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The toxicity of mineral fibers, whether they are natural or man made (MMMF), is usually evaluated in vivo using biopersistence tests in rodents. Development of an in vitro cellular model would be worthwhile in order to reduce, refine and finally replace animal models. For this purpose, we developed an in vitro assay using human monocytic cell line (U-937) to evaluate a new manufactured rock wool fiber (HDN) biodegradation. Experiments on earlier known mineral fibers asbestos (crocidolite) and glass wool fibers (CM44) were also performed. U-937 responded to HDN and CM44 only if they were activated. Among the different activators we used, Escherichia coli living cells as well as FS were the most efficient as evidenced by alterations of HDN and CM44 surface, detected by scanning electron microscopy, and by the measure of silicon released from the rock wool fibers. Asbestos fibers were not degraded when incubated in the presence of living bacteria. The MMMF modifications were function of the fiber composition, the time of exposure to activated cells and the concentration of activators. The pattern of MMMF degradation by our in vitro system was in accordance with those observed in an in vivo study, thus indicating that the fiber degradation by macrophage cells activated by E. coli living cells as well as FS is a valuable system to assess mineral fibers’ biopersistence. [ABSTRACT FROM AUTHOR]

A human monocytes cell line, U-937, incubated in the presence of filtered medium from Escherichia coli culture (FS) has been previously reported to degrade man made mineral fiber and it has been indicated as a good paradigm of in vivo fiber biopersistence evaluation (manuscript accepted for publication). In the present paper, a study is reported aimed to define the molecular modification occurring in the U-937 monocytes during in vitro fiber degradation. The induction of gene expression was investigated in U-937 exposed to rock wool fibers (HDN) in the presence of FS by transcriptome analysis using 20 K DNA microarrays and quantitative RT-PCR. The over-expression of genes related to mobility and cellular adhesion, oxidative stress, immune system stimulation, enzymes, and ions transport protein systems were identified. Among them NCF1 gene, the gene encoding a subunit of NADPH oxidase, over-expression was detected. As the product of this gene allows the formation of superoxide anion that could lead to oxidative stress, HDN fibers were exposed to hydrogen peroxide. Fiber degradation similar to those observed upon incubation with U-937 in the presence of FS was obtained thus suggesting that reactive oxygen species production may be responsible for fiber degradation by U-937 monocytes. [ABSTRACT FROM AUTHOR]

Evaluating the pathogenic potentials of man-made mineral fibers (MMMF) is an important task performed by the European Community. Noting that it has been proposed that the use of laboratory animals for scientific tests should be reduced or phased out, macrophages then become the cells of choice for conducting in vitro studies. We have evaluated the in vitro toxicity of six commercial stonewool fibers (A, B1, B2, C, D, and E) on U-937 cells. The physical interaction between U-937 cells and MMMF was observed using scanning electron microscopy, and the cytotoxicity was evaluated by studying cell viability using MTT assay and cell apoptosis with an ELISA detection kit. Scanning electron microscopy (SEM) analysis has shown that long fibers can be covered by several macrophages, and that a small fiber can be completely engulfed by one cell. With 50 μg/mL of MMMF, a decrease in cell viability appeared after seven days of incubation, whereas 200 μg/mL induced loss of viability and apoptosis after one day. Fiber D, comprising a high proportion of fibers >20 μm in length and a high concentration of MgO, induced the highest loss in viability and the highest rate of apoptosis compared to the other five fibers. Whether this toxic effect is related to either the physical characteristics of the fibers (such as length), or to the high concentration of magnesium is still to be determined. Because the results can be rapidly obtained, the proposed model is suitable for studying the toxicities of mineral components, even if the tested concentrations are far from the ones reached in the lung. [ABSTRACT FROM AUTHOR]

The common observation of Epstein-Barr virus (EBV) viremia in pediatric recipients of renal transplants and the occurrence of an EBV-related pulmonary leiomyoma prompted us to intensify the follow-up of EBV infections from 1995 to October 2000. Follow-up included serology and detection of viral DNA in blood using a semi-quantitative nested polymerase chain reaction (PCR) and later a real-time PCR with higher sensitivity. The aim of this study was the early detection of primary infections or reactivations. We obtained 250 samples from 32 patients. EBV DNA detection was consistently negative in 14 patients. There were 5 patients that were considered at risk for post-transplant lymphoproliferative disease, as they were EBV seronegative and were given a kidney from a positive donor. Of these, 4 had at least one episode of high-level EBV viremia. During these episodes, an absence of noticeable symptoms that could be related to EBV was noted for all but 1 patient. This child presented with severe neutropenia 1 month after grafting and, 28 months later, several nodules of pulmonary leiomyoma, which were found to be EBV related. Four episodes of high-level viremia were observed before the discovery of the leiomyoma. Viral DNA detection is important for the follow-up of such patients that are especially at risk of serious complications of EBV infections. [ABSTRACT FROM AUTHOR]