الفهرس 
Aim of the workOnly 14 pages are availabe for public view
 |  | from | 220 |  |  |
| | | from | 220 | | |
Abstract
One of a neglected parasitic disease is CE caused by E. granulosus. It
has huge socio-economic impact with a worldwide geographical
distribution.The available treatments are surgery and chemotherapy
with ABZ, MBZ or other antihelmintic drugs. Chemotherapeutic
options for echinococcosis are extremely limited and success with
ABZ treatment (the drug of choice) is not guaranteed, therefore, new
drugs are needed.
Among several metal NPs, Ag NPs was found nontoxic substance
with satisfactory antimicrobial efficacy against viruses, bacteria and
parasites. Moreover, it was evaluated as a suitable carrier of various
therapeutic drugs. Another way of treatment is the herbal extracts.
One of these herbs is Punica granatum which is well-known for many
therapeutic and pharmaceutical effects such as anti-cestoidal, antinematoidal
and anti-protozoan, antioxidants and anti-inflammatory
substance that contain polyphenols and tannins.
The present study was conducted in order to evaluate the effects of Ag
NPs and PGE in comparison with ABZ on experimentally infected
mice with E. granulosus. The therapeutic effects of these drugs on
hydatid cyst were assessed through parasitological, histopathological,
TEM ultrastructural studies. Also, serological studies by detection of
IFN-γ in mouse serum samples by ELISA and liver enzymes were
assessed.
Summary and Conclusion
149
The study was carried out on 125 pathogen-free laboratory bred
female albino mice and the experimental animals were divided into
the following seven groups:
group I (GI): Five mice were served as uninfected and untreated
group (negative control).
group II (GII): Twenty mice served as infected untreated group
(positive control).
group III (GIII): Twenty mice served as ABZ-treated group. The
treatment was started three months p.i. ABZ was given at a dose of
200mg/Kg/day for five consecutive days per week orally for two
months.
group IV (GIV): Twenty mice served as Ag NPs treated group. The
treatment was started three months p.i. Ag NPs suspention was given
at a dose of 25mg/Kg/day for five consecutive days per week orally
for two months.
group V (GV): Twenty mice served as ABZ coated with Ag NPs
treated group. The treatment was started three months p.i. ABZ coated
with Ag NPs was given at a dose of 100mg/Kg/day for five
consecutive days per week orally for two months.
group VI (GVI): Twenty mice served as PGE treated group.
Treatment was started two days p.i and the extract was given at a dose
of 0.65 g/kg/day orally for three months.
group VII (GVII): Twenty mice served as combined ABZ with
PGE- treated group. Treatment was started three month p.i. The drugs
were given at a dose of 200mg/Kg/day of ABZ and 0.65 g/kg/day of
PGE for five consecutive days per week orally for two months.
Summary and Conclusion
150
The experiment was terminated five months p.i. All mice overcoming
the infection and treatment period were subjected to evaluate
parasitological, liver histopathological on hydatid cyst using TEM and
also detection of IFN-γ in mouse serum samples by ELISA and
assessment of ALT, AST and ALP liver enzymes activities were
performed.
Concerning mortality rates of experimental animals among the
different studied groups, the highest mortality rate was reported in
group II (infected not treated group) while the lowest mortality rate
was recorded in all the treated groups.
Concerning hydatid cyst sizes, weights and efficacy rates of
treatments results, there were significant differences between the
infected treated groups (GIII, GIV, GV, GVI and GVII) and the
infected untreated group (GII) (P <0.001). The biggest hydatid cyst
size and weight (5.14 ± 1.75 mm, 0.78 ± 0.08 g) were detected in the
infected control group (GII). All the treated groups showed significant
reductions; however, the smallest values were detected in ABZ-loaded
nanoparticles-treated group (GV)(0.39 ± 0.48mm, 0.11 ± 0.14 g)
followed by 0.48 ± 0.47 mm, 0.17 ± 0.18 g which were detected in
PGE+ABZ (GVII). Accordingly, the least efficacy rate of 22.9% was
obtained with ABZ while the highest efficacy rate of 63.9% was
obtained with ABZ-loaded nanoparticles followed by the efficacy rate
of PGE+ABZ (GVII) (56.2%).
Summary and Conclusion
151
The histopathological study revealed the occurrence of granulomatous
reactions (Table 5,oFigo34) associated with liver fibrosis (Table 6, Fig
35), inflammatory infiltration (Table 7, Fig 36), congestion (Table 8,
Fig 37), liver cell necrosis (Table 9, Fig 38), apoptosis (Table 10, Fig
39) and steatosis (Table 11, Fig 40) mainly in the infected control
group and variable degrees of improvements were found in the treated
groups.
The results revealed improvements of the granulomatous reactions
sizes, fibrosis, congestion and the inflammatory infiltrates in all the
treated groups. However, free nanosilver was slightly better than free
ABZ and the best results were obtained with ABZ-loaded nanosilver
treatment followed by PGE+ABZ treatment. Moreover, liver cell
necrosis and steatosis were increased with free ABZ treatment
denoting a hepatotoxic effect of this drug. Conversely, improvements
were observed with PGE (GVI) due to its hepatoprotective effect
followed by ABZ-loaded nanoparticles (GV) indicating the potential
safety of AgNPs in vivo and decreasing ABZ toxic effects.
Concerning the results of TEM, the ultrastructure of the hydatid cysts
obtained from GII revealed intact germinal layer with microtriches
projecting into the laminated layer. Undifferentiated cells with intact
nuclear membranes and non-distorted nucleus were also seen. The
cytoplasm of these cells was intact and rich in glycogen granules.The
hydatid cysts from other treated groups showed slightly vacuolated
cytoplasm, glycogen depletion and heterochromatin (pyknosis) in the
nucleus. The hydatid cysts retrieved from AgNPs and ABZnanoparticle-
treated groups showed destruction of the germinal layer
with the loss of microtriches, separation from the laminated layer and
depletion of glycogen. The nanosilver particles appeared as black rods
inside the nuclei which showed fragmentations.
Summary and Conclusion
152
The cytoplasm showed distortion of the mitochondria, glycogen
depletion and autophagosomes in vacuolated cytoplasm.These
changes were more obvious with ABZ-loaded nanoparticles and
PGE+ABZ treatment.
In the present study, there was a significant difference (P <0.0001)
between the studied groups regarding serum values of IFN-γ. There
were significant increase in the sera IFN-γ in all groups when they
were compared with the normal control group (GI). The highest
significant increase was in ABZ-treated group when it was compared
with GI and GII. Interestingly, with nanosilver treatment either alone
(GIV) or combined with ABZ (GV) and with PGE treatment either
alone (GVI) or combined with ABZ (GVII), there were significant
reductions in serum IFN-γ when they were compared with the infected
control (GII) and ABZ-treated (GIII) groups. Moreover, the most
significant decrease was in ABZ-loaded nanoparticles-treated group
(GV) followed by PGE+ABZ treated group (GVII).
Regarding liver enzymes, there were significant increases (P < 0.001)
of the serum levels of ALT, AST and ALT between the studied
groups. In ABZ-treated group, there were increases in the levels of
ALT, AST and ALP when they were compared with GI. An obvious
improvement of liver enzymes was found in ABZ-loaded Ag NPs
(GV), PGE (GVI) and PGE+ABZ -treated groups (GVII) when it was
compared with the infected control (GII); however, the values were
still significantly higher than the normal control group (GI).
Summary and Conclusion
153
In conclusion, this study demonstrated that ABZ-loaded nanoparticles
showed the best drug efficacy on experimental echinococcosis with
significant reduction of hydatid cyst size, weight and improvement of
the histopathological changes with less hepatic injury in comparison
with ABZ. This was followed by the results obtained with combined
treatment (PGE+ABZ). Therefore, they could be considered as new
potential therapeutic options against E. granulosus infection instead of
using ABZ alone, however, further studies should be done to evaluate
these treatments from other aspects.