PHYTOCHEMICALS OF AEGLE MARMELOS MEDIATED SYNTHESIS AND CHARECTERIZATION OF ANTI BREAST CANCER SILVER NANO PARTICALS FOR MCF-7 CELL LINE.

1. A project Topic On:
PHYTOCHEMICALS OF AEGLE MARMELOS MEDIATED SYNTHESIS
AND CHARECTERIZATION OF ANTI BREAST CANCER SILVER
NANO PARTICALS FOR MCF-7 CELL LINE.
Submitted by:
MAHANTHESHKUMAR G T
2nd year M.Sc in Biotechnology
Dept of biotechnology
Sahyadri science college,
Shivamogga.
Under the guidence of:
DR. PRADEEPA.K
Assistant Professor & Co-ordinator
P G Department of Studies & Research
in Biotechnology.
Sahyadri Science College Shimoga.

2. CONTENTS
1. INTRODUCTION
2. OBJECTIVES
3. METHODOLOGY
4. RESULTS AND DISCUSSION
5. CONCLUSION
6. REFERENCE

3. Breast cancer
Breast cancer is the most common type of cancer and the second leading
cause of death. This disease is the primary cause of mortality among
women aged 45–55 years, and is the second leading cause of cancer-
induced death. The incidence of breast cancer is almost 1-in-8 women,
requiring complete tissue removal, chemotherapy, radiotherapy, and
hormone therapy most of the time.

4.  Skin irritation or dimpling.
 Breast pain.
 Nipple pain or the nipple turning inward.
 Redness, thickening of the nipple or breast skin.
 A nipple discharge other than breast milk.
 A lump in the underarm area.

6. Botanical Classification:
Kingdom - Plantae
Order - Spindales
Family - rutacea
Genus - Aegel
Species -Aeglemarmelos
Aegle marmelos

7. 1.To synthesize silver nanoparticals from the Aegle marmelos plant
extract.
2. To characterize synthesized silver nanoparticals
3. To screen in vitro cytotoxicity property of synthesized silver nano
particals on MCF7 cell line.

8. Collection of plant materials
Synthesis of silver nanoparticles by cold Maceration method
Characterization of silver nanoparticles
UV-VIS Spectroscopy
Fourier Transform Infra-
red Spectroscopy (FTIR)
Atomic Force Microscopy (AFM)
MTT Assay

9. RESULTS AND DISCUSSION
1.Synthesis of silver nanoparticle’s Silver nanoparticles were synthesized using Aegle
marmolis leaves extract and AgNO3 solution.

10. Characterization of silver nano particles using UV spectroscopy, FTIR and AFM.
1:UV-vis spectroscopy.
In the present study, the AgNPs absorbance was measured in UV-vis spectrophotometer
from 270-1060 nm. The surface plasma resonance band of AgNPs appeared at 420nm due
to the presence of components of the plant extract.

11. 2.Atomic Force Microscopy (AFM):
In the present investigation AFM revealed following results
Cursor Position-X-position:4.18µm,Y-position:-546nm,Z-position:83.75pm
Tool Status:Length-707.5nm,DeltaZ-51.46nm,Width-703.1nm,Height-78.12nm
Shape: Triangle

12. 3. Fourier Transform Infra-red Spectroscopy (FTIR)
Sl.No. Wave numbers (cm-1) Functional Groups Bond nature Compounds
1 3440.74 C-H stretch Medium Alkanes
2 2966.80 C-H stretch Medium Alkanes
3 2860.83 C-H stretch Medium Alkanes
4 2422.47 N-O symmetric stretch Medium Nitro compounds
5 1641.46 N-H bend Medium Primary Amines
6 1462.63 C-H bend Medium Alkanes
7 1404.14 C-C stretch in ring Medium Aromatics
8 1268.36 C-H wag Medium Alkyl halides
9 1165.33 C-H wag Medium Alkyl halides
10 1079.80 C-N stretch Medium Aliphatic amines
11 940.52 N-H wag Broad and strong Primary and secondary amines
12 876.86 C-Cl stretch Medium Alkyl halides
13 803.27 C-H stretch (nearest value) Medium Alkanes
14 533.67 C-Br stretch Medium Alkyl halides

13. 4.Cytotoxicity of the synthesized nano particles was determined by
MTT Assay.
MCF-7 Cell line Vs
Sample Am
Test concentrations (in µg/ml)
Blank Untreated Cisplatin
15 µg/ml
10 20 40 80 160
Reading 1 0.017 0.774 0.144 0.754 0.733 0.701 0.412 0.017
Reading 2 0.014 0.776 0.161 0.761 0.77 0.759 0.425 0.02
Mean OD 0.0155 0.775 0.1525 0.7575 0.7515 0.73 0.4185 0.0185
Mean OD-Mean
Blank
0.7595 0.137 0.742 0.736 0.7145 0.403 0.003
Standard deviation 0.0014142 0.012020815 0.00495 0.026163 0.041012 0.009192 0.002121
Standard error 0.001 0.0085 0.0035 0.0185 0.029 0.0065 0.0015
% Standard error 0.1316656 1.11915734 0.460829 2.435813 3.818302 0.855826 0.197498
% Viability 100 18.03818302 97.69585 96.90586 94.07505 53.06122 0.394997
MTT assay results showing cell viability (%) based on concentration

14. Graph showing concentration and viability of Aegle marmolis against MCF-7
cell lines.
Continued…

15. 10 μg Cisplatin
10 μg Cisplatin
Untreated control
Untreated control
The figure shows control and 10µg cisplatin treated cell viability.

16. The figure shows 10µg to 40µg drug treated cell viability.

17. The figure shows 80µg to 160µg drug treated cell viability.

18. CONCLUSION
•The silver nanoparticles synthesized Aegle marmelos, showed positive results for
cytotoxic activity with an IC50 value of 0.9774 µg/ml.
•These positive results confirmed the cytotoxic potential of the Ag nanoparticles
against breast cancer.
•Thus Ag nanoparticles can be used as a potent drug in alternative therapy for treating
the breast cancer patients in the near future.
•Further potential of cytotoxicity against cancer can be enhanced by In vivo studies in
animal models and human volunteers.

19. REFERENCE
1. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J
Clin. 2009-(54-28).
2. Breastcancer.org, Stages of breast cancer.,Chun Christina, What are the sign and symptoms of
Breast Cancer, Healthline.,Swelling of all or part of the breast.
3. .Breastcancer.org, Stages of breast cancer.,MitsukAnzhelika, Breast cancer information for
young women, Turku University of Applied Science
4. A guide for journalists on breast cancer and its treatment-Roche.,American Cancer Society,
Treating Breast Cancer.,American Cancer Society, Treating Breast Cancer
5. A guide for journalists on breast cancer and its treatment-Roche
6. American Cancer Society, Treating Breast Cancer. https://www.cancer.org/cancer/breast-
cancer/treatment.html 23) Breastcancer.org, Treatments and side effects of breast cancer.
7. Breastcancer.org, Treatments and side effects of breast cancer.
8. Rai M, Gupta PN, Dewedi R (1991). Variability in Bael germplasm. Indian J. Plant Genet.
Resour. 40:86-91.