Top

Published in:

Open Access 01-07-2010

Gene Expression and Distribution of Key Bone Turnover Markers in the Callus of Estrogen-Deficient, Vitamin D-Depleted Rats

Authors: Gunhild Melhus, S. H. Brorson, E. S. Baekkevold, G. Andersson, R. Jemtland, O. K. Olstad, F. P. Reinholt

Published in: Calcified Tissue International | Issue 1/2010

Abstract

An experimental rat model was used to test the hypothesis that in osteoporosis (OP) the molecular composition of the extracellular matrix in the fracture callus is disturbed. OP was induced at 10 weeks of age by ovariectomy and a vitamin D₃-deficient diet, and sham-operated animals fed normal diet served as controls. Three months later a closed tibial fracture was made and stabilized with an intramedullary nail. After 3 and 6 weeks of healing, the animals were killed and the fracture calluses examined with global gene expression, in situ mRNA expression, and ultrastructural protein distribution of four bone turnover markers: osteopontin, bone sialoprotein, tartrate-resistant acid phosphatase, and cathepsin K. Global gene expression showed a relatively small number of differently regulated genes, mostly upregulated and at 3 weeks. The four chosen markers were not differently regulated, and only minor differences in the in situ mRNA expression and ultrastructural protein distribution were detected. Gene expression and composition of fracture calluses are not generally disturbed in experimental OP.

Available only for authorised users

Eastelle R (2006) Pathogenesis of postmenopausal osteoporosis. In: Favus M (ed) Primer on the metabolic bone diseases and disorders of mineral metabolism. American Society for Bone and Mineral Research, Washington, DC, pp 259–262

Recker R, Lappe J, Davies K, Heaney R (2000) Characterization of perimenopausal bone loss: a prospective study. J Bone Miner Res 15:1965–1973CrossRefPubMed

Rodan GA (1991) Mechanical loading, estrogen deficiency, and the coupling of bone formation to bone resorption. J Bone Miner Res 6:527–530CrossRefPubMed

Raisz LG (2005) Pathogenesis of osteoporosis: concepts, conflicts, and prospects. J Clin Invest 115:3318–3325CrossRefPubMed

Nikolaou VS, Efstathopoulos N, Kontakis G, Kanakaris NK, Giannoudis PV (2009) The influence of osteoporosis in femoral fracture healing time. Injury 40:663–668CrossRefPubMed

Thompson DD, Simmons HA, Pirie CM, Ke HZ (1995) FDA guidelines and animal models for osteoporosis. Bone 17:125S–133SCrossRefPubMed

Namkung-Matthai H, Appleyard R, Jansen J, Hao Lin J, Maastricht S, Swain M, Mason RS, Murrell GA, Diwan AD, Diamond T (2001) Osteoporosis influences the early period of fracture healing in a rat osteoporotic model. Bone 28:80–86CrossRefPubMed

Kubo T, Shiga T, Hashimoto J, Yoshioka M, Honjo H, Urabe M, Kitajima I, Semba I, Hirasawa Y (1999) Osteoporosis influences the late period of fracture healing in a rat model prepared by ovariectomy and low calcium diet. J Steroid Biochem Mol Biol 68:197–202CrossRefPubMed

Melhus G, Solberg LB, Dimmen S, Madsen JE, Nordsletten L, Reinholt FP (2007) Experimental osteoporosis induced by ovariectomy and vitamin D deficiency does not markedly affect fracture healing in rats. Acta Orthop 78:393–403CrossRefPubMed

10.

Wheeler DL, Eschbach EJ, Montfort MJ, Maheshwari P, McLoughlin SW (2000) Mechanical strength of fracture callus in osteopenic bone at different phases of healing. J Orthop Trauma 14:86–92CrossRefPubMed

11.

Shi HF, Cheung WH, Qin L, Leung AH, Leung KS (2009) Low-magnitude high-frequency vibration treatment augments fracture healing in ovariectomy-induced osteoporotic bone. Bone. doi:10.1016/j.bone.2009.11.028

12.

Lips P, Hosking D, Lippuner K, Norquist JM, Wehren L, Maalouf G, Ragi-Eis S, Chandler J (2006) The prevalence of vitamin D inadequacy amongst women with osteoporosis: an international epidemiological investigation. J Intern Med 260:245–254CrossRefPubMed

13.

Steele B, Serota A, Helfet DL, Peterson M, Lyman S, Lane JM (2008) Vitamin D deficiency: a common occurrence in both high-and low-energy fractures. HSS J 4:143–148CrossRefPubMed

14.

Kaastad TS, Reikeras O, Halvorsen V, Falch JA, Obrant KJ, Nordsletten L (2001) Vitamin D deficiency and ovariectomy reduced the strength of the femoral neck in rats. Calcif Tissue Int 69:102–108CrossRefPubMed

15.

Fu L, Tang T, Miao Y, Hao Y, Dai K (2009) Effect of 1,25-dihydroxy vitamin D₃ on fracture healing and bone remodeling in ovariectomized rat femora. Bone 44:893–898CrossRefPubMed

16.

Young MF IK, Kerr JM, Heegaard AM (1993) Molecular and cellular biology of the major noncollagenous proteins in bone. In: Noda M (ed) Molecular and cellular biology of bone. Academic Press, Orlando, pp 191–234

17.

Oldberg A, Jirskog-Hed B, Axelsson S, Heinegard D (1989) Regulation of bone sialoprotein mRNA by steroid hormones. J Cell Biol 109:3183–3186CrossRefPubMed

18.

Yoon K, Buenaga R, Rodan GA (1987) Tissue specificity and developmental expression of rat osteopontin. Biochem Biophys Res Commun 148:1129–1136CrossRefPubMed

19.

Hultenby K, Reinholt FP, Oldberg A, Heinegard D (1991) Ultrastructural immunolocalization of osteopontin in metaphyseal and cortical bone. Matrix 11:206–213PubMed

20.

Reppe S, Stilgren L, Olstad OK, Brixen K, Nissen-Meyer LS, Gautvik KM, Abrahamsen B (2006) Gene expression profiles give insight into the molecular pathology of bone in primary hyperparathyroidism. Bone 39:189–198CrossRefPubMed

21.

Ek-Rylander B, Bill P, Norgard M, Nilsson S, Andersson G (1991) Cloning, sequence, and developmental expression of a type 5, tartrate-resistant, acid phosphatase of rat bone. J Biol Chem 266:24684–24689PubMed

22.

Lang P, Schultzberg M, Andersson G (2001) Expression and distribution of tartrate-resistant purple acid phosphatase in the rat nervous system. J Histochem Cytochem 49:379–396PubMed

23.

Baekkevold ES, Yamanaka T, Palframan RT, Carlsen HS, Reinholt FP, von Andrian UH, Brandtzaeg P, Haraldsen G (2001) The CCR7 ligand elc (CCL19) is transcytosed in high endothelial venules and mediates T cell recruitment. J Exp Med 193:1105–1112CrossRefPubMed

24.

Gundersen HJ, Bendtsen TF, Korbo L, Marcussen N, Moller A, Nielsen K, Nyengaard JR, Pakkenberg B, Sorensen FB, Vesterby A et al (1988) Some new, simple and efficient stereological methods and their use in pathological research and diagnosis. Acta Pathol Microbiol Immunol Scand 96:379–394

25.

Nordahl J, Andersson G, Reinholt FP (1998) Chondroclasts and osteoclasts in bones of young rats: comparison of ultrastructural and functional features. Calcif Tissue Int 63:401–408CrossRefPubMed

26.

Nordahl J, Hollberg K, Mengarelli-Widholm S, Andersson G, Reinholt FP (2000) Morphological and functional features of clasts in low phosphate, vitamin D-deficiency rickets. Calcif Tissue Int 67:400–407CrossRefPubMed

27.

Hollberg K, Nordahl J, Hultenby K, Mengarelli-Widholm S, Andersson G, Reinholt FP (2005) Polarization and secretion of cathepsin K precede tartrate-resistant acid phosphatase secretion to the ruffled border area during the activation of matrix-resorbing clasts. J Bone Miner Metab 23:441–449CrossRefPubMed

28.

Bianco P, Riminucci M, Silvestrini G, Bonucci E, Termine JD, Fisher LW, Robey PG (1993) Localization of bone sialoprotein (BSP) to Golgi and post-Golgi secretory structures in osteoblasts and to discrete sites in early bone matrix. J Histochem Cytochem 41:193–203PubMed

29.

Raisz LG, Trummel CL, Holick MF, DeLuca HF (1972) 1,25-Dihydroxycholecalciferol: a potent stimulator of bone resorption in tissue culture. Science 175:768–769CrossRefPubMed

30.

Shiraishi A, Takeda S, Masaki T, Higuchi Y, Uchiyama Y, Kubodera N, Sato K, Ikeda K, Nakamura T, Matsumoto T, Ogata E (2000) Alfacalcidol inhibits bone resorption and stimulates formation in an ovariectomized rat model of osteoporosis: distinct actions from estrogen. J Bone Miner Res 15:770–779CrossRefPubMed

31.

Baldock PA, Thomas GP, Hodge JM, Baker SU, Dressel U, O’Loughlin PD, Nicholson GC, Briffa KH, Eisman JA, Gardiner EM (2006) Vitamin D action and regulation of bone remodeling: suppression of osteoclastogenesis by the mature osteoblast. J Bone Miner Res 21:1618–1626CrossRefPubMed

32.

Baldock PA, Need AG, Moore RJ, Durbridge TC, Morris HA (1999) Discordance between bone turnover and bone loss: effects of aging and ovariectomy in the rat. J Bone Miner Res 14:1442–1448CrossRefPubMed

33.

Xiao Y, Fu H, Prasadam I, Yang YC, Hollinger JO (2007) Gene expression profiling of bone marrow stromal cells from juvenile, adult, aged and osteoporotic rats: with an emphasis on osteoporosis. Bone 40:700–715CrossRefPubMed

34.

Kolek OI, Hines ER, Jones MD, LeSueur LK, Lipko MA, Kiela PR, Collins JF, Haussler MR, Ghishan FK (2005) 1alpha,25-Dihydroxyvitamin D₃ upregulates FGF23 gene expression in bone: the final link in a renal–gastrointestinal–skeletal axis that controls phosphate transport. Am J Physiol Gastrointest Liver Physiol 289:G1036–G1042CrossRefPubMed

35.

Yu X, Sabbagh Y, Davis SI, Demay MB, White KE (2005) Genetic dissection of phosphate- and vitamin D-mediated regulation of circulating Fgf23 concentrations. Bone 36:971–977CrossRefPubMed

36.

Goebel S, Lienau J, Rammoser U, Seefried L, Wintgens KF, Seufert J, Duda G, Jakob F, Ebert R (2009) FGF23 is a putative marker for bone healing and regeneration. J Orthop Res 27:1141–1146CrossRefPubMed

37.

Yu JJ, Ruddy MJ, Conti HR, Boonanantanasarn K, Gaffen SL (2008) The interleukin-17 receptor plays a gender-dependent role in host protection against Porphyromonas gingivalis-induced periodontal bone loss. Infect Immun 76:4206–4213CrossRefPubMed

38.

Goswami J, Hernandez-Santos N, Zuniga LA, Gaffen SL (2009) A bone-protective role for IL-17 receptor signaling in ovariectomy-induced bone loss. Eur J Immunol 39:2831–2839CrossRefPubMed

39.

Franzen A, Hultenby K, Reinholt FP, Onnerfjord P, Heinegard D (2008) Altered osteoclast development and function in osteopontin deficient mice. J Orthop Res 26:721–728CrossRefPubMed

40.

Boskey AL, Maresca M, Ullrich W, Doty SB, Butler WT, Prince CW (1993) Osteopontin–hydroxyapatite interactions in vitro: inhibition of hydroxyapatite formation and growth in a gelatin-gel. Bone Miner 22:147–159CrossRefPubMed

41.

Yoshitake H, Rittling SR, Denhardt DT, Noda M (1999) Osteopontin-deficient mice are resistant to ovariectomy-induced bone resorption. Proc Natl Acad Sci USA 96:8156–8160CrossRefPubMed

42.

Duvall CL, Taylor WR, Weiss D, Wojtowicz AM, Guldberg RE (2007) Impaired angiogenesis, early callus formation, and late stage remodeling in fracture healing of osteopontin-deficient mice. J Bone Miner Res 22:286–297CrossRefPubMed

43.

Hultenby K, Reinholt FP, Norgard M, Oldberg A, Wendel M, Heinegard D (1994) Distribution and synthesis of bone sialoprotein in metaphyseal bone of young rats show a distinctly different pattern from that of osteopontin. Eur J Cell Biol 63:230–239PubMed

44.

Chen JJ, Jin H, Ranly DM, Sodek J, Boyan BD (1999) Altered expression of bone sialoproteins in vitamin D-deficient rBSP2.7Luc transgenic mice. J Bone Miner Res 14:221–229CrossRefPubMed

45.

Cao Y, Mori S, Mashiba T, Westmore MS, Ma L, Sato M, Akiyama T, Shi L, Komatsubara S, Miyamoto K, Norimatsu H (2002) Raloxifene, estrogen, and alendronate affect the processes of fracture repair differently in ovariectomized rats. J Bone Miner Res 17:2237–2246CrossRefPubMed

46.

Jeong KS, Lee J, Jeong W, Noh DH, Do SH, Kim YK (2005) Measurement of estrogen effect on bone turnover by 2H₂O labeling. Calcif Tissue Int 76:365–370CrossRefPubMed

47.

Huang J, Wang X, Zhang TL, Wang K (2009) Alterations of ovariectomized rat bone and impact of non-collagenous proteins on mineralization. Joint Bone Spine 76:176–183CrossRefPubMed

48.

Egermann M, Heil P, Tami A, Ito K, Janicki P, Von Rechenberg B, Hofstetter W, Richards PJ (2009) Influence of defective bone marrow osteogenesis on fracture repair in an experimental model of senile osteoporosis. J Orthop Res. doi:10.1002/jor.21041

49.

Gerstenfeld LC, Cullinane DM, Barnes GL, Graves DT, Einhorn TA (2003) Fracture healing as a post-natal developmental process: molecular, spatial, and temporal aspects of its regulation. J Cell Biochem 88:873–884CrossRefPubMed

50.

Palomares KT, Gleason RE, Mason ZD, Cullinane DM, Einhorn TA, Gerstenfeld LC, Morgan EF (2009) Mechanical stimulation alters tissue differentiation and molecular expression during bone healing. J Orthop Res 27:1123–1132CrossRefPubMed

51.

Harvey NES, Cooper C (2006) Epidemiology of osteoporotic fractures. In: Favus MJ (ed) Primer on the metabolic bone diseases and disorders of mineral metabolism. American Society for Bone and Mineral Research, Washington, DC, pp 244–248

52.

McKee MD, Nanci A (1996) Osteopontin at mineralized tissue interfaces in bone, teeth, and osseointegrated implants: ultrastructural distribution and implications for mineralized tissue formation, turnover, and repair. Microsc Res Tech 33:141–164CrossRefPubMed

Title: Gene Expression and Distribution of Key Bone Turnover Markers in the Callus of Estrogen-Deficient, Vitamin D-Depleted Rats
Authors: Gunhild Melhus
S. H. Brorson
E. S. Baekkevold
G. Andersson
R. Jemtland
O. K. Olstad
F. P. Reinholt
Publication date: 01-07-2010
Publisher: Springer-Verlag
Published in: Calcified Tissue International / Issue 1/2010
Print ISSN: 0171-967X
Electronic ISSN: 1432-0827
DOI: https://doi.org/10.1007/s00223-010-9371-2

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Gene Expression and Distribution of Key Bone Turnover Markers in the Callus of Estrogen-Deficient, Vitamin D-Depleted Rats

Abstract

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2010

Analysis of Three Functional Polymorphisms in Relation to Osteoporosis Phenotypes: Replication in a Spanish Cohort

Tissue Restoration After Implantation of Polyglycolide, Polydioxanone, Polylevolactide, and Metallic Pins in Cortical Bone: An Experimental Study in Rabbits

Genotypes and Haplotypes of the Estrogen Receptor Genes, but Not the Retinoblastoma-interacting Zinc Finger Protein 1 Gene, Are Associated with Osteoporosis

Adiponectin and Peak Bone Mass in Men: A Cross-Sectional, Population-Based Study

Is Nitric Oxide a Mediator of the Effects of Low-Intensity Electrical Stimulation on Bone in Ovariectomized Rats?

The Bone Histology Spectrum in Experimental Renal Failure: Adverse Effects of Phosphate and Parathyroid Hormone Disturbances

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page