Nutritional deprivation and LPS exposure as feasible methods for induction of cellular - A methodology to validate for vitro photobiomodulation studies

Autor(es)

Basso, F. G. ^[1] ; Turrioni, A. P. S. ^[1] ; Almeida, L. F. ^[1] ; Soares, D. G. ^[1] ; Oliveira, C. F. ^[2] ; Hebling, J. ^[1] ; de Souza Costa, C. A. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista, Araraquara Sch Dent, Araraquara, SP - Brazil

^[2] Univ Ribeirao Preto, UNAERP, Ribeirao Preto, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY; v. 159, p. 205-210, JUN 2016.
• Citações Web of Science: 2

Resumo

Previous studies have demonstrated that high biostimulation takes place when cells under stress are subjected to phototherapy by laser or light-emitting-diode (LED) devices. Several studies selected nutritional deprivation by reducing the concentration of fetal bovine serum (FBS) in the culture medium or the exposure of cultured cells to lipopolysaccharide (LPS) as an in vitro cellular stress condition. However, there are no data certifying that these stimuli cause stressful conditions for cultured cells. This investigation assessed the induction of cellular stress by decreasing the concentration of FBS or adding LPS to culture medium. Odontoblast-like cells (MDPC-23) were cultured in complete culture medium (DMEM) containing 10% FBS. After a 12-hour incubation period, the DMEM was replaced by fresh medium containing 10% FBS (control), low concentrations of FBS (0, 0.2, 0.5, 2, or 5%) or LPS from Escherichia coli (10 mu g/ml). After an additional 12-hour incubation, cell viability, total cell-counting, total protein production, and gene expression of heat shock protein 70 (HSP70) were assessed. Data were statistically analyzed by ANOVA complemented by the Tukey test, with 5% considered significant. Cell viability was negatively affected only for 0% FBS, while reduced viable cell numbers and total protein production were detected for FBS concentrations lower than 2%. Higher HSP70 gene expression was also observed for FBS concentrations lower than 2% and for cells exposed to LPS. The nutritional deprivation model with culture medium lower than 2% of FBS can be safely used to induce cellular stress for in vitro photobiomodulation studies. (C) 2016 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 13/05879-0 - Efeito da laserterapia de baixa potência e fatores de crescimento sobre a atividade de fibroblastos de gengiva obtidos de pacientes jovens e idosos
• Beneficiário: Carlos Alberto de Souza Costa
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 12/17947-8 - Efeito do laser de baixa intensidade sobre a expressão de citocinas inflamatórias na mucosite oral
• Beneficiário: Fernanda Gonçalves Basso
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado