The effect of warfarin on skeleton development of fetuses’ rats

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Rana. K. Abdulsamad
Faris S. Kata
Alaa A. Sawad


Warfarin, osteocalcin, MGP, skeletal abnormities


The pregnancy period is characterized by the critical time of the pregnant mother's exposure to external impacts, which are most severe during the so-called embryonic period. The study aimed toward study the effects of warfarin in the developing rat fetus. Three-month adult virgin female Wistar albino rats (Rattus norvegicus) were used. They are Left to adapt for one week and then isolated with males by two females with one male for mating determined the zero-gestation day. Pregnant females were arbitrarily divided into three equals with eight rats for each. The first group is control kept without any treatment The second and third groups take 0.1 or 0.05 mg/kg B.t of warfarin respectively, daily from (zero - 15th) day of gestation. The Collection of the blood samples from heart of pregnant rats for biochemical study via using Kit of ELISA technique to estimation the concentration of Osteocalcin and matrix Gla protein. The pregnant animals were sacrificed at (16th) day of gestation The fetuses’ specimens were collected for histological processing. Major anomalies were observed on incomplete ossification of the hind leg and wavy ribs by using double stained Alizarine red and Alcian blue. The histological examination of the vertebral column in the group of treated fetuses reveals degeneration at the vertebrae, damage to the intervertebral tissue, hemorrhage and necrosis. They are a significant decrease in the Matrix Gla protein and Osteocalcin in serum of pregnant rats.

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1. Eason CT, Murphy EC, Wright GRG. and Spurr EB. Assessment of risks of brodifacoum to non-target birds and mammals in New Zea-land. Ecotoxicology 2002; 11(1): 35–48.
2. Fournier-Chambrillon C, Berny PJ, Coiffier, O, Barbedienne, P, Dasse B, Delas, G, Galineau H, Mazet A, Pouzenc P, Rosoux R and Fourni (2004).
3. Wallentin L, Yusuf S, Ezekowitz MD, Alings M, Flather M, Franzosi MG, Pais P, Dans A, Eikelboom J, Oldgren J, Pogue J, Reilly PA, Yang S, Connolly SJ. Efficacy and safety of dabigatran compared with warfarin at different levels of international normalized ratio control for stroke prevention in atrial fibrillation: An analysis of the rely trial. The Lancet 2010; 376 (9745), 975-983.
4. Johnson JA, Gong L, Whirl-Carrillo M, Gage BF, Scott SA, Stein CM, Anderson JL, Kimmel SE, Lee MTM, Pirmohamed M, Wadelius M, Klein TE, Altman RB. Clinical pharmacogenetics implementation consortium guidelines for cyp2c9 and vkorc1 genotypes and warfarin dosing. Clinical Pharmacology and Therapeutics 2011; 90 (4), 625-629.
5. Zoch ML, Clemens TL, Riddle RC. New Insights into the Biology of Osteocalcin. Bone 2016; 82: 42-49.
6. Hoang QQ, Sicheri F, Howard AJ, Yang DS. Bone Recognition Mechanism of Porcine Osteocalcin from Crystal Structure. Nature 2003; 425: 977-980.
7. Roumeliotis S, Roumeliotis A, Dounousi E, Eleftheriadis T, Liakopoulos V. Biomarkers of vascular calcification in serum. Advances in clinical chemistry. 2020; 98, 91-147.
8. AL-Akabi D and Kata FS. Effect of Non –alcoholic fatty liver disease on some of bone biomarker in men. open Access Macedonian journal of medical sciences. 2021; a(A): 924-927.
9. Sawad AA & Al-Asadi FS. Comparative Study on the Time of Appearance of Primary Ossification Centers in the Skeleton of Laboratory Mice Embryo. 2009;
10. ECHA (European Chemical Agency). "Proposal for Harmonized Classification and Labelling, FORMALDEHYDE." (2011).
11. Hussein ZM. Effect of celery seeds extract on the rat fetuses induced with diabetes mellitus 2017;
12. Al-Saeed MH, Al Saeed AH & Jori MM. Study of physiological and histological changes in rabbits induced with hepatic coccidiosis. Journal University of Karbala. 2017;
13. Kim Suvarna S, Christopher Layton, John D. Bancroft: Bancroft’s theory and practice of histological techniques.7th ed., Churchill Livingstone British Library Cataloguing in Publication Data. ISBN: 2013; 978-0-7020-4226-3
14. Menegola E, Broccia ML & Giavini E. Atlas of rat fetal skeleton double stained for bone and cartilage. Teratology, 2001; 64(3), 125-133.
15. Finkelstein Y, Chitayat D, Schechter T, Keating S, and Koren G: Mother risk rounds Warfarin embryopathy following low-dose maternal exposure. J. Obstet. Gynaecol. Can., 2005; 27(7):702-6.
16. Hou J, "Fetal warfarin syndrome", Chang Gung medical journal, 2004; vol. 27, no. 9, pp. 691.
17. Walfisch A, & Koren G. The “warfarin window” in pregnancy: the importance of half-life. Journal of Obstetrics and Gynecology Canada, 2010; 32(10), 988-989.
18. Li X, Zeng Z, Yang Y, Ding W, Wang L, Xu Y & Bi W. Warfarin-related epidural hematoma: a case report. Journal of International Medical Research, 2022; 50(3), 03000605221082891
19. Starling LD, Sinha A, Boyd D & Furck A, "Fetal warfarin syndrome", BMJ case reports, vol. 2012; no. 1, pp. 1-40.
20. Elango K, Javaid A, Khetarpal BK, Ramalingam S, Kolandaivel KP, Gunasekaran K, & Ahsan C, (2021).
21. Sugiyama T, Kugimiya F, Kono S, Kim YT & Oda H. Warfarin use and fracture risk: an evidence-based mechanistic insight. Osteoporosis International, 2015; 26, 1231-1232.
22. Evidence of secondary poisoning of free-ranging riparian mustelids anticoagulant rodenticides in France: Implications for conservation of European mink (Mustela lutreola).
23. Prieto AL, Weber JL,Tracy S, Heeb MJ and Lai C: Gas6, a ligand for the receptor protein-tyrosine kinase Tyro-3, is widely expressed in the central nervous system. Brain Res., 1999; 816: 646–661.
24. Howe, AM and Webster WS: The warfarin embryopathy: a rat model showing maxillonasal hypoplasia and other skeletal disturbances. Teratology., 1992; 46(4): 379-90
25. Sundaram KS, Fan JH, Engelke JA, Foley AL, Suttie JW and Lev M: Vitamin K status influences brain sulfatide metabolism in young mice and rats. J. Nutr., 1996; 126: 2746–2751.