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Pluronics-Formulated Farnesol Promotes Efficient Killing and Demonstrates Novel Interactions with Streptococcus mutans Biofilms.

Mogen AB, Chen F, Ahn SJ, Burne RA, Wang D, Rice KC - PLoS ONE (2015)

Bottom Line: In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms.Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log.Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Florida, 32611, United States of America.

ABSTRACT
Streptococcus mutans is the primary causative agent of dental caries, one of the most prevalent diseases in the United States. Previously published studies have shown that Pluronic-based tooth-binding micelles carrying hydrophobic antimicrobials are extremely effective at inhibiting S. mutans biofilm growth on hydroxyapatite (HA). Interestingly, these studies also demonstrated that non-binding micelles (NBM) carrying antimicrobial also had an inhibitory effect, leading to the hypothesis that the Pluronic micelles themselves may interact with the biofilm. To explore this potential interaction, three different S. mutans strains were each grown as biofilm in tissue culture plates, either untreated or supplemented with NBM alone (P85), NBM containing farnesol (P85F), or farnesol alone (F). In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms. Furthermore, the P85F biofilms formed large towers containing dead cells that were not observed in the other treatment conditions. Tower formation appeared to be specific to formulated farnesol, as this phenomenon was not observed in S. mutans biofilms grown with NBM containing triclosan. Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log. Wild-type biofilms grown in the absence of sucrose or gtfBC mutant biofilms grown in the presence of sucrose did not form towers. However, increased cell killing with P85F was still observed, suggesting that cell killing is independent of tower formation. Finally, repeated treatment of pre-formed biofilms with P85F was able to elicit a 2-log reduction in viability, whereas parallel treatment with F alone only reduced viability by 0.5-log. Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

No MeSH data available.


Related in: MedlinePlus

Effect of gtfBC mutation on S. mutans biofilm architecture and viability in response to P85-formulated farnesol.S. mutans gtfBC mutant was grown for 48 hours in the presence of P85 (A), P85F (B), F alone (C) or untreated (D) in BM media containing 0.25% sucrose and 0.25% glucose. Wells containing adherent biofilm were stained with LIVE/DEAD stain. Biofilm z-stacks of 1 μm cross-sectional images were acquired at 400× magnification by CLSM. Representative orthogonal images of each biofilm are shown. Scale bars = 20 μm. In a parallel experiment (n = 3 biological replicates), the average CFU/ml of each biofilm was also determined (E). All data are representative of 2 independent experiments. Error bars = SEM. *Denotes statistically-significant difference relative to untreated condition (p < 0.05), SNK Test.
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pone.0133886.g008: Effect of gtfBC mutation on S. mutans biofilm architecture and viability in response to P85-formulated farnesol.S. mutans gtfBC mutant was grown for 48 hours in the presence of P85 (A), P85F (B), F alone (C) or untreated (D) in BM media containing 0.25% sucrose and 0.25% glucose. Wells containing adherent biofilm were stained with LIVE/DEAD stain. Biofilm z-stacks of 1 μm cross-sectional images were acquired at 400× magnification by CLSM. Representative orthogonal images of each biofilm are shown. Scale bars = 20 μm. In a parallel experiment (n = 3 biological replicates), the average CFU/ml of each biofilm was also determined (E). All data are representative of 2 independent experiments. Error bars = SEM. *Denotes statistically-significant difference relative to untreated condition (p < 0.05), SNK Test.

Mentions: To independently verify that P85F-induced tower formation does not occur in the absence of sucrose-dependent biofilm matrix, this experiment was repeated on S. mutans gtfBC mutant biofilms grown in the presence of sucrose (Fig 8). As predicted, tower formation was not observed in gtfBC mutant biofilms grown in the presence of sucrose and P85F (Fig 8B), confirming a role for the glucan biofilm matrix in promoting P85F induced tower formation. Growth of either sucrose-independent UA159 biofilms (Fig 6A) or sucrose-dependent gtfBC mutant biofilms (Fig 8A) in the presence of P85 alone did not alter their biofilm structure relative to corresponding untreated biofilms (Figs 6D and 8D).


Pluronics-Formulated Farnesol Promotes Efficient Killing and Demonstrates Novel Interactions with Streptococcus mutans Biofilms.

Mogen AB, Chen F, Ahn SJ, Burne RA, Wang D, Rice KC - PLoS ONE (2015)

Effect of gtfBC mutation on S. mutans biofilm architecture and viability in response to P85-formulated farnesol.S. mutans gtfBC mutant was grown for 48 hours in the presence of P85 (A), P85F (B), F alone (C) or untreated (D) in BM media containing 0.25% sucrose and 0.25% glucose. Wells containing adherent biofilm were stained with LIVE/DEAD stain. Biofilm z-stacks of 1 μm cross-sectional images were acquired at 400× magnification by CLSM. Representative orthogonal images of each biofilm are shown. Scale bars = 20 μm. In a parallel experiment (n = 3 biological replicates), the average CFU/ml of each biofilm was also determined (E). All data are representative of 2 independent experiments. Error bars = SEM. *Denotes statistically-significant difference relative to untreated condition (p < 0.05), SNK Test.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4519314&req=5

pone.0133886.g008: Effect of gtfBC mutation on S. mutans biofilm architecture and viability in response to P85-formulated farnesol.S. mutans gtfBC mutant was grown for 48 hours in the presence of P85 (A), P85F (B), F alone (C) or untreated (D) in BM media containing 0.25% sucrose and 0.25% glucose. Wells containing adherent biofilm were stained with LIVE/DEAD stain. Biofilm z-stacks of 1 μm cross-sectional images were acquired at 400× magnification by CLSM. Representative orthogonal images of each biofilm are shown. Scale bars = 20 μm. In a parallel experiment (n = 3 biological replicates), the average CFU/ml of each biofilm was also determined (E). All data are representative of 2 independent experiments. Error bars = SEM. *Denotes statistically-significant difference relative to untreated condition (p < 0.05), SNK Test.
Mentions: To independently verify that P85F-induced tower formation does not occur in the absence of sucrose-dependent biofilm matrix, this experiment was repeated on S. mutans gtfBC mutant biofilms grown in the presence of sucrose (Fig 8). As predicted, tower formation was not observed in gtfBC mutant biofilms grown in the presence of sucrose and P85F (Fig 8B), confirming a role for the glucan biofilm matrix in promoting P85F induced tower formation. Growth of either sucrose-independent UA159 biofilms (Fig 6A) or sucrose-dependent gtfBC mutant biofilms (Fig 8A) in the presence of P85 alone did not alter their biofilm structure relative to corresponding untreated biofilms (Figs 6D and 8D).

Bottom Line: In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms.Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log.Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Florida, 32611, United States of America.

ABSTRACT
Streptococcus mutans is the primary causative agent of dental caries, one of the most prevalent diseases in the United States. Previously published studies have shown that Pluronic-based tooth-binding micelles carrying hydrophobic antimicrobials are extremely effective at inhibiting S. mutans biofilm growth on hydroxyapatite (HA). Interestingly, these studies also demonstrated that non-binding micelles (NBM) carrying antimicrobial also had an inhibitory effect, leading to the hypothesis that the Pluronic micelles themselves may interact with the biofilm. To explore this potential interaction, three different S. mutans strains were each grown as biofilm in tissue culture plates, either untreated or supplemented with NBM alone (P85), NBM containing farnesol (P85F), or farnesol alone (F). In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms. Furthermore, the P85F biofilms formed large towers containing dead cells that were not observed in the other treatment conditions. Tower formation appeared to be specific to formulated farnesol, as this phenomenon was not observed in S. mutans biofilms grown with NBM containing triclosan. Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log. Wild-type biofilms grown in the absence of sucrose or gtfBC mutant biofilms grown in the presence of sucrose did not form towers. However, increased cell killing with P85F was still observed, suggesting that cell killing is independent of tower formation. Finally, repeated treatment of pre-formed biofilms with P85F was able to elicit a 2-log reduction in viability, whereas parallel treatment with F alone only reduced viability by 0.5-log. Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

No MeSH data available.


Related in: MedlinePlus