Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions

Dias, Kássia de Carvalho; Sousa, Denise Lins de; Barbugli, Paula Aboud; Cerri, Paulo Sérgio; Salih, Vehid; Vergani, Carlos Eduardo

doi:10.1016/j.mimet.2018.07.004

Cited by 20 publications

(14 citation statements)

References 38 publications

(52 reference statements)

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“…Literature reviews describing work from other groups show that the few studies performed mainly report RHG exposed to a single- or dual-species of oral bacteria for maximally 48 hours ( Dongari-Bagtzoglou and Kashleva, 2006 ; Tabatabaei et al., 2020 ). Most of these reports described a disrupted tissue integrity, decreased viability and increase in apoptosis of host tissues within 48 hours ( Andrian et al., 2004 ; Gursoy et al., 2010 ; de Carvalho Dias et al., 2018 ; Morse et al., 2018 ; Beklen et al., 2019 ). One report described a model where a multi-species biofilm was co-incubated with RHG for 3 days without affecting the viability of RHG ( De Ryck et al., 2014 ).…”

Section: Discussionmentioning

confidence: 99%

Stable reconstructed human gingiva–microbe interaction model: Differential response to commensals and pathogens

Zhang

Shang

Roffel

et al. 2022

Front. Cell. Infect. Microbiol.

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BackgroundTo investigate human oral health and disease, models are required which represent the interactions between the oral mucosa and microbiome. Our aim was to develop an organotypic model which maintains viability of both host and microbes for an extended period of time.MethodsReconstructed Human Gingiva (RHG) were cultured air-lifted with or without penicillin-streptomycin (PS) and topically exposed to Streptococcus gordonii (commensal) or Aggregatibacter actinomycetemcomitans (pathogen) for 72 hours in agar. RHG histology, viability and cytokines (ELISA), and bacterial viability (colony forming units) and location (FISH) were assessed.ResultsThe low concentration of topically applied agar did not influence RHG viability. Topically applied bacteria in agar remained localized and viable for 72 hours and did not spill over to infect RHG culture medium. PS in RHG culture medium killed topically applied bacteria. Co-culture with living bacteria did not influence RHG viability (Ki67 expression, MTT assay) or histology (epithelium differentiation, Keratin10 expression). RHG exposed to S. gordonii (with or without PS) did not influence low level of IL-6, IL-8, CCL2, CCL5, CCL20 or CXCL1 secretion. However, all cytokines increased (except CCL2) when RHG were co-cultured with A. actinomycetemcomitans. The effect was significantly more in the presence of living, rather than dead, A. actinomycetemcomitans. Both bacteria resulted in increased expression of RHG antimicrobial peptides (AMPs) Elafin and HBD-2, with S. gordonii exposure resulting in the most Elafin secretion.ConclusionThis technical advance enables living human oral host–microbe interactions to be investigated during a 72-hour period and shows differences in innate immunology triggered by S. gordonii and A. actinomycetemcomitans.

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Section: Discussionmentioning

confidence: 99%

Stable reconstructed human gingiva–microbe interaction model: Differential response to commensals and pathogens

Zhang

Shang

Roffel

et al. 2022

Front. Cell. Infect. Microbiol.

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“…7,34 Association of C. albicans with other microorganisms, like Staphylococcus aureus or S. oralis, could result in deeper invasion into subepithelial collagen matrix. 12,42 Hyphal transformation was not detectable in C. famata; however, its penetration to the lamina propria of the oral mucosa model was reported after 24 h of infection. 41 Invasion of F. nucleatum to collagen matrix is also strain dependent and is enhanced in the biofilm form of F. nucleatum compared to the planktonic form.…”

Section: Survival and Penetration Of Microorganism In Oral Mucosa Modelmentioning

confidence: 95%

“…For this purpose, many researchers use different types of oral mucosa equivalents as a relevant in vitro tool to investigate the interaction of microorganisms with oral mucosa, the process of epithelial layer's damage, and initial steps of infection, as well as treatment approaches. 12 Isolation and expansion of epithelial and fibroblast cells from gingiva, buccal or palatal mucosa, seeding and culture of fibroblast in a suitable substrate, and finally, seeding of epithelial cells onto the engineered connective tissue layer is a common procedure for engineering of oral mucosa models. There are also commercially available oral mucosa models, which can be used for microbiological studies.…”

Section: Introductionmentioning

confidence: 99%

Three-Dimensional In Vitro Oral Mucosa Models of Fungal and Bacterial Infections

Tabatabaei

Moharamzadeh

Tayebi

2020

Tissue Engineering Part B: Reviews

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Oral mucosa is the target tissue for many microorganisms involved in periodontitis and other infectious diseases affecting the oral cavity. Three-dimensional (3D) in vitro and ex vivo oral mucosa equivalents have been used for oral disease modeling and investigation of the mechanisms of oral bacterial and fungal infections. This review was conducted to analyze different studies using 3D oral mucosa models for the evaluation of the interactions of different microorganisms with oral mucosa. In this study, based on our inclusion criteria, 43 articles were selected and analyzed. Different types of 3D oral mucosa models of bacterial and fungal infections were discussed in terms of the biological system used, culture conditions, method of infection, and the biological endpoints assessed in each study. The critical analysis revealed some contradictory reports in this field of research in the literature. Challenges in recovering bacteria from oral mucosa models were further discussed, suggesting possible future directions in microbiomics, including the use of oral mucosa-on-a-chip. The potential use of these 3D tissue models for the evaluation of the effects of antiseptic agents on bacteria and oral mucosa was also addressed. This review concluded that there were many aspects that would require optimization and standardization with regard to using oral mucosal models for infection by microorganisms. Using new technologies-such as microfluidics and bioreactors-could help to reproduce some of the physiologically relevant conditions and further simulate the clinical situation.

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“…However, based on the experience of the cosmetic industry, in vitro-cultured mucosa and skin equivalents may offer a similar condition to in vivo tests with a highly controlled environment, lower cost and rapid procedures (Schmalz & Arenholt-Bindslev, 2009). These equivalents have an interesting perspective, but experiences with dental materials remain very limited (Carvalho et al, 2018;Klausner et al, 2021;Tabatabaei et al, 2020).…”

Section: Implantation Testsmentioning

confidence: 99%

A critical analysis of research methods and experimental models to study biocompatibility of endodontic materials

2022

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Materials used for endodontics and with direct contact to tissues have a wide range of indications, from vital pulpal treatments to root filling materials and those used in endodontic surgery. In principle, interaction with dental materials may result in damage to tissues locally or systemically. Thus, a great variety of test methods are applied to evaluate a materials' potential risk of adverse biological effects to ensure their biocompatibility before commercialization. However, the results of biocompatibility evaluations are dependent on not only the tested materials but also the test methods due to the diversity of these effects and numerous variables involved. In addition, diverse biological effects require equally diverse assessments on a structured and planned approach. Such a structured assessment of the materials consists of four phases: general toxicity, local tissue irritation, pre‐clinical tests and clinical evaluations. Various types of screening assays are available; it is imperative to understand their advantages and limitations to recognize their appropriateness and for an accurate interpretation of their results. Recent scientific advances are rapidly introducing new materials to endodontics including nanomaterials, gene therapy and tissue engineering biomaterials. These new modalities open a new era to restore and regenerate dental tissues; however, all these new technologies can also present new hazards to patients. Before any clinical usage, new materials must be proven to be safe and not hazardous to health. Certain international standards exist for safety evaluation of dental materials (ISO 10993 series, ISO 7405 and ISO 14155‐1), but researchers often fail to follow these standards due to lack of access to standards, limitation of the guidelines and complexity of new experimental methods, which may cause technical errors. Moreover, many laboratories have developed their testing strategy for biocompatibility, which makes any comparison between findings more difficult. The purpose of this review was to discuss the concept of biocompatibility, structured test programmes and international standards for testing the biocompatibility of endodontic material biocompatibility. The text will further detail current test methods for evaluating the biocompatibility of endodontic materials, and their advantages and limitations.

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Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions

Cited by 20 publications

References 38 publications

Stable reconstructed human gingiva–microbe interaction model: Differential response to commensals and pathogens

Stable reconstructed human gingiva–microbe interaction model: Differential response to commensals and pathogens

Three-Dimensional In Vitro Oral Mucosa Models of Fungal and Bacterial Infections

A critical analysis of research methods and experimental models to study biocompatibility of endodontic materials

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