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Antimicrobial nisin acts against saliva derived multi-species biofilms without cytotoxicity to human oral cells.

Shin JM, Ateia I, Paulus JR, Liu H, Fenno JC, Rickard AH, Kapila YL - Front Microbiol (2015)

Bottom Line: Nisin effects on cellular apoptosis and proliferation were evaluated using acridine orange/ethidium bromide fluorescent nuclear staining and lactate dehydrogenase activity assays.Specifically, under biofilm model conditions, nisin interfered with biofilm development and reduced biofilm biomass and thickness in a dose-dependent manner.The treatment of pre-formed biofilms with nisin resulted in dose- and time-dependent disruption of the biofilm architecture along with decreased bacterial viability.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor MI, USA.

ABSTRACT

Objectives: Nisin is a lantibiotic widely used for the preservation of food and beverages. Recently, investigators have reported that nisin may have clinical applications for treating bacterial infections. The aim of this study was to investigate the effects of ultra pure food grade Nisin ZP (>95% purity) on taxonomically diverse bacteria common to the human oral cavity and saliva derived multi-species oral biofilms, and to discern the toxicity of nisin against human cells relevant to the oral cavity.

Methods: The minimum inhibitory concentrations and minimum bactericidal concentrations of taxonomically distinct oral bacteria were determined using agar and broth dilution methods. To assess the effects of nisin on biofilms, two model systems were utilized: a static and a controlled flow microfluidic system. Biofilms were inoculated with pooled human saliva and fed filter-sterilized saliva for 20-22 h at 37°C. Nisin effects on cellular apoptosis and proliferation were evaluated using acridine orange/ethidium bromide fluorescent nuclear staining and lactate dehydrogenase activity assays.

Results: Nisin inhibited planktonic growth of oral bacteria at low concentrations (2.5-50 μg/ml). Nisin also retarded development of multi-species biofilms at concentrations ≥1 μg/ml. Specifically, under biofilm model conditions, nisin interfered with biofilm development and reduced biofilm biomass and thickness in a dose-dependent manner. The treatment of pre-formed biofilms with nisin resulted in dose- and time-dependent disruption of the biofilm architecture along with decreased bacterial viability. Human cells relevant to the oral cavity were unaffected by the treatment of nisin at anti-biofilm concentrations and showed no signs of apoptotic changes unless treated with much higher concentrations (>200 μg/ml).

Conclusion: This work highlights the potential therapeutic value of high purity food grade nisin to inhibit the growth of oral bacteria and the development of biofilms relevant to oral diseases.

No MeSH data available.


Related in: MedlinePlus

Nisin inhibits the formation of multi-species biofilms in a Bioflux controlled flow microfluidic model system. CCS was added, then fed filter sterilized CFS for 20–22 h at 37°C with or without nisin. (A) Confocal microscopy images are represented in the x–y plane. A green signal indicates viable live cells (Syto 9) and a red signal indicates damaged/dead cells (propidium iodide). (B) Biofilm biomass, thickness, and roughness [mean (SD)] were derived from imaging of at least three separate channels (experiments). ∗P < 0.05 and ∗∗P < 0.01: significant differences from the control (nisin-free).
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Figure 3: Nisin inhibits the formation of multi-species biofilms in a Bioflux controlled flow microfluidic model system. CCS was added, then fed filter sterilized CFS for 20–22 h at 37°C with or without nisin. (A) Confocal microscopy images are represented in the x–y plane. A green signal indicates viable live cells (Syto 9) and a red signal indicates damaged/dead cells (propidium iodide). (B) Biofilm biomass, thickness, and roughness [mean (SD)] were derived from imaging of at least three separate channels (experiments). ∗P < 0.05 and ∗∗P < 0.01: significant differences from the control (nisin-free).

Mentions: Under controlled flow microfluidic growth conditions, the anti-biofilm effects of nisin were exerted at concentrations ≥0.5 μg/ml (Figure 3A). The average biofilm biomass and thickness of the control biofilms were 30.88 μm3/μm2 and 31.02 μm (Figure 3B). The average biofilm biomass and thickness of the nisin treated biofilms at 1 μg/ml were 5.06 μm3/μm2 and 7.17 μm (Figure 3B). The formation of biofilms was absent at 4 μg/ml (Figures 3A,B).


Antimicrobial nisin acts against saliva derived multi-species biofilms without cytotoxicity to human oral cells.

Shin JM, Ateia I, Paulus JR, Liu H, Fenno JC, Rickard AH, Kapila YL - Front Microbiol (2015)

Nisin inhibits the formation of multi-species biofilms in a Bioflux controlled flow microfluidic model system. CCS was added, then fed filter sterilized CFS for 20–22 h at 37°C with or without nisin. (A) Confocal microscopy images are represented in the x–y plane. A green signal indicates viable live cells (Syto 9) and a red signal indicates damaged/dead cells (propidium iodide). (B) Biofilm biomass, thickness, and roughness [mean (SD)] were derived from imaging of at least three separate channels (experiments). ∗P < 0.05 and ∗∗P < 0.01: significant differences from the control (nisin-free).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4471743&req=5

Figure 3: Nisin inhibits the formation of multi-species biofilms in a Bioflux controlled flow microfluidic model system. CCS was added, then fed filter sterilized CFS for 20–22 h at 37°C with or without nisin. (A) Confocal microscopy images are represented in the x–y plane. A green signal indicates viable live cells (Syto 9) and a red signal indicates damaged/dead cells (propidium iodide). (B) Biofilm biomass, thickness, and roughness [mean (SD)] were derived from imaging of at least three separate channels (experiments). ∗P < 0.05 and ∗∗P < 0.01: significant differences from the control (nisin-free).
Mentions: Under controlled flow microfluidic growth conditions, the anti-biofilm effects of nisin were exerted at concentrations ≥0.5 μg/ml (Figure 3A). The average biofilm biomass and thickness of the control biofilms were 30.88 μm3/μm2 and 31.02 μm (Figure 3B). The average biofilm biomass and thickness of the nisin treated biofilms at 1 μg/ml were 5.06 μm3/μm2 and 7.17 μm (Figure 3B). The formation of biofilms was absent at 4 μg/ml (Figures 3A,B).

Bottom Line: Nisin effects on cellular apoptosis and proliferation were evaluated using acridine orange/ethidium bromide fluorescent nuclear staining and lactate dehydrogenase activity assays.Specifically, under biofilm model conditions, nisin interfered with biofilm development and reduced biofilm biomass and thickness in a dose-dependent manner.The treatment of pre-formed biofilms with nisin resulted in dose- and time-dependent disruption of the biofilm architecture along with decreased bacterial viability.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor MI, USA.

ABSTRACT

Objectives: Nisin is a lantibiotic widely used for the preservation of food and beverages. Recently, investigators have reported that nisin may have clinical applications for treating bacterial infections. The aim of this study was to investigate the effects of ultra pure food grade Nisin ZP (>95% purity) on taxonomically diverse bacteria common to the human oral cavity and saliva derived multi-species oral biofilms, and to discern the toxicity of nisin against human cells relevant to the oral cavity.

Methods: The minimum inhibitory concentrations and minimum bactericidal concentrations of taxonomically distinct oral bacteria were determined using agar and broth dilution methods. To assess the effects of nisin on biofilms, two model systems were utilized: a static and a controlled flow microfluidic system. Biofilms were inoculated with pooled human saliva and fed filter-sterilized saliva for 20-22 h at 37°C. Nisin effects on cellular apoptosis and proliferation were evaluated using acridine orange/ethidium bromide fluorescent nuclear staining and lactate dehydrogenase activity assays.

Results: Nisin inhibited planktonic growth of oral bacteria at low concentrations (2.5-50 μg/ml). Nisin also retarded development of multi-species biofilms at concentrations ≥1 μg/ml. Specifically, under biofilm model conditions, nisin interfered with biofilm development and reduced biofilm biomass and thickness in a dose-dependent manner. The treatment of pre-formed biofilms with nisin resulted in dose- and time-dependent disruption of the biofilm architecture along with decreased bacterial viability. Human cells relevant to the oral cavity were unaffected by the treatment of nisin at anti-biofilm concentrations and showed no signs of apoptotic changes unless treated with much higher concentrations (>200 μg/ml).

Conclusion: This work highlights the potential therapeutic value of high purity food grade nisin to inhibit the growth of oral bacteria and the development of biofilms relevant to oral diseases.

No MeSH data available.


Related in: MedlinePlus