



@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; 
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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; 
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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., 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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},
	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},
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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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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; 
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. , Doherty PJ et al., editors. Amsterdam: Elsevier; 
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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 {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.}
}
