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