Araujo Heitor Ceolin, Ramírez Carmona Wilmer, Sato Camila, Dos Santos Oliveira Marcelo, Alves Guilherme Dos Santos Gomes, Morato Deborah Nunes, Pessan Juliano Pelim, Monteiro Douglas Roberto
School of Dentistry, Araçatuba, Department of Preventive and Restorative Dentistry, São Paulo State University (Unesp), Araçatuba, São Paulo 16015-050, Brazil.
School of Dentistry, University of Western São Paulo (UNOESTE), Presidente Prudente, São Paulo 19050-920, Brazil.
J Dent. 2022 Oct;125:104246. doi: 10.1016/j.jdent.2022.104246. Epub 2022 Jul 30.
This study assessed the effects of chitosan (CS) on microcosm biofilms derived from saliva of patients with Candida-associated denture stomatitis.
Five removable denture wearers with denture stomatitis were included in the study. The minimum inhibitory concentration (MIC) of CS against clinical isolates of Candida albicans was determined according to the broth microdilution method. Pooled saliva from the donors was used as an inoculum for the formation of biofilms, which were developed during 72 h on acrylic surfaces in the Amsterdam Active Attachment model. The biofilms were then treated with different concentrations of CS, and the antibiofilm effects were evaluated through the quantification of colony-forming units (CFUs), total biomass (TB), metabolic activity (MA), lactic acid production (LAP), and cell viability (by confocal laser scanning microscopy). Chlorhexidine, miconazole, and nystatin were tested as positive controls, while the negative control (NC) was the untreated biofilm. Data were analyzed by 1-way ANOVA and Fischer LSD's post hoc test (α=0.05).
MIC values of CS ranged from 500 to 800 µg/mL. For CFUs, 2500 µg/mL CS was the most effective treatment in reducing total anaerobes, mutans streptococci, and Lactobacillus spp., significantly differing from the controls. For C. albicans CFUs, CS and positive controls did not differ from each other but led to significant reductions compared to NC. Regarding TB, MA, LAP, and cell viability, 2500 µg/mL CS promoted the greatest reductions compared to NC.
CS has similar or superior effects to conventional active principles on important parameters of oral candidiasis microcosm biofilms.
The antibiofilm effects of CS show that this compound has great potential to improve the clinical condition of denture stomatitis patients, and formulations containing this natural polymer could be useful for controlling oral candidiasis.
本研究评估了壳聚糖（CS）对念珠菌相关性义齿性口炎患者唾液来源的微生态生物膜的影响。
本研究纳入了5名患有义齿性口炎的可摘义齿佩戴者。根据肉汤微量稀释法测定CS对白色念珠菌临床分离株的最低抑菌浓度（MIC）。将供体的混合唾液用作生物膜形成的接种物，在阿姆斯特丹主动附着模型中，生物膜在丙烯酸表面上培养72小时。然后用不同浓度的CS处理生物膜，并通过定量菌落形成单位（CFU）、总生物量（TB）、代谢活性（MA）、乳酸产生（LAP）和细胞活力（通过共聚焦激光扫描显微镜）来评估抗生物膜效果。以氯己定、咪康唑和制霉菌素作为阳性对照，而阴性对照（NC）为未处理的生物膜。数据采用单因素方差分析和Fischer LSD事后检验进行分析（α=0.05）。
CS的MIC值范围为500至800μg/mL。对于CFU，2500μg/mL CS是减少总厌氧菌、变形链球菌和乳酸杆菌属最有效的处理方法，与对照组有显著差异。对于白色念珠菌CFU，CS和阳性对照之间没有差异，但与NC相比导致显著减少。关于TB、MA、LAP和细胞活力，与NC相比，2500μg/mL CS促进了最大程度的降低。
在口腔念珠菌病微生态生物膜的重要参数方面，CS与传统活性成分具有相似或更好的效果。
CS的抗生物膜作用表明该化合物具有改善义齿性口炎患者临床状况的巨大潜力，含有这种天然聚合物的制剂可能有助于控制口腔念珠菌病。
