Use of a High-throughput &lt;em&gt;In Vitro&lt;/em&gt; Microfluidic System to Develop Oral Multi-species Biofilms

Samarian, Derek S.; Jakubovics, Nicholas S.; Luo, Ting; Rickard, Alexander H.

doi:10.3791/52467

Cited by 27 publications

(45 citation statements)

References 37 publications

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“…Following the washing step, 100 μ l sterile deionized water was flowed through the channel that contained the biofilm at 20 dyn cm −2 in forward and reverse direction to remove the attached biofilm as described by Samarian et al . ().…”

Section: Methodsmentioning

confidence: 97%

“…CFS was used as a natural nutrient source to mimic in vivo conditions inside the microfluidic biofilm model (Nance et al 2013;Samarian et al 2014). For this, around 30-ml stimulated saliva was collected from the same donors.…”

Section: Patient Samplingmentioning

confidence: 99%

“…Given the bacterial diversity of human oral biofilms, it would conceivably be preferable to use natural inocula to facilitate the development of in vitro biofilms to more broadly represent the in vivo community (Burmolle et al 2014;Kistler et al 2015). Recently, we developed an in vitro microfluidic oral biofilm system that uses filter-sterilized 25% pooled human saliva as the medium and pooled human saliva as the inoculum (Nance et al 2013;Samarian et al 2014;Kolderman et al 2015). Using confocal laser scanning microscopy (CLSM), multispecies biofilms were shown to be architecturally complex and containing predominantly viable cells.…”

Section: Introductionmentioning

confidence: 99%

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The effect of inoculum source and fluid shear force on the development ofin vitrooral multispecies biofilms

Fernández

Aspiras²,

Dodds³

et al. 2017

J Appl Microbiol

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Section: Methodsmentioning

confidence: 97%

Section: Patient Samplingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The effect of inoculum source and fluid shear force on the development ofin vitrooral multispecies biofilms

Fernández

Aspiras²,

Dodds³

et al. 2017

J Appl Microbiol

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“…This protocol has been used previously (Nance et al, 2013; Samarian et al, 2014). Briefly, for the collection of pooled human whole saliva, the protocol required that at least six healthy individuals donate saliva.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2015

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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.

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“…Many models have been developed in an attempt to simulate the oral cavity [55,[60][61][62]. Innovative strategies have been incorporated, such as using a salivary medium and/or teeth as the substratum, to more closely simulate the in vivo environment.…”

Section: Oral Microcosmmentioning

confidence: 99%

Biofilm Models of Polymicrobial Infection

2015

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Interactions between microbes are complex and play an important role in the pathogenesis of infections. These interactions can range from fierce competition for nutrients and niches to highly evolved cooperative mechanisms between different species that support their mutual growth. An increasing appreciation for these interactions, and desire to uncover the mechanisms that govern them, has resulted in a shift from monomicrobial to polymicrobial biofilm studies in different disease models. Here we provide an overview of biofilm models used to study select polymicrobial infections and highlight the impact that the interactions between microbes within these biofilms have on disease progression. Notable recent advances in the development of polymicrobial biofilm-associated infection models and challenges facing the study of polymicrobial biofilms are addressed. Polymicrobial interactions in biofilmsOver the past 2 decades there has been a revolutionary paradigm shift in the field of microbiology with the appreciation that bacteria present in most biological systems exist in biofilms, rather than in a free-living state. This understanding has dramatically changed the way we study bacteria in the laboratory and resulted in the development of many new experimental systems that replicate biofilm environments [1][2][3][4][5]. Studies utilizing these systems have demonstrated time and time again that bacteria behave very differently when in a biofilm than during planktonic growth. In many ways, we now have to relearn everything we thought we knew about bacterial behavior through the view of the biofilm lens.Similarly, the recent explosion of metagenomic studies has significantly increased our appreciation of the complexity of the microbial populations present in biofilms [6,7]. This is also true for infections, most of which are thought to be biofilm related and inherently polymicrobial, including various species of bacteria, fungi and viruses [8]. It is thought that microbes act in concert to establish biofilms, which in turn can increase tolerance to antimicrobials, exacerbate of the host's immune response and increase persistence at the infection site [7,[9][10][11].Genetic diversity of microbes within biofilm communities is thought to increase the fitness of the residing community, making them more equipped to survive environmental stresses. In large part, this is due to an expanded gene pool, which can be more easily shared within the confines of a biofilm community [12]. Community composition and interactions within the community can have huge influences on bacterial behavior. Thus, just as the behavior of planktonic versus biofilm-associated bacteria is dramatically different, so is that of bacteria in single species versus multispecies systems.Interactions between microbes are complex and highly dependent on context. They can range from fierce competition for nutrients and niches, manifested by antagonistic behavior, to highly evolved cooperative mechanisms between different species that support their mutual grow...

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Use of a High-throughput <em>In Vitro</em> Microfluidic System to Develop Oral Multi-species Biofilms

Cited by 27 publications

References 37 publications

The effect of inoculum source and fluid shear force on the development ofin vitrooral multispecies biofilms

The effect of inoculum source and fluid shear force on the development ofin vitrooral multispecies biofilms

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

Biofilm Models of Polymicrobial Infection

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