666666666
11111111112222222222222222222 22222222Redox reaction with biomolecules
pH 7.4
Chromium oxide 90 nm
Alumina, CR 125, 15 nm
Solubility of Chromium oxide and aluminum oxide nanoparticles in water and cell media as function of time.
For detailed theory see : Dissolution kinetics of oxidic nanoparticles: The observation of an unusual behaviour Wolfram Vogelsberger �, Jochen Schmidt, Frank Roelofs, Colloids and Surfaces A: Physicochem. Eng. Aspects 324 (2008) 51–57
Nanoparticle System Parameters 18
19
0 20 40 60 80 100 1200
50
100
150
200
250
300
num
ber w
eigh
ted
mea
nsi
ze [n
m]
time [min]
PEI-SPION R=2 RPMI PEI-SPION R=2 RPMI + FCS PEI-SPION R=2 DMEM PEI-SPION R=2 DMEM + FCS
* for longer times sizes not determinable
*
*
0 20 40 60 80 100 1200
40
80
120
PVA-SPION RPMI PVA-SPION RPMI + FCS PVA-SPION DMEM PVA-SPION DMEM + FCS
num
ber w
eigh
ted
mea
n si
ze [n
m]
time [min]
water
Particles in biological media
Colloidal stability = f(coating and of protein adsorption) Application of DLVO theory is not possible
PEI-SPIONs
NP (SPION) transport in cell media +/- serum
20
0
0.05
0.1
0.15
0.2
0 0.05 0.1 0.15 0.2
mea
sure
d am
ount
of
part
icle
s (g
/m2)
calculated amount of SPION (g/m2)
• circles : cell media without serum,
• squares cell media with serum;
• filled symbol: 1 h incubation time,
• empty symbol 3 h incubation time.
Nanodiara , EPFL, Thesis Sakulkhu Usawadee, EPFL-LTP
In vivo -in vitro comparison 22
molecular weights
Rela
tive
amou
nt %
PVA coated SPION, positive charge
Usawadee Sakulkhu, Lionel Maurizi, Morteza Mahmoudi, Azza Gramun, Marie-Gabrielle Ollivier Beuzelin, Jean-Paul Vallée, Farhad Rezaee, Heinrich Hofmann, Nature Nanotech. Submitted Sept 2013
L. Maurizi , EPFL, 2013
Conclusions
Relationships between particle properties and behaviour in biological media, cell, tissue, circulation are still not known in detail.
This leads to unrealistic demand for detailed particle characterization
More basic knowledge regarding these relationships are key for the development of methods for toxicity assessment
Today, in vitro measurements have still too much uncertainty.
24
25
Assessment of toxicity of oxide NP 26
Zhang et al. ACS Nano VOL. 6 ’ NO. 5 ’ 4349–4368 ’ 2012
27
.
HeLa cells in contact with coated SPION
uncoatedSPIONs (a) negative (b)
neutral (c.) positive (d)