Abstract
Polymicrobial biofilms comprising Candida albicans and Staphylococcus aureus can increase the frequency and severity of oral diseases. This study assessed oral keratinocyte cell death, apoptosis and/or necrosis, promoted by soluble factors from single and dual biofilms of S. aureus and C. albicans. The soluble factors were obtained from the 16-h biofilm growth media. Cell viability was assessed by MTT and cell membrane damage by LDH. SEM was used for morphology changes. Assessment of apoptosis and necrosis was performed using annexin V and propidium iodide and caspases -2, -3, -6, -8 and -9. Statistical analysis was conducted with ANOVA and Tukey tests (α = 5%). Dual biofilms promoted the greatest harmful effect on oral cells, with a viability rate of 31.76%, damage to cell membranes and LDH released. Dual biofilms also induced higher percentages of necrotic cells (24.95%). Apoptosis was associated with caspases -2, -3, -6 and -8 activation.
Author contributions
Kassia de Carvalho Dias: performed assays and analysis. Paula Aboud Barbugli: conception, design of the study, interpretation of results. Carlos Eduardo Vergani: conception, financial support. All authors contributed to the elaboration of the manuscript and approved the submitted version.
Acknowledgement
School of Dentistry (FOAr/UNESP), FAPESP, CAPES, CNPQ.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, n° 304247/2017-0; n° 400658/2012-7).
Data availability statement
The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.