nanoparticle toxicity

1. IMPACT OF OF NANOPARTICLES ON REPRODUCTIVE
SYSTEM
Anju Surendranath
Division of Toxicology

2. Section 1
 Introduction
 Routes of exposure of NPs
 Reproductive toxicity
 Developmental toxicity
 Teratogenicity
 Future advances in research
 References
Section 2
 Research paper
CONTENTS

3. The vast growth of nanotechnologies with all their
far-reaching benefits has fostered concerns about the
potential health risks of nanoparticles (NPs).
It is broadly accepted that the fetus may be more
sensitive to chemical exposures than adults.
Overall, it is plausible that NP may translocate from
the different routes of exposure to the placenta and
fetus, but also that adverse effects may occur
secondarily to maternal inflammatory responses.
INTRODUCTION

5. Malek et al; Curr. Pharm. Biotechnol (2011)

6. Huang et al; Placenta 36 (2015)

7. REPRODUCTIVE TOXICITY
BIOCHEMICA
L RESPONSE
CELLULAR
RESPONSE
TOXICANT
ORGAN
RESPONSE ORGANISM
RESPONSE
IMPAIRED
REPRODUCTION
Adverse Outcome
ROS production
on gonad tissue
Oxidative
stress
Germ Cell
Apoptosis
Molecular
initiating event
Key Events
NPs

8. DEVELOPMENTAL TOXICITY
Yang et al; Sci. Rep. 2 (2012)

9. Pb
Pb
Soares et al; placenta.2011

10. TERATOGENICITY
Thalidomide Disaster

13. COMPLICATIONS OF REPRODUCTIVE TOXICITY
Miscarriage/
Still birth
Low Birth weight Organ
malformation
Birth defects CNS problems Fetal addiction

14. ABNORMALITIES IN
GESTATIONAL AND FOETAL
PARAMETERS DUE TO NPs
EXPOSURE

15. Effect of NP on foetal organ abnormalities

16. Effect of maternal NP exposure on nervous system of foetus

17. Effect of NP on reproductive system of offspring

18. Effect of NP in embryonic and extra embryonic tissues

19. Yang et al; Sci. Rep. 2 (2012)

20. ANALYSIS PATTERNS
• Overall toxicokinetic pattern of nanoparticles.
• Transplacental transfer.
• Human placental perfusion models
• Animal models.
• Potential mechanisms in developmental
toxicity.

21. OECD guidelines for reproductive and
developmental toxicity studies
OECD 421 Reproduction/ Developmental toxicity
screening test.
OECD 422 Combined Repeated Dose Toxicity Study
with Reproduction/ Developmental Toxicity
Screening Test.
OECD 414 Prenatal Development Toxicity Study.
OECD 416 Two-Generation Reproduction Toxicity.
OECD 443 Extended One-Generation Reproductive
Toxicity Study.
OECD 426 Developmental Neurotoxicity Study.

22. Future advances in research
• No in vitro assay can completely recapitulate the
complexities of fetal development or the fetal-
maternal interactions that occur in vivo.
• minimal xenobiotic metabolism occurs in ESCs, it is
likely that most proteratogens will be classified as
false negatives in ESC5 based assays.
• Recently, researchers have utilized placental BeWo
cells to generate toxicokinetic data.

23. RESEARCH PAPER
Topic: Maternal Placental- foetal
Biodistribution of multimodal
polymeric nanoparticles in a
pregnant rat model in mild and late
gestation.

25. Determine the biodistribution of both cationic and anionic
multimodal (fluorescent and paramagnetic) PGMA NPs in
pregnant rat model at two different stages of pregnancy
[embryonic day 10 (ED10) and day 20 (ED20)] using
pharmacological doses for the application of multimodal
polymeric nanoparticles during pregnancy.
Fluorescently-labelled and super paramagnetic
PGMA nanoparticles to evaluate nanomaterial
biodistribution and maternal-fetal exposure.
HYPOTHESIS
AIM

26. Quantitative assessment of
nanoparticle biodistribution.
In vitro and in vivo
assessment of
NP biocompatibility.
Synthesis and characterization
of PGMA and PGMA-PEI NPs.
Placental biodistribution
analysis
Qualitative assessment
using
confocal imaging
Qualitative assessment
using
fluorescence imaging
Experimental Outline

27. RESULTS & DISCUSSION

28. (a) TEM image of PGMA-PEI NPs, scale bar: 100 nm. (b) Excitation (solid line) and emission
(discontinuous line) spectra of PGMA/PGMA-PEI NPs containing P10 fluorophore.
(c)Hydrodynamic size distribution of PGMA (solid) and PGMA-PEI(discontinuous) NPs. (d) Zeta
potentials of PGMA (solid) and PGMA-PEI (discontinuous)
NPs.

29. (a)Hydrodynamic size distribution and (b) zeta potentials of PGMA and PGMA-PEI nanoparticles
after incubation in serum-supplemented media for 4 h. Cytotoxicity assays of PGMA (red bars) and
PGMA-PEI (blue bars) at 10 and 100 µg/ml after (c) 4 h and (d) 24 h incubation.

30. Fluorescence intensity measurements of ED10 and ED20 tissue
homogenates. Data as mean±SD (n=4).

31. (a) Liver of ED10 dams. High levels of fluorescence were observed in liver from both PGMA
and PGMA-PEI treated dams. (b) Other organs of ED10 dams. With the exception of the spleen
of the PGMA-PEI treated dam, none of the organs showed a visible level of P10 fluorescence. (c)
Uterine horns containing conceptuses of ED10 dams. In spite of spectral unmixing, low levels of
autofluorescence was observed in controls and treated dams. (d) Uterine horns containing
placenta and fetuses of ED20 dams. The placentae showed a visible level of auto fluorescence in
theP10 spectral region around the junctional zones in all dams.

32. Representative tissue section of a maternal liver. Higher magnification images
show the accumulation of PGMA and PGMA-PEI nanoparticles in the maternal
liver at ED10 (b & c) and ED20 (d & e). PGMA and PGMA-PEI nanoparticles
were found to be accumulating mainly within Kupffer cells (white arrowheads).

33. (a) Representative tissue section of a maternal spleen. Higher magnification
images show the accumulation of PGMA and PGMA-PEI nanoparticles in the
maternal spleen at ED10 (b & c) and ED20 (d & e). Nanoparticles (white
arrowheads) were observed throughout the red pulp with higher quantities
amassing around the peripheries of white pulp (denoted by dashed lines)

34. (a-f) PGMA and (g-i) PGMA-PEI NPs in the ED10 conceptus , (b ) outline of sections denoting tissue
regions. (c and e) higher magnification images of areas shown in “fig: a”. Even higher magnification images
of areas shown in “fig c and e”. PGMA NPs were found in ectoplacental cone and primary decidual cone.
Fig: (g) represents PGMA-PEI treated dams placenta in ED10 conceptus. (h) outline of sections denoting
tissue regions. (i and k) higher magnification images of areas shown in rectangles in “g”. (j and l) higher
magnification images of panels shown in “i and k” respectively. PGMA-PEI were observed in decidua,
venous sinusoids and tissues close to ectoplacental cone.

35. (a-d) PGMA and (e-h) PGMA-PEI NPs in placenta ED20. (b-d) higher magnification
images of areas depicted in rectangles in “a”. PGMA NPs were observed in chorionic
plate, junctional zone and in labyrinth. (e) representative tissue section from PGMA-PEI
dams placenta. (f-h) higher magnification images of the areas indicated by the rectangles
in “fig: e”. PGMA-PEI NPs were observed in chorionic plate, junctional zone and
labyrinth zone respectively.

36. CONCLUSION
•Biocompatible polymeric materials such as PGMA and PGMA-PEI were
investigated as delivery platforms for therapeutics.
•Despite of techniques such as relaxometry and multispectral imaging, confocal
microscopy using a differential charge based approach provided a relatively sensitive
representation of tissue uptake in the pregnant rat.
•It has been reported that, in general, smaller particles exhibit greater placental
uptake and passage than larger particles; the increased placental uptake of PEI-
PGMA nanoparticles is therefore likely to be not dependant on size.
•This work highlights the advantages and limitations of the use of multimodal
nanoparticles to accurately measure the uptake of polymeric nanoparticles and
subsequently the extent and impact of maternal, fetal and placental exposure.

37. 1. Menezes, V., Malek, A. & Keelan, J. A. Nanoparticulate drug delivery in
pregnancy: placental passage and fetal exposure. Curr.Pharm. Biotechnol. 12,
731–742, doi:10.2174/138920111795471010 (2011).
2. 2. Huang, J.-P. et al. Nanoparticles can cross mouse placenta and induce
trophoblast apoptosis. Placenta 36, 1433–41, doi:10.1016/j. placenta.2015.10.007
(2015).
3. Soares, M. J., Chakraborty, D., Karim Rumi, M. A., Konno, T. & Renaud, S. J.
Rat placentation: An experimental model for investigating the hemochorial
maternal-fetal interface. Placenta 33, 233–243,
doi:10.1016/j.placenta.2011.11.026 (2012).
4. Keelan, J. A., Leong, J. W., Ho, D. & Iyer, K. S. Therapeutic and safety
considerations of nanoparticle-mediated drug delivery in pregnancy. Nanomed. 10,
2229–2247, doi:10.2217/nnm.15.48 (2015).
5. Yang, H. et al. Effects of gestational age and surface modification on materno-
fetal transfer of nanoparticles in murine pregnancy. Sci. Rep. 2, 847–855,
doi:10.1038/srep00847 (2012).
REFERENCES