Analyses of Gingival Adhesion Molecules in Periodontitis: Theoretical In Silico, Comparative In Vivo, and Explanatory In Vitro Models

Gürsoy, Ulvi Kahraman; Zeidán-Chuliá, Fares; Yılmaz, Doğukan; Özdemir, Vural; Mäki-Petäys, Juho; Oliveira, Ben-Hur Neves de; Firatli, Yigit; Güncü, Güliz N.; Çağlayan, Feriha; Könönen, Eija

doi:10.1902/jop.2015.150361

Cited by 10 publications

(8 citation statements)

References 35 publications

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“…Moreover, the proliferation and differentiation of keratinocytes to form multilayers and the synthesis of keratins have been found to be dependent on cell number density (Ryle et al 1989). By combining this information, our group recently demonstrated that gingival keratinocytes can survive, form monolayers, and produce adhesion proteins at least for 24 h when they are cultured at high densities on semi-permeable nitrocellulose membranes at a floating position on a semi-solid growth medium (Gürsoy et al 2016). In the present study, we hypothesized that keratinocyte stratification can be accelerated by culturing them at high cell densities at an air-liquid-solid interface, omitting the submerged growing and air lifting steps of common multilayer cell culture techniques.…”

Section: Introductionmentioning

confidence: 93%

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Gürsoy

Könönen

et al. 2016

Cytotechnology

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In construction of epithelial cells as multilayers, the cells are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted to air to promote their stratification. We recently demonstrated that gingival epithelial cells form uniform monolayers on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows the cells to grow at an airliquid-solid interface from the beginning of the culturing protocol. In this study, the aim was to further develop our previous model to form a multilayered gingival epithelial culture model. Two different epithelial cell lines (HaCaT from skin and HMK from gingiva) were used in all experiments. Both cell lines were grown first as monolayers for 3 days. After that, keratinocytes were trypsinized, counted and seeded on a sterile semi-permeable nitrocellulose membrane placed on the top of a semi-solid growth medium, forming an air-liquid-solid interface for the cells to grow. At days 1, 4, and 7, epithelial cells were fixed, embedded in paraffin, and sectioned for routine Hematoxylin-Eosin staining and immunohistochemistry for cytokeratin (Ck). At day 1, HMK cells grew as monolayers, while HaCaT cells stratified forming an epithelium with two to three layers. At day 4, a stratified epithelium in the HMK model had four to five layers and its proliferation continued up to day 7. HaCaT cells formed a dense and weakly proliferating epithelium with three to four layers of stratification at day 4 but the proliferation disappeared at day 7. At all days, both models were strongly positive for Ck5, Ck7, and Ck 19, and weakly positive for Ck10. Gingival epithelial cells stratify successfully on semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium. This technique allows researchers to construct uniform gingival epithelial cell multilayers at an air-liquid-solid interface, without using collagen gels, resulting in a more reproducible method.

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Section: Introductionmentioning

confidence: 93%

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Gürsoy

Könönen

et al. 2016

Cytotechnology

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“…The more processes are integrated in the form of subnetworks in the interactome, the more accurate will our in silico model be to reflect the molecular pathogenesis of periodontal inflammation. An alternative possibility is, however, that other markers could also act as central members over the threshold upon the integration of additional subnetworks in the model, representing deregulated biological processes in periodontitis (e.g., epithelial cell adhesion; Haapasalmi et al, 1995 ; Gürsoy et al, 2015 ). These newly developed subnetworks might also be considered in the future as part of a systems biology approach utilized in different fields of medicine such as cancer, neurodegenerative, and psychiatric diseases (Rosado et al, 2011 ; Santana-Codina et al, 2013 ; ElRakaiby et al, 2014 ; Podder and Latha, 2014 ; Zeidán-Chuliá et al, 2014b ; Ebhardt et al, 2015 ).…”

Section: Discussionmentioning

confidence: 99%

A Systems Biology Approach to Reveal Putative Host-Derived Biomarkers of Periodontitis by Network Topology Characterization of MMP-REDOX/NO and Apoptosis Integrated Pathways

Zeidán-Chuliá

Gürsoy

Oliveira

et al. 2016

Front. Cell. Infect. Microbiol.

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Periodontitis, a formidable global health burden, is a common chronic disease that destroys tooth-supporting tissues. Biomarkers of the early phase of this progressive disease are of utmost importance for global health. In this context, saliva represents a non-invasive biosample. By using systems biology tools, we aimed to (1) identify an integrated interactome between matrix metalloproteinase (MMP)-REDOX/nitric oxide (NO) and apoptosis upstream pathways of periodontal inflammation, and (2) characterize the attendant topological network properties to uncover putative biomarkers to be tested in saliva from patients with periodontitis. Hence, we first generated a protein-protein network model of interactions (“BIOMARK” interactome) by using the STRING 10 database, a search tool for the retrieval of interacting genes/proteins, with “Experiments” and “Databases” as input options and a confidence score of 0.400. Second, we determined the centrality values (closeness, stress, degree or connectivity, and betweenness) for the “BIOMARK” members by using the Cytoscape software. We found Ubiquitin C (UBC), Jun proto-oncogene (JUN), and matrix metalloproteinase-14 (MMP14) as the most central hub- and non-hub-bottlenecks among the 211 genes/proteins of the whole interactome. We conclude that UBC, JUN, and MMP14 are likely an optimal candidate group of host-derived biomarkers, in combination with oral pathogenic bacteria-derived proteins, for detecting periodontitis at its early phase by using salivary samples from patients. These findings therefore have broader relevance for systems medicine in global health as well.

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“… 9 The network of adhesion molecules in protein–protein interactions in gingival epithelial cells provides a clear picture for understanding the regulation of TJ proteins in periodontitis. 10 …”

Section: Introductionmentioning

confidence: 99%

Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate

et al. 2018

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Tight junctions (TJs) are the most apical intercellular junctions of epithelial cells formed by occludin, claudins, junctional adhesion molecules (JAMs), and zonula occludens (ZO). Tight junction proteins can sense the presence of bacteria and regulate the transcription of target genes that encode effectors and regulators of the immune response. The aim of this study was to determine the impact of TJ proteins in response to Porphyromonas gingivalis (P. gingivalis), P. gingivalis lipopolysaccharide (P. gingivalis LPS), and extracellular adenosine triphosphate (ATP) in the oral epithelial cell culture model. Quantified real time-polymerase chain reaction (RT-PCR), immunoblots, and immunostaining were performed to assess the gene and protein expression in TJs. It was found that P. gingivalis infection led to transient upregulation of the genes encoding occludin, claudin-1, and claudin-4 but not JAM-A, claudin-15, or ZO-1, while P. gingivalis LPS increased claudin-1, claudin-15, and ZO-1 and decreased occludin, JAM-A, and claudin-4. Tight junction proteins showed significant upregulation in the above two groups when cells were pretreated with ATP for 3 h. The findings indicated that P. gingivalis induced the host defence responses at an early stage. P. gingivalis LPS exerted a more powerful stimulatory effect on the disruption of the epithelial barrier than P. gingivalis. ATP stimulation enhanced the reaction of TJ proteins to P. gingivalis invasion and LPS destruction of the epithelium.

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Analyses of Gingival Adhesion Molecules in Periodontitis: Theoretical In Silico, Comparative In Vivo, and Explanatory In Vitro Models

Cited by 10 publications

References 35 publications

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

A Systems Biology Approach to Reveal Putative Host-Derived Biomarkers of Periodontitis by Network Topology Characterization of MMP-REDOX/NO and Apoptosis Integrated Pathways

Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate

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