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JMAU 66 110
Original Article
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Histology Department, Faculty of Medicine, Tanta University, National Institute of Laser Enhanced Sciences (NILES), Cairo University,
Egypt
10 Q3 b Medical Applications of Lasers (MAL) Department, National Institute of Laser Enhanced Sciences (NILES), Cairo University, Egypt
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a r t i c l e
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i n f o
a b s t r a c t
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Article history:
Received 6 February 2015
Received in revised form 21 April 2015
Accepted 2 May 2015
Available online xxx
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Keywords:
Gold nanoparticles (GNPs)
GFAP
Brain
Astrogliosis
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1. Introduction
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http://dx.doi.org/10.1016/j.jmau.2015.05.001
2213-879X/ 2015 Published by Elsevier Ltd. on behalf of Saudi Society of Microscopes.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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2.1. Chemicals
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2.2. Animals
This work was carried out on 20 adult male albino rats
weighing 180200 g. Experimental protocol was approved
by the local ethical Committee of Faculty of Medicine,
Tanta University, Egypt prior to the study and the animal
care was carried out according to the national guidelines
for animal care. Drinking water and conventional food
were provided ad libitum. Animals were divided into two
equal groups. The rst one served as a control group and
kept without treatment throughout the whole period of
the experiment. The second one served as an experimental group and received orally 400 g/kg/day for 8 weeks
of solutions of 110 g of gold NPs/ml by gastric tube
[6]. At the end of the eight weeks, animals were anesthetized and decapitated and the brain was harvested by
careful rapid dissection and was immediately xed in 4%
paraformaldehyde for both histological and immunohistochemical study.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Fig. 1. Showing six layers of cerebral cortex of control group; molecular layer (I), outer granular layer (II), outer pyramidal layer (III), inner granular layer
(IV), inner pyramidal layer (V) and polymorphic layer (VI). These layers showing acidophilic neuropile and rounded open face nuclei with prominent
nucleoli () of the neurons and also of neuroglial cells without properly seen cytoplasm (). H&E, (A) 100; (B) H&E, 400.
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3. Results
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albino rat of control group revealed six distinguished layers with no sharp boundaries in association small blood
vessels in between. These layers were; plexiform layer of
bers traveling parallel to the surface with relatively few
cells, external granular layer, small pyramidal cell layer,
inner granular layer, inner layer of large pyramidal cells
and nally the layer of polymorphic cells (Fig. 1). The
Fig. 2. Brain of control group showing large (A) and medium sized blood vessel (B) and continuous wall of blood capillaries () (C). H&E, 1000.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Fig. 5. Cerebral cortex of gold nanoparticle treated group showing aggregates of gold nanoparticles in the neuropil of the cerebral cortex ()
together with distorted darkly stained neurons with darkly stained nuclei
() and intracytoplasmic vacuoles (V). H&E, 1000.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Fig. 7. Brain of GNPs treated group showing large sized BV (A) and medium sized BV (B) with hyalinization of their walls and GNPs deposition. Notice
disruption of the wall of blood capillaries (C&D) (). H&E, 1000.
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cortex (Fig. 9A) and also in the hippocampus (Fig. 9B). The
choroid showed few localized mildly positive choroidal
epithelial cells for active caspase 3 (Fig. 10). Similarly, gold
nanoparticle treated animals revealed negative caspase 3
reaction in the cerebral cortex and only few positive neurons were seen in the hippocampus (Fig. 11). On the other
hand, choroidal epithelium of the same group showed
strong positive reaction for active caspase-3 in many cells
(Fig. 12).
No signicant difference in the number of caspase 3 positive neurons in the GNPS treated group compared with
control group as regard cerebral cortex and hippocampus.
However, there was signicant increase in the caspase 3
positive immunoreaction of the choroidal epithelium cells
in the GNPS treated group (Table 1). Morphometric analysis revealed that the number of immunoreactive astrocytes
in the control group was more numerous in hippocampus
more than cerebral cortex. Signicant increase in the GFAPimmunoreactive astrocytes was also observed in the brain
of gold nano particles treated group in comparison to control group with specic predominance in the hippocampus
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Fig. 9. Cerebral cortex (A) and hippocampus (B) of brain of control group showing negative immune reaction for caspase-3. Caspase 3 immunostaining,
1000.
Fig. 10. Choroid plexus of control group showing few mildly positive cells
for active caspase-3 in the choroid epithelial cell. Caspase 3 immunostaining, 1000.
Fig. 11. Hippocampus of GNPs treated group showing positive neuron for
active caspase 3 (). Caspase 3 immunostaining, 1000.
Table 1
Mean value of number of caspase-3 positive cells and GFAP positive astrocytes in cerebral cortex and hippocampus of the studied groups.
Measured parameters
Control
0.75
1.26
1.86
7.083
11.75
GNPs group
0.45
0.703
0.351
1.76
2.251
0.833
1.46
5.133
12.028
17.25
0.577
0.516
1.407
3.24
2.712
t value
P value
0.394
0.887
8.72
6.79
4.413
0.698
0.382
0.000
0.003
0.001
Data expressed as mean standard deviation, t = Student (t) test, P value = probability of chance, P < 0.05 is signicant.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Fig. 12. Choroid plexus of GNPs treated group showing many positive
choroidal epithelial cells for active caspase 3 (). Caspase 3 immunostaining, 1000.
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compared to cerebral cortex (Table 1). Similarly the diameters of astrocyte cell bodies, the length and the numbers
of their processes were signicantly increased in the brain
of GNPs treated group compared to control one (Table 2).
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4. Discussion
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The present study demonstrated that gold nanoparticles administration displayed histological alterations in the
Fig. 13. Cerebral cortex (A) hippocampus (B) of control group showing few widely apart small sized GFAP positive astrocytes with few short processes
(). GFAP immunostaining, 1000.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Fig. 14. Cerebral cortex (A) and hippocampus (B) of GNPs treated group showing strongly positive GFAP hypertrophic astrocytes with multiple elongated
processes () (moderate astrogliosis). GFAP immunostaining, 1000.
Fig. 15. Cerebral cortex (A) and hippocampus (B) of GNPs treated group showing strong GFAP reaction and hypertrophic astrocytes (severe astrogliosis).
GFAP immunostaining, 1000.
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Table 2
Mean values of diameter of the astrocytes cell bodies and length and number of their processes in the brain of control and gold nanoparticle treated groups.
Measured parameters
Control
GNPs group
t value
P value
14.63 3.18
17.52 4.98
3.27 0.64
20.74 4.12
32.37 7.32
4.9 0.83
2.370
5.519
5.153
0.033
0.001
0.001
Data expressed as mean standard deviation, t = Student (t) test, P value = probability of chance, P < 0.05 is signicant.
Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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None declared.
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Acknowledgements
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Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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Please cite this article in press as: El-Drieny EAEA, et al. Histological and immunohistochemical study
of the effect of gold nanoparticles on the brain of adult male albino rat. J Microsc Ultrastruct (2015),
http://dx.doi.org/10.1016/j.jmau.2015.05.001
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