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Published in: Head & Face Medicine 1/2021

Open Access 01-12-2021 | Ovariectomy | Research

Effects of estrogen deficiency during puberty on maxillary and mandibular growth and associated gene expression – an μCT study on rats

Authors: Erika Calvano Küchler, Rafaela Mariana de Lara, Marjorie Ayumi Omori, Guido Marañón-Vásquez, Flares Baratto-Filho, Paulo Nelson-Filho, Maria Bernadete Sasso Stuani, Moritz Blanck-Lubarsch, Agnes Schroeder, Peter Proff, Christian Kirschneck

Published in: Head & Face Medicine | Issue 1/2021

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Abstract

Background

Estrogen is a well-known and important hormone involved in skeletal homeostasis, which regulates genes involved in bone biology. Some studies support that estrogen is important for craniofacial growth and development. Therefore this in vivo animal study aimed to investigate, whether and in which way low estrogen levels in the prepubertal period affect craniofacial development in the postpubertal stage and to quantify the gene expression of RANK, RANKL and OPG in cranial growth sites in ovariectomized estrogen-deficient rats during puberty.

Methods

Control (sham-operated, n = 18) and ovariectomy (OVX, n = 18) surgeries were performed on 21-days-old female Wistar rats. Animals euthanized at an age of 45 days (pubertal stage) were used for gene expression analyses (n = 6 per group) and immunohistochemistry of RANK, RANKL and OPG. Animals euthanized at 63 days of age (post-pubertal stage) were used for craniofacial two-dimensional and three-dimensional craniofacial measurements using μCT imaging (n = 12 per group).

Results

In the μCT analysis of the mandible and maxilla many statistically significant differences between sham-operated and OVX groups were observed, such as increased maxillary and mandibular bone length in OVX animals (p < 0.05). Condylar volume was also significantly different between groups (p < 0.05). The sham-operated group showed a higher level of RANK expression in the midpalatal suture (p = 0.036) and the RANKL:OPG ratio levels were higher in the OVX group (p = 0.015).

Conclusions

Our results suggest that estrogen deficiency during the prepubertal period is associated with alterations in the maxillary and mandibular bone length and condylar growth.
Literature
1.
go back to reference Cauley JA. Estrogen and bone health in men and women. Steroids. 2015;99:1115.CrossRef Cauley JA. Estrogen and bone health in men and women. Steroids. 2015;99:1115.CrossRef
3.
go back to reference Khosla S, Melton LJ, Atkinson EJ, O'Fallon WM. Relationship of serum sex steroid levels to longitudinal changes in bone density in young versus elderly men. J Clin Endocrinol Metab. 2001;8:3555–61.CrossRef Khosla S, Melton LJ, Atkinson EJ, O'Fallon WM. Relationship of serum sex steroid levels to longitudinal changes in bone density in young versus elderly men. J Clin Endocrinol Metab. 2001;8:3555–61.CrossRef
6.
go back to reference Wilson CA, Heinrichs C, Larmore KA, Craen M, Brown-Dawson J, Shaywitz S, et al. Estradiol levels in girls with Turner’s syndrome compared to normal prepubertal girls as determined by an ultrasensitive assay. J Pediatr Endocrinol Metab. 2003;1:91–6. Wilson CA, Heinrichs C, Larmore KA, Craen M, Brown-Dawson J, Shaywitz S, et al. Estradiol levels in girls with Turner’s syndrome compared to normal prepubertal girls as determined by an ultrasensitive assay. J Pediatr Endocrinol Metab. 2003;1:91–6.
9.
go back to reference Bachrach LK, Guido D, Katzman D, Litt IF, Marcus R. Decreased bone density in adolescent girls with anorexia nervosa. Pediatrics. 1990;86(3):440–7.PubMed Bachrach LK, Guido D, Katzman D, Litt IF, Marcus R. Decreased bone density in adolescent girls with anorexia nervosa. Pediatrics. 1990;86(3):440–7.PubMed
13.
go back to reference Hsu PY, Tsai MT, Wang SP, Chen YJ, Wu J, Hsu JT. Cortical bone morphological and trabecular bone microarchitectural changes in the mandible and femoral neck of ovariectomized rats; 2016. p. 29. Hsu PY, Tsai MT, Wang SP, Chen YJ, Wu J, Hsu JT. Cortical bone morphological and trabecular bone microarchitectural changes in the mandible and femoral neck of ovariectomized rats; 2016. p. 29.
15.
go back to reference Streicher C, Heyny A, Andrukhova O, Haigl B, Slavic S, Schüler C. Estrogen regulates bone turnover by targeting RANKL expression in bone lining cells. Sci Rep. 2017;7:1–14.CrossRef Streicher C, Heyny A, Andrukhova O, Haigl B, Slavic S, Schüler C. Estrogen regulates bone turnover by targeting RANKL expression in bone lining cells. Sci Rep. 2017;7:1–14.CrossRef
16.
go back to reference Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, et al. Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 1998;17:165–76.CrossRef Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, et al. Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 1998;17:165–76.CrossRef
17.
go back to reference Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, et al. OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 1999;28:315–23.CrossRef Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, et al. OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 1999;28:315–23.CrossRef
18.
go back to reference Li J, Sarosi I, Yan XQ, Morony S, Capparelli C, Tan HL, et al. RANK is the intrinsic hematopoietic cell surface receptor that controls osteoclastogenesis and regulation of bone mass and calcium metabolism. Proc Natl Acad Sci U S A. 2000;15:1566–71.CrossRef Li J, Sarosi I, Yan XQ, Morony S, Capparelli C, Tan HL, et al. RANK is the intrinsic hematopoietic cell surface receptor that controls osteoclastogenesis and regulation of bone mass and calcium metabolism. Proc Natl Acad Sci U S A. 2000;15:1566–71.CrossRef
22.
go back to reference Seifi M, Ashiri M, Hedayati M. Effect of sexual hormone elimination on the changes of craniofacial dimensions in rats. J Dent Sch. 2008;4:365–72. Seifi M, Ashiri M, Hedayati M. Effect of sexual hormone elimination on the changes of craniofacial dimensions in rats. J Dent Sch. 2008;4:365–72.
27.
go back to reference Wei X, Thomas N, Hatch NE, Hu M, Liu F. Postnatal craniofacial skeletal development of female C57BL/6NCrl mice. Front Physiol. 2017;14:697.CrossRef Wei X, Thomas N, Hatch NE, Hu M, Liu F. Postnatal craniofacial skeletal development of female C57BL/6NCrl mice. Front Physiol. 2017;14:697.CrossRef
31.
go back to reference Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001;25:402–8 22.CrossRefPubMed Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001;25:402–8 22.CrossRefPubMed
32.
go back to reference Barreiros D, Pucinelli CM, de Oliveira KM, Paula-Silva WG, Nelson-Filho P, da Silva LAB, et al. Immunohistochemical and mRNA expression of RANK, RANKL, OPG, TLR2 and MyD88 during apical periodontitis progression in mice. J Appl Oral Sci. 2018;26:e20170512.CrossRefPubMedPubMedCentral Barreiros D, Pucinelli CM, de Oliveira KM, Paula-Silva WG, Nelson-Filho P, da Silva LAB, et al. Immunohistochemical and mRNA expression of RANK, RANKL, OPG, TLR2 and MyD88 during apical periodontitis progression in mice. J Appl Oral Sci. 2018;26:e20170512.CrossRefPubMedPubMedCentral
35.
go back to reference Spence J. Methods of studying the skull development of the living rat by serial cephalometric roentgenograms. Am J Orthod. 1940;10:127–39. Spence J. Methods of studying the skull development of the living rat by serial cephalometric roentgenograms. Am J Orthod. 1940;10:127–39.
37.
go back to reference Väänänen HK, Härkönen PL. Estrogen and bone metabolism. Maturitas. 1996;23:65–9.CrossRef Väänänen HK, Härkönen PL. Estrogen and bone metabolism. Maturitas. 1996;23:65–9.CrossRef
40.
go back to reference Tat SK, Amiable N, Pelletier JP, Boileau C, Lajeunesse D, Duval N, et al. Modulation of OPG, RANK and RANKL by human chondrocytes and their implication during osteoarthritis. Rheumatology. 2009;48:1482–90.CrossRef Tat SK, Amiable N, Pelletier JP, Boileau C, Lajeunesse D, Duval N, et al. Modulation of OPG, RANK and RANKL by human chondrocytes and their implication during osteoarthritis. Rheumatology. 2009;48:1482–90.CrossRef
41.
go back to reference Ono T, Hayashi M, Sasaki F, Nakashima T. RANKL biology: bone metabolism, the immune system, and beyond. Inflamm Regen. 2020;40:51–7.CrossRef Ono T, Hayashi M, Sasaki F, Nakashima T. RANKL biology: bone metabolism, the immune system, and beyond. Inflamm Regen. 2020;40:51–7.CrossRef
Metadata
Title
Effects of estrogen deficiency during puberty on maxillary and mandibular growth and associated gene expression – an μCT study on rats
Authors
Erika Calvano Küchler
Rafaela Mariana de Lara
Marjorie Ayumi Omori
Guido Marañón-Vásquez
Flares Baratto-Filho
Paulo Nelson-Filho
Maria Bernadete Sasso Stuani
Moritz Blanck-Lubarsch
Agnes Schroeder
Peter Proff
Christian Kirschneck
Publication date
01-12-2021
Publisher
BioMed Central
Published in
Head & Face Medicine / Issue 1/2021
Electronic ISSN: 1746-160X
DOI
https://doi.org/10.1186/s13005-021-00265-3

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