Raymond B. Garcia1, Czedrick Angelo Nicole U. Aniversario1,
Sarah E. Gaspar1, Leah E. Chico1, Honeylou D. Evasco1,
Henedina A. Maini, RPh1, and Sigfredo R. Solano, MD2
1 College of Pharmacy, Adamson University
2 College of Sciences, Department of Biology, Adamson University
Manila, Philippines
A physiological process
wherein new capillary blood
vessels form from pre-existing
ones.
An important natural function
of the body used in healing
and reproduction.
An extraembryonic membrane formed in birds and reptiles by the apposition of the allantois to the inner face of the chorion.
It mediates the exchange of nutrients and gas within the parts of the egg.
Tumor or neoplasm is an
abnormal mass of tissue
that may is caused by
the progressive,
uncontrollable
proliferation of cells.
Cancer is a class of
diseases characterized
by out-of-control cell.
*Laudico, A.V. et al. (2010). 2010 Philippine Cancer Facts and Estimates. Philippine Cancer Society. Manila, Philippines
The research will lead to the crucial
development of modern medicine, by means
of:
• The study will be beneficial to prevent the further
growth and spread of tumors.
• The research would lead to the identification of
natural sources that can be used as alternative for
the existing treatment.
• It can also be a basis for further research regarding
the anti angiogenic property of other medicinal
plants.
General Objectives
The aim of the study is to evaluate the anti-angiogenic property
of the ethanolic extract of A. muricata L. leaves in chorioallantoic
membrane (CAM) of the duck embryo.
Specific Objectives
• To compare the antiangiogenic property of the ethanolic extract of
A. muricata L. with the standard drug, Docetaxel.
• To identify the dose of the ethanolic extract of the sample that will
elicit the anti-angiogenic property.
• To assess if the ethanolic extract of A. muricata L. can be used as
adjunct to cytotoxic agents used in the treatment of cancer.
Collection and Identification of Annona muricata
L. mature leaves
Washing and Cleansing of the
sample
Air-drying as pre-requisite to the extraction process
Comminution using Nanostar Grinder
Extraction process using 50% Ethanol
Removal of 50% Ethanol from the sample using rotary
evaporator
Preparation of stock solutions in different concentrations
(25%, 50%, 75% and 100% concentrations)
Cleaning of fertilized eggs
Isolation of the fertilized eggs in room temperature
up to Day 3
Administration of several concentrations of extracts
and standard drug using tuberculin syringe
Incubation of eggs up to incubation day 9
Transferring of 3 samples of each group in a glass
plate on incubation day 9, 10, 11 and 12
Photograph and measurement of the developing
vasculature on CAM using ImageJ program
Observations and Data
CONTROL
GROUP
Fertilized Duck Eggs
(Untreated)
Fertilized Duck Eggs
(with Normal Saline
Solution)
Fertilized Duck Eggs
(with Docetaxel)
Fertilized Duck
Eggs
(25%)
Fertilized Duck
Eggs
(50%)
Fertilized Duck
Eggs
(75%)
Fertilized Duck
Eggs
(100%)
TREATED GROUP
(Annona muricata extract)
CONTROL GROUPS
GROUP A
(UNTREATED)
GROUP B
(2 mL 0.9% PNSS)
GROUP C (2 mL Docetaxel)
TREATED GROUPS
GROUP D
(25% Extract)
GROUP E
(50% Extract)
GROUP F
(75% Extract)
GROUP G
(100% Extract)
0.0
500.0
1000.0
1500.0
2000.0
2500.0
3000.0
3500.0
Untreated Control - Control + 25% Con 50% Con 75% Con 100% Con
3029.6
2635.2
179.3
1570.0 1562.4 1696.0
1878.9
Are
a (
mm
2 )
Test Groups
Figure 5. Area of growth of blood vessels of control and treatment after 9 days
of development
The administration of the ethanolic extract of
A. muricata L. in the CAM of the duck embryo provides significant reduction in the growth area of
the blood vessels in the CAM, more specifically, in the
9th day of development of the embryo using the 50%
extract.
Moreover, as the data implies, greater
inhibition of the growth of CAM is not correlative to
the increase in concentration of the extracts in the
test samples.
• The researchers suggest the use of other
assays of angiogenesis to validate the
results given in the study.
• Use of a less toxic standard drug for
exhibiting anti-angiogenic property.
• Make a wider array of concentrations of
the extracts as to determine the
approximate peak of anti-angiogenic
activity.
