In vitro cytotoxicity of one-step dentin bonding systems.

The cytotoxicity of resinous monomers may vary on mixing, calling into question the cytotoxicity of the new dentin bonding agents that are mixed in a single vial. The cytotoxicity of fourth and fifth generation dentin bonding agents was compared in vitro. All-Bond 2, One-Step, Scotchbond Multi-Purpose, Scotchbond One, Syntac, Syntac Single Component, Tenure, and Tenure Quick were tested uncured. The cytotoxicity of several dilutions of fourth generation dentin primer, dentin bonding agent, dentin primer + dentin bonding agent, and fifth generation single component products diluted at 10(-1) to 10(-8) in culture medium was recorded with a MTT assay on L929 fibroblasts. Only one synergistic (increased cytotoxicity after mixing primer and bonding agent) cytotoxic effect was observed with Tenure at 10(-5) dilution. An antagonistic effect (decreased cytotoxicity after mixing primer and bonding agent) was observed with All-Bond 2 between 10(-4) and 10(-8) dilutions. Scotchbond and Syntac fifth generation dentin bonding agents were less cytotoxic than their fourth generation counterparts. All-Bond and Tenure fifth generation dentin bonding agents were more cytotoxic than their fourth generation counterparts only in low dilutions. Clinical studies should confirm these good results, because no dramatic synergistic cytotoxic effect could be detected.

Concentration of eugenol apically released from zinc oxide-eugenol-based sealers.

This study investigated the amount of eugenol released from a zinc oxide-eugenol-based sealer at the apex of teeth filled according to two techniques: the single-cone and the Thermafil. The crown of 10 maxillary central incisors was removed, and the canal was prepared with ProFile to a size 30 under NaOCl irrigation. The patency of the apex was checked with a #8 K-file between each ProFile. Five roots were filled with a Thermafil #30 and 0.03 g of Sealite; five roots were filled using a Lentulo with 0.07 g of sealer and a gutta-percha cone #30. The powder/liquid ratio of the sealer was of 5/1. The concentration of eugenol released in phosphate-buffered saline was spectrofluorimetrically determined immediately after sealing, after 1 day, and after 1 month of storage. The roots filled with the single-cone technique released significantly more eugenol than these filled with Thermafil immediately after sealing (p = 0.002); but, after 1-day or 1-month storage, there was no difference. For both techniques, eugenol concentration decreased over time (p = 0.01): the immediate concentration was higher than the 1-day concentration (p = 0.04). Eugenol concentration after 1-month storage was undetectable. The results of this work show that the level of eugenol released from a zinc oxide-eugenol-based sealer beyond the apex is very low and decreases over time.

Helium-neon laser irradiation is not a stressful treatment: a study on heat-shock protein (HSP70) level.

BACKGROUND AND OBJECTIVE: Helium-neon (He-Ne) laser irradiation has been clinically used to reduce chemotherapy-induced mucositis. This work was designed to find out if this treatment is stressful at the cellular level by studying its effects on the level of the stress-inducible heat shock proteins. STUDY DESIGN/MATERIALS AND METHODS: Human desmodontal and mouse L929 fibroblasts were irradiated using a 60 mW laser by a single application of 1.5 and 3J/cm2 in continuous mode. Heat shock protein level was studied by gel electrophoresis and Western blotting using monoclonal antibodies. RESULTS: He-Ne treatment does not induce heat shock protein synthesis in human desmodontal nor in mouse fibroblasts at the energy densities used in this study. CONCLUSIONS: These results indicate that the treatment is not stressful at the cellular level.

Effects of ethanol on intercellular communications and polarization of hepatocytes in short-term culture.

The formation of intracellular lumina with apical differentiation is observed in several cancerous epithelial cell lines including human hepatocarcinoma. This disorder of cell polarization can be induced by the inhibition of cell-cell communication, a known factor of carcinogenesis. This work was designed to study the effects of ethanol on the differentiation of hepatocytes in short-term culture. Isolated hepatocytes were plated on plastic culture dishes that were 35 mm in diameter (10(6) cells/dish). Three hours after plating, the hepatocytes were incubated in the presence of 20 mmol/L ethanol for 1 hr. Treated cells were compared with controls using morphometric methods after conventional treatment for ultramicroscopy and by measuring cellular dye coupling by the fluorescent Lucifer Yellow CH transfer method. Bile canaliculi formation decreased in alcohol-treated cells (6.5% vs. 9.9%, 2p less than 0.05), whereas intracellular lumina incidence increased (3.1% vs. 0.5%, 2p less than 0.01). In parallel, the dye-coupling capacity decreased significantly when hepatocytes were treated with alcohol (2p less than 0.01). This work shows that short-term ethanol treatment induces significant disturbances of cell polarization and inhibits the reestablishment of cell-cell communication in cultured hepatocytes. These disorders could, at least in part, explain the carcinogenic effects of ethanol.

The effects of monensin on the transport of horseradish peroxidase into intracellular lumina in cultured rat hepatocytes.

A kinetic study of horseradish peroxidase transport was carried out on short-term cultured adult rat hepatocytes. After 4 hr of plating, cells were preincubated with monensin (1 microM) for 1 hr before their incubation with horseradish peroxidase for different times. Monensin treatment resulted in the dilatation of the Golgi apparatus and caused the appearance of numerous intracellular lumina lined with microvilli in associated as well as isolated hepatocytes but did not modify newly formed bile canaliculi. The frequency of their appearance increased to 28% in cells pretreated with monensin compared to 2% in controls. Intracellular lumina membranes had the morphological features of apical membranes and were stained with horseradish peroxidase more frequently than those of newly formed bile canaliculi. This work therefore provides a model for studying bile secretion in cultured hepatocytes. Our results also suggest that the biliary transport of horseradish peroxidase does not involve the Golgi apparatus, since horseradish peroxidase was never observed in the Golgi stacks even after monensin treatment.

[Intracellular lumen in cultured hepatocytes of rats. Study of an original cytoplasmic infrastructure]. / La lumière intracellulaire dans les hépatocytes de rat en culture. Etude d'une infrastructure cytoplasmique originale.

A few hours after plating, isolated rat hepatocytes reassociate into clusters and differentiate intercellular cavities bordered by junctional complexes. These structures show a great resemblance to bile canaliculi seen in vivo. Intracellular lumens surrounded by microvilli are observed in the cytoplasm of some cultured hepatocytes. The formation of these structures, which contain an osmiophilic substance, probably results from modifications in the functioning of the secretory apparatus. It can be speculated that these intracellular lumens may contribute to the formation of new canaliculi differentiated between reassociated cells.