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1.
J Biomed Mater Res A ; 80(2): 276-82, 2007 Feb.
Article in English | MEDLINE | ID: mdl-16958052

ABSTRACT

Medical devices manufactured for implantation into humans must be free of any contamination with viable bacteria. However, remnants of dead bacteria and bacterial components alone may induce an inflammatory immune response. Pyrogen tests for such inflammatory contaminations are generally performed either by determining the content of lipopolysaccharide in rinsing solutions of batch samples by limulus amoebocyte lysate assay, by injecting the rinsing solutions into rabbits or by implanting batch samples into rabbits and measuring change of body temperature. In this study, we show that the in vitro pyrogen test (IPT), which measures the release of the inflammatory cytokine IL-1beta in fresh or cryopreserved human whole blood, can be used to assess the pyrogenic contamination of implantable medical devices. This test was used to check neurosurgical implants, namely aneurysm clips, as a proof of principle. Owing to the direct contact of the test material with the blood cells, this test does not require rinsing procedures, which have variable efficacy. The use of human blood ensures the detection of all substances that are pyrogenic for humans and reflects their relative potency. The safety of the products as delivered could be confirmed. The effects of sterilization and depyrogenization procedures on intentional pyrogenic contaminations of samples could be followed. This new application of the already internationally validated method promises to replace further rabbit pyrogen tests. It generates extremely sensitive results with an extended range of detectable pyrogenic contaminants.


Subject(s)
Equipment and Supplies/microbiology , Prostheses and Implants/microbiology , Pyrogens/analysis , Equipment and Supplies/standards , Humans , Interleukin-1beta/blood , Methods , Microbiological Techniques/methods , Prostheses and Implants/standards , Sterilization , Surgical Instruments/microbiology
2.
Gene ; 318: 83-9, 2003 Oct 30.
Article in English | MEDLINE | ID: mdl-14585501

ABSTRACT

It is widely recognized that metal compounds may modify gene expression. In this context, we have searched for genes whose expression may be affected by cadmium and platinum ions within the context of a cell culture system. Cadmium is well known for its carcinogenic potential while platinum is destined to become more and more interesting because of its increasing use in the automotive industries. By applying differential display to cultures of mouse fibroblast, we have identified two transcripts (acute lymphoblastic leukemia-1, All-1, and a novel gene named metal-responsive gene, MERE-1) that were responsive to platinum and cadmium ions. Moreover, further experiments with a panel of metal compounds have shown that MERE-1 was strongly induced also by La(NO3)2 and Cr(NO3)3, and to a lesser extent, by Na2CrO4 and (NH4)2TeCl6.


Subject(s)
Cadmium/pharmacology , Fibroblasts/drug effects , Gene Expression Regulation/drug effects , Platinum/pharmacology , Animals , DNA, Complementary/chemistry , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Fibroblasts/cytology , Fibroblasts/metabolism , Metals/pharmacology , Mice , Mice, Inbred BALB C , Molecular Sequence Data , NIH 3T3 Cells , RNA, Messenger/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
3.
Altern Lab Anim ; 30(2): 209-17, 2002.
Article in English | MEDLINE | ID: mdl-11971756

ABSTRACT

The results are reported of the second stage in a programme for a systematic in vitro study on the carcinogenic potential of metal compounds with Balb/3T3 clone A31-1-1 mouse fibroblasts. Nineteen metal compounds that exhibited a strong cytotoxic effect during a previous screening run with a 100 microM fixed dose were tested with a 72-hour exposure over a wide range of concentrations (from 0.1 microM to 1000 microM), to produce dose-effect curves to permit extrapolation of the 50% inhibition concentration (IC50) values for each metal compound. This allows the establishment of a suitable range of doses for individual metal species, for use in the subsequent Balb/3T3 assay based on a two-stage concurrent cytotoxicity and morphological transformation protocol. Another 13 metal compounds were also tested, to determine whether the Balb/3T3 cell transformation assay is really a valuable in vitro model in relation to the problem of metal speciation. Of the metal compounds assayed, 26 showed a dose related cytotoxic response with calculated IC50 values ranging from 0.25 microM (CH3HgCl) to 140 microM [(C5H5)2TiCl2], whereas six metal compounds, namely (NH4)6Mo7O24*4H2O, CH3AsO(OH)2, C2H6AsNaO2(3H2O, KBr, CrCl3*6H2O and (NH4)2[TiO(C2O4)2]*H2O, displayed no observable cytotoxicity or low cytotoxicity at all the doses tested. The determination of IC50 values permits a ranking of the cytotoxicity responses of metal compounds with the highest cytotoxicities. Dose-effect curves and IC50 values of different chemical forms of individual metal compounds of As, Br, Cr, Hg, Ir, Pt, Te, Ti and V (cationic/anionic inorganic or organometallic species) showed clearly how the chemical nature of the metal strongly influences the toxic response. This confirms that the Balb/3T3 cell line is a valuable in vitro model with respect to the problem of metal speciation. This is a fundamental aspect to be considered when incorporating the results from in vitro cell transformation assays of the carcinogenic potential of metal compounds into regulatory testing schemes. In this context, the choice of test metal species for the development and validation of such assays cannot disregard the possibility that humans will be exposed to specific chemical forms of individual metal compounds (different oxidation states, and inorganic or organometallic natures) that can profoundly affect their toxicity.


Subject(s)
3T3 Cells/drug effects , Carcinogenicity Tests , Carcinogens/toxicity , Metals/toxicity , Animal Testing Alternatives/methods , Animals , Dose-Response Relationship, Drug , Inhibitory Concentration 50 , Mice , Mice, Inbred BALB C , Predictive Value of Tests
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