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Alcohol intake during pregnancy reduces offspring bone epiphyseal growth plate chondrocyte proliferation through transforming growth factor β-1 inhibition in the Sprague Dawley rat humerus

  • Diana Pillay (Department of Human Anatomy and Histology, School of Medicine, Sefako Makgatho Health Sciences University) ;
  • Vaughan Perry (Department of Human Anatomy and Histology, School of Medicine, Sefako Makgatho Health Sciences University) ;
  • Robert Ndou (Department of Human Anatomy and Histology, School of Medicine, Sefako Makgatho Health Sciences University)
  • 투고 : 2023.12.29
  • 심사 : 2024.04.22
  • 발행 : 2024.09.30

초록

Intrauterine alcohol exposure delays bone maturation and intensifies osteoporosis and fracture risk. As most studies emphasize the neurological aspects of intrauterine alcohol exposure, there is a lack of research on the implications pertaining to osseous tissue. Previous studies investigated these effects in fetuses, with limited studies on postnatal life. Postnatal studies are crucial since peak bone growth occurs during adolescence. This study aimed at assessing the effects of prenatal alcohol exposure on the humerus proximal and distal growth plate chondrocytes in 3-week-old rats. Sprague Dawley rats (n=9) were assigned to either the ethanol group (n=3), saline (n=3), and untreated (n=3) group and time-mated. Once pregnant, as confirmed by the presence of a copulation plug, the former 2 groups were treated with 0.015 ml/g of 25.2% ethanol and 0.9% saline. The untreated group received no treatment. The left humeri belonging to 6 pups per group were used. Serial sections were cut with a microtome at 5 ㎛ thickness. These sections were stained with haematoxylin and eosin for assessment of normal morphology or immunolabeled with anti-Ki-67 and transforming growth factor β-1 (TGFβ-1) antibody. Prenatal alcohol exposure adversely effected the growth plate sizes and the number of cells in the proliferative zone. Fewer TGFβ-1 immunopositive and proliferative chondrocytes were found using the anti-Ki-67 antibody. This may explain the growth retardation in offspring exposed to gestational alcohol, showing that gestational alcohol exposure inhibits cell proliferation, aiding the diminished stature.

키워드

과제정보

Ms. Hasiena Ali provided technical assistance whereas the staff of the University of the Witwatersrand CAS assisted with animal husbandry.

참고문헌

  1. Pielage M, El Marroun H, Odendaal HJ, Willemsen SP, Hillegers MHJ, Steegers EAP, Rousian M. Alcohol exposure before and during pregnancy is associated with reduced fetal growth: the Safe Passage Study. BMC Med 2023;21:318. 
  2. Simpson M, Duggal S, Keiver K. Prenatal ethanol exposure has differential effects on fetal growth and skeletal ossification. Bone 2005;36:521-32. 
  3. Snow ME, Keiver K. Prenatal ethanol exposure disrupts the histological stages of fetal bone development. Bone 2007;41:181-7. 
  4. Abel EL, Dintcheff BA. Effects of prenatal alcohol exposure on growth and development in rats. J Pharmacol Exp Ther 1978;207:916-21. 
  5. Chaudhuri JD. Alcohol and the developing fetus--a review. Med Sci Monit 2000;6:1031-41. 
  6. Day NL, Jasperse D, Richardson G, Robles N, Sambamoorthi U, Taylor P, Scher M, Stoffer D, Cornelius M. Prenatal exposure to alcohol: effect on infant growth and morphologic characteristics. Pediatrics 1989;84:536-41. 
  7. Miralles-Flores C, Delgado-Baeza E. Histomorphometric analysis of the epiphyseal growth plate in rats after prenatal alcohol exposure. J Orthop Res 1992;10:325-36. 
  8. Kanaan RA, Kanaan LA. Transforming growth factor beta1, bone connection. Med Sci Monit 2006;12:RA164-9. 
  9. Bismar H, Kloppinger T, Schuster EM, Balbach S, Diel I, Ziegler R, Pfeilschifter J. Transforming growth factor beta (TGF-beta) levels in the conditioned media of human bone cells: relationship to donor age, bone volume, and concentration of TGF-beta in human bone matrix in vivo. Bone 1999;24:565-9. 
  10. Kumar M, Dandapat S, Sinha MP, Kumar A, Raipat BS. Different blood collection methods from rats: a review. Balneo Res J 2017;8:46-50. 
  11. Parasuraman S, Raveendran R, Kesavan R. Blood sample collection in small laboratory animals. J Pharmacol Pharmacother 2010;1:87-93. Erratum in: J Pharmacol Pharmacother 2017;8:153. 
  12. Tiscione NB, Alford I, Yeatman DT, Shan X. Ethanol analysis by headspace gas chromatography with simultaneous flame-ionization and mass spectrometry detection. J Anal Toxicol 2011;35:501-11. 
  13. Balcombe JP, Barnard ND, Sandusky C. Laboratory routines cause animal stress. Contemp Top Lab Anim Sci 2004;43:42-51. 
  14. Kelly RR, McDonald LT, Jensen NR, Sidles SJ, LaRue AC. Impacts of psychological stress on osteoporosis: clinical implications and treatment interactions. Front Psychiatry 2019;10:200. 
  15. Chen G, Deng C, Li YP. TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci 2012;8:272-88. 
  16. Pillay DS, Ndou R. Intrauterine alcohol exposure delays growth and disturbs trabecular morphology in 3-week-old Sprague - Dawley rat femur. J Anat Soc India 2022;71:93-101. 
  17. Dlamini GF, Ndou R. Osteoblastogenesis and osteolysis in the Zucker Diabetic Sprague Dawley rat humerus head. Anat Cell Biol 2023;56:552-61. 
  18. Pillay D, Ndou R. Intrauterine alcohol exposure disturbs trabecular morphology in the Sprague Dawley rat humerus epiphysis up to 3-weeks postnatally. Int J Morphol 2021;39:1436-42. 
  19. Ndou R, Bello NK, Perry V, Pillay D. Distal tibial trabecular morphometry in a Sprague Dawley rat model of fetal alcohol syndrome: a micro focus X-ray computed tomography case-control study. Pan Afr Med J 2023;46:35. 
  20. Nwaogu IC. Foetal alcohol syndrome: growth rate of bones in rats. J Appl Anim Res 2002;22:249-53. 
  21. Janssens K, ten Dijke P, Janssens S, Van Hul W. Transforming growth factor-beta1 to the bone. Endocr Rev 2005;26:743-74.