A Screening Model described by Dr. Debasish Pradhan, Pharmacy Dept. (UDPS) Utkal University.

3. • Silver Nanoparticles: Nanoparticles (NPs) are tiny
materials with size varying from 1 to 100 nm. NPs
have attracted increasing interest from each branch of
medicine in their ability to deliver drugs in the
optimal dosage range. Silver nanoparticles is
basically used in this work for targeting the cancer
cells.
• Metastasis: Metastasis is a widely used terminology
in which cancer cells invade to the neighboring
tissues and secondary organs and finally colonize in
these organs.
• Biofilm: Biofilms are a collection of one or more
forms of microorganisms that can grow on several
different surfaces.

4. • Determination of the proliferative activities of
MCF-7 breast cancer cells in response to each
plant extract treatment.
• Determination of percent viability of MCF-7
breast cancer cells in response to each plant
extracts treatment.
• Determination of anti-microbial activities
using the bacterial strains of K. pneumonia, E.
coli and S. aureus.

5. • The water chestnut is an annual aquatic herb
(Trapa natans) of the family, Lythraceae. It grows
best in areas where annual daytime temperatures
are within the range 15 - 25°C.
• In India it is well known as singhara or paniphal
(eastern India) and is widely cultivated in
freshwater lakes.
• In India, the plant has different regional names,
such as siṅghāḍā (Hindi: स िंघाडा), pānī-
phal (Hindi: पानीफल), pani singada (Odia:
ପାଣିସିଙ୍ଗଡା).

6. • Preparation of Plant Extract:
• Biosynthesis of Silver Nanoparticles:
• Characterization of Biosynthesized Silver Nanoparticles.
 UV-Vis Spectral Analysis
 FTIR Spectroscopy
 XRD Analysis
 Microscopic Study
• In vitro Study
 MTT Assay
 NRU Assay
• Antibiofilm Study

7. Preparation of Plant Extract
Washed, Peels
removed
Shed dried and
Crushed

8. Soxhlet
extraction
Drying

10. Solubility

11. Thin Layer Chromatography (TLC)

12. Preparation of Silver Nanoparticles
Aqueous Extract of
Trapa natans
0.01 M Silver nitrate
24hr
Dark
room
Yellowish Brown indicates
the formation of
nanoparticles

13. UV-Vis Spectroscopy

14. Fourier transform infrared spectroscopy
(FTIR)

15. X-ray diffraction (XRD) Analysis

16. Scanning Electron Microscopy (SEM)

17. Transmission Electron Microscopy (TEM)

18. MTT Assay
0
10
20
30
40
50
60
70
80
90
100 Control
1
μg/ml
2
μg/ml
5
μg/ml
10
μg/ml
25
μg/ml
50
μg/ml
100
μg/ml
NPs
Extract
Doxorubicin
Cell
Viability
(%)
NP Concentration μg/ml

19. Neutral Red Uptake Assay
0
10
20
30
40
50
60
70
80
90
100
NP Concentration μg/ml
Cell
Viability
(%)

20. Reactive Oxygen Species (ROS)
0
20
40
60
80
100
120
140
160
NPs
Extract
Docetaxel
ROS
Generation
(%)

21. Flow Cytometry Analysis

22. Anti-Biofilm Activity
70 75 80 85 90
E. coli
K. pneumoniae
S. aureus
12.5
25
50
100
Biofilm inhibition (%)

23. Characterization Result
UV-Vis Spectroscopy AgNPs exhibit strong absorption peak in the 420–
500 nm range due to Surface Plasmon Resonance.
FTIR analysis Phenol and alcohol compounds (at 3447 cm-1),
carbonyl groups (at 1631 cm-1), alcoholic groups (at
1384 cm−1) and polysaccharides (at 1093 cm-1).
XRD Analysis Highly crystalline in nature
SEM Diameter of AgNPs ranges to be 30-90 nm
TEM Size of AgNPs range to be 15 ± 2 nm

24. NP Concentration μg/ml of AgNPs
Control 01
μg/ml
02
μg/m
l
05
μg/m
l
10
μg/m
l
25
μg/m
l
50
μg/m
l
100
μg/m
l
Cell Viability (%) 100 90 80 60 50 30 26 25
Calculation of MTT Assay
NP Concentration μg/ml of AgNPs
Control 1
μg/m
l
2
μg/m
l
5
μg/m
l
10
μg/m
l
25
μg/m
l
50
μg/m
l
100
μg/m
l
Cell Viability (%) 100 98 90 60 50 40 42 38
Calculation of NRU Assay

25. IC50 of T.natans AgNPs, T.natans Extract and Doxorubicin.
Cell Stage Cell %
Control 12hr 24hr 48hr
Viable 94.4± 0.35 80.55± 0.67 78.25± 0.25 74.3± 0.35
Early Apoptosis 5.55± 0.75 7.57*± 0.95 8.85*± 0.35 11.7*± 0.45
Late Apoptosis/
Necrosis
3.92± 0.20 8.85*± 0.95 10.85*± 0.75 12.3*± 0.20
Flow cytometry analysis of FITC conjugated annexin V/ PI staining
Cell line IC50 μg/mL
T.natans AgNPs T.natans Extract Doxorubicin
MCF-7 26 ± 3.14 μg/mL 34 ± 2.04 μg/mL 47 ± 1.09 μg/mL

26. Bacteria Concentration of Ag-NPs (µg/ml)
12.5 µg/ml 25 µg/ml 50 µg/ml 100 µg/ml
E.coli 82.32 84.84 86.86 88.42
K.pneumoniae 81.90 82.32 83.52 84.86
S.aureus 78.90 79.62 80.82 81.84
Antibiofilm activity of the biosynthesized AgNPs

27. • T.natans-AgNPs exhibited strong cytotoxic activity
against MCF-7 breast cancer cell line.
• Green synthesized AgNPs significantly inhibited
proliferation of MCF-7 breast cancer cells.
• T.natans-AgNPs is cost-effective and eco-friendly
approach towards synthesis
• This shows potential biomedical applications and
alternative therapy, design of new antibiofilm agents
and multi drug resistant bacterial infections.
• May be developed as a template for other anti-cancer
actions.

28. • Future work may be aimed at extensive research
up to the molecular level.
• Toxicity study should be done to find out if the
drug is showing any adverse effect on long term
use.
• The active constituent investigation can be carried
out to elucidate the exact mechanism of
antimetastasis and antibiofilm effect of Trapa
natans.
• In vivo studies should be carried out in order to
confirm the effect of Trapa natans against
metastasis.

29. 1. Bandana B, Debasish P, Adyasa S, Gitanjali T and Shakti P; Multifunctional Green
Synthetic Hematite (α-Fe2O3) Nanoparticles Mediated by Camellia Sinensis (Indian
Tea Plant) Extracts towards Antioxidant and Antiproliferative Activities against MCF-7
Cell Lines; Journal of Pharmaceutics and Therapeutics; ISSN: 2639-0221, Vol-6(1):
page 355-367 (2020)
2. Debasish P, Bandana B, Shakti P and Adyasa S; Screening of Cytotoxic Activity of
Hematite (α-Fe2O3) Nanoparticles from Butea monosperma on MCF-7 Cells;
Pharmacognosy Research; ISSN: 0976-4836, Vol-12(3).pg 260-266, July-September,
2020.
3. Debasish P, Gitanjali T, Shakti P, Bandana B and Adyasa S; The Emergence of Novel
SARS-CoV-2 (COVID-19) a Triple Reassortant Corona Virus Immunogenicity and
Reactogenicity in Human; JCDR; ISSN: 0973-709X , Vol-14(6): LE01-LE04; June
2020.
4. Debasish P, Bandana B, Shakti P and Adyasa S; Antiproliferative and cytotoxic activity
of Hematite (α-Fe2O3) nanoparticles from Butea monosperma on MCF-7 Cells; AJPP;
ISSN: 1996-0816, Vol. 14: Issue 2; February 2020.
5. Debasish P, Bandana B, Shakti P and Adyasa S; Toxicity study of Hematite (Α-Fe2O3)
nanoparticles synthesized from aqueous flower extract of Butea monosperma on MCF-
7 human breast cancer cell line; IJPCM; ISSN: 2576-4772, Vol. 3: Issue 4; 2019; Dec.
2019.

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