This document summarizes a study on using nanocurcumin to mitigate toxicities caused by the chemotherapy drug cisplatin in rats. The study synthesized nanocurcumin particles between 10-20nm in size and tested their ability to reduce cisplatin-induced damage in brain, bone marrow, sperm, and blood cells of treated rats. Results showed nanocurcumin was effective in reducing oxidative stress, genetic damage and abnormalities caused by cisplatin, demonstrating its potential for managing cisplatin side effects. Further analysis of liver, kidney, heart and histopathological effects is ongoing.

1. By:
Nasreen Ahmad
12PGD538
Adv. PG Diploma in Nanotechnology
Centre of Excellence in Materials Science
(Nanomaterials)
Dept. of Applied Physics, AMU, Aligarh.
Pt
NH3
NH3
H
Mitigation of Cisplatin induced toxicities by
Nanocurcumin in male wistar rats
Under Supervision of:
Prof. Alim Hussain Naqvi
Co-ordinator,Centre of Excellence in Materials
Science (Nanomaterials)
Dept. of Applied Physics,
AMU, Aligarh.
Under co-supervision of:
Prof. M. Mobarak Hossain
Director, Interdisciplinary Brain Research
Centre,
Faculty of Medicine,
AMU, Aligarh.

2. Plan of Talk
What is Cisplatin?
What is
Nanocurcumin?
How to synthesize
nanocurcumin?
Is it really
Nanocurcumin?
How did we do it?-
Experimental
design.
Is it really
effective?-our
findings.

3. Pt
NH3
NH3
H
Cisplatin
 It is a chemotherapeutic drug.
[molecular formula- H6Cl2N2Pt,
molar mass-301.1g/mol]
 It is used to treat various
types of cancers.
 It is accompanied by various toxicity
including, Neurotoxicity, Genotoxicity,
Ototoxicity, Nephrotoxicity and
Hepatotoxicity
Image showing enlarged rat
stomach on cisplatin
exposure

4. Pt
NH3
NH3
H Hepatocellular carcinoma in rats
H
ATP
ADP
P
t
N
H
3
N
H
3
H

5. Adenine Guanine Cytosine Thymine
H
Pt
N
N
H
NN
H
2Cl

6. Adenine Guanine Cytosine Thymine
Pt
N
N
H
NN
APOPTOSIS

7. H3CO
H3C
OH
OH
OH
O
Nanocurcumin (diferuloylmethane)
 antioxidant, anti-inflammatory, antimicrobial
 hepato and nephro-protective , myocardial infarction protection
A natural product extracted from curcuma longa
 Soluble in water while curcumin was not soluble.
 More bioavailability due to small sizes.

8. Synthesis of Nanocurcumin
Dried Turmeric Ethanol:water::4:1
Shake vigorously for 10-15
min to obtain the shown
image
Left on standing overnight.

9. {The upper clear liquid was
collected and this process was
repeated until a clear solution
was obtained}
(dropwise
in to water
at 95 ºC)

10. (Nanocurcumin)
Dried &stored for further
characterisation
Kept in liquid form for
oral dosing to subject
rats.

11. Experimental Design
Brain taken
for
Biochemical
& Protein
Estimations
Blood Sample
collected for
Hematological
Estimations
Femur (Bone marrow)
collected for
Genotoxicity
Estimations
Epididymis collected for
the assessment of
Sperm Abnormalities

12. ExperimentalProcedures
TISSUE IS HOMOGENIZED TO
MAKE 10 % HOMOGENATE
(W/V)
POST MITOCHONDRIAL
SUPERNATANT (PMS) IS PREPARED
BY CENTRIFUGATION AT 10000rpm
FOR 20 MINUTES
CONTROL
Cisplatin GRP
Nanocurcumin
GRP 1 (0.5mg/kg)
Treatment GRP
2(NC-1+cis)
Rat Brain
Nanocurcumin
GRP 2 (1mg/kg)
Treatment GRP
1(NC-0.5+cis)

13. Assessment Parameters
Non-Enzymatic
assays
• Non-Protein thiol
• GSH Assay
• TSH Assay
Biochemical
Estimation
 Lipid
Peroxidation
Antioxidant
Enzyme Assay
 Glytathion-S-
Transferase
estimation
Reproductive
toxcity
Hematological
Estimations
Genotoxicity
Estimations
 Micronucleus Test

14. UV-Vis Spectroscopy
 Absorbance maxima
at 420nm

15. Fourier Transmission Infrared
Microscopy
 peaks at 3415 cm−1
corresponds to OH
stretching vibrations
 at 1,608 cm−1 were
attributed to carboxylate
OCO asymmetric stretching
 at 1,588 cm−1 was assigned
to C–OH deformation
vibration with contribution of
O–C–O symmetric
stretching vibration of
carboxylate group

16. (A)Figure shows the TGA of nanocurcumin and fig (B)shows
the DSC of nanocurcumin
Differential Scanning Calorimetry
& Thermogravimetric Analysis
 first phase
transition at
100.934ºC and the
second phase
transition at
440.699ºC
 air dehydrated
from 49 to 140 °C
and the free
curcumin in the
product melted at
~ 250 °C

17.  TEM images show
sizes in the range of
10-20nm.
Transmission Electron
Microscopy

18. GSH
Effect of different
concentration of
Nanocurcumin on
cisplatin altered GSH
level in brain of Male
Wistarrats.Values are
expressed as mean ±
standard error (SE,
n=6). It is measured as
µmoles GSH/gm tissue.
Significant difference
are indicated by
(*)when compared with
control and (#) when
compared with cisplatin
group.

19. Effect of different
concentration of
Nanocurcumin on cisplatin
induced depletion of non-
protein thiols in brain of Male
Wistarrats.Values are
expressed as mean ±
standard error (SE, n=6). It
is measured as µmoles NP-
SH/gmtissue.Significant
difference are indicated by
(*)when compared with
control and (#) when
compared with cisplatin
group.
Non-Protein Thiol

20. Total Thiol
Effect of different
concentration of
Nanocurcumin on cisplatin
altered T-SH level in brain
of Male Wistarrats.Values
are expressed as mean ±
standard error (SE, n=6).
It is measured as mmoles
TSH/gm tissue. Significant
difference are indicated by
(*)when compared with
control and (#) when
compared with cisplatin
group

21. Lipid Peroxidation
Effect of different
concentration of
Nanocurcumin on
cisplatin induced lipid
peroxidation (LPO) in
brain of Male
Wistarrats.Values are
expressed as mean ±
standard error (SE,
n=6). LPO is measured
as µmoles TBARS
formed/hr/gm tissue.

22. Glutathion-S-Transferase
Effect of different
concentration of
Nanocurcumin on
cisplatin altered GST
activity in brain of Male
Wistarrats.Values are
expressed as mean ±
standard error (SE,
n=6). It is measured as
nmoles of CDNB
conjugated/min/mg
protein.

23. (E)
(D)(C)
(F)
(A) (B)
Micronucleus Test
(A) Represent normal bone
marrow cells while (B) shows
the micronucleated cells, (C&D)
shows the normal cells,and
(E&F)shows decline in MNs
count with protection of
nanocurcumin.

24. Dose Dependent Comparative
representation of Micronucleus
Dose-dependent
profile of
Micronucleated
Polychromatic
Erythrocytes by
Cisplatin and
Nanocurcumin in
Rattus norvegicus.

25. Total Sperm Abnormality
Fig (A) showing the
normal sperm,
(B,C,D) coiled tail,
amorphous head,
broken head, reversed
head position,
and coiled sperms
showing the abnormality
caused by the cisplatin.
(A) (B) (C)
(D)

26. Dose Dependent Comparative
representation of Sperm
Abnormalities
Frequency
distribution of
sperm
abnormalities in
cisplatin and
Nanocurcumin
treated R.
norvegicus

27. Hematological Assessments
(A, C& D) showing the normal
blood cells and (B,E&F) shows
various kind of anomalies in
blood cells of rat by the treatment
of cisplatin.
(A) (B)
(C) (D)
(F)(E)

28. Inference From This Study??
 Small sizes of Nanocurcumin enhanced its bioavailability.
 Proved to be a potent candidate for the management of cisplatin
related side effects.
And Much More…
 Further we are also investigating liver, kidney and heart samples for
different analysis.
 Also samples are being investigated for histopathological effects.
 Comet assays are also being done.