Graduate Program in Dentistry. Federal University of Paraíba. Cidade Universitária, João Pessoa, Paraiba, Brazil.
Department of Clinic and Social Dentistry. Federal University of Paraíba. Cidade Universitária, João Pessoa, Paraiba, Brazil.
Braz Dent J. 2023 May-Jun;34(3):73-81. doi: 10.1590/0103-6440202305420.
Experimental models that consider host-pathogen interactions are relevant for improving knowledge about oral candidiasis. The aim of this study was to assess the epithelial immune responses, Candida penetration of cell monolayers, and virulence during mixed species culture infections. Single species cultures of Candida albicans and mixed cultures (C. albicans, Streptococcus mutans, and Streptococcus sanguinis) were used to infect monolayers of HaCaT and FaDu ATCC HTB-43 cells for 12 h. After infection, IL-18 and IL-34 gene expression was measured to assess epithelial cell immune responses, and lactate dehydrogenase (LDH) activity was measured as an indicator of cell damage. Microscopy determined C. albicans morphology and penetration of fungal cells through the keratinocyte monolayer. Monolayers devoid of infection served as controls. Data were analyzed by an ANOVA one-way test followed by Tukey's post-hoc test (α = 0.05). The results found that IL-18 and IL-34 gene expression and LDH activity were significantly (p < 0.05) upregulated for both cell lines exposed to mixed species cultures compared with C. albicans alone. Candida albicans yeast and hyphae were evident in C. albicans only infections. In contrast, monolayers infected by C. albicans, S. mutans, and S. sanguinis exhibited higher microbial invasion with several hyphal aggregates detected. The presence of streptococci in C. albicans infection enhances the virulence and pathogenicity of the fungus with associated increased immune responses and tissue damage. Extrapolation of these findings to oral infection would indicate the added potential benefit of managing bacterial components of biofilms during treatment.
实验模型考虑宿主-病原体相互作用对于提高对口腔念珠菌病的认识是相关的。本研究的目的是评估上皮免疫反应、念珠菌穿透细胞单层和混合物种培养感染期间的毒力。使用白色念珠菌的单种培养物和混合培养物(白色念珠菌、变形链球菌和血链球菌)感染 HaCaT 和 FaDu ATCC HTB-43 细胞单层 12 小时。感染后,测量 IL-18 和 IL-34 基因表达以评估上皮细胞免疫反应,并用乳酸脱氢酶(LDH)活性作为细胞损伤的指标。显微镜确定白色念珠菌形态和真菌细胞穿透角质形成细胞单层。未感染的单层作为对照。通过单因素方差分析测试,然后进行 Tukey 事后检验(α=0.05)对数据进行分析。结果发现,与单独的白色念珠菌相比,两种细胞系暴露于混合物种培养物时,IL-18 和 IL-34 基因表达和 LDH 活性均显著(p<0.05)上调。仅白色念珠菌感染中可观察到白色念珠菌酵母和菌丝。相比之下,由白色念珠菌、变形链球菌和血链球菌感染的单层显示出更高的微生物入侵,检测到几个菌丝聚集。链球菌存在于白色念珠菌感染中会增强真菌的毒力和致病性,伴随相关的免疫反应和组织损伤增加。将这些发现外推到口腔感染,表明在治疗过程中管理生物膜中细菌成分可能会带来额外的益处。