



@article {2568,
	title = {Histological effects of iron accumulation on mice liver and spleen after administration of a metallic solution},
	journal = {BiomaterialsBiomaterials},
	volume = {20},
	year = {1999},
	note = {Cited By (since 1996): 15Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/S0142-9612(99)00124-6PubMed ID: 10555088Language of Original Document: EnglishCorrespondence Address: Pereira, M.C.; Departamento de Engenharia Quimica, Faculdade de Engenharia, Universidade do Porto, Rua dos Bragas, 4050-123 Porto, Portugal; email: mcsp@fe.up.ptChemicals/CAS: Iron, 7439-89-6; SolutionsReferences: Rheinold, J.B., Trace elements - A selective survey (1975) Clin Chem, 21, p. 476;Crichton, R.R., Ward, R.J., Structure and molecular biology of iron binding-proteins and the regulation of free iron pools (1992) Iron and Human Diseases, pp. 23-75. , R.B. Lauffer. Boca Raton: CRC Press Inc; 
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Britton, S.R., Ramm, G.A., Olynyk, J., Singh, R., O{\textquoteright}Neill, R., Bacon, B.R., Pathophysiology of iron toxicity (1994) Progress in Iron Research, pp. 239-253. , C. et al. Hershko. New York: Plenum Press
},
	month = {1999///},
	pages = {2193 - 2198},
	publisher = {Elsevier Science Ltd},
	abstract = {Special attention has been focused on the toxicity of some metallic species released from implanted materials, which accumulate in vital organs over long periods of time. A set of experiments with mice was designed to investigate the individual effects caused by iron in the liver and spleen. Histological features of these organs were evaluated and slight morphological changes were observed during the treatment time suggesting a negative correlation with the duration of the iron treatment. In addition, to associate the histological changes in the organs with iron accumulation an electrochemical method, adsorptive stripping voltammetry, was chosen to quantify the iron levels in these mentioned organs. The accuracy of the proposed method was checked by atomic absorption spectrometry. Both organs showed elevated concentrations of iron, nearly twofold, 7 days after iron administration compared to control organs.Special attention has been focused on the toxicity of some metallic species released from implanted materials, which accumulate in vital organs over long periods of time. A set of experiments with mice was designed to investigate the individual effects caused by iron in the liver and spleen. Histological features of these organs were evaluated and slight morphological changes were observed during the treatment time suggesting a negative correlation with the duration of the iron treatment. In addition, to associate the histological changes in the organs with iron accumulation an electrochemical method, adsorptive stripping voltammetry, was chosen to quantify the iron levels in these mentioned organs. The accuracy of the proposed method was checked by atomic absorption spectrometry. Both organs showed elevated concentrations of iron, nearly twofold, 7 days after iron administration compared to control organs.
},
	keywords = {Absorption spectroscopy, accuracy, Adsorption, Adsorptive stripping voltametry, Adsorptive stripping voltammetry, animal experiment, animal model, animal tissue, Animalia, Animals, article, atomic absorption spectrometry, Atomic spectroscopy, bioaccumulation, biomaterial, controlled study, Corrosion, electrochemistry, histology, histopathology, Implants (surgical), Injections, Subcutaneous, Iron, iron metabolism, Kinetics, Liver, liver toxicity, male, metal implantation, Mice, morphology, mouse, nonhuman, Organ Size, potentiometry, priority journal, quantitative assay, Solutions, Spleen, Stainless steel, tissue distribution, toxicity},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0032855084\&partnerID=40\&md5=2e7274d8d0372ff55881c2d9de94c54b},
	author = {Pereira, M.C. and Pereira, M.L. and Sousa, J.P.}
}
@article {2577,
	title = {In vitro osteoblastic differentiation of human bone marrow cells in the presence of metal ions},
	journal = {Journal of Biomedical Materials ResearchJ. Biomed. Mater. Res.},
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	year = {1999},
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},
	month = {1999///},
	pages = {176 - 190},
	publisher = {John Wiley \& Sons Inc},
	abstract = {For periods up to 21 days human bone marrow was cultured in control conditions that favor the proliferation and differentiation of osteoblastic cells. The effect of AISI 316L corrosion products and the corresponding major separate metal ions (Fe, Cr, and Ni) were studied in three different phases of the culture period in order to investigate the effects of metal ions in cell populations representative of osteoblastic cells in different stages of differentiation. Toxicity consequences of the presence of metal ions in bone marrow cultures were evaluated by biochemical parameters (enzymatic reduction of MTT, alkaline phosphatase activity, and total protein content), histochemical assays (identification of ALP-positive cells and Ca and phosphates deposits), and observation of the cultures by light and scanning electron microscopy. Culture media were analyzed for total and ionized Ca and P and also for metal ions (Fe, Cr, and Ni). The presence of AISI 316L corrosion products and Ni salt in bone marrow cultures during the first and second weeks of culture significantly disturbs the normal behavior of these cultures, interfering in the lag phase and exponential phase of cell growth and ALP expression. However, the presence of these species during the third week of culture, when expression of osteoblastic functions occurs (mineralization process), did not result in any detectable effect. Fe salt also disturbs the behavior of bone marrow cell cultures when present during the lag phase and proliferation phase, and a somewhat compromised response between the normal pattern (control cultures) and intense inhibition (AISI 316L corrosion products and Ni salt-added cultures) was observed. Fe did not affect the progression of the mineralization phase. Osteogenic cultures exposed to Cr salt (Cr                        3+) presented a pattern similar to the controls, indicating that this element does not interfere, in the concentration studied, in the osteoblastic differentiation of bone marrow cells. Quantification of metal ions in the culture media showed that Cr (originated from AISI 316L corrosion products but from not Cr
                        3+ salt) and Ni (originated from AISI 316L corrosion products and Ni salt) appear to be retained by the bone marrow cultures.
For periods up to 21 days human bone marrow was cultured in control conditions that favor the proliferation and differentiation of osteoblastic cells. The effect of AISI 316L corrosion products and the corresponding major separate metal ions (Fe, Cr, and Ni) were studied in three different phases of the culture period in order to investigate the effects of metal ions in cell populations representative of osteoblastic cells in different stages of differentiation. Toxicity consequences of the presence of metal ions in bone marrow cultures were evaluated by biochemical parameters (enzymatic reduction of MTT, alkaline phosphatase activity, and total protein content), histochemical assays (identification of ALP-positive cells and Ca and phosphates deposits), and observation of the cultures by light and scanning electron microscopy. Culture media were analyzed for total and ionized Ca and P and also for metal ions (Fe, Cr, and Ni). The presence of AISI 316L corrosion products and Ni salt in bone marrow cultures during the first and second weeks of culture significantly disturbs the normal behavior of these cultures, interfering in the lag phase and exponential phase of cell growth and ALP expression. However, the presence of these species during the third week of culture, when expression of osteoblastic functions occurs (mineralization process), did not result in any detectable effect. Fe salt also disturbs the behavior of bone marrow cell cultures when present during the lag phase and proliferation phase, and a somewhat compromised response between the normal pattern (control cultures) and intense inhibition (AISI 316L corrosion products and Ni salt-added cultures) was observed. Fe did not affect the progression of the mineralization phase. Osteogenic cultures exposed to Cr salt (Cr
                        3+) presented a pattern similar to the controls, indicating that this element does not interfere, in the concentration studied, in the osteoblastic differentiation of bone marrow cells. Quantification of metal ions in the culture media showed that Cr (originated from AISI 316L corrosion products but from not Cr
                        3+ salt) and Ni (originated from AISI 316L corrosion products and Ni salt) appear to be retained by the bone marrow cultures.
},
	keywords = {alkaline phosphatase, Alloys, article, Batch cell culture, Biocompatible Materials, biotechnology, Bone, Bone cells, bone marrow cell, Bone Marrow Cells, bone mineralization, bone prosthesis, Calcium, Cell culture, cell differentiation, Cell Division, cell population, Cells, Cultured, Chromium, controlled study, Corrosion, Corrosion products, Cytotoxicity, Differentiation, human, human cell, Human osteoblast-like cells, Humans, Implants (surgical), Ions, Iron, joint prosthesis, materials testing, metal ion, Metal ions, Metals, Nickel, osteoblast, Osteoblasts, phosphorus, Prostheses and Implants, Proteins, Stainless steel, Steel corrosion, Tetrazolium Salts, Thiazoles},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0033080559\&partnerID=40\&md5=721fda55da342bbc327b233d73789014},
	author = {Morais, S. and Dias, N. and Sousa, J.P. and Fernandes, M.H. and Carvalho, G.S.}
}
@article {2570,
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	year = {1999},
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},
	month = {1999///},
	pages = {275 - 280},
	abstract = {To study the accumulation and the histological effects in mice organs caused by hexavalent chromium, one of the corrosion products released from AISI 316L stainless steel implants, mice groups were subcutaneously injected with a metallic solution of chromium during a certain period of time. Similar injections were made with HBSS (Hank{\textquoteright}s Balanced Salt Solution) in other groups to be used as controls. The levels of chromium found in the liver, kidney and spleen of the control and the treated animals were obtained by atomic absorption spectrometry (AAS) and were compared to those obtained by AdSV (adsorptive stripping voltammetry) to test the accuracy of the results. During the experimental period, the liver and spleen showed a progressive and significant accumulation of chromium whereas in the kidney the significant accumulation found after the first week practically remained unchanged during the four weeks. Apparently, the histological analysis of these tissues did not evidence any relevant morphological alteration induced by the chromium accumulations during the four weeks of treatment.},
	keywords = {animal experiment, animal tissue, Animalia, Animals, article, atomic absorption spectrometry, Biocompatible Materials, biomaterial, body weight, Chromium, controlled study, Corrosion, histology, histopathology, implant, Injections, Subcutaneous, Kidney, Liver, liver level, male, Mice, Mice organs, Mice, Inbred Strains, mouse, nonhuman, potentiometry, Prostheses and Implants, Spleen, Stainless steel, tissue distribution},
	isbn = {09660844 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0032840626\&partnerID=40\&md5=e81da6070ec455e49604d3773033d58d},
	author = {Pereira, M.D.C. and Pereira, M.D.L. and Sousa, J.P.}
}
@article {2588,
	title = {Adsorptive stripping measurements of iron accumulation in mice kidney using microelectrodes and histological features},
	journal = {Journal of Trace Elements in Medicine and BiologyJ. Trace Elem. Med. Biol.},
	volume = {12},
	year = {1998},
	note = {Cited By (since 1996): 5Export Date: 21 November 2012Source: ScopusCODEN: JTEBFPubMed ID: 9638613Language of Original Document: EnglishCorrespondence Address: Sousa, J.P.; INEB-Inst. de Engenharia Biomedica, Departamento de Engenharia Quimica, Faculdade Engenharia Univ. do Porto, Praca do Coronel Pacheco, 1, 4050 Porto, PortugalChemicals/CAS: Iron, 7439-89-6; Mercury, 7439-97-6References: Traisnel, M., Le Maguer, D., Hildebrand, H.F., Iost, A., Corrosion of surgical implants (1990) Clin. Mater., 5, pp. 309-318;Sutow, E.J., Pollack, S.R., The biocompatibility of certain stainless steels (1981) Biocompatibility of Clinical Implant Materials, 1, pp. 77-98. , Williams, D.F. eds CRC Press, Boca Raton, Florida; 
Ferguson Jr., A.B., Laing, P.G., Hodge, E.S., The ionization of metal implants in living tissues (1960) J. Bone Joint Surg., 42 A, pp. 77-90; 
Winter, G.D., Wear and corrosion products in tissues and the reactions they provoke (1976) Biocompatibility of Implant Materials, pp. 28-54. , Williams, D.F. eds London; 
Lux, F., Zeisler, R., Investigations of the corrosive deposition of components of metal implants and the behaviour of biological trace elements in metallosis tissue by means of instrumental multi-element activation analysis (1974) J. Radioanal. Chem., 19, pp. 289-297; 
Merritt, K., Brown, S.A., Sharkey, N.A., Blood distribution of nickel, cobalt, and chromium following intramuscular injection into hamsters (1984) J. Biomed. Mater. Res., 18, pp. 991-1004; 
Merritt, K., Brown, S.A., Biological effects of corrosion products from metals (1985) Corrosion and Degradation of Implant Materials: Second Symposium, pp. 195-207. , Fraker, A.C., Griffin, C.D. eds American Society for Testing and Materials, Philadelphia; 
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Morais, S., Carvalho, G.S., Sousa, J.P., Iron determination in osteoblast-like cell cultures medium by adsorptive cathodic stripping voltammetry with mercury microelectrodes (1997) Electroanalysis, 9; 
Wang, J., Mannino, S., Application of adsorptive stripping voltammetry to the speciation and determination of iron (III) and total iron in wines (1989) Analyst, 114, pp. 643-645; 
Smith, G.K., Black, J., Estimation of in vivo type 316L stainless steel corrosion rate from blood transport and organ accumulation data (1985) Corrosion and Degradation of Implant Materials: Second Symposium, pp. 223-247. , Fraker, A.C. and Griffin. Eds. American Society for Testing Materials; 
Pereira, M.C., Sousa, J.P., Determination of total iron in biological sample solutions with mercury microelectrodes (1996) Portugaliae Electrochimica Acta, 14, pp. 273-278; 
Pereira, M.C., Pereira, M.L., Sousa, J.P., Adsorptive stripping voltammetric measurements of chromium accumulation in mice organs using mercury microelectrodes (1997) Electroanalysis, 9, pp. 941-944
},
	month = {1998///},
	pages = {50 - 55},
	abstract = {The toxic effects caused by iron in kidney was performed on experimental studies with mice following administration of a metallic solution of this specie to simulate the iron corrosion products of a metallic implant. To quantify the total levels of iron present in this organ, an electrochemical method was chosen based on the application of square wave voltammetry using adsorptive collection of iron-catechol on a mercury film microelectrode (MFM). The optimal working conditions to produce a very stable and reproducible iron peak in the digested kidney samples were found to be pH=7.2 provided by 8.0 mmol/L PIPES buffer, catechol concentration of 3.0x10},
	keywords = {, biomaterials, Adsorptive stripping, animal experiment, animal model, animal tissue, Animals, article, atomic absorption spectrometry, biomaterial, controlled study, electrochemical analysis, electrochemistry, histology, Iron, iron absorption, Kidney, male, Mercury, metal implantation, Mice, microelectrode, Microelectrodes, mouse, nonhuman, potentiometry, priority journal, Toxicology},
	isbn = {0946672X (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0031746863\&partnerID=40\&md5=f85b47bb916eab32df5be2219498793f},
	author = {Pereira, M.C. and Pereira, M.L. and Sousa, J.P.}
}
@article {2584,
	title = {Chromium determination in osteoblast-like cell culture medium by catalytic cathodic stripping voltammetry with a mercury microelectrode},
	journal = {Journal of Trace Elements in Medicine and BiologyJ. Trace Elem. Med. Biol.},
	volume = {12},
	year = {1998},
	note = {Cited By (since 1996): 7Export Date: 21 November 2012Source: ScopusCODEN: JTEBFPubMed ID: 9760419Language of Original Document: EnglishCorrespondence Address: Morais, S.; Inst. Engenharia Biomedica (INEB), Praca Coronel Pacheo 1, 4050 Porto, PortugalChemicals/CAS: Acetates; Chromium, 7440-47-3; Culture Media; Mercury, 7439-97-6; NitratesReferences: Ferguson Jr., A.B., Laing, P.G., Hodge, E.S., The ionization of metal implants in living tissues (1960) J. Bone Joint Surg., 42 A, pp. 77-90;Ferguson Jr., A.B., Akahoshi, Y., Laing, P.G., Hodge, E.S., Trace metal ion concentration in the liver, kidney, spleen and lung of normal rabbits (1962) J Bone Joint Surg., 44 A, pp. 317-322; 
Ferguson Jr., A.B., Akahoshi, Y., Laing, P.G., Hodge, E.S., Characteristics of trace ions released from embedded metal implants in the rabbit (1962) J. Bone Joint Surg. 1962, 44 A, pp. 323-336; 
Lux, F., Ziesler, R., Investigations of the corrosive deposition of components of metal implants and of the behaviour of biological trace elements in metallosis tissue by means of instrumental multi-elemental activation analysis (1974) J. Radioanal. Chem., 19, pp. 289-297; 
Smith, G.K., Black, J., Estimation of in vivo type 316L stainless steel corrosion rate from blood transport and organ accumulation data (1985) Corrosion and Degradation of Implant Materials: Second Symposium, pp. 223-247. , ASTM STP 859 (Fraker, A. C. and Griffin, C. D. ,Eds.). American Society for Testing and Materials, Philadelphia; 
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Merritt, K., Brown, S.A., Biological effects of corrosion products from metals (1985) Corrosion and Degradation of Implant Materials: Second Symposium, pp. 195-207. , ASTM STP859 (Fraker, A. C. and Griffin, C. D., Eds.), American Society for Testing and Materials, Philadelphia; 
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Morais, S., Carvalho, G.S., Sousa, J.P., Determination of nickel in osteoblast-like cell culture medium by adsorptive cathodic stripping voltammetry with a mercury microelectrode (1997) Electroanalysis, 9, pp. 422-426; 
Morais, S., Carvalho, G.S., Sousa, J.P., Potentiometric determination of total and ionised calcium in osteoblast-like cell culture medium (1996) Electroanalysis, 8, pp. 1174-1178; 
Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., In vitro biomineralisation by osteoblast-like cells: Part I- Retardation of tissue mineralisation by metal salts (1998) Biomaterials, 19, pp. 13-21; 
Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., Bruijn, J.D., Van Blitterswijk, C.A., Decreased consumption of Ca and P during in vitro biomineralisation and biologically-induced deposition of Ni and Cr in the presence of stainless steel corrosion products (1998) J. Biomed. Mater. Res., , in press; 
Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., In vitro osteoblastic differentiation of human bone marrow cells in the presence of metal ions (1998) J. Biomed. Mater. Res., , in press; 
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},
	month = {1998///},
	pages = {101 - 108},
	abstract = {A catalytic cathodic stripping voltammetric procedure for the determination of total chromium in osteoblast-like cell culture medium using a mercury film microelectrode (MFM) was optimised. The method is based on the pre-concentration of the Cr(III)-diethylenetriaminepentaacetic acid (DTPA) complex by adsorption at the potential of -1.00 V (vs. Ag/AgCl) in the presence of 10 x 10},
	keywords = {Acetates, animal cell, Animalia, Animals, article, atomic absorption spectrometry, Bone Marrow Cells, catalysis, Cell culture, Cells, Cultured, Chromium, complex formation, controlled study, Culture Media, culture medium, drug determination, electrochemistry, Electrodes, Hydrogen-Ion Concentration, in vitro study, Kinetics, Mercury, microelectrode, Microelectrodes, Nitrates, nonhuman, osteoblast, Osteoblasts, potentiometry, priority journal, Rabbits, Reproducibility of Results, sensitivity and specificity, Stromal Cells, technique},
	isbn = {0946672X (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-7344255754\&partnerID=40\&md5=16bbb746d7dcdd04b95a01edcc764771},
	author = {Morais, S. and De Carvalho, G.S. and Sousa, J.P.}
}
@article {2579,
	title = {Decreased consumption of Ca and P during in vitro biomineralization and biologically induced deposition of Ni and Cr in presence of stainless steel corrosion products},
	journal = {Journal of Biomedical Materials ResearchJ. Biomed. Mater. Res.},
	volume = {42},
	year = {1998},
	note = {Cited By (since 1996): 22Export Date: 21 November 2012Source: ScopusCODEN: JBMRBdoi: 10.1002/(SICI)1097-4636(199811)42:2<199::AID-JBM5>3.0.CO;2-KPubMed ID: 9773816Language of Original Document: EnglishCorrespondence Address: Fernandes, M.H.; Faculdade de Medicina Dentaria, Universidade do Porto, Rua Manuel Pereira da Silva, 4200 Porto, PortugalChemicals/CAS: Biocompatible Materials; Calcium, 7440-70-2; Chromium, 7440-47-3; Nickel, 7440-02-0; Phosphorus, 7723-14-0; Steel, 12597-69-2References: Ferguson Jr., B., Akahoshi, Y., Laing, P.G., Hodge, E.S., Trace metal ion concentration in the liver, kidney, spleen and lung of normal rabbits (1962) J. Bone Joint Surg., 44 A, pp. 317-322;Ferguson Jr., A.B., Akahoshi, Y., Laing, P.G., Hodge, E.S., Characteristics of trace ions released from embedded metal implants in the rabbit (1962) J. Bone Joint Surg., 44 A, pp. 323-336; 
Ferguson Jr., A.B., Laing, P.G., Hodge, E.S., The ionization of metal implants in living tissues (1960) J. Bone Joint Surg., 42 A, pp. 77-90; 
Lux, F., Ziesler, R., Investigations of the corrosive deposition of components of metal implants and of the behavior of biological trace elements in metallosis tissue by means of instrumental multi-elemental activation analysis (1974) J. Radioanal. Chem., 19, pp. 289-297; 
Smith, G.K., Black, J., Estimation of in vivo type 316L stainless steel corrosion rate from blood transport and organ accumulation data (1985) Corrosion and Degradation of Implant Materials: Second Symposium, ASTM STP 859, pp. 223-247. , A. C. Fraker and C. D. Griffin (eds.), American Society for Testing and Materials, Philadelphia, Pennsylvania; 
Merritt, K., Brown, S.A., Biological effects of corrosion products from metals (1985) Corrosion and Degradation of Implant Materials: Second Symposium, ASTM STP 859, pp. 195-207. , A. C. Fraker and C. D. Griffin, (eds.), American Society for Testing and Materials, Philadelphia, Pennsylvania; 
Smith, G.K., Black, J., Models for systemic effects of corrosion products (1977) Retrieval and Analysis of Orthopedics Implants, pp. 23-28. , A. Weinstein D. Gibbons, S. A. Brown, and A. W. Ruff (eds.), NBS Special Publication 472, U.S. Government Printing Office, Washington, D.C; 
Deutman, R., Mulder, T.J., Brian, R., Nater, J.P., Metal sensitivity before and after total hip arthroplasty (1977) J. Bone Joint Surg., 59 A, pp. 862-865; 
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Pereira, M.L., Abreu, A.M., Sousa, J.P., Carvalho, G.S., Chromium accumulation and ultrastructural changes in the mouse liver caused by stainless steel corrosion products (1995) J. Mater. Sci. Mater. Med., 6, pp. 523-527; 
Tracana, R.B., Pereira, M.L., Abreu, A.M., Sousa, J.P., Carvalho, G.S., Stainless steel corrosion products cause alterations on mouse spleen cellular populations (1995) J. Mater. Sci. Mater. Med., 6, pp. 56-61; 
Bravo, I., Sousa, S.R., Carvalho, G.S., Barbosa, M., Metals electrochemically dissolved in human serum inhibit T-lymphocyte function (1992) Advances in Biomaterials, pp. 7-11. , P. J. Doherty et al. (eds.), Amsterdam, Elsevier; 
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Betts, F., Wright, T., Salvati, E.A., Boskey, A., Mansal, M., Cobalt alloy metal debris in periarticular tissues from total hip revision arthoplasties (1992) Clin. Orthop., 278, pp. 75-82; 
Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., In vitro biomineralization by osteoblast-like cells: Part 1-Retardation of tissue mineralization by metal salts (1998) Biomaterials, 19, pp. 13-21; 
Morais, S., Carvalho, G.S., Sousa, J.P., Potentiometric determination of total and ionized calcium in osteoblast-like cell culture medium (1996) Electroanalysis, 8, pp. 1174-1178; 
Morais, S., Carvalho, G.S., Sousa, J.P., Iron determination in osteoblast-like cell culture medium by adsorptive cathodic stripping voltammetry with a mercury microelectrode (1997) Electroanalysis, 9, pp. 791-795; 
Morais, S., Carvalho, G.S., Sousa, J.P., Electrochemical characterization of iron species in osteoblast-like cell culture medium (1995) Port. Electrochim. Acta, 13, pp. 483-487; 
Morais, S., Carvalho, G.S., Sousa, J.P., Determination of nickel in osteoblast-like cell culture medium by adsorptive cathodic stripping voltammetry with mercury microelectrodes (1997) Electroanalysis, 9, pp. 422-426; 
Morais, S., Carvalho, G.S., Sousa, J.P., Optimization study for nickel quantification in cell culture medium using mercury film microelectrodes (1996) Port. Electrochim. Acta, 14, pp. 267-272; 
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Aronow, M.A., Gerstenfeld, L.C., Owen, T.A., Tassinari, M.S., Stein, G.S., Lian, J.B., Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells (1990) J. Cell Physiol., 143, pp. 213-221; 
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Cheng, S.L., Yang, J.W., Rifas, L., Zhang, S.F., Avioli, L.V., Differentiation of human bone marrow osteogenic stromal cells in vitro. Induction of the osteoblast phenotype by dexamethasone (1994) Endocrinology, 134, pp. 277-286; 
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Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., De Bruijn, J.D., Van Blitterswijk, C.A., Effects of AISI 316L corrosion products in in vitro bone formation (1998) Biomaterials, 19, pp. 999-1007; 
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},
	month = {1998///},
	pages = {199 - 212},
	publisher = {John Wiley \& Sons Inc},
	abstract = {The purpose of this study was to investigate the effects of 316L stainless steel (SS) corrosion products on the in vitro biomineralization process, because tissue necrosis, bone loss, impaired bone mineralization, and loosening of orthopedic implants are associated with ions and debris resulting from biodegradation. Rat bone marrow cells were cultured in experimental conditions that favored the proliferation and differentiation of osteoblastic cells and were exposed to SS corrosion products obtained by electrochemical means for periods ranging from 1 to 21 days. Quantification of total and ionized Ca and P, as well as Fe, Cr, and Ni, ions in the culture media of control and metal added cultures during the incubation period was performed to study the influence of corrosion products on the Ca and P consumption that occurs during the mineralization process. Control cultures and metal effects on cultures were evaluated concerning DNA content, enzymatic reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity. Histochemical detection of ALP, Ca, and phosphate deposition, and examination of the cultures by scanning and transmission electron microscopy (SEM and TEM) were also performed. The presence of SS corrosion products resulted in impairment of the normal behavior of rat bone marrow cultures. Levels of Cr and Ni in the medium of cultures exposed to 316L SS corrosion products decreased throughout the incubation period, suggesting a regular deposition of these species; these results were supported by TEM observation of the cultures. Cultures exposed to the corrosion products presented lower DNA content, MTT reduction, and ALP activity and failed to form mineralized areas. These cultures showed negative staining on histochemical reactions for the identification of calcium and phosphate deposition and SEM and TEM examination did not show mineral globular structures or mineralization foci, respectively, which is characteristic of cultures grown in control conditions. These results suggest that metal ions associated with 316L SS are toxic to osteogenic cells, affecting their proliferation and differentiation.The purpose of this study was to investigate the effects of 316L stainless steel (SS) corrosion products on the in vitro biomineralization process, because tissue necrosis, bone loss, impaired bone mineralization, and loosening of orthopedic implants are associated with ions and debris resulting from biodegradation. Rat bone marrow cells were cultured in experimental conditions that favored the proliferation and differentiation of osteoblastic cells and were exposed to SS corrosion products obtained by electrochemical means for periods ranging from 1 to 21 days. Quantification of total and ionized Ca and P, as well as Fe, Cr, and Ni, ions in the culture media of control and metal added cultures during the incubation period was performed to study the influence of corrosion products on the Ca and P consumption that occurs during the mineralization process. Control cultures and metal effects on cultures were evaluated concerning DNA content, enzymatic reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity. Histochemical detection of ALP, Ca, and phosphate deposition, and examination of the cultures by scanning and transmission electron microscopy (SEM and TEM) were also performed. The presence of SS corrosion products resulted in impairment of the normal behavior of rat bone marrow cultures. Levels of Cr and Ni in the medium of cultures exposed to 316L SS corrosion products decreased throughout the incubation period, suggesting a regular deposition of these species; these results were supported by TEM observation of the cultures. Cultures exposed to the corrosion products presented lower DNA content, MTT reduction, and ALP activity and failed to form mineralized areas. These cultures showed negative staining on histochemical reactions for the identification of calcium and phosphate deposition and SEM and TEM examination did not show mineral globular structures or mineralization foci, respectively, which is characteristic of cultures grown in control conditions. These results suggest that metal ions associated with 316L SS are toxic to osteogenic cells, affecting their proliferation and differentiation.
},
	keywords = {3 (4,5 dimethyl 2 thiazolyl) 2,5 diphenyltetrazolium bromide, alkaline phosphatase, animal cell, Animal cell culture, Animals, article, Biocompatible Materials, Biodegradation, biomineralization, Bioprosthesis, Bone, Bone and Bones, bone atrophy, bone marrow, Calcification, Physiologic, Calcium, calcium ion, Catalyst activity, Cells, Cultured, Chromium, controlled study, Corrosion, Corrosion products, Enzymes, Implants (surgical), metal implantation, mineralization, Nickel, nonhuman, osteoblast, Osteoblast-like cells, phosphate, phosphorus, prosthesis loosening, rat, Rats, Scanning electron microscopy, Stainless steel, Stainless steel corrosion products, Steel, Steel corrosion, tissue necrosis, transmission electron microscopy},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0032210768\&partnerID=40\&md5=5275a87eebeeda6fdce8b3382dc3eaa5},
	author = {Morais, S. and Sousa, J.P. and Fernandes, M.H. and Carvalho, G.S. and De Bruijn, J.D. and Van Blitterswijk, C.A.}
}
@article {2585,
	title = {Effects of AISI 316L corrosion products in in vitro bone formation},
	journal = {BiomaterialsBiomaterials},
	volume = {19},
	year = {1998},
	note = {Cited By (since 1996): 38Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/S0142-9612(97)00234-2PubMed ID: 9692798Language of Original Document: EnglishCorrespondence Address: Morais, S.; Universidade do Porto, Porto, PortugalChemicals/CAS: Alkaline Phosphatase, EC 3.1.3.1; Calcium Phosphates; Calcium, 7440-70-2; Phosphorus, 7723-14-0; Stainless Steel, 12597-68-1References: Spector, M., Cease, C., Tong-Li, X., The local tissue response to biomaterials (1989) Crit Rev Biocompat, 5, pp. 269-295;Van Blitterswijk, C.A., Bakker, D., Hesseling, S.C., Koerten, H.K., Reactions of cells at implant surfaces (1991) Biomaterials, 12, pp. 187-193; 
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Pules, D.A., Huh, W.W., Acute toxicity of metal ions in cultures of osteogenic cells derived from bone marrow stromal cells (1995) J Appl Biomater, 6, pp. 109-116; 
Thompson, G.J., Pules, D.A., Effects of sublethal metal ion concentrations on osteogenic cells derived from bone marrow stromal cells (1995) J Appl Biomater, 6, pp. 249-258; 
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Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., In vitro biomineralisation by osteoblast-like cells: Part 1 - Retardation of tissue mineralisation by metal salts Biomaterials 1998; 
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Tracana, R.B., Pereira, M.L., Abreu, A.M., Sousa, J.P., Carvalho, G.S., Stainless steel corrosion products cause alterations on mouse spleen cellular populations (1995) J Mater Sci Mater Med, 6, pp. 56-61; 
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McGee-Russell, S.M., Histochemical methods for calcium (1958) J Histochem Cytochem, 6, pp. 22-42; 
Morais, S., Carvalho, G.S., Sousa, J.P., Potentiometric determination of total and ionized calcium in osteoblast-like cell culture medium (1996) Electroanalysis, 8, pp. 1174-1178; 
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De Bruijn, J.D., Cpat, K., De Groot, K., Van Blitterswijk, C.A., The ultrastructure of the one-hydroxyapatite interface in vitro (1992) J Biomed Mat Res, 26, pp. 1365-1382; 
Davies, J.E., Chernecky, R., Lowenberg, B., Shiga, A., Deposition and resorption of calcified matrix in vitro by rat marrow cells (1991) Cells and Mat, 1, pp. 3-15; 
Bellows, C.G., Aubin, J.E., Heersche, J.N.M., Physiological concentrations of glucocorticoids stimulte formation of bone nodules from isolated rat calvaria cells in vitro (1987) Endocrinology, 121, pp. 1985-1992; 
Aronow, M.A., Gerstenfeld, L.C., Owen, T.A., Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells (1990) J Cell Phys, 143, pp. 213-221; 
Gotoh, Y., Hiraiwa, K., Narajama, M., In vitro mineralization of osteoblastic cells derived from human bone (1990) Bone Min, 8, pp. 239-250; 
Cheng, S.L., Yang, J.W., Riffins, L., Differentiation of human bone marrow osteogenic stromal cells in vitro. Induction of the osteoblast phenotype by dexamethasone (1994) Endocrinology, 134, pp. 277-286; 
Gundle, R., Joyner, C.J., Triffit, J.T., Human bone tissue formation in diffusion chamber culture in vivo by bone derived cells and marrow stromal fibroblastic cells (1995) Bone, 16, pp. 597-601; 
Bellows, C.G., Heersche, J.N.M., Aubin, J.E., Inorganic phosphate added exogenously or released from β-glycerophosphate initiates mineralization of osteoid nodules in vitro (1992) Bone Min, 17, pp. 15-29; 
Boskey, A.L., Stiner, D., Doty, S.B., Binderman, I., Levoy, P., Studies of mineralization in tissue culture: Optimal conditions for cartilage calcification (1992) Bone Min, 16, pp. 11-36; 
Bellows, C.G., Aubin, J.E., Heersche, J.N.M., Initiation and progression of mineralization of bone nodules formed in vitro: The role of alkaline phosphatase and organic phosphate (1991) Bone Min, 14, pp. 27-40; 
Chung, C.H., Golub, E.E., Forbes, E., Mechanisms of action of β-glycerophosphate on bone cell mineralization (1992) Calcif Tissue Int, 51, pp. 305-311
},
	month = {1998///},
	pages = {999 - 1007},
	publisher = {Elsevier Sci Ltd},
	abstract = {Rat bone marrow cells were cultured in experimental conditions that favour the proliferation and differentiation of osteoblastic cells (i.e., 2.52 x 10},
	keywords = {Absorption spectroscopy, alkaline phosphatase, animal cell, Animal cell culture, Animals, article, ascorbic acid, atomic absorption spectrometry, Bioassay, Biomaterials, bone development, bone marrow cell, Bone Marrow Cells, Calcium, Calcium phosphates, Cell culture, Cell Death, Cells, Cultured, Corrosion, Corrosion products, culture medium, dexamethasone, electrochemical analysis, electrochemistry, Enzymes, glycerophosphate, histochemistry, Histocytochemistry, in vitro study, male, metal ion, Mineralisation, mineralization, nonhuman, ossification, osteoblast, Osteoblast-like cells, Osteoblasts, phosphate, Phosphate minerals, phosphorus, priority journal, rat, Rats, Rats, Wistar, Scanning electron microscopy, Slurries, Stainless steel, Stainless steel 316L, Stainless-steel corrosion products, Steel corrosion},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0032102774\&partnerID=40\&md5=1b60be114642b919f64414bb11673a19},
	author = {Morais, S. and Sousa, J.P. and Fernandes, M.H. and Carvalho, G.S. and De Bruijn, J.D. and Van Blitterswijk, C.A.}
}
@article {2587,
	title = {Evaluation of nickel toxicity on liver, spleen, and kidney of mice after administration of high-dose metal ion},
	journal = {Journal of Biomedical Materials ResearchJ. Biomed. Mater. Res.},
	volume = {40},
	year = {1998},
	note = {Cited By (since 1996): 27Export Date: 21 November 2012Source: ScopusCODEN: JBMRBdoi: 10.1002/(SICI)1097-4636(199804)40:1<40::AID-JBM5>3.0.CO;2-RPubMed ID: 9511097Language of Original Document: EnglishCorrespondence Address: Sousa, J.P.; Instituto de Engenharia Biomedica, Faculdade de Engenharia, Universidade do Porto, Praca Coronel Pacheco, 1, 4050 Porto, PortugalChemicals/CAS: Nickel, 7440-02-0References: Cigada, A., Rondelli, G., Vicentini, B., Brunella, F., Dallaspezia, G., Corrosion behavior of high nitrogen stainless steels for biomedical applications (1994) Compatibility of Biomedical Implants, pp. 185-195. , P. Kovacs and N. S. Istephanous (eds.), The Electrochemical Society, Inc., Pennington, New Jersey;Bumgardner, J.D., Lucas, L.C., An evaluation of the corrosion properties and cellular compatibility of nickel-based dental casting alloys (1994) Compatibility of Biomedical Implants, pp. 241-252. , P. Kovacs and N. S. Istephanous (eds.), The Electrochemical Society, Inc., Pennington, New Jersey; 
Williams, D.F., (1990) Encyclopedia of Medical and Dental Materials, , Pergamon Press, Oxford; 
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Tracana, R.B., Pereira, M.L., Abreu, A.M., Sousa, J.P., Carvalho, G.S., Stainless steel products cause alterations on mouse spleen cellular populations (1995) J. Mater. Sci. Mater. Med., 6, pp. 56-61; 
Tracana, R.B., Sousa, J.P., Carvalho, G.S., Mouse inflammatory response to stainless steel corrosion products (1994) J. Mater. Sci. Mater. Med., 5, pp. 596-600; 
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Panigutti, M.A., Merrit, K., Bruner, R.J., Kraay, M.J., Brown, S.A., Correlation of allergy, metal levels, implant alloy, and implant damage in patients undergoing revision joint arthroplastics (1992) Trans. Soc. Biomater., 15, p. 7; 
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},
	month = {1998///},
	pages = {40 - 47},
	publisher = {John Wiley \& Sons Inc},
	abstract = {The toxic effects caused by nickel (Ni) per si were explored by performing in vivo studies on mice following subcutaneous administration of a metallic solution of nickel at 1, 2, 3, and 4 weeks to evaluate the side effects of this metal ion when released from stainless steel implants. Other groups were similarly injected with HBSS and used as controls. Accumulation of Ni ions on liver, spleen, and kidney was assessed by an electrochemical method, adsorptive stripping voltammetry (AdSV) using microelectrodes, and atomic absorption spectrometry (AAS). Alterations of those organs induced by Ni ions were studied, showing that several histological changes had been induced. Chemical analysis and histological features indicate that Ni is partially accumulated in the study organs.The toxic effects caused by nickel (Ni) per si were explored by performing in vivo studies on mice following subcutaneous administration of a metallic solution of nickel at 1, 2, 3, and 4 weeks to evaluate the side effects of this metal ion when released from stainless steel implants. Other groups were similarly injected with HBSS and used as controls. Accumulation of Ni ions on liver, spleen, and kidney was assessed by an electrochemical method, adsorptive stripping voltammetry (AdSV) using microelectrodes, and atomic absorption spectrometry (AAS). Alterations of those organs induced by Ni ions were studied, showing that several histological changes had been induced. Chemical analysis and histological features indicate that Ni is partially accumulated in the study organs.
},
	keywords = {Absorption spectroscopy, Adsorptive stripping voltammetry, animal experiment, animal tissue, Animals, article, atomic absorption spectrometry, biomaterial, body weight, controlled study, Dose-Response Relationship, Drug, electrochemistry, Evaluation Studies, histology, Implants (surgical), Kidney, Liver, liver toxicity, Living systems studies, Mice, Microelectrodes, mouse, nephrotoxicity, Nickel, nonhuman, potentiometry, Spectrophotometry, Atomic, Spleen, spleen injury, Stainless steel, toxicity},
	isbn = {00219304 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0032053959\&partnerID=40\&md5=b1b91f83e841302830f88702c7146ae0},
	author = {Pereira, M.C. and Pereira, M.L. and Sousa, J.P.}
}
@article {2594,
	title = {In vitro biomineralization by osteoblast-like cells I. Retardation of tissue mineralization by metal salts},
	journal = {BiomaterialsBiomaterials},
	volume = {19},
	year = {1998},
	note = {Cited By (since 1996): 39Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/S0142-9612(97)00149-XPubMed ID: 9678845Language of Original Document: EnglishCorrespondence Address: Carvalho, G.S.; IEC, Universidade do Minho, Largo do Paco, 4719 Braga Codex, PortugalChemicals/CAS: Alkaline Phosphatase, EC 3.1.3.1; Calcium, 7440-70-2; Cations; Chromium, 7440-47-3; Iron, 7439-89-6; Metals; Nickel, 7440-02-0; Phosphates; Stainless Steel, 12597-68-1; Tetrazolium Salts; Thiazoles; thiazolyl blue, 298-93-1References: Evans, E.M., Freeman, M.A.R., Miller, A.J., Vernon-Roberts, B., Metal sensitivity as a cause of bone necrosis and loosening of the prosthesis in total joint replacement (1974) J Bone Jt Surg, 56 B, pp. 626-642;Deutman, R., Rulder, T.J., Brian, R., Nater, J.P., Metal sensitivity before and after total hip arthroplasty (1977) J Bone Jt Surg, 59 A, pp. 862-865; 
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},
	month = {1998///},
	pages = {13 - 21},
	publisher = {Elsevier Sci Ltd},
	abstract = {The cytocompatibility of stainless steel 316L (SS 316L) corrosion products was investigated with particular focus on the dose- and time-effect of electrochemically dissolved SS and the corresponding separate metal ions on osteogenic bone marrow derived cells. Type AISI 316L stainless steel (Fe 63.9\%, Cr 18.0\%, Ni 12.5\%, Mo 2.8\%, Si 1.2\%, Mn 1.6\% and C 0.025\%, weight for weight) was anodically dissolved in Hank{\textquoteright}s Balanced Salt Solution (HBSS) and diluted to the following concentrations: 500 μgml},
	keywords = {Absorption spectroscopy, alkaline phosphatase, animal cell, Animalia, Animals, article, ascorbic acid, atomic absorption spectrometry, balanced salt solution, Bioassay, Biocompatibility, biomaterial, Biomaterials, biomineralization, bone marrow cell, bone mineralization, Bovinae, Calcification, Physiologic, Calcium, calcium phosphate, Carbon, Cations, Cattle, Cell Division, cell proliferation, cell survival, cell viability, Cells, Cultured, Chromium, controlled study, Corrosion, dexamethasone, electrochemistry, enzyme activity, Enzyme kinetics, glycerol 2 phosphate, histochemistry, Iron, Kinetics, manganese, metal ion, Metal ions, Metals, molybdenum, Nickel, nonhuman, Oryctolagus cuniculus, osteoblast, Osteoblast cells, Osteoblast-like cells, Osteoblasts, Phosphates, priority journal, rabbit, Rabbits, serum, Silicon, Stainless steel, Steel corrosion, Tetrazolium Salts, Thiazoles, Tissue culture},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0031812308\&partnerID=40\&md5=946525340a0c386c984ce614d56ae081},
	author = {Morais, S. and Sousa, J.P. and Fernandes, M.H. and Carvalho, G.S.}
}
@article {2593,
	title = {In vitro biomineralization by osteoblast-like cells II. Characterization of cellular culture supernatants},
	journal = {BiomaterialsBiomaterials},
	volume = {19},
	year = {1998},
	note = {Cited By (since 1996): 7Export Date: 21 November 2012Source: ScopusCODEN: BIMADdoi: 10.1016/S0142-9612(97)00150-6PubMed ID: 9678846Language of Original Document: EnglishCorrespondence Address: Carvalho, G.S.; IEC, Universidade do Minho, Largo do Paco, 4719 Braga Codex, PortugalChemicals/CAS: Calcium, 7440-70-2; Chromium, 7440-47-3; Culture Media; Iron, 7439-89-6; Metals; Nickel, 7440-02-0; Phosphorus, 7723-14-0; Stainless Steel, 12597-68-1References: Pereira, M.L., Silva, A., Tracana, R., Carvalho, G.S., Toxic effects caused by stainless steel corrosion products on mouse seminiferous cells (1994) Cytobios, 77, pp. 73-80;Pereira, M.L., Abreu, A.M., Sousa, J.P., Carvalho, G.S., Chromium accumulation and ultrastructural changes in the mouse liver caused by stainless steel corrosion products (1995) J Mater Sci Mater Med, 6, pp. 523-527; 
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Morais, S., Sousa, J.P., Fernandes, M.H., Carvalho, G.S., In vitro biomineralisation by osteoblast like cells. I. Retardation of tissue mineralisation by metal salts (1998) Biomaterials, 19, pp. 13-21; 
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},
	month = {1998///},
	pages = {23 - 29},
	publisher = {Elsevier Sci Ltd},
	abstract = {The quantification of total calcium, phosphorus, iron, chromium and nickel in cell culture medium by electrochemical or spectroscopic means may require digestion of samples. Nevertheless, when pH adjustment is performed for values higher than about 6.5, the formation of two phases occurs: a white precipitate and a clear solution. Analysing both phases using microelectrodes, atomic absorption spectrometry (AAS), diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. X-ray dispersive (XRD) analysis, scanning electron microscopy (SEM) and energy dispersive spectroscopic (EDS) analysis, it was oberved that iron, chromium and nickel are not co-precipitating with the white solid phase. If quantification of calcium, phosphorus and magnesium is intended, a ten-fold dilution at least, must be performed to avoid most of these elements going into the precipitate. This knowledge is crucial if a mineralization study is going to be made.The quantification of total calcium, phosphorus, iron, chromium and nickel in cell culture medium by electrochemical or spectroscopic means may require digestion of samples. Nevertheless, when pH adjustment is performed for values higher than about 6.5, the formation of two phases occurs: a white precipitate and a clear solution. Analyzing both phases using microelectrodes, atomic absorption spectrometry (AAS), diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, X-ray dispersive (XRD) analysis, scanning electron microscopy (SEM) and energy dispersive spectroscopic (EDS) analysis, it was observed that iron, chromium and nickel are not co-precipitating with the white solid phase. If quantification of calcium, phosphorus and magnesium is intended, a ten-fold dilution at least, must be performed to avoid most of these elements going into the precipitate. This knowledge is crucial if a mineralization study is going to be made.
},
	keywords = {Absorption spectroscopy, Animals, article, atomic absorption spectrometry, Biomaterials, bone mineralization, Calcification, Physiologic, Calcium, Cell culture medium, Cells, Cultured, Chromium, Culture Media, culture medium, Diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, electrochemistry, Energy dispersive spectroscopic analysis, Fourier transform infrared spectroscopy, Hydrogen-Ion Concentration, infrared spectroscopy, Iron, Magnesium, Metals, microelectrode, Microelectrodes, Nickel, nonhuman, osteoblast, Osteoblast cells, Osteoblasts, pH effects, phosphorus, precipitation, Precipitation (chemical), priority journal, Rabbits, Scanning electron microscopy, Spectrophotometry, Atomic, Spectroscopic techniques, Spectroscopy, Stainless steel, supernatant, Supernatants, Tissue culture, X ray analysis, X ray dispersive analysis},
	isbn = {01429612 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0031841533\&partnerID=40\&md5=62f6b7869003505ce8adfdf96ca2a9f0},
	author = {Morais, S. and Carvalho, G.S. and Faria, J.L. and Gomes, H.T. and Sousa, J.P.}
}
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},
	month = {1997///},
	pages = {941 - 944},
	abstract = {Chromium is the metallic species of most concern with the use of stainless steel implants. To study the distribution of chromium in mice organs with time, several animals were subcutaneously injected with 2.70 {\texttimes} 10},
	keywords = {Adsorptive stripping voltammetry, Biomaterials, Chromium, Microelectrodes},
	isbn = {10400397 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0000379196\&partnerID=40\&md5=9521717895db74058cd9402ada6394b3},
	author = {Pereira, M.C. and Pereira, M.L. and Sousa, J.P.}
}
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},
	month = {1997///},
	pages = {791 - 795},
	abstract = {Mercury film microelectrodes were used to determine iron in osteoblast-like cell culture medium by performing adsorptive cathodic stripping voltammetry in the presence of catechol. The optimized analytical conditions are a catechol concentration of 5.00{\texttimes}10},
	keywords = {Cell culture medium, Iron, Mercury film microelectrode},
	isbn = {10400397 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0001767651\&partnerID=40\&md5=b6108f6d2f2d7bbe73818c78d82a74eb},
	author = {Morais, S. and Carvalho, G.S. and Sousa, J.P.}
}
@article {2608,
	title = {Nickel Determination in Osteoblast-Like Cell Culture Medium by Adsorptive Cathodic Stripping Voltammetry with a Mercury Microelectrode},
	journal = {ElectroanalysisElectroanalysis},
	volume = {9},
	year = {1997},
	note = {Cited By (since 1996): 16Export Date: 21 November 2012Source: ScopusCODEN: ELANELanguage of Original Document: EnglishCorrespondence Address: Sousa, J.P.; Inst. de Engenharia Biomedica, Depto. de Engenharia Qu{\'\i}mica, Universidade do Porto, Pra{\c c}a Coronel Pacheco, 1, 4050 Porto, PortugalReferences: Williams, D.F., (1981) Biomaterials and Biocompatibility: An Introduction, in Fundamental Aspects of Biocompatibility, 1. , CRC Press, Boca Raton, FL;Tsalev, D.L., Zaprianov, Z.K., (1983) Atomic Absorption Spectrometry in Occupational and Environmental Health Practice, 1. , CRC Press, Boca Raton, FL; 
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},
	month = {1997///},
	pages = {422 - 426},
	abstract = {The purpose of this study is to investigate the influence of nickel, which is an alloying element in commonly used metallic biomaterials, on the biomaterials mineralization process. An electrochemical method was developed to quantify this metal ion in osteoblast-like cell culture medium (OST) by performing adsorptive cathodic stripping voltammetry (CSV) with dimethylglyoxime (DMG) at a mercury film microelectrode (MFM). The optimized analytical conditions and the square-wave CSV parameters for the analysis are: DMG concentration: 5.00 {\texttimes} 10},
	keywords = {Cell culture medium, Mercury film microelectrode, Nickel},
	isbn = {10400397 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0010434908\&partnerID=40\&md5=2547683b89d88a2385cea47a77bed5e8},
	author = {Morais, S. and Carvalho, G.S. and Sousa, J.P.}
}
@article {2613,
	title = {Nickel Quantification in Mice Organs by Adsorptive Cathodic Stripping Voltammetry Using Mercury Microelectrodes},
	journal = {ElectroanalysisElectroanalysis},
	volume = {9},
	year = {1997},
	note = {Cited By (since 1996): 8Export Date: 21 November 2012Source: ScopusCODEN: ELANELanguage of Original Document: EnglishCorrespondence Address: Sousa, J.P.; Inst. de Engenharia Biomedica, Depto. de Engenharia Qu{\'\i}mica, Faculdade Engenharia da Univ. do P., Pra{\c c}a do Coronel Pacheco, 1, P-4050 Porto, PortugalReferences: Mark H.B., Jr., (1964) J. Electroanal. Chem., 7, p. 276;Crow, D.R., Rose, M.E., (1979) Electrochim. Acta, 24, p. 41; 
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},
	month = {1997///},
	pages = {150 - 154},
	abstract = {Mercury film microelectrodes (MFMs) were used to quantify trace levels of nickel accumulated in mice organs samples after dosing the mice with a physiological solution containing 150 ppm of Ni ions over a period of time, by performing adsorptive cathodic stripping square wave voltammetry (AdSWV). The residues obtained after digestion of the biological organs with nitric acid which affects the electrochemical response of Ni were investigated by XRD and IR spectrometry and found to be mainly composed by p-nitrobenzoic acid. Electrochemical conditions for the quantification of Niwith MFMs in the biological solutions were optimized. The electrochemical data compared favorably with that obtained by atomic absorption spectrometry.},
	keywords = {Adsorptive stripping voltammetry, Mice organs, Microelectrodes, Nickel},
	isbn = {10400397 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0000122410\&partnerID=40\&md5=634a3532a71c048970c98a2652cca21b},
	author = {Pereira, M.C. and Faria, J.L. and Reis, M.L. and Sousa, J.P.}
}
@article {2626,
	title = {Inhibition and stimulation of enzymatic activities of human fibroblasts by corrosion products and metal salts},
	journal = {Journal of Materials Science: Materials in MedicineJ. MATER. SCI. MATER. MED.},
	volume = {7},
	year = {1996},
	note = {Cited By (since 1996): 9Export Date: 21 November 2012Source: ScopusCODEN: JSMMELanguage of Original Document: EnglishCorrespondence Address: Carvalho, G.S.; INEB-Instituto Engenharia Biomedica, Portugal Universidade do Minho, Praca Coronel Pacheco, 1, 4000 Porto, PortugalChemicals/CAS: 3 (4,5 dimethyl 2 thiazolyl) 2,5 diphenyltetrazolium bromide, 298-93-1; acid phosphatase, 9001-77-8, 9025-88-1; chromium, 16065-83-1, 7440-47-3; DNA, 9007-49-2; iron, 14093-02-8, 53858-86-9, 7439-89-6; neutral red, 553-24-2; nickel, 7440-02-0; stainless steel, 12597-68-1References: Pohler, O.E.M., (1983) Biomaterials in Reconstructive Surgery, p. 158. , C.V. Mosby, St. Louis;Galante, J.O., Lemons, J., Spector, M., Wilson Jr., P.D., Wright, T.M., (1991) J. Orthop. Res., 9, p. 760; 
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},
	month = {1996///},
	pages = {77 - 83},
	publisher = {Chapman \& Hall Ltd},
	abstract = {Fibroblasts from normal human skin were cultured for a period of 21 days in the absence or in the presence of metal ions. The effects of stainless steel (SS) corrosion products were compared to the effects of iron, chromium and nickel ions used either separately (Fe, Cr, or Ni solutions) or combined (Fe+Cr+Ni solution). At several periods of time (4, 7, 14 and 21 days) the cell cultures were analysed for the following parameters: (a) metal ion accumulation by atomic absorption spectrometry; (b) cell morphology and viability by the neutral red assay; (c) cell proliferation by DNA assessment, and enzyme activity by both (d) MTT reduction and (e) acid phosphatase activity. Results showed that SS-corrosion products and the corresponding metal ions combined at the same concentrations, Fe+Cr+Ni solution, had opposite effects on fibroblast cultures. In fact, SS-corrosion products caused no apparent effects on cell morphology nor on cell proliferation whereas Fe+Cr+Ni solution stimulated both neutral red uptake and cell proliferation. The enzymatic assays showed that SS-corrosion products caused inhibition of both MTT reduction and acid phosphatase activity in contrast to Fe+Cr+Ni solution which stimulated their activity. Furthermore, in all biological parameters studied, a strong association was observed between the effects of Fe+Cr+Ni solution and Cr alone, suggesting that Cr was the metal ion mostly involved in the stimulatory effects of the combined solution.Fibroblasts from normal human skin were cultured for 21 days in the absence, or presence, of metal ions. The effects of stainless steel (SS) corrosion products were compared to the effects of iron, chromium, and nickel ions used either separately or combined. At different periods of time, the cell cultures were analyzed for the following parameters: metal ion accumulation by atomic absorption spectrometry; cell morphology and viability by the neutral red assay; cell proliferation by DNA assessment; and enzyme activity by both MTT reduction and acid phosphatase activity. Results show that SS-corrosion products and the corresponding metal ions combined at the same concentrations, Fe+Cr+Ni solution, have opposite effects on fibroblast cultures.
},
	keywords = {3 (4,5 dimethyl 2 thiazolyl) 2,5 diphenyltetrazolium bromide, Absorption spectroscopy, Acid phosphatase, article, atomic absorption spectrometry, Atomic spectroscopy, Bioassay, Cell culture, Cell morphology, cell proliferation, cell structure, cell viability, Chromium, controlled study, Corrosion, Corrosion products, cytology, DNA, Enzymatic assays, Enzyme Activation, enzyme activity, Enzyme inhibition, fibroblast, human, human cell, Ions, Iron, metal, Metal ion accumulation, Metal salts, Metals, neutral red, Nickel, priority journal, Salts, Skin, skin fibroblast, Stainless steel, Steel corrosion},
	isbn = {09574530 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0030087165\&partnerID=40\&md5=a26cd67a43c1fef54a5df943014fd7c6},
	author = {Carvalho, G.S. and Castanheira, M. and Diogo, I. and Abreu, A.M. and Sousa, J.P. and Loon, J.A. and Van Blitterswijk, C.A.}
}
@article {2616,
	title = {Potentiometric Determination of Total and Ionized Calcium in Osteoblast-Like Cell Culture Medium},
	journal = {ElectroanalysisElectroanalysis},
	volume = {8},
	year = {1996},
	note = {Cited By (since 1996): 6Export Date: 21 November 2012Source: ScopusCODEN: ELANELanguage of Original Document: EnglishCorrespondence Address: Sousa, J.P.; Inst. de Engenharia Biom{\'e}dica, Faculdade de Engenharia, Universidade do Porto, Pra{\c c}a Coronel Pacheco, 1, P-4050 Porto, PortugalReferences: Nernst, W., (1886) Z. Phys. Chem., 2, p. 613;Nernst, W., (1889) Z. Phys. Chem., 4, p. 129; 
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Ringbom, A., (1963) Complexation in Analytical Chemistry, , Interscience, New York; 
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},
	month = {1996///},
	pages = {1174 - 1178},
	abstract = {The effect of the ionic strength and composition of solutions in the direct potentiometric determination of total and ionized calcium content using a calcium selective electrode was investigated. In the presence of a constant complexation buffer (CCB), the addition of sodium chloride and sodium bicarbonate causes a decrease in the limit of detection of the potentiometric response. When the procedure was tested in osteoblast-like cell culture medium no electrode interferences or method interferences were observed by the presence of proteins, i.e., the total and ionized calcium in this medium can be measured by potentiometric means.},
	keywords = {Cell culture medium, potentiometry, Total and ionized calcium},
	isbn = {10400397 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0000007614\&partnerID=40\&md5=e2532b414ac34fdd4bb8b71ac5cbabcd},
	author = {Morais, S. and Carvalho, G.S. and Sousa, J.P.}
}
@article {2636,
	title = {Stainless steel corrosion products cause alterations on mouse spleen cellular populations},
	journal = {Journal of Materials Science: Materials in MedicineJ. MATER. SCI. MATER. MED.},
	volume = {6},
	year = {1995},
	note = {Cited By (since 1996): 19Export Date: 21 November 2012Source: ScopusCODEN: JSMMEdoi: 10.1007/BF00121249Language of Original Document: EnglishCorrespondence Address: Tracana, R.B.; Natl. Inst. Biomedical Engineering, INEB, Praca Coronel Pacheco 1, 4000 Porto, Portugal},
	month = {1995///},
	pages = {56 - 61},
	publisher = {Chapman \& Hall Ltd},
	abstract = {Stainless steel is a metallic biomaterial commonly used in orthopaedic surgery. In this study we looked at the effects of stainless steel corrosion products on spleen, in order to evaluate their potential immunotoxicological effects. For this purpose stainless steel, type AISI 316L, was electrochemically dissolved in a physiological salt solution. The final solution, containing 490 μg/ml Fe, 224 μg/ml Cr and 150 μg/ml Ni, was injected subcutaneously in mice. After several periods of time (4, 10 and 14 days) spleens were removed and analysed for: (a) their contents in Fe, Cr and Ni, (b) histological alterations, (c) enumeration of cellular populations. Results showed that stainless steel corrosion products accumulated in spleen, caused histological alterations, and induced changes in cellular populations. When compared to chromium and nickel, iron was the metal ion preferentially accumulated in spleen. The increase of the multinucleated giant cell population was accompanied by depletion of lymphocyte populations. This study indicates that stainless steel corrosion products can spread systemically and accumulate in spleen inducing histological and cellular alterations which may give rise to immunotoxicological consequences.Stainless steel is a metallic biomaterial commonly used in orthopaedic surgery. In this study we looked at the effects of stainless steel corrosion products on spleen, in order to evaluate their potential immunotoxicological effects. For this purpose stainless steel, type AISI 316L, was electrochemically dissolved in a physiological salt solution. The final solution, containing 490 μg/ml Fe, 224 μg/ml Cr and 150 μg/ml Ni, was injected subcutaneously in mice. After several periods of time (4, 10 and 14 days) spleens were removed and analysed for: (a) their contents in Fe, Cr and Ni, (b) histological alterations, (c) enumeration of cellular populations. Results showed that stainless steel corrosion products accumulated in spleen, caused histological alterations, and induced changes in cellular populations. When compared to chromium and nickel, iron was the metal ion preferentially accumulated in spleen. The increase of the multinucleated giant cell population was accompanied by depletion of lymphocyte populations. This study indicates that stainless steel corrosion products can spread systematically and accumulate in spleen inducing histological and cellular alterations which may give rise to immunotoxicological consequences.
},
	keywords = {animal cell, animal experiment, animal tissue, Animalia, article, Biocompatibility, biomaterial, cell population, Cells, Chromium, Chronoamperometry, controlled study, Corrosion, Corrosion products, Dissolution, electrochemistry, giant cell, histology, immunology, immunotoxicity, Immunotoxicological effects, Ions, Iron, Lymphocyte, lymphocyte subpopulation, male, mouse, Nickel, nonhuman, orthopedic surgery, priority journal, sodium chloride, Spleen, spleen cell, Stainless steel, Steel corrosion, subcutaneous drug administration, toxicity},
	isbn = {09574530 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0028883275\&partnerID=40\&md5=0d4737517419b9f7ccafa996ef5e1bb1},
	author = {Tracana, R.B. and Pereira, M.L. and Abreu, A.M. and Sousa, J.P. and Carvalho, G.S.}
}
@article {2638,
	title = {Mouse inflammatory response to stainless steel corrosion products},
	journal = {Journal of Materials Science: Materials in MedicineJ. MATER. SCI. MATER. MED.},
	volume = {5},
	year = {1994},
	note = {Cited By (since 1996): 15Export Date: 21 November 2012Source: ScopusCODEN: JSMMEdoi: 10.1007/BF00120337Language of Original Document: EnglishCorrespondence Address: Tracana, R.B.; Nat. Inst. Biomedical Engineering, Praca Coronel Pacheco 14000 Porto, Portugal},
	month = {1994///},
	pages = {596 - 600},
	publisher = {Chapman \& Hall Ltd},
	abstract = {Corrosion occurs regularly following long-term implantation of stainless steel. Little is known about the inflammatory and immunological potential of stainless steel corrosion products. AISI 316L stainless steel was anodically dissolved in a physiologically solution, HBSS, through a chronoamperometric process by imposing an external constant current of 0.5 mA. The solution, containing 245 μg of Fe, 112 μg of Cr, 75 μg of Ni and 13 μg of Mo, was injected in the peritoneal cavity of male C57BL/6 mice. Five animals were used per survival period of time: 4, 16, 24, and 48 h, 1, 2, 4, 8 and 16 weeks. Three control mice per survival period of time were injected with HBSS. For each assay, peritoneal samples were analysed not only for the total number of cells but also for the percentages of macrophages, lymphocytes and polymorphonuclear (PMN) cells, which were estimated by differential counting on Wright-stained cytocentrifuge preparations. Our follow-up study showed that stainless steel corrosion products induced an acute inflammatory response for a period of one week as demonstrated by the influx of PMN cells and macrophages. In contrast neither a chronic inflammation nor an immune response was observed indicating that the stainless steel solution caused a minor tissue response.A follow-up study of the changes to mouse peritoneal cell populations, succeeding intraperitoneal injection of stainless steel corrosion products, in order to assess the inflammatory response to such metals was presented. AISI stainless steel was anodically dissolved in a physiologically solution, Hank{\textquoteright}s balanced salt solution (HBSS), which was injected in the peritoneal cavity of male mice. Five animals were designated per survival time interval of 4, 16, 24, and 48 h, 1, 2, 4, 8 and 16 weeks. Three control mice per survival period of time were treated with HBSS. For each assay, peritoneal samples were tested for total number of cells and percentages of macrophages, lymphocytes and polymorphonuclear (PMN) cells. The follow-up study revealed that stainless steel corrosion products induced an acute inflammatory response for a duration of one week as shown by the influx of PMN cells and macrophages. In contrast, neither a chronic inflammation nor an immune response was noted suggesting that stainless steel solution caused a minor tissue response.
},
	keywords = {animal cell, animal experiment, Animalia, article, Assays, Cell Count, Cells, Chromium, Chronoamperometry, controlled study, Corrosion, Dissolution, follow up, immune response, immunology, Inflammation, Inflammatory response, Iron, Lymphocyte, Lymphocytes, macrophage, Macrophages, male, metal implantation, Mice (animal), molybdenum, mouse, Nickel, nonhuman, Peritoneal cavity, Polymorphonuclear, polymorphonuclear cell, priority journal, Stainless steel, time, Tissue, Wright stained cytocentrifuge},
	isbn = {09574530 (ISSN)},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-0028500732\&partnerID=40\&md5=f1f2116e247beed68534321b6ad1f423},
	author = {Tracana, R.B. and Sousa, J.P. and Carvalho, G.S.}
}
