الفهرس 
TitleOnly 14 pages are availabe for public view
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Abstract
Breast cancer is the most common female malignancy accounting for 22.9% and
37.7% of all female cancers worldwide and in Egypt, respectively. Breast cancer in Egypt
carries an unfavorable prognosis with 29% mortality and 1: 3.7 incidences to mortality ratio.
Breast cancer accounts for 16% of all cancer deaths among women globally, according to the
report by the World Health Organization. It is the most common solid tumor diagnosed in
women. Although the incidence of breast cancer increases with age, certain lifestyle and
environmental factors play an important role on breast cancer risk. In Egypt, breast cancer is
the most common cancer among women, representing 18.9% of total cancer cases (35.5% in
women and 2.2% in men) among the Egypt National Cancer Institute (NCI) series of 10,556
patients during the year 2002-2003.
Breast cancer and cancer related diseases have been treated using surgery,
chemotherapy, and radiation therapy, or a combination of these. But despite these therapeutic
options, cancer remains associated with high mortality. This is basically due to difficulties in
early diagnosis, exorbitant cost of treatment, with the often late presentation of breast cancer
that generally characterizes cancer diagnosis among Egypt and other African women. Owing
to these several shortcomings, there is a need for better therapeutic options which will
increase the chances of survival of breast cancer patients with minimal or no side effects of
treatment.
Photodynamic laser therapy with exposure to ultrasound is a breakthrough and a
modern technology to eliminate tumor and infected tissues without affecting the healthy tissue
adjacent to and away from the tumor. This requires the use of a so-called laser-ultrasound
sensor, which is highly concentrated in the tumor area, which responds ideally to photonic
energy and ultrasonic waves, which works to stop tumor growth and elimination, which is a
promising technique to eliminate superficial and deep cancer tumors.
The present work aims to cure Ehrlich ascites carcinoma tumor implanted to group of
mice as an experimental animals using a sonophotodynamic modality in combination with
chlorophyll as a sonophoto sensitizer drug. Two sources of energy were used; namely infrared
laser at two frequency levels (4000 and 7000 Hz) and Ultrasound (pulsed and continuous
wave mode) for 3 min.
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A total of 140 male Swiss albino mice Tumor bearing animals were divided into sub
groups each of 10 mice as follows:
Control mice: 10 mice served as control group, i.e., free of tumor.
Tumor- bearing mice were divided into the following groups:
group I: (20 mice)
a) 10 mice: tumor bearing mice without treatment.
b) 10 mice: Tumor bearing mice treated with (CHL) as sonophotosensitizer only.
group II: (20 mice, laser irradiated group)
a) 10 mice: Exposed to Infra-Red Laser, 4000Hz, for 3 minutes.
b) 10 mice: Exposed to Infra-Red Laser, 7000Hz, for 3 minutes.
group III: (20 mice, ultrasound group)
a) 10 mice: Exposed to pulsed ultrasound for 3 minutes.
b) 10 mice: Exposed to continuous ultrasound for 3 minutes.
group IV: (20 mice, CHL, laser group), the mice of this group were injected intraperitoneally
(IP) with (CHL), then the tumor site were irradiated to laser light at same conditions of group II.
group V: (20 mice, CHL, ultrasound group), the mice of this group were injected (IP) with
(CHL), then were divided into 2 sub-groups. The tumor site was irradiated to ultrasound at
same conditions of group III.
group VI: (30 mice, combined treatment groups)
a) 10 mice: The tumor site was irradiated to laser light for 3 minutes, followed by ultrasound
for 3 minutes.
b) 10 mice: Injected (IP) with (CHL). The tumor site was irradiated to laser light for 3 min,
followed by ultrasound for 3 minutes.
c) 10 mice: Injected (IP) with (CHL + Vitamin B12). The tumor site was irradiated to laser
light for 3 minutes, followed by ultrasound for 3 minutes.
The treatment effects evaluation:
The dimensions of the tumor, e.g., the length, the width and the height of were
measured using a slide digital caliper every day before any treatment modality. The
tumor volume was calculated.
Tumor mass inhibition ratio and tumor volume growth ratio were also calculated.
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Biochemical examinations were appliedto detect serum levels of ALT, AST, urea and
creatinine to detect the effect of CHLinjection on liver and kidney functions.
The concentrations of selenium (Se), iron (Fe), nickel (Ni), chromium (Cr), cadmium
(Cd), lead (Pb), zinc (Zn) and copper (Cu) were estimated using Atomic absorption
spectrophotometry.
Activities of some antioxidants were measured, namely; glutathione-S-transferase
activity (GST), super oxide dismutase activity (SOD), glutathione reductase activity
(GR), catalase activity (Cat), total antioxidant activity (TAC) and malondialdehyde
(MDA).
Measurements of the dielectric properties of the tumor tissue and its surrounding at the
end of treatment period to monitor the efficiency of the different treatment modality.
Evaluating thehistopathological changes in the tumor tissues following the different
treatment methodsusing Hematoxylin and Eosin (H&E) stainusing light microscope.
Molecular detection of P21, P16 and P53 gene expression by RT-PCR.
Results of thestudy:
The injection of the photosensitizer (CHL) has no effect on the tumor volume.
The effect of exposing the tumor to IR laser as a photodynamic therapy increased with
increasing the laser energy resulted in decreasing the tumor volume, tumor growth rate
and inhibition ratio. These effects were observed either on using infrared laser alone
(with its two frequencies) or in the presence of the CHL.
The inhibition in thetumor volume has minimum value on using 7000 Hz IR laser in
the presence of the CHL.
The effect of exposure to continuous ultrasound wave was more than that in case of
using pulsed ultrasound wavebut with high mortality rate in mice as a small model.
Similar variations occurred in case of using ultrasound exposure only or in the
presence of the CHL, with minimum effect occurred on using ultrasound in the
presence of the CHL.
Combined treatment of IR laser at 7000 Hz and pulsed ultrasound wave in the
presence of CHL was more effective than either IR laser or ultrasound alone.
The presence of Vit. B12 have no significant effect in the combined treatment.
In the group of mice carrying the tumor only, a significant increase in the levels of
MDA as compared to the control group of animals. The combination of photodynamic
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andsonodynamic therapy in the presences of chlorophyll decreasedsignificantly the
levels of MDA.
In all Ehrlich bearing mice groups decreased activities of enzymatic antioxidants
(SOD, CAT, GR, GST and TAC) in comparison with normal group were observed.
Significant increases in the enzymatic and non-enzymatic antioxidant guard were
observed in the groupssubjected to combination of photodynamic andsonodynamic
therapy in the presences of CHL.
A statistically significant elevation of heavy metals and trace elements (lead,
cadmium, chromium, nickel and iron) concentrations was detected in serum of mice
group bearing the tumor compared to the control group.
A significant decrease in theheavy metals and trace elements was observed in
thegroup of mice subjected tocombination of photodynamic andsonodynamic therapy
in the presence of CHL.
It was observed that treatment with CHLameliorated the levels of serum creatinine and
urea which is an indication of renal protection. This also confirms the protective role
of CHL against renal toxicity. Also treatment with CHLprotected against increase in
serum levels of ALT, AST, and GGT, which is an indication of hepatoprotection by
CHL. This also confirms the protective role of CHL against hepatotoxicity.
In the group of animals subjected to combination of photodynamic andsonodynamic
therapy in the presences of CHL, large foci distinct necrosis areas were appeared.
Relative permittivity of the tumor tissue decreased exponentially with increasing the
applied frequency. Relative permittivity of treated tumor tissue with different
treatment modalities decreased than that of untreated tumor. The real conductivity
increased exponentially with increasing applied frequency for all the modes of
treatments, while decreased in treated tumor than untreated tumor in the absence of the
sensitizer but coincidence occurred in the presence of sensitizer. The variation of the
imaginary permittivity with frequency showed sharp increase at low frequency
followed by exponential decreased with all methods of treatments used.
P53 and p16 gene expressions significantly higher in mice groups treated with
sonophoto therapy (in presence of sensitizer) than those treated with photo- or sonotherapy
only (in presence of sensitizer alone) followed by photo- or sono-therapy only
(in absence of sensitizer alone) while the lowest expression was among untreated
cancerous group. In case of p21 gene expressions significantly lower in mice groups
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treated with sonophoto therapy (in presence of sensitizer) than those treated with
photo- or sono-therapy only (in presence of sensitizer alone) followed by photo- or
sono-therapy only (in absence of sensitizer alone) while the highest expression was
among untreated cancerous group.
The histological evaluation revealed that all tumors from the group of mice bearing the
tumor included highly malignant cells and none of the tumors showed necrosis.
Tumors excised from animals receiving treatment (CHL) showed significant areas of
necrosis compared to groups without sensitizer.