1. In The Name
Of
GOD
1

2.  Evelyn Assadian*1,2, Jalal Pourahmad1, Mohammad
hadi Zarei1
 Hamid Degampanah2, Ali Ghanadzadeh gilani2
 1Faculty of Pharmacy, Shahid Beheshti University of
Medical Sciences, Tehran, Iran
 2 Department of Chemistry, Faculty of Science
,Guilan University,Gilan, Iran
2

3. Toxicity of Copper oxide (CuO)
nanoparticles on human blood
lymphocytes
3

4. 4

5. Copper oxide (CuO) nanoparticles have different
application in various part of human life such as
medicine, engineering and technology.
In this study we investigated the cytotoxicity of CuO
nanoparticle on human lymphocytes and then the
effect of CuO nanoparticles on different cellular and
subcellular factor was assessed.
Our results showed CuO Nanoparticles induced
cytotoxicity on human blood lymphocytes was
associated with significant increase at intracellular ROS
level and loss of mitochondrial membrane potential and
lysosomal membrane leakage.
5

6. Copper oxide (CuO) nanoparticles have a lot of use in
different human life sectors such as medicine,
engineering and technology.
These nanoparticles were used as catalysts,
semiconductors, sensors, gaseous and solid ceramic
pigments, magnets rotatable devices, electrode
materials, hydrophilic materials and super conductors.
Furthermore, CuO nanoparticles were used in the
pharmaceutical industry and in the production of anti-
microbial fabric treatment or prevention of infections
caused by Escherichia coli and methicillin-resistant
Staphylococcus aureus (Ren, Hu et al. 2009).
6

7. Possible routes of exposure to Cuo
nanoparticles are through inhalation and skin
exposure.
Toxicity of this nanoparticles have been
reported in various studies, but no study so far
investigated complete cellular mechanisms
involved in CuO nanoparticles toxicity on
human cells (Chang, Zhang et al. 2012).
In this study we aimed to assess the
cytotoxicity of Cuo nanoparticles on human
blood lymphocytes.
7

8. There is evidence that these substances affect the
immune system, but no comprehensive
mechanistic research has been done about them.
 Extensive contact with nanoparticles of copper
in its applications in industry, medicine and even
textiles
 The critical role of lymphocytes in the immune
system
 Increase in diseases related to the immune
system
8

9. Blood lymphocytes were isolated from healthy
male volunteers’ blood, using Ficoll polysaccharide
followed by gradient centrifugation.
Cell viability, reactive oxygen species (ROS)
formation, lipid peroxidation, glutathione levels,
and damage to mitochondria and lysosome were
determined in blood lymphocytes after 6-h
incubation with different concentrations of CuO
Nanoparticle (Salimi, Vaghar -Moussavi et al. 2016).
9

10. Böyum A, 1996
2 h 4 h 6 h 12 Hour
EC50
10

11. Nontoxic
Toxicity
threshold
Toxic
Before 12 hour 12 hour After 12 hour
Mechanistic
Study
EC50 Cytotoxic Study
June 29, 2015 10
RP
TC RP
TC
11

12. Measurement of cytotoxicity using
trypan blue and determination
EC50
• Pourahmad J et al, 2012
Measuring reactive oxygen radicals
with the reagent dichlorofluorescein
diacetate
• Chignell CF et al, 2003
Measurement GSH,GSSG with
Hissin-Hilf method
Hissin PJ et al, 1976
12

13. Evaluation of lipid peroxidation by
measuring(MDA)
Ohkawa H et al, 1979
Measurement of mitochondrial
membrane potential with Cations
Rhodamine 123
Andersson B et al, 1987
Measurement of membrane damage
of lysosomes with using acridine
orange reagent
Pourahmad J et al, 2001
13

14. ‫آناليز‬‫نتايج‬‫توسط‬‫ن‬‫مو‬‫ز‬‫آ‬ANOVA‫يک‬‫طرفه‬‫و‬‫دوطرفه‬‫و‬‫به‬‫ترتیب‬
‫با‬‫پست‬‫تست‬Tukey‫و‬Bonferroni‫انجام‬‫می‬‫گيرد‬.‫جهت‬
‫ی‬ ‫س‬‫ر‬‫بر‬‫نيسيته‬‫ژ‬‫همو‬‫يانس‬‫ر‬‫وا‬‫در‬‫يک‬،‫ه‬‫و‬‫گر‬‫از‬‫تست‬Leven
‫استفاده‬‫کرده‬‫و‬‫نتايج‬‫ت‬‫ر‬‫بصو‬‫ميانگين‬‫سه‬‫مايش‬‫ز‬‫آ‬‫جداگانه‬
mean±SD‫ش‬‫ر‬‫ا‬‫ز‬‫گ‬‫خواهد‬‫شد‬.
14

15. 15

16. 16

17. 17

18. 18

19. Our results showed that CuO nanoparticle, in
particular, decreased cell viability in a
concentration-dependent manner and 382µM
CuO nanoparticles caused 50% decrease in
human lymphocytes viability (Fig. 1).
CuO nanoparticles induced cytotoxicity on
human blood lymphocytes was associated with
significant increase at intracellular ROS level
(Fig. 2) and loss of mitochondrial membrane
potential and lysosomal membrane leakage.
19