Evaluation of apoptosis/necrosis and cytokine release provoked by three root canal sealers in human polymorphonuclears and monocytes.

AIM: To evaluate the in vitro cytotoxicity and cytokine release of three fresh root canal sealers and to determine the type of cell death they induce. METHODOLOGY: The sealers tested were Sealer 26 (S26), AH Plus (AHP), and Endosequence BC Sealer (END). Fresh sealers were cultivated in contact with monocytes and polymorphonuclears (PMNs) obtained from the peripheral blood of humans. Cell viability, apoptosis and necrosis were analysed at 4 h (PMNs) or 24 h (monocytes) using Annexin-V and propidium iodide in a cytometer. The supernatants were used to quantify Interleukin (IL)-4, IL-6, IL-10, IL-12 and tumour necrosis factor-α (TNF-α) in monocytes and IL-8 in PMNs by ELISA. One-way ANOVA and the Tukey post-test were used to compare data for cytotoxicity, and the multiple T-test was used to determine the differences between sealers in the release of cytokines that were statistically significant. RESULTS: After 4 h of treatment, S26 was associated with greater cell viability than the other sealers (P < 0.05) in the PMN culture and had similar values of necrosis as END (P > 0.05). After 24 h of treatment, AHP and END had greater monocyte cell viability than S26 (P < 0.05), which had more necrosis (P < 0.05). END had the lowest levels of IL-12 compared to the other sealers (P < 0.05) and higher levels of IL-6 compared to S26 (P < 0.05). The tested sealers did not differ in the release of IL-8, IL-10, TNF-α and IL-4 (P > 0.05). CONCLUSIONS: The effect of toxic agents released varied depending on the cell type studied. The composition of the sealers appeared to alter the form of self-regulation in the production of these cytokines by cells.

Morphological changes and EGF expression in the granular convoluted tubule cells of submandibular glands of Trypanosoma cruzi infected rats.

We have previously demonstrated in rats that Chagas' disease affects the salivary glands, by promoting an enlargement of the submandibular gland. In order to further investigate possible functional alterations on infected submandibular glands, the objective of the present study was to analyze epidermal growth factor (EGF) expression on rat submandibular glands during Trypanosoma cruzi infection. Results demonstrated that infected rats presented lower levels of testosterone, and morphological changes in the granular convoluted tubule (GCT) cells of the submandibular glands, along with acinar enlargement and delayed ductal maturation at the developing granular ducts. Immunohistochemistry analysis additionally showed that only few cells immunolabelled with anti-EGF on infected rats during the acute phase of Chagas' disease, while after 64 and 90 days (chronic phase) of infection, EGF expression was similar to non-infected rats. The present findings suggest that at the acute phase of Chagas' disease, lower levels of testosterone may lead to a delayed maturation of GCT, which positively correlates with decreased EGF production by submandibular glands cells.