Méndez Daniela Alejandra Cusicanqui, Gutierrez Eliezer, Dionísio Evandro José, Oliveira Thaís Marchini, Buzalaf Marília Afonso Rabelo, Rios Daniela, Machado Maria Aparecida Andrade Moreira, Cruvinel Thiago
Department of Pediatric Dentistry, Orthodontics and Public Health, Bauru School of Dentistry, University of São Paulo, Alameda Dr. Octávio Pinheiro Brisolla, 9-75, Vila Universitária, Bauru, SP, 17012-901, Brazil.
Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.
Lasers Med Sci. 2018 Apr;33(3):479-487. doi: 10.1007/s10103-017-2379-3. Epub 2017 Nov 8.
Antimicrobial photodynamic therapy (aPDT) has been proposed as an adjuvant treatment of dental caries, although there are no well-defined protocols to its clinical application. This study aimed to evaluate the influence of aPDT on the viability of microorganisms, vitality of biofilms, and lactic acid production of dentin caries microcosms. Biofilms were grown on bovine dentin discs in anaerobic conditions at 37 °C for 5 days, inoculating infected carious dentin in modified McBain medium plus 1% sucrose. The biofilms were treated by the combination of deionized water or 100 mg L methylene blue (MB) with 0, 37.5, or 75 J cm LED at 630 nm. The counts of total microorganisms, total streptococci, mutans streptococci, and total lactobacilli were determined by colony-forming units (CFU). The vitality of microbial cells in intact biofilms was measured by confocal laser scanning microscope (CLSM). The lactic acid production was analyzed by enzymatic spectrophotometry at 340 nm. Statistical analysis was conducted by Kruskal-Wallis and post hoc Dunn's tests (P < 0.05). MB and 37.5 J cm LED alone did not interfere in the viability of microorganisms, unlike 75 J cm LED alone that decreased the total microorganism and lactobacillus counts. The combination of MB and 75 J cm LED reduced the viability of all microorganisms and the vitality of intact biofilms. The production of lactic acid was statistically lower in all treatment groups in comparison with that of the control (no treatment), except for MB alone. Therefore, the MB-mediated aPDT was effective in controlling the viability, vitality and the acidogenicity of dentin caries microcosms.
尽管目前尚无明确的临床应用方案,但抗菌光动力疗法(aPDT)已被提议作为龋齿的辅助治疗方法。本研究旨在评估aPDT对微生物活力、生物膜活性以及牙本质龋微观模型中乳酸生成的影响。生物膜在37℃厌氧条件下于牛牙本质圆盘上生长5天,将感染的龋坏牙本质接种于改良的 McBain 培养基加1%蔗糖中。生物膜分别用去离子水或100 mg/L亚甲蓝(MB)与波长630 nm、能量密度为0、37.5或75 J/cm²的发光二极管(LED)联合处理。通过菌落形成单位(CFU)测定总微生物、总链球菌、变形链球菌和总乳酸菌的数量。利用共聚焦激光扫描显微镜(CLSM)测量完整生物膜中微生物细胞的活性。通过340 nm处的酶促分光光度法分析乳酸生成情况。采用Kruskal-Wallis检验和事后Dunn检验进行统计学分析(P < 0.05)。单独使用MB和37.5 J/cm²的LED对微生物活力无影响,而单独使用75 J/cm²的LED可降低总微生物和乳酸菌数量。MB与75 J/cm²的LED联合使用可降低所有微生物的活力以及完整生物膜的活性。与对照组(未处理)相比,除单独使用MB外,所有处理组的乳酸生成在统计学上均较低。因此,MB介导的aPDT在控制牙本质龋微观模型的活力、活性和产酸能力方面是有效的。