EFFECT OF SILVER NANOPARTICLES ON

SWISS ALBINO MICE AND THEIR FOETUSES

7th
PROGRESS REPORT

JYOTI PRAKASH PANI

SUPERVISOR-:DR ROYANA SINGH
 INTRODUCTION
In nanotechnology
nanoparticle is defined
as a small object that
behaves as a whole unit
in the term of transport
and properties.
Measures 100 to 1nm
range either less or
more. It shows optical,
magnetic and electrical
properties. Medically it
also shows antibacterial
and antifungal nature.
 Why silver nanoparticle is important?
Silver nanoparticle is important because
of application on catalysis, optics,
electronics, magnetics and medical
field which includes diagnostic and
therapeutics.

DIAGNOSTIC

THERAPEUTICS
NANO POLLUTION
 Toxicity
• Exposure to silver nanoparticles
have been associated with
inflammatory, oxidative, genotoxic
& cytotoxic response .
• It causes adverse health effect in
the respiratory tract as well as in
extra pulmonary organs.
• It causes harm to plants.
• It also interferes with nitrogen
fixing bacteria and plant host and
also prevents transmission of signal
between the two.
• Biochemically it also causes severe
alteration of enzymes of vital
organs at molecular level and alters
the blood parameters.
 WORK DONE DURING THE PERIOD OF 30TH
MAY 2015 TO 29TH SEPT. 2015

• FATE OF APOPTOTIC AND NECROTIC CELLS

ESTIMATED.
• ATTENDED 7TH INTERNATIONAL SYMPOSIUM OF
CLINICAL AND APPLIED ANATOMY,BRATISLAVA
SLOVAKIA AND ORALLY PRESENTED PAPER
ENTITLED “ PARTICLE SIZE DEPENDANT
TERATOGENICITY EFFECT OF SILVER
NANOPARTICLES IN MICE.
ESTIMATION OF LIVE AND DEAD CELLS IN LIVER TISSUE OF CONTROL AND TREATED GROUP FETUSES AFETR REPEATED NANOSILVER COLLOIDAL SOLUTION ORAL GAVAGES THERAPY DONE BY

• IMAGE-JC
• CON FOCAL MICROSCOPY
A
CONT.. A/O S.P. 0.5.. A/O S.P. 1.. A/O S.P.

5.. A/O S.P. 10.. A/O S.P. 15.. A/O S.P. 20.. A/O S.P.
CONT.. DAPI S.P. 0.5. DAPI S.P. 1.. DAPI S.P.

5. DAPI S.P. 10. DAPI S.P. 15.. DAPI S.P. 20.. DAPI S.P.
CONT.. E/B S.P. 0.5.. E/B S.P. 1.. E/B S.P.

5.. E/B S.P. 10.. E/B S.P. 15.. E/B S.P. 20.. E/B S.P.
 IMAGE-J ESTIMATION OF APOPTOTIC AND NECROTIC CELL
(CONTROL GROUP AND 0.5, 1,5, 10, 15, 20MG/KG/DAY
AgNPs TREATED GROUPS)
APOPTOTIC AND NECROTIC LIVE CELL ESTIMATION FROM
FRESH TISSUE
CONTROL GROUP AND TREATED GROUPS
Slice Count Total Area Average Size Mean Mode
Ctrl 6254 246568.0 1887.446 64.463 58.807(AO)
0.5 6180 237699.0 1821.876 62.456 57.962(AO)
1 6172 224759.0 1799.776 60.356 56.862(AO)
5 4993 161608.0 1737.720 59.450 35.473(AO)
10 3988 161303.0 1707.720 54.450 31.473(AO)
15 2983 161003.0 1697.720 49.450 26.473(AO)
20 1978 156003.0 1647.720 44.450 21.473(AO)
Slice Count Total Area Average Size Mean Mode
Ctrl 2311 71888.00 71.107 56.876 254.811 (DAPI)
0.5 1885 67169.00 70.084 55.803 249.094(DAPI)
1 1875 67069.00 69.984 54.703 248.994(DAPI)
5 405 11617.00 13.127 41.500 236.777(DAPI)
10 300 11612.00 8.127 36.500 231.777(DAPI)
15 175 11212.00 5.127 31.500 226.777(DAPI)
20 69 11012.00 2.127 26.500 221.777(DAPI)
Slice Count Total Area Average Size Mean Mode
Ctrl 96 4097 1042.677 40.392 57.681 (EB)
0.5 93 4722 921.742 37.453 40.430 (EB)
1 83 4688 918.742 36.453 40.030(EB)
5 28 1978.00 872.786 34.392 39.888(EB)
10 23 1973.00 862.286 32.392 34.888(EB)
15 18 1968.00 791.686 30.392 29.888(EB)
20 13 1963.00 781.186 29.392 24.888(EB)
7000

6254
6180 6172

6000

4993
5000

3988
4000

2983
3000

2331

1978
1885 1875
2000

1000

405
300
175
96 93 83 28 23 69 13
0 0.5 1 5 10 15 18 20
0
1 2 3 4 5 6 7

GROUP AO DAPI EB
 GROUP

1% 10%
2%

39%

19%

1
2
3
4
5
6
7

29%
 DISCUSSION

During the study we observed that the number of live cell after Acridine orange, DAPI
and Ethidium Bromide staining in 0.5,1 ,5,10,15&20 mg/kg b.w. AgNPs oral
gavages treated group was less than control. AO, DAPI and EB is used to visualize
nuclear morphology hence less number of live cells in 0.5,1 ,5,10,15&20 mg/kg b.w.
AgNPs oral gavages treated group determine that in 0.5,1 ,5,10,15&20 mg/kg b.w.
AgNPs oral gavages treated group affected mice more number of cell must have
undergone apoptosis and necrosis leading to fragmentation of cell
which does not occur in normal cell.
 CONCLUSION
-IN AgNPs COLLOIDAL SOLUTION REPEATED ORAL GAVAGES TESTING
EXPERIMENTATION ON PREGNANT SWISS ALBINO MICE SHOWS
RATIONAL EFFECTS OF SMALLER AND BIGGER SIZE SILVERPARTICLES
ON TISSUE(LIVER TISSUE) LEVEL OF FETUSES.
- SMALLER PARTICLES SHOWS MORE INJURY BUT BIGGER PARTICLES
SHOWS LESS INJURY. AND HIGHER DOSE SHOWS MORE INJURY ON CELLULAR
ORGANELLES INCLUDING DEATH,DEGENERATION AND FRAGMENTATION.THERE
IS AN RATIONAL EFFECTS OF APOPTOSIS AND NECROSIS.
-THE LIVE CELL ESTIMATION SHOWS CHRONOLOGICAL DECREASE OF NUMBER AS
DOSE OF SILVER NANOPARTICLES INCREASES.
-THE DEAD CELL ESTIMATION SHOWS CHRONOLOGICAL INCREASE OF PERCENTAGES
AS DOSE OF SILVER NANOPARTICLES INCREASES.
 REFERENCES

• Kim YS, Kim JS, Cho HS, Rha DS, Kim JM, Park JD, et al. Twenty-eight-day oral
toxicity, genotoxicity, and gender-related tissue distribution of silver
nanoparticles in Sprague-Dawley rats. Inhal Toxicol. 2008; 20(6): 575-583.
• Sardari RRR, Zarchi SR, Talebi A, Nasri S, Imani S, Khoradmehr A, et al.
Toxicological effects of silver nanoparticles in rats. Afr J Microbiol Res. 2012;
6(27): 5587-5593.
• Yousef J, Hendi H, Hakami FS, Awad MA, Alem AF, Hendi AA, et al. Toxicity of
Silver Nanoparticles after Injected Intraperitoneally in Rats. J Am Sci. 2012; 8(3):
589-593.
• Korani M, Rezayat S, Gilani K, Bidgoli SA, Adeli S. Acute and sub chronic dermal
toxicity Heydarnejad MS, et al Nanomed. J., Vol. 1, No. 3, Spring 2014 197 of
nanosilver in guinea pig. Int J Nanomedicine. 2011; 6(1): 855–862.
• S.D. Solomon, M. Bahadory, A.V. Jeyarajasingham, S.A. Rutkowsky, C. Boritz, and
L. Mulfinger, Journal of Chemical Education.2007; 84, 322-325.
• Schneider CA, Rasband WS, Eliceiri KW (2012). "NIH Image to Image-J: 25 years of
image analysis". Nat Methods 9 (7): 671–675.
 ACKNOWLEDGEMENT
I SINCERELY PAY MY TRIBUTE & HOMAGE TO DEPARTMENT OF ANATOMY,INSTITUTE
MEDICAL SCIENCES,BANARAS HINDU UNIVERSITY FOR HELPING ME FROM SITU
LEVEL TO CARRY OUT PRACTICAL AND THEORITICAL PART OF MY Ph.D. PROJECT
ENTITLED “ EFFECT OF SILVER NANOPARTICLE ON SWISS ALBINO MICE AND THEIR
FETUSES”.
I SINCERELY AND OPEN HEARTLY EXPRESS MY HONOUR,RESPECT TO MY
SUPERVISOR DR ROYANA SINGH ASSOCIATE PROFESSOR, DEPARTMENT OF
ANATOMY ,INSTITUTE OF MEDICAL SCIENCES,BANARAS HINDU UNIVERSITY TO GIVE
ME PROPER GUIDANCE AND WHOSE IMMENSE HELP AND CONSTURCTIVE CRITICISM
CAUSE IN TRIGGERING TOWARDS INCREASE IN VALUABILITY OF MY PRESENT PhD
WORK.
I ALSO EXPRESS MY HONOUR AND GRATEFULLNESS TOWARDS DR SATYA
NARAYAN SHAMAL HON,BLE H.O.D. DEPARTMENT OF ANATOMY FOR HIS VALUABLE
GUIDE AND SUPPORT FROM TIME TO TIME.
I SINCERELY PAY MY RESPECT AND HONOUR TO DR CHHANDAMAYEE
MOHANTY & PROF.DR BRAJA KISHORE DAS FOR THEIR VALUABLE GUIDE IN THE
INITIAL PHASE OF MY PhD WORK WHICH HELP ME TO REACH UPTO THIS END STAGE.

CONTINUE
 I ALSO EXPRESS MY HONOUR AND RESPECT TO MY CO-SUPERVISOR DR SURENDRA
PRATAP MISHRA THEIR CO LAB WORKERS AND THEIR PG(M.D.) STUDENTS DR BEDABRATA
MUKHOPADHYAY,DR ROSHNI GAVEL AND ALL THE SUMMER TRAINEES WHO HELPED ME OPEN
HEARTLY AT THE TIME OF NEED BY WHICH I AM ABLE TO FINISH THE BIOCHEMISTRY PART OF
MY PhD WORK IN TIME. I AM NOT ABLE TO FORGET THEIR IMMENSE HELP.
I AM THANKFULL TO VALUABLE SUPPORT AFFORDED BY -:
TEACHING STAFF DEPARTMENT OF ANATOMY, IMS,BHU -: PROF.DR S.N.SHAMAL(H.O.D. DEPT.
OF ANATOMY),PROF.DR SHASHIKANT PANDEY,PROF.DR CHHANDAAMAYEE MOHANTY, DR
ROYANA SINGH,DR ANAND MISHRA,DR RAGHUNATH S. MORE
NON-TEACHING STAFF DEPARTMENT OF ANATOMY, IMS,BHU-:SMT TESSY PAUL P., SHREE
NARESH KUMAR , SHREE SHIB PUJAN, ,SHREE MAHESH PRASAD,SHREE GULAB CHAND,SHREE
D.N.BISWAKARMA ,SHREE MUNALAL,SHREE RAM SAGAR YADHAV, SHREE NAND LAL,SHREE
SOURAV SAMEER,SHREE SINDHU RAJ, SHREE AMIT SINGH, SHREE SHASHIKANT, SHREE
BANARASILAL AND SHREEMATI ASMA RANI.
POST GRADUATE STUDENT DEPARTMENT OF ANATOMY, IMS, BHU-: DR GUNJAN RAI,DR
SUBHANGI YADHAV, DR SOUMYA KHANNA, DR KAPIL KUMAR MALAVIYA,DR PRATIBHA
SHAKYA, DR PRASENJIT BOSE,DR DEEPSIKHA SINGH,DR SAMATA TIWARI,DR NIDHI SUNHARE.
PhD STUDENT DEPARTMENT OF ANATOMY,IMS,BHU AND CO LAB WORKERS-: SUSHREE PREETI
KUMARI,MISS RASHMI GUPTA, , MR JANARDHAN CHOUDHURY ,MR SUNIL KUMAR RAI, MRS
NEETURANI DHIMAN, , MR MANEESH MISHRA, SMT. JAYA DIKHIT,SMT SHARADA SINGH,
SUSHREE BARKHA SINGH, MRS SUPRIYA CHOUDHURY, DR SANGEETA RAI.