Chronic anaemia causes degenerative changes in trophoblast cells of the rat placenta

Year 2017, Volume: 11 Issue: 2, 72 - 78, 20.08.2017

Omer Awad Shem J. Ochieng Abdel Malek Julius Ogeng’o

Abstract

Objectives: Iron deficiency anaemia causes adverse pregnancy outcome. There are few studies on effects of anaemia on the structure of trophoblastic cells which are important in placental function. These data are important for understanding the function and disorders of the placenta. The aim of this study was to investigate the ultrastructural cellular changes associated with iron deficiency anaemia in rat placenta.

Methods: Forty-nine female Sprague-Dawley rats were randomly separated into experimental and control groups. The experimental group was rendered anaemic by removing 1.5 ml of blood per bleed on five alternate days, and the placentas were collected on gestational days 17, 19 and 21. For light microscopy, five cubic millimeter segments were fixed in 10% buffered formaldehyde solution; dehydrated in ethanol and embedded in paraffin wax. Five micron thick sections were cut, deparaffinized and stained with Hematoxylin and Eosin. For transmission electron microscopy, 1 mm3 sections were fixed in 2.5% phosphate buffered glutaraldehyde, post fixed in 2% osmium tetroxide, dehydrated in ethanol, cleared in propylene and embedded in epon resin. Ultrathin sections stained with uranyl acetate and lead citrate were examined with JEOL electron microscope.

Results: Cytotrophoblast, syncytiotrophoblast and giant trophoblastic cells of placentas of anaemic rats showed cytoplasmic and nuclear vacuolation with loss of cell margins. In addition, there was atrophy of microvilli on the cell surface, as well nuclear chromatolysis, nucleolar degeneration and appearance of dark bodies.

Conclusion: Chronic anaemia causes trophoblastic cell degeneration. This may undermine the functional integrity of the cells and constitute part of the mechanism for poor fetal outcome.

Keywords

anaemia, cytotrophoblast, degeneration, giant cells, placenta, syncytiotrophoblast

References

• 1. Mayhew TM. A stereological perspective on placental morphology in normal and complicated pregnancies. J Anat 2009;215:77–90.
• 2. Soni R, Nair S. Study of histological changes in placenta of anaemic mothers. IOSR Journal of Dental and Medical Sciences 2013;9:42–6.
• 3. Cline JM, Dixon D, Ernerudh J, Faas MM, Göhner C, Häger JD, Markert UR, Pfarrer C, Svensson-Arvelund J, Buse E. The placenta in toxicology. Paper III: Pathological assessment of the placenta. Toxicol Pathol 2014;42:339–44.
• 4. Furukawa S, Kuroda Y, Sugiyama A. A comparison of the histological structure of the placenta in experimental animals. J Toxicol Pathol 2014;27:11–8.
• 5. Allen L. Anemia and iron deficiency: effects on pregnancy outcome. Am J Clin Nutr 2000;71:1280S–4S.
• 6. Noronha JA, Bhaduri A, Vinod H, Kamath A. Maternal risk factors and anaemia in pregnancy: a prospective retrospective cohort study. Obstet Gynaecol 2010;30:132–6.
• 7. Kiran N, Zubair A, Khalid H, Zafar A. Morphometrical analysis of intervillous space and villous membrane thickness in maternal anaemia. J Ayub Med Coll Abbottabad 2014;26:207–11.
• 8. Sabina S, Iftequar S, Zaheer Z, Khan M, Khan S. An overview of anemia in pregnancy. Journal of Innovations in Pharmaceuticals and Biological Sciences 2015;2:144–51.
• 9. Agarwal KN, Gupta V, Agarwal S. Effect of maternal iron status on placenta, fetus and newborn. International Journal of Medicine and Medical Sciences 2013;5:391–5.
• 10. Biswas S, Meyur R, Adhikari A, Bose K, Kundu P. Placental changes associated with maternal anaemia. Eur J Anat 2014;18:165–9.
• 11. Crocker IP, Tansinda DM, Jones CJ, Baker PN. The influence of oxygen and tumor necrosis factor-α on the cellular kinetics on term placental villous explants in culture. J Histochem Cytochem 2004;52:749–57.
• 12. Hu D, Cross JC. Development and function of trophoblast giant cells in the rodent placenta. Int J Dev Biol 2010;54:341–54.
• 13. Hung TH, Skepper JN, Charnock-Jones DS, Burton GJ. Hypoxiareoxygenation: a potent inducer of apoptotic changes in the human placenta and possible etiological factor in preeclampsia. Circ Res 2002;90:1274–81.
• 14. Markovic SD, Milosevic M, Dordevic N, Ognjanovic B, Stajn AS, Zorica S. Time course of hematological parameters in bleeding induced anemia. Archives of Biological Science Belgrade 2009;61: 165–70.
• 15. Hunter E. Practical electron microscopy: a beginner’s illustrated guide. 2nd ed. Cambridge (NY): Cambridge University Press; 1993.
• 16. Bozzola JJ, Russell LD. Electron microscopy. 2nd ed. Sudbury (MA): Jones and Bartlett Publishers; 1999.
• 17. de Luca Brunori I, Battini L, Brunori E, Lenzi P, Paparelli A, Simonelli M, Valentino V, Genazzani AR. Placental barrier breakage in pre- eclampsia: ultrastructural evidence. Eur J Obstet Gynecol Reprod Biol 2005;118:182–9.
• 18. Salgado SS, Salgado MKR. Structural changes in pre-eclamptic and eclamptic placentas – an ultrastructural study. J Coll Physicians Surg Pak 2011;21:482–6.
• 19. Selim ME, Elshmry NG, Rashed EA. Electron scanning microscopic observations on the syncitiotrophoblast microvillous membrane contribution to preeclampsia in early placental rats. J Blood Disord Transfus 2013;4:137.
• 20. Judson JP, Fun LP, Nadarajah VD, Nalliah S, Chakravathi S, Thanikhacalam P, Santhanaraj L. Ultrastructural and immunofluorescence studies of placental tissue in hypertensive diseases of pregnancy. Research Journal of Biological Sciences 2010;5:155–63.
• 21. Crocker IP, Barratt S, Kaur M, Baker PN. The in-vitro characterization of induced apoptosis in placental cytotrophoblasts and syncytiotrophoblasts. Placenta 2001;22:822–30.
• 22. Serin IS, Ozçelik B, Basbug M, Kiliç H, Okur D, Erez R. Predictive value of tumor necrosis factor alpha (TNF-alpha) in preeclampsia. Eur J Obstet Gynecol Reprod Biol 2002;100:143–5.
• 23. Grill S, Rusterholz C, Zanetti-Dällenbach R, Tercanli S, Holzgreve W, Hahn S, Lapaire O. Potential markers of preeclampsia–a review. Reprod Biol Endocrinol 2009;7:70.
• 24. Linton EA. Human trophoblast syncytialization: a cornerstone of placental function. Endocrine Abstracts 2005;10:515.
• 25. Battistelli M, Burattini S, Pomini F, Scavo M, Caruso A, Falcieri E. Ultrastuctural study on human placenta from intrauterine growth cases. Microsc Res Tech 2004;65:150–8.
• 26. James JL, Stone PR, Chainley LW. Cytotrophoblast differentiation in the first trimester of pregnancy: evidence for separate progenitors of extravillous trophoblasts and syncytiotrophoblast. Reproduction 2005;130:95–103.
• 27. Ain R, Canham LN, Soares MJ. Gestation stage-dependent intrauterine trophoblast cell invasion in the rat and mouse: novel endocrine phenotype and regulation. Dev Biol 2003;260:176–90.
• 28. Soares MJ, Konno T, Alam SM. The prolactin family: effectors of pregnancy-dependent adaptations. Trends Endocrinol Metab 2007; 18:114–21.
• 29. Padmanabhan R, Shafiullah M. Intrauterine growth retardation in experimental diabetes: possible role of the placenta. Arch Physiol Biochem 2001;109:260–71.
• 30. Padmanabhan R, Al-Menhali NM, Ahmed I, Kataya HH, Ayoub MA. Histological, histochemical and electron microscopic changes of the placenta induced by exposure to hyperthermia in the rat. Int J Hyperthermia 2005;21:29–44.
• 31. Kosif R, Akta G, Öztekin A. Microscopic examination of placenta of rats prenatally exposed to aloe barbadensis: A preliminary study. Int J Morphol 2008;26:275–81.
• 32. Tait S, Tassinari R, Maranghi F, Mantovani A. Bisphenol A affects placental layers morphology and angiogenesis during early pregnancy phase in mice. J Appl Toxicol 2015;35:1278–91.
• 33. Omar AR, El-Din EYS, Abdelrahman HA. Implications arising from the use of cymbopogen proximus; proximal on placenta of pregnant albino rats. Brazilian Archives of Biology and Technology 2016;59: e16160165.
• 34. Rebelato HJ, Esquisatto MA, de Sousa Righi EF, Catisti R. Gestational protein restriction alters cell proliferation in rat placenta. J Mol Histol 2016;47:203–11.
• 35. Adelman DM, Gertsenstein M, Nagy A, Simon MC, Maltepe E. Placental cell fates are regulated in vivo by HIF-mediated hypoxia responses. Genes Dev 2000;14:3191–203.