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Antiinflamatory effect of materials based on gold and silver nanoparticles and Sambucus Nigra L. on carrageenan-
induced rat paw edema Adriana Filipa, Doina Daicoviciua, Nicoleta Deceaa, Pompei Bolfab, Simona Clichicia, Adriana Muresana, Luminita Davidc, Liliana Olenicd
aDepartment of Physiology, „Iuliu Hatieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania; bDepartment of Morphopathology, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania; cFaculty of Chemistry and Chemical Engineering, Babes-Bolyai –University, Cluj-Napoca, Romania; dNational Institute for Research and Development of Isotopic and Molecular Technologies, Cluj-Napoca, Romania
22nd EADV, October 2-6, 2013 Istanbul, Turkey
This work was elaborated in the frame project no 147/2012, through the program “Partnerships in priority areas-PN II”
Introduction
The biological synthesis of silver and gold nanoparticles using plants or fruits extracts plays an important role in the field of nanotechnology and nanomedicine as it offers alternative therapeutic options which are safe, free of side effects and effective for a variety of diseases.
Inflammation is a complex biological response involved in pathogenesis of various diseases.
Although steroids and NSAIDs are the main therapeutic agents used in the treatment of inflammation they cause serious side effects. Therefore, the development of new materials with comparable results and no side effects is needed.
AimAimto investigate the biological activity of materials based on gold to investigate the biological activity of materials based on gold (Au-SN) and silver (Ag-SN) nanoparticles and natural (Au-SN) and silver (Ag-SN) nanoparticles and natural compounds extracted from European black elderberry - compounds extracted from European black elderberry - SambucusSambucus nigra L.,nigra L., (SN) on acute inflammation model in Wistar (SN) on acute inflammation model in Wistar rats.rats.
Materials and Methods the inflammation was induced by intraplantar injection of 100 μl 1%
carrageenan SN extract (15 mg/kg b.w.) and the materials based on gold (Au-SN) and
silver (Ag-SN) nanoparticles functionalized with SN extract (0.3 mg/kg b.w.) were administrated orally during 4 days before the injection of carageenan
the response was compared to indomethacin (5 mg/b.w.) as positive control and saline solution as negative control.
at 2h, 24h and 48h after carageenan injection were evaluated: the paw edema the immunohistochemical expression of iNOS and COX-2 in the paw
tissue the malondialdehyde (MDA), glutathione reduced/glutathione oxidized
ratio (GSH/GSSG), IL-1α, IL-1β and IL-6 levels in soft plantar tissue
Comparisons were made by one-way ANOVA, with Tukey multiple comparisons test using a GraphPad Prism software program, version 5.0 (GraphPad, San Diego, Ca, USA).
p<0,05 was considered as significant.
Malondialdehyde level (a), GSH/GSSG ratio (b) and paw edema (c) in soft tissue at 2h, 24h and 48h after carrageenan injection
GSH/GSSG 24h
Indom ctrl
SNAu-SN
Ag-SN0
2
4
6
8
**ratio
GSH
/GSS
G
MDA 48h
ctrl
Indom SNAu-SN
Ag-SN0.0
0.1
0.2
0.3
0.4
*
nmol
es/m
g pr
otei
n
MDA 2h
Indom ctr
lSN
Au-SNAg-SN
0.0
0.5
1.0
1.5
* *nmol
es/m
g P
MDA 24h
Indom ctr
lSN
Au-SNAg-SN
0.0
0.2
0.4
0.6
0.8 ***
nmol
es/m
g P
GSH/GSSG 2h
Indom ctr
lSN
Au-SNAg-SN
0
1
2
3
4
5
ratio
GSH
/GSS
G
GSH/GSSG 48h
Indom ctrl
SNAu-S
NAg-S
N0
5
10
15
20 **
ratio
GSH
/GSS
G
Paw edema
0 10 20 30 40 500.0
0.5
1.0
1.5
ctrl
Ag-SNSNAu-SN
Indom
time (hour)
paw
thic
knes
s (m
m)
a
b
c
Proinflammatory cytokines levels in soft plantar tissue at 2h, 24h and 48h after carrageenan injection in animals pretreated with materials based on gold and silver nanoparticles and natural extracts
IL-1 24h
ctrl
Ind SNAU-S
NAg-S
N0
200
400
600
800M
FI
IL-1 2h
Indom ctrl
SN
Au-SNAg-SN
0
500
1000
1500
2000
* ** **
MFI
IL-1 24h
ctrl
Indom SN
Au-SNAg-SN
0
200
400
600
MFI
IL-1 48h
ctrl
Indom SN
Au-SNAg-SN
0
1000
2000
3000
MFI
IL-1 , 2h
ctrl
Ind SNAu-SN
Ag-SN0
500
1000
1500
*****
MFI
IL-1 48h
ctrl
Ind SNAu-S
NAg-S
N0
2000
4000
6000
MFI
iNOS 2h COX-2, 2h
COX-2, 24hiNOS, 24h
IL-6 2h
ctrl
Indom SNAu-S
NAg-S
N0
50
100
150
*****
MFI
IL-6 24h
0
10
20
30
MFI
IL-6 48h
ctrl
Indom SNAu-SN
Ag-SN0
50
100
150
200
**MFI
Conclusions SN significantly inhibited edema formation and MDA SN significantly inhibited edema formation and MDA
generation in carrageenan-induced paw edema model, generation in carrageenan-induced paw edema model, particularly at 2h. particularly at 2h.
Silver nanoparticles functionalized with SN extract Silver nanoparticles functionalized with SN extract increased GSH/GSSG ratio at 48h after induction of increased GSH/GSSG ratio at 48h after induction of inflammation. inflammation.
Both SN extract and silver nanopartciles based on SN Both SN extract and silver nanopartciles based on SN diminished IL-1diminished IL-1αα, IL-1, IL-1ββ and IL-6 levels and reduced iNOS and IL-6 levels and reduced iNOS and COX-2 expressions at 2h after carrageean injection.and COX-2 expressions at 2h after carrageean injection.
These findings suggested that both silver nanoparticles These findings suggested that both silver nanoparticles functionalized with SN as well as SN extract may be useful functionalized with SN as well as SN extract may be useful agents in the treatment of inflammatory conditions.agents in the treatment of inflammatory conditions.
HE, 48h COX-2, 48hiNOS 48h