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白色念珠菌生物膜胞外多糖基质的共聚焦分析

Confocal analysis of the exopolysaccharide matrix of Candida albicans biofilms.

作者信息

Gonçalves Letícia M, Del Bel Cury Altair A, de Vasconcellos Andréa A, Cury Jaime A, da Silva Wander J

机构信息

Department of Prosthodontics and Periodontology, Piracicaba Dental School, University of Campinas, Piracicaba, São Paulo, Brazil.

Department of Physiological Sciences, Piracicaba Dental School, University of Campinas, Piracicaba, São Paulo, Brazil.

出版信息

J Investig Clin Dent. 2015 Aug;6(3):179-85. doi: 10.1111/jicd.12093. Epub 2014 Mar 7.

Abstract

AIM

Confocal laser-scanning microscopy (CLSM) was carried out to investigate the exopolysaccharide matrix of Candida albicans (C. albicans) biofilms developed on denture material under dietary carbohydrate exposure.

METHODS

Biofilms were developed on poly(methyl methacrylate) discs in culture media without (control) or with supplementation by glucose or sucrose for 72 h. For the CLSM analysis, biofilms were labeled with concanavalin A (ConA) during its development. Afterwards, biofilms were also labeled with SYTO-9. To confirm the results, the matrix was investigated by the phenol-sulfuric method. Data were analyzed by anova, followed by Tukey's test, with the level of significance set at 5%.

RESULTS

The use of ConA during biofilm development provided effective labeling of the exopolysaccharide matrix. The exposure to sucrose resulted in biofilms with the highest exopolysaccharide matrix biovolume (P < 0.05). The characterization obtained by CLSM was confirmed by the phenol-sulfuric method.

CONCLUSION

Confocal laser-scanning microscopy was found to be an effective tool for investigating the exopolysaccharide matrix of C. albicans biofilms, and exposure to sucrose resulted in increased matrix production.

摘要

目的

采用共聚焦激光扫描显微镜(CLSM)研究在膳食碳水化合物暴露条件下,白色念珠菌在义齿材料上形成的生物膜的胞外多糖基质。

方法

在聚甲基丙烯酸甲酯圆盘上,于不含(对照)或添加葡萄糖或蔗糖的培养基中培养生物膜72小时。对于CLSM分析,在生物膜形成过程中用伴刀豆球蛋白A(ConA)进行标记。之后,生物膜还用SYTO - 9进行标记。为证实结果,采用苯酚 - 硫酸法对基质进行研究。数据采用方差分析,随后进行Tukey检验,显著性水平设定为5%。

结果

在生物膜形成过程中使用ConA可有效标记胞外多糖基质。暴露于蔗糖导致生物膜具有最高的胞外多糖基质生物体积(P < 0.05)。CLSM获得的表征结果通过苯酚 - 硫酸法得到证实。

结论

共聚焦激光扫描显微镜是研究白色念珠菌生物膜胞外多糖基质的有效工具,暴露于蔗糖会导致基质产量增加。

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