Transcript MedicalApplications of SNP

Applications of Silver
Nanoparticles in Medicine
Arinola Awomolo
BioE 494
11/27/2007
Background Leading up to Resurgence
of Interest in Silver
•
Silver has been reported to be in use as far back as before Christ
– It has been used:
•
•
•
•
•
By Greeks and Roman empire in wine and water storage to maintain freshness
By the wealthy during the middle ages to prevent food spoilage
By early Americans. Silver coins were placed in drinking water and also milk
In 1800’s doctors began using silver in surgical wounds
What lead to the reduction of Silver Use?
– Mostly due to the advent of antibiotics
•
What has led to renewed interest?
– Antibiotics resistant bacteria
– Silver is effective against most types of microbes
http://www.silver-colloids.com/Pubs/history-silver.html
Properties of Silver Nanoparticles
•
Silver is a good conductor
•
They have an intense light scattering
and absorption
“Using Solution-Phase Nanoparticles, Surface-Confined Nanoparticle
Arrays and Single Nanoparticles as Biological Sensing Platforms” A.
Haes, et al, Journal of Fluorescence, 14,359-367, (2004)
•
They have antimicrobial properties
“Polysilazane-derived antibacterial silver-ceramic nanocomposite”,
V. Bakumov, et al Journal of the European ceramic Society, 27, 3287-3292,(2007)
Method for Making Silver
Nanoparticles
•
AgNO3 solution and reducing agents
typically ammonia, bacteria like K.
pneumoniae , NaBH4 .
– Various reducing agents and
combinations are used in order to
control formation of clusters,
biocompatibility, and morphology
– Color change to yellow or brown is
observed
•
Using UV-Vis spectroscopy the peak
is usually around 430 nm for
spherical Ag nanoparticles
“Interaction of silver nanoparticles with HIV-1”, Y.
Elechiguerra, et al, Journal of Nanobiotechnology, 3, 6,
(2005)
Applications
1. Antimicrobial
a. Medical device polymers / Clothing fabrics
b. Microbes including antibiotic and multi-drug
resistant strains
2. Wound Healing
3. Biosensor/Probes
Antimicrobial Applications – Medical Polymers
and Fabrics
• Experiment involved forming Poly (vinyl
Alcohol) (PVA) nanofibers using Electrospinning
technique that contained SNP.
• PVA/AgNO3 mixture was electrospun.
– Electorspinning involves melting a polymer into a
syringe (mm sized nozzle) and applying a high
voltage that forces the solution out the syringe.
• The nanofiber webs formed were between two hot
plates at 155 ºC and 1 cm apart . This cross-linked
the PVA and also formed some SNP
• Ultra-violet lamp was used for the reduction of
Ag+ to SNP.
“Preparation of antimicrobial poly(vinyl alcohol) nanofibers containing silver nanoparticles”, Hong K, et al,Journal
of Polymer Science: Part B: Polymer Physics, 2468-2474, (2006)
Antimicrobial Applications – Medical Polymers
and Fabrics
“Preparation of antimicrobial poly(vinyl alcohol) nanofibers containing silver nanoparticles”, Hong K, et al,Journal
of Polymer Science: Part B: Polymer Physics, 2468-2474, (2006)
Summary of Medical Device Polymers/ Fabric
Applications
Medical Polymer
Synthesis
Impregnation
Microbe tested
Result
Paper
Date
Application
Ceramic:
poly(methylvinyl)
silazane Ceraset®
VL20
Silver acetate
was dissolved in
oleylamine and
toluene and
heated to 110 C
and refluxed.
Acetone was
added to the
final solution,
SNP was
centrifuged out
of the solution
SNP produced was
added into boiling
polymer solution
Staphylococcus
aureus,
Escherichia coli
Antibacterial
activity was
observed against
both tested species
1
2007
Bone
replacement,
Food industry
Filling an organic
polymer with dilute
organometallic
precursors using
supercritical CO2 at
4000 psi, 40 C for 24
hr. The organometallic
precursors were then
decomposed by using
H2 gas at 1500 psi, 40
C for 24 hr.
Staphylococcus
No bacteria
adhered to
impregnated
silicone discs
washed with water
after 1 hr exposure
even when plasma
was applied to
disc. SNP diffusion
continued to occur
in the discs.
9
2004
Implant
devices
Size: 5 – 7 nm
Silicone
During
Impregnation
Size: 10-100 nm
epidermidis
Summary of Medical Device Polymers/ Fabric
Applications
Medical Polymer
Synthesis
Impregnation
Microbe
tested
Result
Paper
Date
Application
Cotton (weighing 109
g/ml), polyester (89
g/ml), polyester/ cotton
blend (69/35 ratio,
weighing 80 g/ml),
polyester/spandex
blended (92/8 ratio, 85
g/ml)
N/A
Padding
S. aureus, K.
Antibacterial activity
remained after 20
cycles of washing.
Antibacterial activity
was also observed
when using knitted
stretchable single
span fabrics and
printable fabrics
12
2003
Clothing
Polymethymethacrylate
(PMMA)
N/A
Mixing with
Polymethymethacry
late
S. epidermidis,
methicillinresistant S.
epidermidis ,
methicillinresistant S.
aureus
No cytotoxicity
effects. SNP treated
PMMA was effective
against all types of
bacteria
13
2004
Anchoring
artificial
joints
Poly(ethylene
terephthalate)(PET),
chitosan
During
Impregnation
Size: 10-40 nm
Alternately
immersing PET
films in chitosan
containing AgNO3
and heparin. and
ascorbic acid.
AgNO3 was reduced
by ascorbic acid to
form SNP
E. coli
Low cell cytotoxicity,
good anticoagulation
activity, and
antibacterial activity
14
2006
Cardiovascul
ar implants
Pneumoniae
Size: N/A
Summary of Medical Device Polymers/ Fabric
Applications
Medical Polymer
Synthesis
Impregnation
Microbe tested
Result
Paper
Date
Application
Diamondlike
carbon–silver,
diamondlike
carbon–platinum,
diamondlike
carbon–silver–
platinum
composite films
N/A
Pulsed laser deposition
technique
Staphylococcus
warneri
Diamondlike
carbon–silver
composite film
gave high
hardness,
corrosion
resistance, and
good
antimicrobial
activity
15
2006
Cardiovascular,
orthopedic,
MEMS
devices,
Biosensors, etc
Polyethylene
N/A
Ion Beam deposition
technique
Staphylococcus
epidermis
Reduction in
adhered bacteria
was observed
16
2002
Short-term
medical
devices
Immersing in colloid
solution and shaking at
600 rpm
S. aureus
No growth of
bacteria was
observed on the
treated fabric.
17
2007
Clothing
Cotton
Fusarium
Oxysporum (F.
oxysporum) was
grown and used
in making the
silver
nanoparticles
Size: 2-5 nm
Summary of Medical Device Polymers/ Fabric
Applications
Medical Polymer
Synthesis
Impregnation
Microbe tested
Result
Paper
Date
Application
Nylon, Silk
SNPs were
produced by
photo induced
reduction under
UV lamp of
AgNO3/poly(me
thacrylic acid)
solution
Size: <100 nm
During synthesis
Staphylococcus
aureus
Reduction in
bacterial activity
was observed
18
2006
Clothing,
Water
treatment
Polyvinyl alcohol
(PVA)
Polyvinyl
alcohol
(PVA)/AgNO3
solution was
electrospun.
Ultra-violet
lamp was used
for the reduction
of Ag+ to SNP
During synthesis
Staphylococcus
aureus and
Klebsiella
pneumoniae
Growth of both
bacteria was
inhibited by the
presence of SNP in
the electrospun
PVA
19
2006
Wound
Dressing
Plasma-enhanced
deposition of silver
onto surfaces
Five strains of
Listeria
Monocytogens
No viable were
detected after 1218 hr of all species
on the SNP coated
silicone surfaces
20
2004
Medical
surfaces
Size: 5.9-6.3 nm
Silicone Rubber
surfaces
N/A
Use of SNP as Antimicrobial- HIV study
•
The interaction of silver nanoparticles (SNPs) synthesized
using three different capping materials with HIV-1 virus
was studies
– foamy carbon (FC), poly (N-vinyl-2-pyrrolidone)
(PVP), and bovine serum albumin (BSA)
• Results
– Only particles between 1-10 nm bounded the virus
– The viruses did not attach randomly but regular
spatial arrangements among groups of three
– they assumed that the spatial arrangement was due
to silver nanoparticles attaching the specific
disulfide bond location of gp 120 thereby
explaining why larger particles do not interact with
the virus
– The highest inhibition was observed when foamy
carbon was used as capping agent
“Interaction of silver nanoparticles with HIV-1”, Y. Elechiguerra, et al, Journal of Nanobiotechnology, 3, 6, (2005)
Use of SNP as Antimicrobial- HIV study
Results
Spatial arrangement is
observed in BSA-conjugated
SNP showing interaction with
gp 120
gp 120 that binds to CD4
receptors on host cells
protruded more on the outside
and is therefore more open to
attack by SNP
“Interaction of silver nanoparticles with HIV-1”, Y. Elechiguerra, et al, Journal of Nanobiotechnology, 3, 6, (2005)
How Silver Nanoparticles (SNPs)
interferes with Microbe
• (Transmission Electron Microscope) TEM Image Studies of SNP Interaction
with E. Coli Bacteria. (a) SNP congregate at certain areas on the cell wall.
(b) SNP enter the cell wall. (c) SNP anchor at various sites causing
perforations in membrane.
“Characterization of enhanced antibacterial effects of novel silver nanoparticles”, S. Shrivastava, et al, Nanotechnology 18 (2007) 225103-225111
How Silver Nanoparticles (SNPs)
interferes with Microbe
• Note:
– Gram negative have lipopolysaccharide cell wall that are negative. This
negative charge attracts the weak positive charge of SNP.
– Gram positive have peptidoglycan cell walls that are more rigid and contains
extended cross-link providing fewer attachment site for SNPs and less
penetration
• Free radical formation
• Phosphotyrosine profile studies
– Dephosphorylation of two peptides was observed in gram negative bacteria.
Tyrosine phosphorylation in bacteria leads to activation of RNA polymerase
sigma factors and other important growth enzymes.
Characterization of enhanced antibacterial effects of novel silver nanoparticles”, S. Shrivastava, et al, Nanotechnology 18 (2007) 225103-225111
Cytotoxicity of SNP to Eukaryotic Cell
• SNP is not toxic to eukaryotic cell at low
concentrations
• Why?
– Eukaryotic cells are larger
– Higher structural and functional redundancy
• Electron transport chain are intracellular
• Lots of mitochondria
“An in vitro assessment of the antibacterial properties and cytotoxicity of nanoparticulate silver bone cement”, V. Alt, et al, Biomaterials,25,4383-4391,(2004)
Summary of Antimicrobial Applications
SNP Synthesis
Microbe
Application
Method
Result
Paper
Year
K.Pneumonia in silver
nitrate solution
Size: 5 - 32 nm
S. aureus, E. coli
Added into
agar plates in
addition to
antibioticspenicillin G,
amoxicillin,
erythromycin,
clindamycin,
and
vancomycin
In the presence of SNP,
Increased antimicrobial
activity of antibioticspenicillin G, amoxicillin,
erythromycin,
clindamycin, and
vancomycin was observed
2
2007
Dissolving silver
nitrate in water and
adding NH3 and
adding D-glucose,
hydrazine, to increase
reaction rate
Non-resistant Escherichia
Coli, ampicillin resistant
Escherichia Coli,
chloramphenicol,
amoxicillin, and
trimethoprim resistant
Salmonella typhus,
Staphylococcus aureus
Added into
agar
plates
Antibacterial effect of SNP
was independent of the
type of resistant acquired
by the bacteria. SNP was
more effective against
gram negative bacteria.
Increasing concentration of
SNP produced better
inhibition of gram negative
bacterial growth. Higher
concentrations of SNP
were required to inhibit
growth in gram positive
bacteria
3
2007
Size: 10 – 15 nm
Summary of Antimicrobial Applications
SNP Synthesis
Microbe
Application
Method
Result
Paper
Year
by mixing cold
solutions of AgNO3
with NaBH4
E. coli, yeast, S. aureus
Added into
agar plates
containing
Itraconazol
for yeast and
gentamicin
in E. coli
Minimum inhibitory
concentration was
determined to be greater
than 6.6 nm, 33 nm and 3.3
nm in yeast, E. coli, and S.
aureus respectively.
Specificity for silver was
observed
10
2007
SNPs were synthesized
using three different
capping materials i.e.
foamy carbon (FC),
poly (N-vinyl-2pyrrolidone) (PVP),
and bovine serum
albumin (BSA).
Size:
FC –16.19 ± 8.69 nm
PVP-6.53 ± 2.41 nm
BSA-3.12 ± 2.00 nm
HIV-1
During
Synthesis
Only particles between 1-10
nm interacted with HIV-1.
The interaction was
spatially dependent due
distribution of glycoprotein
knobs (gp 120) on the viral
cell that are vulnerable to
attack by SNP.
6
2005
Biosensor Applications
• The study involves the use of silver
nanoparticle (SNP) to enhance the
sensitivity of glucose biosensors.
– Glucose Oxidase (GOx) extracted from
Aspergillus niger was mixed with
silver nanoparticles was coated onto a
platinum electrode using a sol-gel
process
– Current measurements were used in
determining the sensitivity of the
glucose biosensor
– When current values remained
steady, the different concentrations of
β-D glucose were added.
How SNP affected the Sensor
• An alternate pathway for the
flow of electrons was created
in the presence of SNP
• The response time was
quicker by using SNP and the
sensing is improved
“Using silver nanoparticle to enhance current response
of
biosensor”, R. Xiangling ,et al , Biosensors and bioelectronics,
21, 433-437, (2005)
Summary of Biosensor Applications
Spectroscopic
method
Synthesis
Application
Result
Paper
Year
Surface
Enhanced
Raman
Spectroscopic
(SERS)
The produced silica
spheres were dispersed
in 30 mM AgNO3
ethylene glycol solution
with 50 μL ammonium
hydroxide added; the
stirred solution was left
for 10 hr at 50 C
Surface-enhanced
Raman spectroscopic
tagging material (SER
dots) containing silica
spheres surrounded by
silver nanoparticles and
organic Raman labels
for targeting cellular
cancer in living cells
Images showed the
distribution of specific
antibodies to the cells
tested. Controls yielded
no results.
5
2006
Glucose Detection by
current measurement
Sensitivity and
response time improved
by the application of
SNP.
7
2005
Size: 4-5 nm
Glucose
Biosensor
SNPs were synthesized
by reduction of AgNO3
by NaBH4. Glucose
Oxidase extracted from
Aspergillus Glucose
Oxidase extracted from
Aspergillus niger was
mixed with silver
nanoparticles and was
coated onto a platinum
electrode using a solgel process.
Size: 4.8 nm
Summary of Biosensor Applications
Spectroscopic
method
Synthesis
Application
Result
Paper
Year
SERS
By depositing 9 nm
thick silver on a
cleaned glass surface
with 1 nm thick
chromium layer by
physical vapor
deposition (PVD) and
then dipping in several
organic mixtures.
Gene Detection
Spatial and
Spectroscopic chemical
analysis could be done
using this procedure.
The analysis could be
done over a wide range
of spectral regions
simultaneously.
8
2005
Localized
Surface
Plasmon
Resonance
Spectroscopy
SNP was dropped on
glass substrate and used
for other steps in
experiment
Bimolecular Detection
The potential use of the
device was demonstrated
using streptavadin/biotin
system.
11
2003
Wound Healing Application of Silver
Nanoparticle
• Burn wounds were caused on mice
– SNP, silver sulfadiazine (SSD) was applied to
wound site
– Wounds treated with silver nanoparticles were
compared antibiotics amoxicillin and
metronidazole treated wounds
Results of Experiment on Wound Healing
They were able to show that silver nanoparticles both reduce scar appearance and
speeds up recovery
“Topical Delivery of Silver Nanoparticles Promotes Wound Healing” J.Tian, K.Wong, et al, ChemMedChem,2,129-136, (2007)
Silver Nanoparticles (SNPs) Play a greater role in Wound
Healing not just antibacterial
•
Cytokine Modulation
was detected in
samples treated with
SNPs
– SNP Controls
Inflammation
i.e. decreased
inflammation is
observed
Dark Pink
Spots
represent
Neutrophils
“Topical Delivery of Silver Nanoparticles Promotes Wound Healing” J.Tian, K.Wong, et al, ChemMedChem,2,129-136, (2007)
Summary
•
SNP has a wide range of current and potential applications as antibacterial,
biosensor, and wound healing component in medicine.
•
SNP is typically produced by reducing Ag+ in aqueous solution using chemical
reducing agent or microbes.
•
SNP kills bacteria by entering into the cell, interfering with the various cellular
processes, and is not yet well understood. It is more effective against gram
negative.
•
SNP not only kills bacteria during wound healing but has been shown to also
reduce inflammation through cytokine modulations
•
It is not only effective against bacteria! It can act on viruses and fungi as well.
•
Since SNP so far has been shown to be non-toxic to eukaryotic cell, their
application against drug resistant bacteria, viruses, and fungi seem very promising.
Reference
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»Thanks
• Questions/Comments