scholarly journals Saliva-Derived Commensal and Pathogenic Biofilms in a Human Gingiva Model

2017 ◽  
Vol 97 (2) ◽  
pp. 201-208 ◽  
Author(s):  
J.K. Buskermolen ◽  
M.M. Janus ◽  
S. Roffel ◽  
B.P. Krom ◽  
S. Gibbs
Keyword(s):  
Oral Mucosa ◽  
Bacterial Species ◽  
Human Saliva ◽  
In Vitro Models ◽  
Antimicrobial Protein ◽  
Operational Taxonomic Units ◽  
Oral Biofilms ◽  
Human Gingiva ◽  
Pathogenic Biofilms

In vitro models that closely mimic human host-microbiome interactions can be a powerful screening tool for antimicrobials and will hold great potential for drug validation and discovery. The aim of this study was to develop an organotypic oral mucosa model that could be exposed to in vitro cultured commensal and pathogenic biofilms in a standardized and scalable manner. The oral mucosa model consisted of a tissue-engineered human gingiva equivalent containing a multilayered differentiated gingiva epithelium (keratinocytes) grown on a collagen hydrogel, containing gingiva fibroblasts, which represented the lamina propria. Keratinocyte and fibroblast telomerase reverse transcriptase–immortalized cell lines were used to overcome the limitations of isolating cells from small biopsies when scalable culture experiments were required. The oral biofilms were grown under defined conditions from human saliva to represent 3 distinct phenotypes: commensal, gingivitis, and cariogenic. The in vitro grown biofilms contained physiologic numbers of bacterial species, averaging >70 operational taxonomic units, including 20 differentiating operational taxonomic units. When the biofilms were applied topically to the gingiva equivalents for 24 h, the gingiva epithelium increased its expression of elafin, a protease inhibitor and antimicrobial protein. This increased elafin expression was observed as a response to all 3 biofilm types, commensal as well as pathogenic (gingivitis and cariogenic). Biofilm exposure also increased secretion of the antimicrobial cytokine CCL20 and inflammatory cytokines IL-6, CXCL8, and CCL2 from gingiva equivalents. This inflammatory response was far greater after commensal biofilm exposure than after pathogenic biofilm exposure. These results show that pathogenic oral biofilms have early immune evasion properties as compared with commensal oral biofilms. The novel host-microbiome model provides an ideal tool for future investigations of gingiva responses to commensal and pathogenic biofilms and for testing novel therapeutics.

scholarly journals Human Saliva-Mediated Hydrolysis of Eugenyl-β-D-Glucoside and Fluorescein-di-β-D-Glucoside in In Vivo and In Vitro Models

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 172
Author(s):  
Mariusz Dziadas ◽  
Adam Junka ◽  
Henryk Jeleń
Keyword(s):  
Oral Cavity ◽  
Control Sample ◽  
Rapid Test ◽  
Human Saliva ◽  
In Vitro Models ◽  
Microbial Hydrolysis ◽  
Amoxicillin Trihydrate ◽  
Potassium Clavulanate

Eugenyl-β-D-glucopyranoside, also referred to as Citrusin C, is a natural glucoside found among others in cloves, basil and cinnamon plants. Eugenol in a form of free aglycone is used in perfumeries, flavourings, essential oils and in medicinal products. Synthetic Citrusin C was incubated with human saliva in several in vitro models together with substrate-specific enzyme and antibiotics (clindamycin, ciprofloxacin, amoxicillin trihydrate and potassium clavulanate). Citrusin C was detected using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Citrusin C was completely degraded only when incubated with substrate-specific A. niger glucosidase E.C 3.2.1.21 (control sample) and when incubated with human saliva (tested sample). The addition of antibiotics to the above-described experimental setting, stopped Citrusin C degradation, indicating microbiologic origin of hydrolysis observed. Our results demonstrate that Citrusin C is subjected to complete degradation by salivary/oral cavity microorganisms. Extrapolation of our results allows to state that in the human oral cavity, virtually all β-D-glucosides would follow this type of hydrolysis. Additionally, a new method was developed for an in vivo rapid test of glucosidase activity in the human mouth on the tongue using fluorescein-di-β-D-glucoside as substrate. The results presented in this study serve as a proof of concept for the hypothesis that microbial hydrolysis path of β-D-glucosides begins immediately in the human mouth and releases the aglycone directly into the gastrointestinal tract.

scholarly journals Novel Model for Multispecies Biofilms That Uses Rigid Gas-Permeable Lenses

2011 ◽  
Vol 77 (10) ◽  
pp. 3413-3421 ◽  
Author(s):  
Rebecca Peyyala ◽  
Sreenatha S. Kirakodu ◽  
Jeffrey L. Ebersole ◽  
Karen F. Novak
Keyword(s):  
Laser Scanning ◽  
Bacterial Species ◽  
Disease Process ◽  
Laser Scanning Microscopy ◽  
Host Cells ◽  
Confocal Laser ◽  
Content Type ◽  
Oral Biofilms ◽  
Multispecies Biofilms

ABSTRACTOral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system to study initial stages of interaction between oral biofilms and the host cells that contribute to the disease process, we developed a novelin vitromodel system to grow biofilms on rigid gas-permeable contact lenses (RGPLs), which enable oxygen to permeate through the lens material. Bacterial species belonging to early- and late-colonizing groups were successfully established as single- or three-species biofilms, with each group comprisingStreptococcus gordonii,Streptococcus oralis, andStreptococcus sanguinis;S. gordonii,Actinomyces naeslundii, andFusobacterium nucleatum; orS. gordonii,F. nucleatum, andPorphyromonas gingivalis. Quantification of biofilm numbers by quantitative PCR (qPCR) revealed substantial differences in the magnitude of bacterial numbers in single-species and multispecies biofilms. We evaluated cell-permeable conventional nucleic acid stains acridine orange, hexidium iodide, and Hoechst 33258 and novel SYTO red, blue, and green fluorochromes for their effect on bacterial viability and fluorescence yield to allow visualization of the aggregates of individual bacterial species by confocal laser scanning microscopy (CLSM). Substantial differences in the quantity and distribution of the species in the multispecies biofilms were identified. The specific features of these biofilms may help us better understand the role of various bacteria in local challenge of oral tissues.

Nutritional Influences on Biofilm Development

1997 ◽  
Vol 11 (1) ◽  
pp. 81-99 ◽  
Author(s):  
G.H.W. Bowden ◽  
Y.H. Li
Keyword(s):  
Oral Cavity ◽  
Bacterial Species ◽  
Oral Bacteria ◽  
Carbon Limitation ◽  
Cell Surfaces ◽  
Nutritional Influences ◽  
Oral Biofilms ◽  
Biofilm Development

The amounts and types of nutrients in the environment influence the development and final bacterial and chemical composition of biofilms. In oligotrophic environments, organisms respond to nutrient stress by alterations in their cell morphology and cell surfaces, which enhance adherence. Little is known of the responses to stress by bacteria in the animal oral cavity. The environment in the oral cavity is less extreme, and saliva provides a constant source of nutrients. Catabolic cooperation among oral bacteria allows carbon and nitrogen from salivary glycoproteins to be utilized. Modification of growth environments of oral bacteria can influence their cell surfaces and adhesion. Studies in experimental animals have shown that feeding either glucose or sucrose diets or fasting has little effect on the initial stages of development of oral biofilms. However, diet can influence the proportions of different bacterial species later in biofilm development. Studies of competition among populations in communities of oral bacteria in vitro and in vivo have shown the significance of carbon limitation and excess and changes in environmental pH. Relatively few studies have been made of the role of a nitrogen metabolism in bacterial competition in biofilms. In keeping with biofilms in nature, oral biofilms provide a sequestered habitat, where organisms are protected from removal by saliva and where interactions among cells generate a biofilm environment, distinct from that of saliva. Oral biofilms are an essential component in the etiologies of caries and periodontal disease, and understanding the biology of oral biofilms has aided and will continue to aid in the prevention and treatment of these diseases.

scholarly journals Experimental Models of Oral Biofilms Developed on Inert Substrates: A Review of the Literature

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Lopez-Nguyen Darrene ◽  
Badet Cecile
Keyword(s):  
Biofilm Formation ◽  
Bacterial Species ◽  
Experimental Models ◽  
Pathogenic Microorganisms ◽  
Review Of The Literature ◽  
Complex Environment ◽  
Oral Biofilm ◽  
Mucosal Surfaces ◽  
Oral Biofilms

The oral ecosystem is a very complex environment where more than 700 different bacterial species can be found. Most of them are organized in biofilm on dental and mucosal surfaces. Studying this community is important because a rupture in stability can lead to the preeminence of pathogenic microorganisms, causing dental decay, gingivitis, or periodontitis. The multitude of species complicates biofilm analysis so its reproduction, collection, and counting are very delicate. The development of experimental models of dental biofilms was therefore essential and multiplein vitrodesigns have emerged, each of them especially adapted to observing biofilm formation of specific bacteria within specific environments. The aim of this review is to analyze oral biofilm models.

scholarly journals Role of Lipopolysaccharide, Derived from Various Bacterial Species, in Pulpitis—A Systematic Review

Biomolecules ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 138
Author(s):  
Aniela Brodzikowska ◽  
Monika Ciechanowska ◽  
Michał Kopka ◽  
Albert Stachura ◽  
Paweł K. Włodarski
Keyword(s):  
Dental Pulp ◽  
Bacterial Species ◽  
Cell Types ◽  
In Vitro Models ◽  
Pulp Inflammation ◽  
Pulp Cells ◽  
Specific Receptors ◽  
Meta Analyses ◽  
Different Cell Types

Lipopolysaccharide (LPS) is widely used for induction of inflammation in various human tissues, including dental pulp. The purpose of this study was to summarize current medical literature focusing on (1) cell types used by researchers to simulate dental pulp inflammation, (2) LPS variants utilized in experimental settings and how these choices affect the findings. Our study was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). We searched for studies reporting outcomes of lipopolysaccharide application on dental pulp cells in vitro using electronic databases: MEDLINE, Web of Science and Scopus. Having gathered data from 115 papers, we aimed to present all known effects LPS has on different cell types present in dental pulp. We focused on specific receptors and particles that are involved in molecular pathways. Our review provides an essential foundation for further research using in vitro models of pulpitis.

scholarly journals Precision-guided antimicrobial peptide as a targeted modulator of human microbial ecology

2015 ◽  
Vol 112 (24) ◽  
pp. 7569-7574 ◽  
Author(s):  
Lihong Guo ◽  
Jeffrey S. McLean ◽  
Youngik Yang ◽  
Randal Eckert ◽  
Christopher W. Kaplan ◽  
...  
Keyword(s):  
Antimicrobial Peptide ◽  
Therapeutic Potential ◽  
Bacterial Species ◽  
Recovery Period ◽  
Human Microbiome ◽  
Human Saliva ◽  
Oral Microbiome ◽  
Individual Species ◽  
Significant Shift

One major challenge to studying human microbiome and its associated diseases is the lack of effective tools to achieve targeted modulation of individual species and study its ecological function within multispecies communities. Here, we show that C16G2, a specifically targeted antimicrobial peptide, was able to selectively kill cariogenic pathogenStreptococcus mutanswith high efficacy within a human saliva-derived in vitro oral multispecies community. Importantly, a significant shift in the overall microbial structure of the C16G2-treated community was revealed after a 24-h recovery period: several bacterial species with metabolic dependency or physical interactions withS. mutanssuffered drastic reduction in their abundance, whereasS. mutans’ natural competitors, including health-associated Streptococci, became dominant. This study demonstrates the use of targeted antimicrobials to modulate the microbiome structure allowing insights into the key community role of specific bacterial species and also indicates the therapeutic potential of C16G2 to achieve a healthy oral microbiome.

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