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M C Jaurand

Publications and source records attributed to M C Jaurand.

At least 73 records · Page 4Linked to original sources

Growth of normal and neoplastic rat pleural mesothelial cells in the presence of conditioned medium from neoplastic mesothelial cells.

Several transformed cells have been demonstrated to secrete growth factors. We studied the effect of conditioned medium from neoplastic rat pleural mesothelial cells on normal and neoplastic mesothelial cell growth. The results showed that the concentrated conditioned medium stimulated neoplastic mesothelial cell growth but inhibited reversibly normal mesothelial cell growth.

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Pleural carcinogenic potency of mineral fibers (asbestos, attapulgite) and their cytotoxicity on cultured cells.

The carcinogenicity of several samples of mineral fibers was tested following injection of 20 mg in the pleural cavity of noninbred Sprague-Dawley rats. Three samples of chrysotile asbestos (mean length: 3.2, 2.1, and 1.2 micron) induced mesotheliomas at a rate of 48, 52, and 19%, respectively. The first sample was acid leached prior to intrapleural injection; in that group, the percentage of mesotheliomas was reduced to 25%. Treatment with amosite and crocidolite resulted in the occurrence of 57 and 56% of mesotheliomas. Acid-treatment of amphiboles did not significantly modify the percentage of mesotheliomas. When the Stanton's fiber dimensions were taken into consideration to correlate with mesothelioma incidence, the observed number of mesotheliomas in the chrysotile-treated animals was much lower than that expected, suggesting that other fiber parameters (chemistry, physicochemistry) play a role in the carcinogenicity. Attapulgite fibers (mean length: 0.77 micron) did not induce tumor, and the mean survival time was of the same order as that observed in the control groups. The injection of quartz resulted in no mesothelioma but did result in 6 malignant histiocytic lymphomas (17%) and 2 malignant schwannomas (6%). In vitro experiments did not show strong correlation between cytotoxicity and the carcinogenic potency of these minerals, but the qualitative cellular responses might give some indications on the fiber's potency. In addition, the in vitro effects of the fibers seem to be modulated by their size.

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The interactions between asbestos fibers and metaphase chromosomes of rat pleural mesothelial cells in culture. A scanning and transmission electron microscopic study.

Rat pleural mesothelial cells (PMCs) in culture at the exponential growing phase were exposed to 5 micrograms/ml of chrysotile (CH) or crocidolite (CR) asbestos fibers: the cells and their chromosomes were studied 48 hours thereafter by light, scanning, and transmission electron microscopy (LM, SEM, TEM). PMCs phagocytized both CH and CR. Mild vacuolar cytoplasmic changes by LM and a few small surface blebbings by SEM were present, mainly in cells treated with CH. Metaphase chromosomes were well separated and retained surface details by SEM in the control group. Chromosomes were frequently entangled with, adherent to, and severed or pierced by long and thin curvilinear CH with occasional chromatin fibers threading over the partly severed asbestos. Similar chromosomal changes were much less frequently found in CR-treated cells; TEM confirmed the same findings. CH and CR have different physicochemical properties and also appear to have direct, intricate, but different interactions with chromosomes, as well as the cytoplasm, of PMCs.

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Chromosomal changes induced by chrysotile fibres or benzo-3,4-pyrene in rat pleural mesothelial cells.

The induction of chromosomal aberrations in rat pleural mesothelial cells (RPMC) following in vitro treatment with chrysotile fibres has been demonstrated. The production of chromosomal aberrations was also observed after treatment of the cells with benzo-3,4-pyrene (BP). The yield of abnormal metaphases was dose-dependent and reached 58% at a BP dose of 2 micrograms/ml. Chrysotile fibres at 7 micrograms/ml induced 21% abnormal metaphases and the frequency decreased with further increases in fibre concentration. Their decline is possibly related to a lethal effect. Chrysotile-induced chromosomal aberrations were primarily of the chromatid type and included breaks and fragments. BP induced chromosome exchanges which were not seen following chrysotile treatment. Minutes and double minutes were detected in BP-treated RPMC and occasionally found after chrysotile application. These results confirm that chrysotile fibres are clastogenic for some cultured cells and demonstrate that the fibres induce chromosome damage in target RPMC.

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In vitro growth characteristics of rat mesothelioma cells in culture.

The study reports morphological growth characteristics and chromosome analysis of neoplastic rat pleural mesothelial cells (RPMC) isolated from a mesothelioma-bearing rat. The pleural mesothelioma was induced by intrapleural injection of chrysotile fibers. Neoplastic RPMC were cultured by the standard methods used for normal RPMC. Neoplastic RPMC cultures had a population doubling time of 19 hr versus 30 hr for the normal cells. Plating efficiency in liquid medium was almost 100%. Cultures of neoplastic RPMC were anchorage-independent since 70% of the seeded cells formed colonies after one week; after the second week, colony size was enhanced but colony recovery was not. The serum dependence of neoplastic cells was less than that of the normal cells. 79 out of 100 metaphase cells analyzed had 41 to 43 chromosomes, and the modal number was 42 (38%). A large metacentric chromosome was observed in 77 of the 100 neoplastic metaphase cells analyzed, but not in any normal metaphase cells. In nude mice, the neoplastic RPMC were tumorigenic.

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In vitro transformation of rat pleural mesothelial cells by chrysotile fibres and/or benzo[a]pyrene.

Rat pleural mesothelial cells (RPMC) were treated, in vitro, in a two-stage model of carcinogenesis. RPMC obtained from passage 12th were treated once with 1 microgram/ml of benzo-[a]pyrene (BP) and from passage 13th to 40th with 0.4 microgram/cm2 of chrysotile fibres (Chr). Transformation was determined by the observation of the colonies formed when the cells were plated at low density. Colonies were classified into four classes according to the level of lack in contact inhibition between cells (0 to III). BP as well as chrysotile fibres were potent in inducing a high proportion of type III colonies which were not observed in control series. In addition, there was no synergistic effect between BP and Chr. These results would indicate that chrysotile fibres do not act as a promoter on rat pleural mesothelial cells in culture, but induce morphologically transformed colonies.

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In vitro biodegradation of chrysotile fibres by alveolar macrophages and mesothelial cells in culture: comparison with a pH effect.

The modification of the chemistry of asbestos chrysotile fibres (Mg3(Si2O5)(OH)4) after their ingestion by cultured cells has been studied. Two types of cells involved in asbestos related pulmonary disease were used, rabbit alveolar macrophages (AM), recovered by bronchoalveolar lavage, and pleural mesothelial cells (PMC) obtained from the rat parietal pleura. Chemical characterisation of intracellular fibres was performed on unstained ultrathin sections by electron probe microanalysis. The results showed a progressive leaching of Mg, characterised by a time dependent decrease of Mg/Si. AM were more efficient than PMC at leaching intracellular chrysotile fibres since it took longer to obtain the same proportion of leached fibres with PMC than with AM. As in vitro Mg-leaching can be obtained by acid treatment, chrysotile fibres were incubated, either untreated or pretreated with cell membranes, at pH 4 or 7 for various times. The data show that the kinetic of leaching by AM was comparable with leaching at pH 4. The leaching by PMC was of the same order as leaching at pH 7. When membranes were adsorbed on to the fibres, a delayed leaching was observed. The results indicate that the solubilisation of chrysotile by AM could be an intraphagolysosomal event due to a pH effect. With PMC, however, it is not possible to draw this conclusion since nothing is known about the intracellular pH.

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Role of pleural mesothelial cells in the production of the submesothelial connective tissue matrix of lung.

The pleura is comprised of a single layer of mesothelial cells resting on a complex layer of connective tissue. The ability of mesothelial cells to produce the components of this connective tissue was investigated using cultured rat mesothelial cells. These cells produced several components of extracellular matrix, including 6.8 +/- 0.2 X 10(5) collagen pro-alpha-chains per cell per hour, which represented 3.09 +/- 0.05% of all proteins synthesized by these cells. Chemical and immunologic criteria were used to demonstrate that these collagen chains included those of collagen types I, III, and IV. In addition, these cells produced elastin, as well as the connective tissue glycoproteins laminin and fibronectin. Moreover, electron microscopic studies revealed that lung mesothelial cells were capable of organizing these components into complex structures that resembled components of the extracellular matrix (thick collagen fibers, the amorphous component of elastic fibers, and basement membranelike structures), and restricted the formation of these structures to the basal region below the cells in culture. Thus, pleural mesothelial cells are active sources of a variety of connective tissue macromolecules found beneath mesothelial cells in situ, and can assemble these components into structures resembling the pleural extracellular matrix.

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Early cellular and biochemical alveolar responses following intra-tracheal inoculation with low dose of asbestos and quartz.

The study of broncho-alveolar lavage harvested from rats intratracheally dusted with chrysotile (0.5 mg), leached chrysotile (0.5 mg), crocidolite (0.5 mg) and quartz (0.5 and 5 mg) indicated: 1. A cytotoxic lysis of alveolar macrophages in relation to phagocytosis of dust particles in the following decreasing order: quartz, crocidolite, chrysotile. 2. Regarding cell intensity and duration while asbestos, even leached chrysotile, gave merely an early and transient response. This cell recruitment concerned mostly PMN leukocytes and at a less extent alveolar macrophage. 3. Cell recruitment was associated with an increased protein and phospholipids alveolar content. The increase of proteins came probably mostly from an inflammatory serum exudation. However, an increase synthesis of complement C3 and phospholipids is not excluded.

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Biological effects of New Caledonia nickel ore samples on red blood cells, alveolar macrophages and pleural mesothelial cells.

To examine the toxicity of three nickeliferous ores (Noumea Mines, Société le Nickel, in vitro tests were carried out with mammalian cells: haemolysis of human red blood cells (RBC), release of enzymes by rabbit alveolar macrophages (AM) for studying inflammation, and study of rat pleural mesothelial cells (PMC) to determine the effect on cellular growth and genotoxic effects.

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Effect of chrysotile and crocidolite on the morphology and growth of rat pleural mesothelial cells.

The effects of the UICC asbestos fibers chrysotile A and crocidolite on the morphology and growth of rat pleural mesothelial cells were examined. The response was different according to the fiber used. Within 4 hr of treatment with 5, 10, 20, or 50 micrograms/ml of chrysotile fibers, cell morphology showed intense vacuolization, in both logarithmic and confluent cells. Following treatment of growing cells with 20 or 50 micrograms/ml of chrysotile fibers, cell spreading was also observed. Four hours after treatment of cells with 5, 10, or 20 micrograms/ml of crocidolite fibers, no such changes were seen. Vacuolization appeared later and was much less marked than for chrysotile fibers. The mesothelial cell population-doubling time of about 30 hr was not significantly altered by addition of 5 to 20 micrograms/ml of crocidolite fibers, but a concentration of 50 micrograms/ml lengthened doubling time to about 90 hr. Treatment with 5 or 10 micrograms/ml chrysotile fibers usually prolonged this time, but 20 or 50 micrograms/ml caused cell lysis. After 48 hr of incubation with both types of fiber, the proportion of mitosis was the same as in control cultures, whatever the fiber concentration, and asbestos fibers were often seen inside dividing cells.

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Mechanism of haemolysis by chrysotile fibres.

Haemolysis by chrysotile fibres may be related to an adsorption of the red blood cell (RBC) membranes on the fibres and not to an interaction between magnesium (Mg) from the fibres and sialic acid from RBC. This was demonstrated using neuraminidase (NANASE)-treated RBC and Mg-depleted chrysotile fibres. The mechanism may be extended to other minerals since a relationship can be obtained between the ability of various minerals to adsorb liposomes of dipalmitoyl phosphatidylcholine (DPPC) and their haemolytic activity.

Adsorption↗

Incidence of various types of thoracic malignancy induced in rats by intrapleural injection of 2 mg of various mineral dusts after inhalation of 222Ra.

After inhalation of 222Ra at equilibrium with radon daughters, male Sprague-Dawley rats were inoculated intrapleurally with 2 mg of unleached or acid-leached asbestos fibres or glass fibres or quartz. The additive co-carcinogenic effects of this type of insult were demonstrated by the increased incidence of malignant thoracic tumours. In rats given mineral materials, bronchopulmonary carcinomas and mixed carcinomas were observed, as well as typical mesotheliomas and combined pulmonary pleural tumours, whereas in control rats inhaling radon alone, only bronchopulmonary carcinomas occurred. No significant differences in tumour incidence were observed between the groups of animals given the different types of dusts, but statistically significant differences were noted in survival times, according to the histological type of tumour. The co-carcinogenic effects of an insult of the pleura by mineral dusts following inhalation of radon are discussed in relation to a possible tumour-promoting effect by these agents.

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Comparative toxicities of different forms of asbestos on rat pleural mesothelial cells.

The effects of UICC crocidolite and chrysotile A, either oxalic acid-leached or unleached, on the viability, morphology and growth characteristics of rat pleural mesothelial cells (PMC) were examined; DQ12 quartz particles were also used. When asbestos fibers were added for 48 hr at the beginning of exponential growth, 20 or 50 micrograms/mL of chrysotile fibers were cytotoxic and no growth occurred; with 5 or 10 micrograms/mL a latent period was observed, and the mean population doubling time was increased. Chrysotile ingestion was associated with morphological changes (spreading, intense vacuolation); moreover, a large proportion of the cells was binucleated (more than 30% with 10 micrograms/mL). The oxalic acid-leached chrysotile inhibited growth at a concentration of 50 micrograms/mL; with 5 or 10 micrograms/mL, no spreading occurred, but a shrinkage of some cells was observed. A few large vacuoles were seen in the cytoplasm of the cells; there were fewer binucleated cells. Addition of 5 or 10 micrograms/mL of crocidolite, either leached or unleached, did not significantly change the growth rate, in spite of the presence of a large number of fibers inside the cells which persisted when the cells reached confluency. With 20 or 50 micrograms/mL, the mean population doubling time was increased in a dose-dependent manner. A slight vacuolation of the cells occurred. The sample of quartz did not modify the parameters studied in this report. The results confirm the different in vitro reactivities of the two kinds of unleached asbestos fibers. Leaching of chrysotile fibers decreased their reactivity; alternatively, leaching of crocidolite increased the effects on PMC.

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Studies on in vitro chrysotile-pleural mesothelial cell interaction: morphological aspects and metabolism of benzo-3,4-pyrene.

Cultures of rat pleural mesothelial cells (PMC) were exposed to nonlethal doses of UICC chrysotile A. The morphology was studied by optical and electron microscopy. The consequences of chrysotile ingestion on the rate of pinocytosis of horseradish peroxidase (HPR) metabolism and benzo-3-4-pyrene (BP) were studied. Nonlethal doses of chrysotile (5 micrograms/mL) induced a time-dependent vacuolation of PMC; a dose-dependent inhibition of the vacuolation was observed when PMC were pretreated with DMSO. The origin of the vacuoles is not clear, but some features of autophagy and lysosomal storage were observed. Chrysotile fibers did not modify the rate of pinocytosis of HRP. Similarly, the metabolism of BP was unchanged when BP and chrysotile were both added to the culture medium or when PMC were preincubated with the fibers 24 hr prior to the addition of BP.

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