Abstract
Objective. Lipopolysaccharide (LPS) from cariogenic microorganisms and resin monomers like HEMA (2-hydroxyethyl methacrylate) included in dentin adhesive are present in a clinical situation in deep dentinal cavity preparations. Here, cell survival, expression of proteins related to redox homeostasis, and viability of mouse macrophages exposed to LPS and HEMA were analyzed with respect to the ...
Abstract
Objective. Lipopolysaccharide (LPS) from cariogenic microorganisms and resin monomers like HEMA (2-hydroxyethyl methacrylate) included in dentin adhesive are present in a clinical situation in deep dentinal cavity preparations. Here, cell survival, expression of proteins related to redox homeostasis, and viability of mouse macrophages exposed to LPS and HEMA were analyzed with respect to the influence of oxidative stress. Methods. Cell survival of RAW264.7 mouse macrophages was determined using a crystal violet assay, protein expression was detected by Western blotting, and HEMA- or LPS-induced apoptosis (cell viability) was analyzed by flow cytometry. Cells were exposed to HEMA (0-8 mM), LPS (0.1 mu g/ml) or combinations of both substances for 24 h. The influence of mitogen-activated protein kinases (MAPK) was analyzed using the specific inhibitors PD98059 (ERK1/2), 5B203580 (p38) or SP600125 (JNK), and oxidative stress was identified by the antioxidant N-acetylcysteine (NAC). Results. Cell survival was reduced by HEMA. LPS, however, increased cell survival from 29% in cultures exposed to 8 mM HEMA, to 46% in cultures co-exposed to 8 mM HEMA/LPS. Notably, LPS-induced apoptosis was neutralized by 4-6 mM HEMA but apoptosis caused by 8 mM HEMA was counteracted by LPS. Expression of NOS (nitric oxide synthase), p47(Phox) and p67(Phox) subunits of NADPH oxidase, catalase or heme oxygenase (HO-1) was associated with HEMA- or LPS-induced apoptosis. While no influence of MAPK was detected, NAC inhibited cytotoxic effects of HEMA. Significance. HEMA- and LPS-triggered pathways may induce apoptosis and interfere with physiological tissue responses as a result of the differential formation of oxidative stress. (C) 2016 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.
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