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Advantages and disadvantages of the animal models v. in vitro studies in iron metabolism: a review

Published online by Cambridge University Press:  21 June 2013

Y. García  and
J. Díaz-Castro
Y. García
Affiliation:
Restorative Laboratory, National Centre of Biological Products, Mayabeque 6048, Cuba
J. Díaz-Castro*
Affiliation:
Department of Physiology and Institute of Nutrition and Food Technology, University of Granada, E-18071 Granada, Spain
*
E-mail: javierdc@ugr.es
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Abstract

Iron deficiency is the most common nutritional deficiency in the world. Special molecules have evolved for iron acquisition, transport and storage in soluble, nontoxic forms. Studies about the effects of iron on health are focused on iron metabolism or nutrition to prevent or treat iron deficiency and anemia. These studies are focused in two main aspects: (1) basic studies to elucidate iron metabolism and (2) nutritional studies to evaluate the efficacy of iron supplementation to prevent or treat iron deficiency and anemia. This paper reviews the advantages and disadvantages of the experimental models commonly used as well as the methods that are more used in studies related to iron. In vitro studies have used different parts of the gut. In vivo studies are done in humans and animals such as mice, rats, pigs and monkeys. Iron metabolism is a complex process that includes interactions at the systemic level. In vitro studies, despite physiological differences to humans, are useful to increase knowledge related to this essential micronutrient. Isotopic techniques are the most recommended in studies related to iron, but their high cost and required logistic, making them difficult to use. The depletion–-repletion of hemoglobin is a method commonly used in animal studies. Three depletion–-repletion techniques are mostly used: hemoglobin regeneration efficiency, relative biological values (RBV) and metabolic balance, which are official methods of the association of official analytical chemists. These techniques are well-validated to be used as studies related to iron and their results can be extrapolated to humans. Knowledge about the main advantages and disadvantages of the in vitro and animal models, and methods used in these studies, could increase confidence of researchers in the experimental results with less costs.

Keywords

experimental modelsiron metabolismin vitro studiesanimal models
Type
Physiology and functional biology of systems
Information
animal , Volume 7 , Issue 10 , October 2013 , pp. 1651 - 1658
Copyright
Copyright © The Animal Consortium 2013 

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References

Aggett, PJ 1999. Iron, copper, and zinc absorption and turnover; the use of stable isotopes. European Journal of Pediatrics 156, S29–S34.Google Scholar
Andrews, NC 1999. Disorders of iron metabolism. New English Journal of Medicine 341, 1986–1995.Google Scholar
Arredondo, M, Kloosterman, J, Núñez, S, Segovia, F, Candia, V, Flores, S, Le Blanc, S, Olivares, M, Pizarro, F 2008. Heme iron uptake by Caco-2 cells is a saturable, temperature sensitive and modulated by extracellular pH and potassium. Biological Trace Element Research 125, 109–119.Google Scholar
Beach, SB, Hansen, M, Bukhave, K, Jensen, M, Sorensen, SS, Kristensen, L, Purslow, PP, Skibsted, LH, Sandstrom, B 2003. Non heme iron absorption from a phytate rich meal is increased by the addition of small amounts of pork meat. American Journal of Clinical Nutrition 77, 173–179.CrossRefGoogle Scholar
Beard, J 2007. Recent evidence from human and animal studies regarding iron status and infant development. Journal of Nutrition 137, 524S–530S.CrossRefGoogle ScholarPubMed
Bothwell, TH, Charlton, RW, Cook, JD, Finch, CA 1979. Iron metabolism in man. Blackwell Scientific, Oxford, UK.Google Scholar
Casanueva, E, Viteri, FE 2003. Iron and oxidative stress in pregnancy. Journal of Nutrition 133, 1700S–1708S.Google Scholar
Conrad, ME, Umbreit, JN, Moore, EG, Rodning, CR 1992. Newly identified iron-binding protein in human duodenal mucosa. Blood 79, 244–247.CrossRefGoogle ScholarPubMed
Donovan, A, Lima, CA, Pinkus, JL, Pinkus, GS, Zon, LI, Robine, S, Andrews, NC 2005. The iron exporter ferroportin/slc40a1 is essential for iron homeostasis. Cell Metabolism 1, 191–200.Google Scholar
Fleming, MD, Romano, MA, Su, MA, Garrick, LM, Garrick, MD, Andrews, NC 1998. Nramp2 is mutated in the anemic Belgrade (b) rat: evidence of a role for Nramp2 in endosomal iron transport. Proceedings of the National Academy of Sciences of the United States of America 95, 1148–1153.CrossRefGoogle ScholarPubMed
Fiorito, V, Geninatti, SC, Silengo, L, Altruda, F, Aime, S, Tolosano, E 2012. Assessment of iron absorption in mice by ICP-MS measurements of 57Fe levels. European Journal of Nutrition 51, 783–789.CrossRefGoogle ScholarPubMed
Galy, B, Ferring-Appel, D, Kaden, S, Grone, HJ, Hentze, MW 2008. Iron regulatory proteins are essential for intestinal function and control key iron absorption molecules in the duodenum. Cell Metabolism 7, 79–85.Google Scholar
Glahn, RP, Van Campen, DR 1997. Iron uptake is enhanced in caco-2 cell monolayers by cysteine and reduced cysteinyl glycine. Journal of Nutrition 127, 642–647.Google Scholar
Goddard, WP, Coupland, K, Smith, JA, Lon, RG 1997. Iron uptake by isolated human enterocyte suspensions in vitro is dependent on body iron stores and inhibited by other metal cations. Journal of Nutrition 127, 177–183.CrossRefGoogle ScholarPubMed
Golub, MS, Hogrefe, CE, Germann, SL, Capitanio, JP, Lozoff, B 2006. Behavioral consequences of developmental iron deficiency in infant rhesus monkeys. Neurotoxicology and Teratology 28, 3–17.CrossRefGoogle ScholarPubMed
González-Iglesias, H, Fernández-Sánchez, ML, López-Sastre, J, Sanz-Medel, A 2012. Nutritional iron supplementation studies based on enriched (57) Fe, added to milk in rats, and isotope pattern deconvolution-ICP-MS analysis. Electrophoresis 33, 2407–2415.CrossRefGoogle ScholarPubMed
Gordon, DT, Godber, JS 1989. The enhancement of nonheme iron bioavailability by beef protein in the rat. Journal of Nutrition 119, 446–452.Google Scholar
Gunshin, H, Fujiwara, Y, Custodio, AO, Direnzo, C, Robine, S, Andrews, NC 2005. Slc11a2 is required for intestinal iron absorption and erythropoiesis but dispensable in placenta and liver. Journal of Clinical Investigation 115, 1258–1266.Google Scholar
Hartmann, F, Bissel, M 1982. Journal of Clinical Investigation 70, 23–29.Google Scholar
Heaton, SJ, Eady, J, Parker, ML, Gotts, KL, Dainty, JR, Fairweather-Tait, SJ, McArdle, HJ, Srai, KS, Elliott, RM 2008. The use of BeWo cells as an in vitro model for placental iron transport. American Journal of Physiology – Cell Physiology 295, C1445–C1453.Google Scholar
Hentze, MW, Muckenthaler, MU, Andrews, NC 2004. Balancing acts: molecular control of mammalian iron metabolism. Cell 117, 285–297.Google Scholar
Hoppler, M, Schonbachler, A, Meile, L, Hurrell, RF, Walczyk, T 2008. Ferritin-iron is released during boiling and in vitro gastric digestion. Journal of Nutrition 138, 878–884.Google Scholar
Huang, FW, Pinkus, JL, Pinkus, GS, Fleming, MD, Andrews, NC 2005. A mouse model of juvenile hemochromatosis. Journal of Clinical Investigation 115, 2187–2191.CrossRefGoogle ScholarPubMed
Jin, F, Frohman, C, Thannhauser, TW, Welch, RM, Glahn, RP, Jin, F 2009. Effects of ascorbic acid, phytic acid and tannic acid on iron bioavailability from reconstituted ferritin measured by an in vitro digestion-Caco-2 cell model. British Journal of Nutrition 101, 972–981.Google Scholar
Knutson, MD, Vafa, MR, Haile, DJ, Wessling-Resnick, M 2003. Iron overloading and erythrophagocytosis increase ferroportin 1 (FPN1) expression in J774 macrophages. Blood 102, 4191–4197.CrossRefGoogle Scholar
Kolachala, VL, Sesikeran, B, Nair, KM 2007. Evidence for a sequential transfer of iron amongst ferritin, transferrin and transferrin receptor during duodenal absorption of iron in rat and human. World Journal of Gastroenterology 13, 1042–1052.Google Scholar
Krause, A, Neitz, S, Magert, HJ, Schulz, A, Forssmann, WG, Schulz-Knappe, P, Adermann, K 2000. LEAP-1, a novel highly disulfide-bonded human peptide, exhibits antimicrobial activity. Febs Letter 480, 147–150.CrossRefGoogle ScholarPubMed
Lane, DJR, Lawen, A 2008. Non-transferrin iron reduction and uptake are regulated by transmembrane ascorbate cycling in k562 cells. Journal of Biological Chemistry 283, 12701–12708.CrossRefGoogle ScholarPubMed
Latunde-Dada, GO 2009. Iron metabolism: microbes, mouse, and man. BioEssays 31, 1309–1317.CrossRefGoogle ScholarPubMed
Latunde-Dada, GO, Pires-Bianchi, ML, Dutra-Oliveira, JE 1998. On the methods for studying the mechanism and bioavailability of iron. Nutrition Reviews 3, 76–80.Google Scholar
Lönnerdal, B 2009. Soybean ferritin: implications for iron status of vegetarians. Amercian Journal of Clinical Nutrition 89, 1680S–1685S.CrossRefGoogle ScholarPubMed
Lonnerdal, B, Bryant, A, Liu, X, Theil, E 2006. Iron absorption from soybean ferritin in non anemic woman. American Journal of Clinical Nutrition 83, 103–107.Google Scholar
Lynch, S 2007. Iron metabolism. In Nutritional anemia. (ed. K Kraemer and MB Zimmermann), pp. 66–67. Sight and Life Press, Basel, Switzerland.Google Scholar
Lynch, SR, Stoltzfus, RJ 2003. Iron and ascorbic acid: proposed fortification levels and recommended iron compounds. Journal of Nutrition 133, 2978S–2984S.Google Scholar
Mastrogiannaki, M, Matak, P, Keith, B, Simon, MC, Vaulont, S, Peyssonnaux, C 2009. HIF-2alpha, but not HIF-1alpha, promotes iron absorption in mice. Journal of Clinical Investigation 119, 1159–1166.Google Scholar
McLean, E, Egli, I, de Benoist, B, Wojdyla, D, Cogswell, M 2007. Worldwide prevalence of anemia in preschool aged children, pregnant women and non-pregnant women of reproductive age. In Nutritional anemia (ed. K Kraemer and MB Zimmermann), pp. 108–109. Sight and Life Press, Basel, Switzerland.Google Scholar
Meynard, D, Kautz, L, Darnaud, V, Canonne-Hergaux, F, Coppin, H, Roth, MP 2009. Lack of the bone morphogenetic protein BMP6 induces massive iron overload. Nature Genetics 41, 478–481.Google Scholar
Miller, DD, Schricker, BR, Rasmussen, RR, Van Campen, D 1981. An in vitro method for estimation of iron availability from meals. American Journal of Clinical Nutrition 34, 2248–2256.Google Scholar
Miller, ER, Ullrey, DE 1987. The pig as a model for human nutrition. Annual Review of Nutrition 7, 361–382.Google Scholar
Morais, MB, Feste, A, Miller, RG, Lifschitz, CH 1996. Effect of resistant and digestible starch on intestinal absorption of calcium, iron and zinc in infant pigs. Pediatrics Research 39, 872–876.CrossRefGoogle ScholarPubMed
Moretti, D, Zimmermann, MB, Wegmüller, R, Walczyk, T, Zeder, C, Hurrell, RF 2006. Iron status and food matrix strongly affect the relative bioavailability of ferric pyrophosphate in humans. American Journal of Clinival Nutrition 83, 632–638.Google Scholar
Morgan, EH, Oates, PS 2002. Mechanisms and regulation of intestinal iron absorption. Blood Cell Molecular and Disease 29, 384–399.Google Scholar
Moshtaghie, AA 2006. Investigation of manganese and iron absorption by rat everted gut sac. Pakistan Journal of Biological Sciences 9, 1346–1349.Google Scholar
Moshtaghie, AA, Taher, M 1993. Aluminium interference with iron absorption by everted gut sac. Journal of Islamic World Academic Science 6, 277–281.Google Scholar
Nadadur, SS, Srirama, K, Mudipalli, A 2008. Iron transport and homeostasis mechanisms: their role in health and disease. Indian Journal of Medical Resesearch 8, 533–544.Google Scholar
Nagababu, E, Gulyani, S, Earley, CJ, Cutler, RG, Mattson, MP, Rifkind, JM 2008. Iron deficiency anemia enhanced red blood cell oxidative stress. Free Radical Research 42, 824–829.Google Scholar
Nemeth, E, Ganz, T 2006. Hepcidin and iron-loading anemias. Haematologica 91, 727–732.Google Scholar
Nicolas, G, Bennoun, M, Devaux, I, Beaumont, C, Grandchamp, B, Kahn, A, Vaulont, S 2001. Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2) knockout mice. Proceedings of the National Academy of the Sciences USA 98, 8780–8785.Google Scholar
Nicolas, G, Chauvet, C, Viatte, L, Danan, JL, Bigard, X, Devaux, I, Beaumont, C, Kahn, A, Vaulont, S 2002. The gene encoding the iron regulatory peptide hepcidin is regulated by anemia, hypoxia, and inflammation. Journal of Clinical Investigation 110, 1037–1044.Google Scholar
Patel, N, Masaratana, P, Díaz-Castro, J, Latunde-Dada, GO, Qureshi, A, Lockyer, P, Jacob, M, Arno, M, Matak, P, Mitry, RR, Hughes, RD, Dhawan, A, Patterson, C, Simpson, RJ, McKie, AT 2012. BMPER protein is a negative regulator of hepcidin and is up-regulated in hypotransferrinemic mice. Journal of Biological Chemistry 287, 4099–4106.Google Scholar
Patterson, JK, Lei, XG, Millar, DD 2008. The pig as an experimental model for elucidating the mechanisms governing dietary influence on mineral absorption. Experimental Biology and Medicine 233, 651–664.CrossRefGoogle ScholarPubMed
Pearson, WN, Reich, M 1965. In vitro studies of 59Fe absorption by everted intestinal sacs of the rat. Journal of Nutrition 87, 117–124.Google Scholar
Pizarro, F, Olivares, M, Hertrampf, E, Mazariegos, DI, Arredondo, M, Letelier, A, Gidi, V 2002. Iron bis-glycine chelate competes for the nonheme-iron absorption pathway. American Journal of Clinical Nutrition 76, 577–581.Google Scholar
Qiu, A, Jansen, M, Sakaris, A, Min, SH, Chattopadhyay, S, Tsai, E, Sandoval, C, Zhao, R, Akabas, MH, Goldman, ID 2006. Identification of an intestinal folate transporter and the molecular basis for hereditary folate malabsorption. Cell 127, 917–928.Google Scholar
Quintero, GA, González, RG, Sánchez, MJ 2008. Bioavailability of heme iron of biscuit filling using piglets as an animal model for human. International Journal of Biological Sciences 4, 58–62.Google Scholar
Roberts, SK, Henderson, RW, Young, GP 1993. Modulation of uptake of heme by rat small intestinal mucosa in iron deficiency. American Journal of Physiology 265, G712–G718.Google ScholarPubMed
Sanchez, M, Galy, B, Muckenthaler, MU, Hentze, MW 2007. Iron-regulatory proteins limit hypoxia-inducible factor-2alpha expression in iron deficiency. Nature Structural & Molecular Biology 14, 420–426.Google Scholar
Simpson, RJ, Raja, KB, Peters, TJ 1986. Fe2+ uptake by mouse intestinal mucosa in vivo and by isolated intestinal brush-border membrane vesicles. Biochimica et Biophysica Acta 860, 229–235.Google Scholar
Smith, SR, Cooperman, S, Lavaute, T, Tresser, N, Ghosh, M, Meyron-Holtz, E, Land, W, Ollivierre, H, Jortner, B, Switzer, R 3rd, Messing, A, Rouault, TA 2004. Severity of neurodegeneration correlates with compromise of iron metabolism in mice with iron regulatory protein deficiencies. Annals of The New York Academy of Sciences 1012, 65–83.Google Scholar
Srigiridhar, K, Nair, M 2000. Supplementation with α-tocopherol or a combination of α-tocopherol and ascorbic acid protects the gastrointestinal tract of iron-deficient rats against iron-induced oxidative damage during iron repletion. British Journal of Nutrition 84, 165–173.Google Scholar
South, PK, Lei, S, Miller, D 2000. Meat enhances nonheme iron absorption in pigs. Nutrition Research 20, 1749–1759.Google Scholar
Swindle, M, Smith, AC, Laberlaid, K, Duncan, ML 1994. Swine in biomedical research: management and models. In Farm animals in biological research. Part one (ed. M Swindle, AC Smith, K Laberlaid and ML Duncan), pp. 1–8. ILAR news, Washington.Google Scholar
Tako, E, Rutzke, MA, Glahn, RP 2010. Using the domestic chicken (Gallus gallus) as an in vivo model for iron bioavailability. Poultry Science 89, 514–521.Google Scholar
Theurl, I, Aigner, E, Theurl, M, Nairz, M, Seifert, M, Schroll, A, Sonnweber, T, Eberwein, L, Witcher, DR, Murphy, AT, Wroblewski, VJ, Wurz, E, Datz, C, Weiss, G 2008. Regulation of iron homeostasis in anemia of chronic disease and iron deficiency. Blood 113, 5277–5286.Google Scholar
Troost, FJ, Saris, HW, Haenen, GR, Bast, A, Brummer, RJ 2003. New method to study oxidative damage and antioxidants in the human small bowel: effect on iron application. American Journal of Physiology – Gastrointestinal and Liver Physiology 285, G354–G359.CrossRefGoogle ScholarPubMed
Vaghefi, N, Nedjaoum, F, Guillochon, D, Bureau, F, Arhan, P, Bougle, D 2005. Iron absorption from concentrated hemoglobin hydrolysate by rat. Journal of Nutritional Biochemistry 16, 347–352.Google Scholar
Walczyk, T, Tuntipopipat, S, Zeder, C, Sirichakwal, P, Wasantwisut, E, Hurrell, RF 2005. Iron absorption by human subjects from different iron fortification compounds added to Thai fish sauce. European Journal of Clinical Nutrition 59, 668–674.Google Scholar
West, AR, Oates, PS 2008. Mechanisms of heme iron absorption: current questions and controversies. World Journal of Gastroenterology 14, 4101–4110.Google Scholar
Zhu, L, Glahn, RP, Yeung, CK, Miller, DD 2006. Iron uptake by Caco-2 cells from NaFeEDTA and FeSO4: effects of ascorbic acid, pH, and a Fe(II) chelating agent. Journal of Agriculture and Food Chemistry 54, 7924–7928.Google Scholar
Zinn, KR, Chaudhuri, TR, Mountz, JM, van den Berg, GJ, Gordon, DT, Johanning, GL 1999. 59Fe is retained from an elemental 59Fe powder supplement without effects on 65Zinc, 47Calcium and 67Copper in young pigs. Journal of Nutrition 129, 181–187.Google Scholar