Abstract
This study investigated whether 17β-estradiol (E2) may have different effects on osteoblasts derived from estrogen-deficient ovariectomized (OVX) mice compared to sham-operated normal animals. We studied the specific effects of 17β-estradiol on the differentiation and function of cultured osteoblasts derived from these groups of animals, with or without estrogen replacement treatment. One-month-old mice were ovariectomized or sham-operated, and treated (every second day) for 4 weeks with 0.5 mg/kg 17β-estradiol or with vehicle alone. At the end of the experiment, bones were removed for primary osteoblast cultures or for morphological and chemical evaluation. In cells from untreated OVX animals, alkaline phosphatase (ALP) specific activity was reduced, while collagen production and mineralization were unchanged when compared to cells from controls. In vivo estrogen pretreatment of the OVX mice elevated ALP activity and mineralization of the cells, while collagen production was reduced. The addition of 17β-estradiol to the culture medium increased ALP activity, collagen production, and mineralization by all cultured osteoblasts, except in those derived from sham-operated, estrogen-pretreated mice, where these features remained unchanged. Osteocalcin production was unchanged. Addition of testosterone or 1,25(OH)2D3 to the culture medium induced changes that differed among the groups depending on the source of the cultured cells. It seems that ovariectomy in mice prior to culture affected the phenotype of the cultured osteoblasts and their response to estradiol, testosterone, and 1,25(OH)2D3, depending on whether animals were pretreated with estradiol or not. These results imply that the animal’s estrogen status prior to culture can influence the response to estrogens; this finding may have important implications for hormone replacement therapy (HRT) in postmenopausal women.
Similar content being viewed by others
References
FP Chen N Lee K-C Wang YK Soong KE Huang (2002) ArticleTitleEffects of estrogen and 1α,25(OH)2, -vitamin D3 on the activity and growth of human primary osteoblast-like cells in vitro Fertil Steril 77 1038–43 Occurrence Handle10.1016/S0015-0282(02)03065-0 Occurrence Handle12009364
LG Rao LJ Liu TM Murray E McDermott X Zhang (2003) ArticleTitleEstrogen added intermittently, but not continuously, stimulates differentiation and bone formation in SaOS-2 cells Biol Pharm Bull 26 936–45 Occurrence Handle10.1248/bpb.26.936 Occurrence Handle12843614
HJJ Verhaar CA Damen Duursma BAA Scheven (1994) ArticleTitleA comparison of action of progestins and estrogen on the growth and differentiation of normal adult human osteoblast-like cells in vitro Bone 15 307–11 Occurrence Handle10.1016/8756-3282(94)90293-3 Occurrence Handle8068452
Q Qu M Perala-Heape A Kapanen J Dahllund J Salo HK Vaananen P Harkonen (1998) ArticleTitleEstrogen enhances differentiation of osteoblasts in mouse bone marrow culture Bone 22 201–9 Occurrence Handle10.1016/S8756-3282(97)00276-7 Occurrence Handle9514212
R Okazaki D Inoue M Shibata M Saika S Kido H Ooka H Tomiyama Y Sakamoto T Matsumoto (2002) ArticleTitleEstrogen promotes early osteoblast differentiation and inhibits adipocyte differentiation in mouse bone marrow stromal cell lines that express estrogen receptor (ER) alpha or beta Endocrinology 143 2349–56 Occurrence Handle10.1210/en.143.6.2349 Occurrence Handle12021200
Z Schwartz T Neubauer M Goldstein S Adi WA Soskolne A Ornoy (1991) ArticleTitleDirect and sex specific enhancement of bone formation and calcification by sex steroids in fetal mice long bone in vitro (biochemical and morphometric study) Endocrinology 129 1167–74 Occurrence Handle1874163
M Morishita T Yamamura A Shimazu AH Bachchu Y Iwamoto (1999) ArticleTitleEstradiol enhances the production of mineralized nodules by human periodontal ligament cells J Clin Periodontol 26 748–51 Occurrence Handle10.1034/j.1600-051X.1999.t01-7-261101.x Occurrence Handle10589811
A Samuels MJ Perry JH Tobias (1999) ArticleTitleHigh-dose estrogen induces de novo medullary bone formation in female mice J Bone Miner Res 14 178–86 Occurrence Handle9933470
PE Keeting RE Scott DS Colvard IK Han TC Spelsberg BL Riggs (1991) ArticleTitleLack of a direct effect of estrogen on proliferation and differentiation of normal human osteoblast-like cells J Bone Miner Res 6 297–304 Occurrence Handle2035356
KE McDougall MJ Perry RL Gibson SM Colley KS Korach JH Tobias (2003) ArticleTitleEstrogen receptor-α dependency of estrogen’s stimulatory actin on cancellous bone formation in male mice Endocrinology 144 1994–9 Occurrence Handle10.1210/en.2002-0074 Occurrence Handle12697707
L Cao R Bu JI Oakley SE Kalla HC Blair (2003) ArticleTitleEstrogen receptor-β modulates synthesis of bone matrix proteins in human osteoblast-like MG63 cells J Cell Biochem 89 152–64 Occurrence Handle10.1002/jcb.10486 Occurrence Handle12682916
BL Riggs LJ Melton Suffix3rd (1986) ArticleTitleInvolutional osteoporosis N Engl J Med 314 1676–86 Occurrence Handle3520321
A Ornoy S Giron R Aner M Goldstein BD Boyan Z Schwartz (1994) ArticleTitleGender dependent effect of testosterone and 17β-estradiol on bone growth and modeling in young mice Bone Miner 24 43–54 Occurrence Handle8186733
N Patlas Y Zadik P Yaffe Z Schwartz A Ornoy (2000) ArticleTitleOophorectomy-induced osteopenia in rats in relation to age and time postoophorectomy Cells Tissues Organs 166 267–74 Occurrence Handle10.1159/000016740 Occurrence Handle10765022
S Katzburg M Lieberherr A Ornoy BY Klein D Hendel D Somjen (1999) ArticleTitleIsolation and hormonal responsiveness of primary cultures of human bone-derived cells: gender and age differences Bone 25 667–73 Occurrence Handle10.1016/S8756-3282(99)00225-2 Occurrence Handle10593411
S Katzburg A Ornoy D Hendel M Lieberherr AM Kaye D Somjen (2000) ArticleTitleAge and gender specific stimulation of creatine kinase specific activity by gonadal steroids in human bone derived cells in culture J Endocrinol Invest 24 166–72
X Wang Z Schwartz P Yaffe A Ornoy (1999) ArticleTitleThe expression of TGF-β and IL-1β mRNA and response to 1,25(OH)2D3, 17β -estradiol and testosterone is age dependent in primary cultures of mouse-derived osteoblasts in vitro Endocrine 11 13–22 Occurrence Handle10.1385/ENDO:11:1:13 Occurrence Handle10668636
JP Bretaudierre T Spillman (1984) Alkaline phosphatase HU Bergmeyer (Eds) Methods of enzymatic analysis EditionNumber9th ed. Verlag Chemica Weinhein 75–92
B Pewterkofsky R Diegelmann (1971) ArticleTitleUse of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins Biochemistry 10 988–94 Occurrence Handle10.1021/bi00782a009 Occurrence Handle4323854
DC Ireland S Bord SR Beavan JE Compston (2002) ArticleTitleEffects of estrogen on collagen synthesis by cultured human osteoblasts depend on the rate of cellular differentiation J Cell Biochem 86 251–7 Occurrence Handle10.1002/jcb.10228 Occurrence Handle12111994
G Holzer TA Einhorn RJ Majeska (2002) ArticleTitleEstrogen regulation of growth and alkaline phosphatase expression by cultured human bone marrow stromal cells J Orthop Res 20 281–8 Occurrence Handle10.1016/S0736-0266(01)00092-4 Occurrence Handle11918307
GS Stein JB Lian (1993) ArticleTitleMolecular mechanisms mediating proliferation / differentiation interrelationships during progressive development of the osteoblast phenotype Endocrine Rev 14 424–42 Occurrence Handle10.1210/er.14.4.424
RT Turner P Backup PJ Sherman E Hill GL Evans TC Spelsberg (1992) ArticleTitleMechanism of action of estrogen on intramembranous bone formation: regulation of osteoblast differentiation and activity Endocrinology 131 883–9 Occurrence Handle10.1210/en.131.2.883 Occurrence Handle1639030
MS Kung Sutherland LG Rao SA Muzaffar JN Wylie MM Wong RJ McBroom TM Murray (1995) ArticleTitleAge-dependent expression of osteoblastic phenotypic markers in normal human osteoblasts cultured long-term in the presence of dexamethasone Osteoporosis Int 5 335–43 Occurrence Handle10.1007/BF01622255
JR Farley JL Ivey DJ Baylink (1980) ArticleTitleHuman skeletal alkaline phosphatase J Biol Chem 255 4680–6 Occurrence Handle7372601
KH Wlodavsky AH Reddi (1986) ArticleTitleAlkaline phosphatase as a marker of osteoinductive cells Calcif Tissue Int 39 382–5 Occurrence Handle3100001
CJ Rosen CR Kessenich (1997) ArticleTitleThe pathophysiology and treatment of postmenopausal osteoporosis. An evidence-based approach to estrogen replacement therapy Endocrinol Metab Clin North Am 26 295–311 Occurrence Handle10.1016/S0889-8529(05)70248-6 Occurrence Handle9193885
B Lees JC Stevenson (2000) ArticleTitleThe prevention of osteoporosis using sequential low-dose hormone replacement therapy with estradiol 17β and dydrogesterone Osteoporosis Int 12 251–8 Occurrence Handle10.1007/s001980170113
DJ Torgerson SEM Bell-Syer (2001) ArticleTitleHormone replacement therapy and prevention of non-vertebral fractures: a meta-analysis of randomized trials JAMA 285 2891–7 Occurrence Handle10.1001/jama.285.22.2891 Occurrence Handle11401611
S Zhou Y Zilberman K Wassermann SD Bain Y Sadovsky D Gazit (2001) ArticleTitleEstrogen modulates estrogen receptor alpha and beta expression, osteogenic activity, and apoptosis in mesenchymal stem cells (MSCs) of osteoporotic mice J Cell Biochem 81S 144–55 Occurrence Handle10.1002/jcb.1096
RA Mason HA Morris (1997) ArticleTitleEffects of dihydrotestosterne on bone biochemical markers in sham and oophorectomized rats J Bone Miner Res 12 1431–7 Occurrence Handle9286759
P van Kesteren P Lips LJ Gooren H Asscheman J Megens (1998) ArticleTitleLong-term follow-up of bone mineral density and bone metabolism in transsexuals treated with cross-sex hormones Clin Endocrinol 48 347–54 Occurrence Handle10.1046/j.1365-2265.1998.00396.x
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Patlas, N., Zadik, Y., Yaffe, P. et al. The response to sex steroid hormones and vitamin D of cultured osteoblasts derived from ovariectomized mice with and without 17β-estradiol pretreatment. Odontology 93, 16–23 (2005). https://doi.org/10.1007/s10266-005-0051-z
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10266-005-0051-z