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Development of an in vitro system to study oral biofilms in real time through impedance technology: validation and potential applications.

ABSTRACT: Background and objectives: We have developed a standardized, easy-to-use in vitro model to study single- and multiple-species oral biofilms in real time through impedance technology, which elucidates the kinetics of biofilm formation in 96-well plates, without the requirement for any further manipulation. Design and Results: Using this system, biofilms of Streptococcus mutans appear to be sugar-dependent and highly resistant to amoxicilin, an antibiotic to which this oral pathogen is highly sensitive in a planktonic state. Saliva, tongue and dental plaque samples were also used as inocula to form multiple-species biofilms. DNA isolation and Illumina sequencing of the biofilms showed that the multi-species biofilms were formed by tens or hundreds of species, had a similar composition to the original inoculum, and included fastidious microorganisms which are important for oral health and disease. As an example of the potential applications of the model, we show that oral biofilms can be inhibited by amoxicilin, but in some cases they are induced by the antibiotic, suggesting the existence of responders and non-responders to a given antibiotic. Conclusions: We therefore propose the system as a valid in vitro model to study oral biofilm dynamics, including their susceptibility to antibiotics, antiseptics or anti-adhesive compounds.

SUBMITTER: Mira A

PROVIDER: S-EPMC6507917 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Bacteria found in diverse ecosystems grow in a community of aggregated cells that favors their survival and colonization. Different extracellular polymeric substances are used to entrap this multispecies community forming a biofilm, which can be associated to biotic and abiotic surfaces. This widespread and successful way of bacterial life, however, can lead to negative effects for human activity since many pathogen and spoiling bacteria form biofilms which are not easy to eradicate. Therefore, the search for novel anti-biofilm bio-active molecules is a very active research area for which simple, reliable, and fast screening methods are demanded. In this work we have successfully validated an impedance-based method, initially developed for the study of adherent eukaryotic cells, to monitor the formation of single-species biofilms of three model bacteria in real time. The xCelligence real time cell analyzer (RTCA) equipment uses specific microtiter E-plates coated with gold-microelectrodes that detect the attachment of adherent cells, thus modifying the impedance signal. In the current study, this technology allowed the distinction between biofilm-producers and non-producers of Staphylococcus aureus and Staphylococcus epidermidis, as well as the formation of Streptococcus mutans biofilms only when sucrose was present in the culture medium. Besides, different impedance values permitted discrimination among the biofilm-producing strains tested regardless of the nature of the polymeric biofilm matrix. Finally, we have continuously monitored the inhibition of staphylococcal biofilm formation by the bacteriophage phi-IPLA7 and the bacteriophage-encoded endolysin LysH5, as well as the removal of a preformed biofilm by this last antimicrobial treatment. Results observed with the impedance-based method showed high correlation with those obtained with standard approaches, such as crystal violet staining and bacteria enumeration, as well as with those obtained upon other abiotic surfaces (polystyrene and stainless steel). Therefore, this RTCA technology opens new opportunities in the biofilm research arena and its application could be further explored for other bacterial genera as well as for different bio-active molecules.

Project description:The study objective was to evaluate whether a novel global position system (GPS)-like position-sensing technology will enable accurate co-registration of images between imaging modalities. Co-registration of images obtained by different imaging modalities will allow for comparison and fusion between imaging modalities, and therefore has significant clinical and research implications. We compared ultrasound (US) and magnetic resonance imaging (MRI) scans of carotid endarterectomy (CEA) specimens using a novel position-sensing technology that uses an electromagnetic (EM) transmitter and sensors mounted on a US transducer. We then evaluated in vivo US-US and US-MRI co-registration.Thirteen CEA specimens underwent 3.0 Tesla MRI, after which images were uploaded to a LOGIQ E9 3D (GE Healthcare, Wauwatosa, WI) US system and registered by identifying two to three common points. A similar method was used to evaluate US-MRI co-registration in patients with carotid atherosclerosis. For carotid intima-media thickness (C-IMT) measurements, 10 volunteers underwent bilateral carotid US scans co-registered to three-dimensional US maps created on the initial visit, with a repeat scan 2 days later.For the CEA specimens, there was a mean of 20 (standard error [SE] 2.0) frames per MRI slice. The mean frame difference, over 33 registration markers, between MRI and US scans for readers 1 and 2 was -2.82 ± 19.32 and 2.09 ± 14.68 (mean ± 95% CI) frames, respectively. The US-MRI intraclass correlation coefficients (ICCs) for the first and second readers were 0.995 and 0.997, respectively. For patients with carotid atherosclerosis, the mean US frames per MRI slice (9 [SE 2.3]) was within range of that observed with CEA specimens. Inter-visit, intra-reader, and inter-reader reproducibility of C-IMT measurements were consistently high (side-averaged ICC >0.9).Accurate co-registration between US and other modalities is feasible with a GPS-like technology, which has significant clinical and research applicability.

Project description:Study objectivesReal-time monitoring of upper airway collapse during sleep could be instrumental for studies in biomechanics of obstructive sleep apnea (OSA) and selecting individualized treatment modalities. Although some imaging techniques are used under sedated sleep, none are available during the entire natural sleep process. We hypothesized that electrical impedance tomography (EIT) can be used for noninvasive continuous imaging of the upper airway during natural sleep and quantifying upper airway collapse in terms of its size.MethodsAfter determining surface landmarks to attach the electrodes for monitoring the retroglossal airway, EIT was conducted in 10 healthy participants. As a feasibility test of EIT in detecting upper airway collapse, transient airway closure was induced by the swallowing maneuver. These EIT images were confirmed by simultaneous magnetic resonance imaging (MRI) scans. Subsequently, EIT scans were conducted in 7 healthy participants and 10 patients with OSA under nonsedated sleep to determine whether it could identify upper airway narrowing or collapse. Respiratory events were identified by concurrent polysomnography (PSG).ResultsSwallowing-induced airway closure was identified successfully in all 10 participants on simultaneous EIT and MRI scans. Sizes and positions of the upper airway closures in reconstructed EIT images were well correlated with those in magnetic resonance images. Obstructive hypopnea and apnea were detected successfully by EIT in 10 patients with OSA, and no significant changes in EIT data were observed in 7 healthy participants during concurrent EIT and PSG tests. Additionally, conductivity changes in the airway were greater during obstructive apnea than during hypopnea (64.3% versus 26.3%, respectively; P < .001) compared with those during baseline respiration.ConclusionsEIT could be a useful real-time monitoring device for detecting upper airway narrowing or collapse during natural sleep in patients with OSA. Currently, changes in the upper airway size can be estimated with good accuracy, but shape estimation needs future improvements in the EIT image quality.

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Development of an in vitro system to study oral biofilms in real time through impedance technology: validation and potential applications.

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