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.

摘要

包含白色念珠菌和金黄色葡萄球菌的多微生物生物膜可以增加口腔疾病的频率和严重程度。本研究评估了由来自金黄色葡萄球菌和白色念珠菌的单一和双重生物膜的可溶性因子促进的口腔角质形成细胞死亡、凋亡和/或坏死. 可溶性因子从 16 小时的生物膜生长培养基中获得。通过 MTT 评估细胞活力，通过 LDH 评估细胞膜损伤。SEM用于形态变化。使用膜联蛋白 V 和碘化丙啶和半胱天冬酶 -2、-3、-6、-8 和 -9 进行细胞凋亡和坏死的评估。使用 ANOVA 和 Tukey 检验 (α = 5%) 进行统计分析。双生物膜对口腔细胞的危害最大，成活率为31.76%，细胞膜受损，LDH释放。双生物膜还诱导更高百分比的坏死细胞 (24.95%)。细胞凋亡与半胱天冬酶-2、-3、-6 和-8 激活有关。