Confocal Raman microscopy to identify bacteria in oral subgingival biofilm models

Kriem, Lukas Simon; Wright, Kevin J.; Ccahuana-Vásquez, Renzo Alberto; Rupp, Steffen

doi:10.1371/journal.pone.0232912

Cited by 24 publications

(21 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We detected various bacterial taxa typically present in the oral (Streptococcus dentisani [ 60 ]; Streptococcus intermedius [ 61 ]; Fusobacterium nucleatum [ 62 ]; Prevotella spp. [ 63 ]) and intestinal microflora (Barnesiella intestinihominis [ 64 ]; Porphyromonadaceae [ 65 ]). In addition, we observed the genus Phenylobacterium, which was reported to be present in the middle ear [ 66 ], putatively being a commensal of the upper aerodigestive tract.…”

Section: Resultsmentioning

confidence: 99%

Neutrophil Extracellular Traps Promote the Development and Growth of Human Salivary Stones

Mantsopoulos

Koch

Weidner

et al. 2020

Cells

View full text Add to dashboard Cite

Salivary gland stones, or sialoliths, are the most common cause of the obstruction of salivary glands. The mechanism behind the formation of sialoliths has been elusive. Symptomatic sialolithiasis has a prevalence of 0.45% in the general population, is characterized by recurrent painful periprandial swelling of the affected gland, and often results in sialadenitis with the need for surgical intervention. Here, we show by the use of immunohistochemistry, immunofluorescence, computed tomography (CT) scans and reconstructions, special dye techniques, bacterial genotyping, and enzyme activity analyses that neutrophil extracellular traps (NETs) initiate the formation and growth of sialoliths in humans. The deposition of neutrophil granulocyte extracellular DNA around small crystals results in the dense aggregation of the latter, and the subsequent mineralization creates alternating layers of dense mineral, which are predominantly calcium salt deposits and DNA. The further agglomeration and appositional growth of these structures promotes the development of macroscopic sialoliths that finally occlude the efferent ducts of the salivary glands, causing clinical symptoms and salivary gland dysfunction. These findings provide an entirely novel insight into the mechanism of sialolithogenesis, in which an immune system-mediated response essentially participates in the physicochemical process of concrement formation and growth.

show abstract

Section: Resultsmentioning

confidence: 99%

Neutrophil Extracellular Traps Promote the Development and Growth of Human Salivary Stones

Mantsopoulos

Koch

Weidner

et al. 2020

Cells

View full text Add to dashboard Cite

show abstract

“…We detected various bacterial taxa typically present in the oral [Streptococcus dentisani 55 ; Streptococcus intermedius 56 ; Fusobacterium nucleatum 57 ; Prevotella spp. 58 ] and intestinal micro ora [Barnesiella intestinihominis 59 ; Porphyromonadaceae 60 ]. In addition, we observed the genus Phenylobacterium, reported to be present in the middle ear 61 , putatively being a commensal of the upper aerodigestive tract.…”

Section: Resultsmentioning

confidence: 99%

Development and Growth of Human Salivary Stones by Neutrophil Extracellular Traps

Mantsopoulos

Koch

Weidner

et al. 2020

Preprint

View full text Add to dashboard Cite

Salivary gland stones, or sialoliths, are the most common cause of the obstruction of salivary glands. Symptomatic sialolithiasis has a prevalence of 0.45% in the general population, is characterized by recurrent painful periprandial swelling of the affected gland and often results in sialadenitis with the need for surgical intervention. The mechanism behind the formation of sialoliths has been elusive. Here we show that neutrophil extracellular traps (NETs) initiate the formation and growth of sialoliths. The deposition of neutrophil granulocyte extracellular DNA around small crystals results in their dense aggregation, and the subsequent mineralization creates alternating layers of dense mineral, predominantly calcium salt deposits and DNA. Further agglomeration and appositional growth of these structures promotes the development of macroscopic sialoliths that finally occlude the efferent ducts of the salivary glands, causing clinical symptoms and salivary gland dysfunction. These findings provide an entirely novel insight into the mechanism of sialolithogenesis in which an immune system mediated response essentially participates in the physico-chemical process of concrement formation and growth.

show abstract

“…In a previous publication by the same authors, a ThermoFisher Scientific DXR2xi instrument was used for acquisition. Renishaw inVia Qontor was used in this research because of its increased specificity and signal count compared to the ThermoFisher Scientific DXR2xi, thus being able to show minor bands (Kriem et al, 2020). The instrument was equipped with a 532 nm filter and a 532 nm laser to capture a spectral range of 282.8-2016.2 cm −1 and 1,015 collection points.…”

Section: Raman Data Acquisition Processing and Microscopic Imagingmentioning

confidence: 99%

“…Bacteria in subgingival biofilms occupy the same habitat and, therefore, show similar chemical compositions. We previously demonstrated that differentiation between species can be successfully achieved using spectral fingerprint patterns and multivariate statistical models for five subgingival species from four different "Socransky complexes" (Socransky et al, 1998;Kriem et al, 2020). Additionally, we successfully determined and differentiated mono-species biofilms based on their spectral fingerprint region.…”

Section: Introductionmentioning

confidence: 99%

“…One of these techniques, confocal Raman microscopy (CRM), measures scattered radiation and energy shifts from samples after they were excited with a laser beam. Previously, we demonstrated the use of CRM to identify different oral microbes in pure culture (Kriem et al, 2020). Key advantages of this technique beyond microbial identification are: (1) very minimal sample preparation, (2) affordable measurements, and (3) the opportunity of continuous evaluation of the same location due to the technology's non-destructive nature of measuring a sample.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Mapping of a Subgingival Dual-Species Biofilm Model Using Confocal Raman Microscopy

et al. 2021

Self Cite

View full text Add to dashboard Cite

Techniques for continuously monitoring the formation of subgingival biofilm, in relation to the determination of species and their accumulation over time in gingivitis and periodontitis, are limited. In recent years, advancements in the field of optical spectroscopic techniques have provided an alternative for analyzing three-dimensional microbiological structures, replacing the traditional destructive or biofilm staining techniques. In this work, we have demonstrated that the use of confocal Raman spectroscopy coupled with multivariate analysis provides an approach to spatially differentiate bacteria in an in vitro model simulating a subgingival dual-species biofilm. The present study establishes a workflow to evaluate and differentiate bacterial species in a dual-species in vitro biofilm model, using confocal Raman microscopy (CRM). Biofilm models of Actinomyces denticolens and Streptococcus oralis were cultured using the “Zürich in vitro model” and were analyzed using CRM. Cluster analysis was used to spatially differentiate and map the biofilm model over a specified area. To confirm the clustering of species in the cultured biofilm, confocal laser scanning microscopy (CLSM) was coupled with fluorescent in vitro hybridization (FISH). Additionally, dense bacteria interface area (DBIA) samples, as an imitation of the clusters in a biofilm, were used to test the developed multivariate differentiation model. This confirmed model was successfully used to differentiate species in a dual-species biofilm and is comparable to morphology. The results show that the developed workflow was able to identify main clusters of bacteria based on spectral “fingerprint region” information from CRM. Using this workflow, we have demonstrated that CRM can spatially analyze two-species in vitro biofilms, therefore providing an alternative technique to map oral multi-species biofilm models.

show abstract

Confocal Raman microscopy to identify bacteria in oral subgingival biofilm models

Cited by 24 publications

References 45 publications

Neutrophil Extracellular Traps Promote the Development and Growth of Human Salivary Stones

Neutrophil Extracellular Traps Promote the Development and Growth of Human Salivary Stones

Development and Growth of Human Salivary Stones by Neutrophil Extracellular Traps

Mapping of a Subgingival Dual-Species Biofilm Model Using Confocal Raman Microscopy

Contact Info

Product

Resources

About