Abstract
Uranium is widely spread in the environment due to its natural and anthropogenic occurrences, hence the importance of understanding its impact on human health. The skeleton is the main site of long-term accumulation of this actinide. However, interactions of this metal with biological processes involving the mineralized extracellular matrix and bone cells are still poorly understood. To get a better insight into these interactions, we developed new biomimetic bone matrices containing low doses of natural uranium (up to 0.85 µg of uranium per cm2). These models were characterized by spectroscopic and microscopic approaches before being used as a support for the culture and differentiation of pre-osteoclastic cells. In doing so, we demonstrate that uranium can exert opposite effects on osteoclast resorption depending on its concentration in the bone microenvironment. Our results also provide evidence for the first time that resorption contributes to the remobilization of bone matrix-bound uranium. In agreement with this, we identified, by HRTEM, uranium phosphate internalized in vesicles of resorbing osteoclasts. Thanks to the biomimetic matrices we developed, this study highlights the complex mutual effects between osteoclasts and uranium. This demonstrates the relevance of these 3D models to further study the cellular mechanisms at play in response to uranium storage in bone tissue, and thus better understand the impact of environmental exposure to uranium on human bone health.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analyzed during this study are included in this published article [and its supplementary information files] or are available from the corresponding author on reasonable request.
References
Arruda-Neto JDT, Guevara MVM, Nogueira GP et al (2004) Long-term accumulation of uranium in bones of Wistar rats as a function of intake dosages. Radiat Prot Dosim 112:385–393. https://doi.org/10.1093/rpd/nch405
ATSDR (2013) ATSDR-toxicological profile: uranium. http://www.atsdr.cdc.gov/toxprofiles/TP.asp?id=440&tid=77. Accessed 22 Jul 2016
Basset C, Averseng O, Ferron P-J et al (2013) Revision of the biodistribution of uranyl in serum: is fetuin-A the major protein target ? Chem Res Toxicol 26:645–653. https://doi.org/10.1021/tx400048u
Bostick WD, Jarabek RJ, Conca JL (1999) Phosphate-induced metal stabilization: Use of apatite and bone char for the removal of soluble radionuclides in authentic and simulated DOE groundwater. In: Air and Waste 92nd annual meeting and exhibition proceedings
Bourgeois D, Burt-Pichat B, Le Goff X et al (2015) Micro-distribution of uranium in bone after contamination: new insight into its mechanism of accumulation into bone tissue. Anal Bioanal Chem 407:6619–6625. https://doi.org/10.1007/s00216-015-8835-7
Bozal CB, Martinez AB, Cabrini RL, Ubios AM (2005) Effect of ethane-1-hydroxy-1,1-bisphosphonate (EHBP) on endochondral ossification lesions induced by a lethal oral dose of uranyl nitrate. Arch Toxicol 79:475–481. https://doi.org/10.1007/s00204-005-0649-5
Díaz Sylvester PL, López R, Ubios AM, Cabrini RL (2002) Exposure to subcutaneously implanted uranium dioxide impairs bone formation. Arch Environ Health 57:320–325. https://doi.org/10.1080/00039890209601415
Ellender M, Haines JW, Harrison JD (1995) The distribution and retention of plutonium, americium and uranium in CBA/H mice. Hum Exp Toxicol 14:38–48. https://doi.org/10.1177/096032719501400109
Eriksen EF (2010) Cellular mechanisms of bone remodeling. Rev Endocr Metab Disord 11:219–227. https://doi.org/10.1007/s11154-010-9153-1
Faruqi AF, Rao H, Causer J, Beltzer JP (2011) Corning osteo assay surface for the study of bone resorption. Bone 48:S47. https://doi.org/10.1016/j.bone.2010.10.132
Fukuda S, Ikeda M, Chiba M, Kaneko K (2006) Clinical diagnostic indicators of renal and bone damage in rats intramuscularly injected with depleted uranium. Radiat Prot Dosim 118:307–314. https://doi.org/10.1093/rpd/nci350
Fuller CC, Bargar JR, Davis JA, Piana MJ (2002) Mechanisms of uranium interactions with hydroxyapatite: implications for groundwater remediation. Environ Sci Technol 36:158–165. https://doi.org/10.1021/es0108483
Fuller CC, Bargar JR, Davis JA (2003) Molecular-scale characterization of uranium sorption by bone apatite materials for a permeable reactive barrier demonstration. Environ Sci Technol 37:4642–4649. https://doi.org/10.1021/es0343959
Gigliotti CL, Boggio E, Clemente N et al (2016) ICOS-ligand triggering impairs osteoclast differentiation and function in vitro and in vivo. J Immunol 197:3905–3916. https://doi.org/10.4049/jimmunol.1600424
Gritsaenko T, Pierrefite-Carle V, Lorivel T et al (2017) Natural uranium impairs the differentiation and the resorbing function of osteoclasts. Biochim Biophys Acta-Gen Subj 1861:715–726. https://doi.org/10.1016/j.bbagen.2017.01.008
Gritsaenko T, Pierrefite-Carle V, Creff G et al (2018) Methods for analyzing the impacts of natural uranium on in vitro osteoclastogenesis. J Vis Exp. https://doi.org/10.3791/56499
Guglielmotti MB, Ubios AM, de Rey BM, Cabrini RL (1984) Effects of acute intoxication with uranyl nitrate on bone formation. Experientia 40:474–476. https://doi.org/10.1007/BF01952392
Guglielmotti MB, Ubios AM, Cabrini RL (1985) Alveolar wound healing alteration under uranyl nitrate intoxication. J Oral Pathol Med 14:565–572. https://doi.org/10.1111/j.1600-0714.1985.tb00530.x
Hauge EM, Qvesel D, Eriksen EF et al (2001) Cancellous bone remodeling occurs in specialized compartments lined by cells expressing osteoblastic markers. J Bone Miner Res 16:1575–1582. https://doi.org/10.1359/jbmr.2001.16.9.1575
Hurault L, Creff G, Hagège A et al (2019) Uranium effect on osteocytic cells in vitro. Toxicol Sci 170:199–209. https://doi.org/10.1093/toxsci/kfz087
Huynh T-NS, Vidaud C, Hagège A (2016) Investigation of uranium interactions with calcium phosphate-binding proteins using ICP/MS and CE-ICP/MS. Metallomics 8:1185–1192. https://doi.org/10.1039/c6mt00147e
Kim HM, Rey C, Glimcher MJ (1995) Isolation of calcium-phosphate crystals of bone by non-aqueous methods at low temperature. J Bone Miner Res 10:1589–1601. https://doi.org/10.1002/jbmr.5650101021
Kurttio P, Komulainen H, Leino A et al (2005) Bone as a possible target of chemical toxicity of natural uranium in drinking water. Environ Health Persp 113:68–72. https://doi.org/10.1289/ehp.7475
Larivière D, Tolmachev SY, Kochermin V, Johnson S (2013) Uranium bone content as an indicator of chronic environmental exposure from drinking water. J Environ Radioact 121:98–103. https://doi.org/10.1016/j.jenvrad.2012.05.026
Leggett RW (1994) Basis for the ICRP’s age-specific biokinetic model for uranium. Health Phys 67:589–610. https://doi.org/10.1097/00004032-199412000-00002
Locock AJ, Burns PC (2003) The crystal structure of synthetic autunite, Ca[(UO2)(PO4)]2(H2O)11. Am Mineral 88:240–244. https://doi.org/10.2138/am-2003-0128
Lutter A-H, Hempel U, Wolf-Brandstetter C et al (2010) A novel resorption assay for osteoclast functionality based on an osteoblast-derived native extracellular matrix. J Cell Biochem 109:1025–1032. https://doi.org/10.1002/jcb.22485
Mehta VS, Maillot F, Wang Z et al (2016) Effect of reaction pathway on the extent and mechanism of uranium(VI) immobilization with calcium and phosphate. Environ Sci Technol 50:3128–3136. https://doi.org/10.1021/acs.est.5b06212
Milgram S, Carrière M, Thiebault C et al (2008) Cytotoxic and phenotypic effects of uranium and lead on osteoblastic cells are highly dependent on metal speciation. Toxicology 250:62–69. https://doi.org/10.1016/j.tox.2008.06.003
Neuman WF, Neuman MW (1949) The deposition of uranium in bone; adsorption studies in vitro. J Biol Chem 179:325–333
Ng PY, Brigitte Patricia Ribet A, Pavlos NJ (2019) Membrane trafficking in osteoclasts and implications for osteoporosis. Biochem Soc Trans 47:639–650. https://doi.org/10.1042/BST20180445
Pierrefite-Carle V, Santucci-Darmanin S, Breuil V et al (2016) Effect of natural uranium on the UMR-106 osteoblastic cell line: impairment of the autophagic process as an underlying mechanism of uranium toxicity. Arch Toxicol 91:1903–1914. https://doi.org/10.1007/s00204-016-1833-5
Priest ND, Howells GR, Green D, Haines JW (1982) Uranium in bone: metabolic and autoradiographic studies in the rat. Hum Toxicol 1:97–114. https://doi.org/10.1177/096032718200100202
Qi L, Basset C, Averseng O et al (2014) Characterization of UO2(2+) binding to osteopontin, a highly phosphorylated protein: insights into potential mechanisms of uranyl accumulation in bones. Metallomics 6:166–176. https://doi.org/10.1039/c3mt00269a
Ravel B, Newville M (2005) ATHENA, ARTEMIS, HEPHAESTUS: data analysis for X-ray absorption spectroscopy using IFEFFIT. J Synchrotron Radiat 12:537–541. https://doi.org/10.1107/S0909049505012719
Rehr JJ, Kas JJ, Vila FD et al (2010) Parameter-free calculations of X-ray spectra with FEFF9. Phys Chem Chem Phys 12:5503–5513. https://doi.org/10.1039/b926434e
Rodrigues G, Arruda-Neto JDT, Pereira RMR et al (2013) Uranium deposition in bones of Wistar rats associated with skeleton development. Appl Radiat Isotopes 82:105–110. https://doi.org/10.1016/j.apradiso.2013.07.033
Sitaud B, Solari PL, Schlutig S et al (2012) Characterization of radioactive materials using the MARS beamline at the synchrotron SOLEIL. J Nucl Mater 425:238–243. https://doi.org/10.1016/j.jnucmat.2011.08.017
Su X, Sun K, Cui FZ, Landis WJ (2003) Organization of apatite crystals in human woven bone. Bone 32:150–162. https://doi.org/10.1016/S8756-3282(02)00945-6
Tasat DR, Orona NS, Mandalunis PM et al (2007) Ultrastructural and metabolic changes in osteoblasts exposed to uranyl nitrate. Arch Toxicol 81:319–326. https://doi.org/10.1007/s00204-006-0165-2
Thakur P, Moore RC, Choppin GR (2009) Sorption of U(VI) species on hydroxyapatite. Radiochim Acta 93:385–391. https://doi.org/10.1524/ract.2005.93.7.385
Ubios AM, Guglielmotti MB, Steimetz T, Cabrini RL (1991) Uranium inhibits bone formation in physiologic alveolar bone modeling and remodeling. Environ Res 54:17–23. https://doi.org/10.1016/S0013-9351(05)80191-4
Vidaud C, Bourgeois D, Meyer D (2012) Bone as target organ for metals: the case of f-elements. Chem Res Toxicol 25:1161–1175. https://doi.org/10.1021/tx300064m
Wade-Gueye NM, Delissen O, Gourmelon P et al (2012) Chronic exposure to natural uranium via drinking water affects bone in growing rats. Biochim Biophys Acta-Gen Subj 1820:1121–1127. https://doi.org/10.1016/j.bbagen.2012.04.019
Wellman DM, Gunderson KM, Icenhower JP, Forrester SW (2007) Dissolution kinetics of synthetic and natural meta-autunite minerals, X3−n(n)+ [(UO2)(PO4)]2·xH2O, under acidic conditions. Geochem Geophys Geosy. https://doi.org/10.1029/2007GC001695
Acknowledgements
The authors would like to thank Chantal Cros and Colette Ricort for helpful technical assistance. The authors acknowledge the MARS beamline of SOLEIL synchrotron (Gif sur Yvette, France) that was used to perform XAS experiments and the IRCAN's Molecular and Cellular Core Imaging (PICMI) Facility which is supported by grants from the Ministère de l’Enseignement Supérieur, the Région PACA, the Conseil Départemental des Alpes Maritimes, INSERM, the FEDER, the GIS IBiSA, the Canceropole PACA and the foundation ARC.
Funding
The author’s lab work was funded by Université Côte d’Azur (UCA) and grants from the CEA (“Programme Transversal de Toxicologie Nucléaire”) and the ANR (ANR-16-CE34-0003-01). CCMA electron microscopy equipments have been funded by the Région Sud PACA, the Conseil Départemental des Alpes Maritimes and the GIS-IBiSA.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Rights and permissions
About this article
Cite this article
Gritsaenko, T., Pierrefite-Carle, V., Creff, G. et al. Low doses of uranium and osteoclastic bone resorption: key reciprocal effects evidenced using new in vitro biomimetic models of bone matrix. Arch Toxicol 95, 1023–1037 (2021). https://doi.org/10.1007/s00204-020-02966-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-020-02966-1