Tomé Fernanda Malagutti, Paula Ramos Lucas De, Freire Fernanda, Pereira Cristiane Aparecida, de Oliveira Ingrid Christine Barbosa, Junqueira Juliana Campos, Jorge Antonio Olavo Cardoso, Oliveira Luciane Dias de
Department of Biosciences and Oral Diagnosis, São Paulo State University (Unesp), Institute of Science and Technology, São José dos Campos, Francisco José Longo 777, São Dimas, São José dos Campos, SP, 12245-000, Brazil.
Lasers Med Sci. 2017 Aug;32(6):1237-1243. doi: 10.1007/s10103-017-2201-2. Epub 2017 Apr 7.
This study has evaluated the effects of photodynamic inactivation (PDI) using erythrosine as photosensitizer and green light-emitting diode (LED) on biofilms of Candida albicans alone and in combination with Enterococcus faecalis and Streptococcus mutans. We have also evaluated the effect of sucrose on biofilm formation and bacterial growth and sensitivity to PDI. Biofilms were formed in suspension of 10 cells/ml on plates before being grown in broth culture with and without sucrose and incubated for 48 h. Next, the treatment was applied using erythrosine at a concentration of 400 μM for 5 min and green LED (532 ± 10 nm) for 3 min on biofilms alone and in combination. The plates were washed and sonicated to disperse the biofilms, and serial dilutions were carried and aliquots seeded in Sabouraud agar before incubation for 48 h. Next, the colony-forming units per milliliter (CFU/ml; log) were counted and analyzed statistically (ANOVA, Tukey test, P ≤ 0.05). Results show that S. mutans favors the growth of C. albicans in biofilms with sucrose, with treatment not being effective. However, when the biofilm was grown without sucrose, we found a reduction in biofilm formation and a significant decrease in the PDI treatment (P < 0.0001). In conclusion, both growth and sensitivity to PDI in biofilms of C. albicans are strongly influenced by bacterial combination, and the presence of sucrose affected directly the growth and sensitivity of the biofilm to PDI as sucrose is the substrate for construction of the exopolysaccharide matrix.
本研究评估了以赤藓红为光敏剂、绿色发光二极管(LED)进行的光动力灭活(PDI)对白色念珠菌生物膜单独作用以及与粪肠球菌和变形链球菌联合作用的效果。我们还评估了蔗糖对生物膜形成、细菌生长以及对PDI敏感性的影响。生物膜在平板上以10个细胞/毫升的悬浮液形成,然后在添加和不添加蔗糖的肉汤培养基中培养48小时。接下来,对单独及联合的生物膜使用浓度为400μM的赤藓红处理5分钟,并用绿色LED(532±10纳米)照射3分钟。将平板冲洗并超声处理以分散生物膜,进行系列稀释并取等分试样接种于沙氏琼脂中,然后培养48小时。接着,计数每毫升菌落形成单位(CFU/毫升;对数)并进行统计学分析(方差分析、Tukey检验,P≤0.05)。结果表明,在含蔗糖的生物膜中,变形链球菌有利于白色念珠菌的生长,处理无效。然而,当生物膜在无蔗糖条件下生长时,我们发现生物膜形成减少,且PDI处理后显著下降(P<0.0001)。总之,白色念珠菌生物膜的生长及对PDI的敏感性均受到细菌组合的强烈影响,并且蔗糖的存在直接影响生物膜的生长及对PDI的敏感性,因为蔗糖是胞外多糖基质构建的底物。