Antibacterial and Antiadhesion Effects of Psidium guajava Fractions on a Multispecies Biofilm Associated with Periodontitis

Abstract Objective: To assess the antibacterial activity of Psidium guajava fractions and their effects on adhesion of a multispecies biofilm consisting of Streptococcus gordonii, Fusobacterium nucleatum, and Porphyromonas gingivalis in vitro. Material and Methods: Guava leaves were obtained from the mountains of northern Peru, where they grow wild and free of pesticides. The antimicrobial activity of 25 mg/mL petroleum ether, 25 mg/mL dichloromethane and 25 mg/mL methanol fractions of P. guajava was evaluated by measuring inhibition halos, as well as the effect on the adhesion of multispecies biofilms at 4, 7 and 10 days of growth by measuring the optical density. In addition, antimicrobial susceptibility was compared using the Kruskal-Wallis test and its multiple comparison tests, and differences in mean biofilm adhesion between each fraction were assessed by repeated measures analysis and the Tukey multiple comparison test. Results: The rank-based Kruskal-Wallis test highlighted differences in the effects of the fractions on the zone of inhibition for each oral bacterium, including S. gordonii (p=0.000), F. nucleatum (p=0.000), and P. gingivalis (p=0.000), the Tukey test showed that the group treated with 0.12% chlorhexidine exhibited the least amount of adhesion, followed by the group treated with the 1.56 mg/mL methanol fraction. Conclusion: The methanol fraction of P. guajava had an antibacterial effect on S. gordonii and P. gingivalis, and the 1.56 mg/mL methanol fraction decreased biofilm adhesion.

Identification of Proteins Associated with the Formation of Oral Biofilms

ABSTRACT Objective: To identify proteins associated with the formation of Streptococcus gordonii and Fusobacterium nucleatum biofilms. Material and Methods: Biofilms composed of two bacterial species, S. gordonii and F. nucleatum, were cultured for 1, 4, 7, and 10 days. The presence of both species was confirmed via amplification of the srtA and radD genes using real-time PCR. The concentrations of proteins associated with the biofilms and individual species were quantified using Western blotting. Results: The protein profiles of S. gordonii and F. nucleatum from individual cultures determined using one-dimensional electrophoresis revealed proteins found in S. gordonii and in F. nucleatum. Ct and reciprocal Ct values were determined for the exposed S. gordonii and F. nucleatum biofilms. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was detected in biofilms and F. nucleatum, whereas HSP40 protein was present only in biofilms after 7 and 10 days of formation. Conclusion: HSP40 was detected only in the formed biofilms; thus, HSP40 is an essential proteins for adhesion.