a nanotechnology platform for biomedical image …mams.rmit.edu.au/6c6adu7fgb8k.pdfa nanotechnology...
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A Nanotechnology Platform for Biomedical Image Enhancement
Ning GuJiangsu Laboratory for Biomaterials and Devices
State Key Laboratory of BioelectronicsSoutheast University, Nanjing, China
Email: [email protected]://www.lmbe.seu.edu.cn/nano/
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Location
Southeast University, Nanjing
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Southeast University, founded in 1902
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Main ContentsIntroduction
Micro CT and Au nanoparticles
Superparamagnetic iron oxide nanoparticle as
MRI contrast agents
FeFe33OO44 NPNP-- inclusion inclusion MicrobubblesMicrobubbles as Bi-
modality contrast agents for MRI and US
The action of The action of microbubblesmicrobubbles to cells under to cells under
ultrasound exposureultrasound exposure
Conclusions
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The development of medical imagingD
iagn
ostic
tech
nolo
gy
Tissue and cell level Molecular and gene level
1892-2002Single modaltiy
NowadaysStructure and function imaging
FutureDiagnostic combined with therapeutic
1895 X-ray MRI, PET/CT, US
Nanomedicine
Time
anatomical structure level
EarlyClearSee
Nanomaterials and Molecular assembly
Molecular imagingImaging
Micro-MRI, Micro-PET/CT, US
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NanotechnologyNanomaterial
Nanodevice or nano-scale functional structure
Nano-characterization and measurement
Biomedicine: Nanobiotechnology and Nanomedicine
Interaction and Bioeffects
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Copyright ©Radiological Society of North America, 2001
Weissleder, R. et al. Radiology 2001, 219: 316-333
Schematics show prerequisites to in vivo molecular imaging
分子影像学
Molecular imaging refers to the characterization and measurement of biological processes at the cellular and/ or molecular level.
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Micro CT and Au nanoparticles
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Home made Micro X-CTwith high-speed algorithms software
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The head of a Small Mouse
Chinese 1 yuan
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Mouse 3D structure by Micro CT
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Drug Analysis and Quality ControlThe CT images of the example pill
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Au Nanoparticles
J Phys Chem C,2007; J Phys Chem C,2008
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Au Nanoparticles: with diamater as 10nm, 20nm, 50nm
Synthesis of Au Nanomaterials (with various size and shapes)
Au NanoRods with 10 nm in diameter and AR:
3, 5, 7, 18
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(1) DI-Water
(2) 5mg/ml 20nm Au Nanoparticles
• 5mg/ml 50nm Au Nanoparticles
• 5mg/ml Injection Iopromidi.
1234
44 KV 55 KV 66KV
139152156149
61696966
20242422
1234
1234
55 KV1
2
3
83
85
84
(1) 10mg/ml 20nm Au Nanoparticles
(2) 10mg/ml Au Rods(AR=3)
(3) 10mg/ml Injection Iopromidi.
Low concentration level
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Superparamagnetic Iron OxidNanoparticles (MNPs) as MRI
Contrast Agents
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Magnetic nanomaterials Diagnosis and therapy
Magnetic nanomaterialsNano-Devices based on Magnetic nanomaterialsCharacterization and Mesurment for Magnetic nanostructure
Detection and early diagnosis
Effective therapy:
•Carriers for Thermal and Chemical Therapy
•Magnetic target probe
•Monitors after treatments
•Combination?Eg., To Cancer and so on
•Sensor•Contrast
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Solvent-thermal preparation method
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D=12.5±1.1 nmD=12.5±1.1 nm D=22.5±2.2nmD=22.5±2.2nm
Size and size distribution
10 11 12 13 14 150.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
Per
cent
age
diameter (nm)14 15 16 17 18 19 20 21
0.0
0.1
0.2
0.3
0.4
0.5
0.6Pe
rcen
tage
diameter (nm)
19 20 21 22 23 24 25 260.0
0.1
0.2
0.3
0.4
0.5
0.6
Pece
ntag
e
Diameter (nm)
D=17.3±1.6 nmD=17.3±1.6 nm
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The size of MNPs can influence the MRI signal
R2 relaxation rate increases with the increase of MNPs size
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Surface modification of Fe3O4 nanoparticles with DMSA
+DMSA
Aleate-Fe3O4 nanoparticlesDMSA-Fe3O4 nanoparticles
Double exchange
DMSA:2,3-dimercaptosuccinnic acid
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PEG coating of MNPs
TEM images of 12.5 and 22.5 nm MNPs after PEG coating
Poly (ethylene glycol): PEG
EDC (1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride)
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PEG modified MNPs influence the cellular uptake
Iron uptake by RAW264.7
12.5nm MNPs 22.5nm MNPs
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Chitosan (CS) modified MNPs
a) DMSA@MNPs(17nm)
b) CS@MNPs(17nm)
c) CS-DMSA@MNPsaggregates(100nm)
a-COO-
OH
OH
SH
HS
O
O
b -NH4+
O
OH
O
OH
AcHN
O O
HONHAc
HO
O
OH
O
OH
AcHN
O O
HONHAc
HOc
OH
OH
SH
HS
O
O
-COO-
Co-precipitation method
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Modified MNPs influence the cancer cellular uptake
0 20 40 60 80 100 120 1400
10
20
30
40
50
60
KB cell 8h
iron
per c
ell/p
g
concentration(ug/ml)
DMSA@MNPs CS@MNPs CS-DMSA@MNPs
0 20 40 60 80 1000
10
20
30
40
50
60SMMC-7721 8h
iron
per c
ell/p
g
concentration(ug/ml)
DMSA@MNPs CS@MNPs CS-DMSA@MNPs
Determination of the various MNPs for interaction with KB and SMMC-7721 cells. The cells were incubated with different
concentration of MNPs for 8h. Results are shown as mean±SD.
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In vitro MRI experiments
T2* imaging of different cell numbers when labeled with
DMSA@MNPs, CS@MNPs, CS-DMSA@MNPs.
CS-DMSA@MNPs has the strongest MRI T2
* signal intensity.
T2* singal enhanced1. The amount by cell uptake2. The aggregation of MNPS
enhance the MRI signal
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Higher positive charge
SI of T2*(2Χ105
cell)
31.8550.2
15.2263.7
++++14.55.188.585.7CS_DMSA@MNPs
+++10.24.981.016.5CS@MNPs
+-31.45.188.517.3DMSA@MNPs
Iron uptakeZeta potential/mV (pH 7.4)
Aggregate index
Hydrodynamic size/nm
Mean size/nm
Higher aggregation
More uptaken MNPsby cancer cells
Better MRI signal detection
Summary of Magnetic NP for MRI
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Anti-Sperm Protein 17 Immunomagnetic Nanoparticles
J. Nanosci. Nanotechnol. 2008, 8, 2341–2346
The anti-Sp17 mAb was grafted on the surface of carboxylated PEG coated MNPs by
covalent bonds.
TEM image of the carboxylated PEG coated MNPs
Magnetization curve of (a) naked MNPs,
and (b) carboxylated PEG coated MNPs
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Magnetic Microbubbles as MRI and US contrast agents
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Ultrasound images ~ MRI、 X-ray CT
Low cost when compared with X-CT and MRInon-invasiveness Real-time imaging featuresmore and more widely used in clinical applications
.
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Rationale of US imaging
Leen E, et al. Eur Radiol 2004;Miller D L, et al. Ultrasound Medicine, 2008
MicrobubblesMicrobubblesUltrasoundBiological
tissue
Biological effectsthermal
cavitation
radiation
US therapy
……
Imaging
brighter
Aggregation
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The preparation of FeThe preparation of Fe33OO44 NPNP-- inclusion inclusion microbubblesmicrobubbles
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Microscopy observation of mcirobubble
Microscopy images:(a)Non-Fe3O4 inclusion microbubbles, (b- d) Fe3O4-inclusion microbubbles
5μm
a
5μm
b
5μm
c
5μm
d
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The ultrasound imaging in the different samples in vitro (A), Control;(B) the multiple emulsion bubbles without SPIO; (C ) the multiple emulsion bubbles with SPIO
Tab.1 The mean grey scale within ROI measured by
using an ultrasound imaging system.
93.4±4.757.9±3.829.2±4.1mean grey scale
bubbles withSPIO
bubbles withoutSPIO
degassed and deionized
waterSamples
Ultrasound Imaging in vitro: Statistic
ROI
ROI
Materials Letters, 2008, 62: 121-124
ROI
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In vivo US imaging: Rabbit Liver
Ultrasound image of rabbit liver showing the enhanced contrast by using SPIO microbubbles (a) Conventional ultrasound image pre- injection of the microbubble contrast agent and (b) Post-injection( after 2, 6, 10min)of microbubble contrast agent.
d
ROI
b
ROI
a
ROI
c
ROI
The substantial contrast enhancement was observed with the injected contrast agent (microbubbles with SPIO) compared to the image of the control case obtained pre-injection of the contrast agent. After intravenous injection of the multiple emulsion microbubbles in the rabbit, the brightness in the liver increased over the time.
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It was found that within the first 1min after injection, the grey scale was increased rapidly. Then the brightness of images increased gradually, then decreased. The SPIO- inclusion bubbles have better image enhancement. Therefore the SPIO indeed had the contributions to the ultrasound imaging enhancement.
Mean grey scale in rabbit liver vs time after injecting microbubbles
Phys. Med. Biol, 2008, 53: 6129-6141
With SPIO
Without SPIO
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The influence of Fe3O4nanoparticles in the shell
Different NPs- embedded shell viscoelastic characterization Different US imaging effects
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The visoelastic parameter values
With the increasing concentration of Fe3O4 nanoparticles in the shell, the scattering cross-sections increase at first and then decrease. Therefore, by depositing appropriate solid nanoparticles on the surface of microbubbles, the echo character of the microbubble response can be changed significantly.
189.54532.02E-011.85E+01180.23
257.24731.81E-011.73E+01145.24
423.60881.63E-011.61E+01122.85
594.84771.58E-011.56E+01105.69
608.86281.51E-011.31E+0186.47
594.84771.50E-011.25E+0154.23
456.62671.47E-011.27E+0133.14
409.45321.32E-011.32E+0112.06
356.64641.28E-011.55E+015.73
265.59761.87E-011.70E+010
Scattering cross section(μm2)
Viscosity parameter μs(Pa)
Elastic parameterGs(MPa)Fe3O4 concentration(μg/ml)
increasedecrease
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Microbubbles concentration influences the MRI
SPIOBubbleTotal RRR 222 +≈
R2 Total contributed by SPIO Fe3O4 nanoparticles embedded in EMBs is greater than that contributed by the free SPIO Fe3O4 nanoparticles in the solution of the same concentration when volume fraction is greater.
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In vivo MRI imaging
Although the T2 signal negative enhancement of MRI immediately starts to appear when the two types of microbubbles are injected, the SNR time-course of SPIO-inclusion microbubbles has longer negative enhancement than mcirobubbles non-SPIO-inclusion.
0min 10min 30min 60min
120min100min80min70min
ROI1
ROI4 ROI3
ROI2
Biomaterials, 2009, 30: 3882-3890
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The action of The action of microbubblesmicrobubbles to cells to cells under ultrasound exposureunder ultrasound exposure
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Microbubbles as ultrasound imaging and drug delivery systems
Microbubbles structure:
N2 as ultrasound imaging mediam
NBD- cholesterol as model drug
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Setups of the in vitro ultrasound exposure:
different acoustic pressure to interact microbubbleswith cells
The schematic diagram of ultrasound exposure apparatus
A focusing transducer(radius=9.2 mm and focal length=8 mm) of 1 MHz was used.
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Different ultrasound energy exposure leads to different uptake efficiency
The mean fluorescence intensity initially increases as psp increases and
reaches the maximum when psp = 0.25 MP and then decreases.
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Cell observation by SEM
A B
D
FE
C
2μm(A)control, without micobubble, US;(B)control, without microbubble, with 0.25MPa/40S US;(C)-(F)
with only microbubble,,Psp=0.19,0.25,0.38 and 0.48MPa/40S/US
A B
C D
FE
500nm
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In drug delivery applications of ultrasound, the
choice of adequate acoustic pressure amplitude is
important. If psp is too small, sonoporation will not be
induced. If psp is too great, nonreparable sonoporation
may be induced; the disruption of cell membranes can
be too great to be repaired by self-sealing.
The membrane repair
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ConclusionsNew approaches to realize the designed micro- or nano-structure to be the nanoprobe for imaging enhancement;
The Action of functional nanoparticles to Bio-object, such as DNA, protein, cells;
The microbubbles structure embedded nanoparticles in the shell can be used as multimodality or multifunction carriers.
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