Characterisation and optimisation of organotypic oral mucosal models to study Porphyromonas gingivalis invasion.
Porphyromonas gingivalis is a Gram-negative, keystone pathogen in periodontitis that leads to tissue destruction and ultimately tooth loss. The organism is able to infect oral epithelial cells and two-dimensional (monolayer) cultures have been used to investigate this process. However, recently there has been interest in the use of three-dimensional, organotypic mucosal models to analyse infection. These models are composed of collagen-embedded fibroblasts overlain with multilayers of oral epithelial cells. In this study we report for the first time significant differences in the response of oral mucosal models to P. gingivalis infection when compared to monolayer cultures of oral epithelial cells. Intracellular survival (3-fold) and bacterial release (4-fold) of P. gingivalis was significantly increased in mucosal models compared with monolayer cultures, which may be due to the multi-layered nature and exfoliation of epithelial cells in these organotypic models. Furthermore, marked differences in the cytokine profile between infected organotypic models and monolayer cultures were observed, particularly for CXCL8 and IL6, which suggested that degradation of cytokines by P. gingivalis may be less pronounced in organotypic compared to monolayer cultures. These data suggest that use of oral mucosal models may provide a greater understanding of the host responses to P. gingivalis invasion than simple monolayer cultures.
Integrated Biosciences, University of Sheffield, School of Clinical Dentistry, 19 Claremont Crescent, Sheffield S10 2TA, UK.
Abigail
A
Pinnock
Integrated Biosciences, University of Sheffield, School of Clinical Dentistry, 19 Claremont Crescent, Sheffield S10 2TA, UK.
Craig
C
Murdoch
Integrated Biosciences, University of Sheffield, School of Clinical Dentistry, 19 Claremont Crescent, Sheffield S10 2TA, UK.
Keyvan
K
Moharamzadeh
Integrated Biosciences, University of Sheffield, School of Clinical Dentistry, 19 Claremont Crescent, Sheffield S10 2TA, UK.
Simon
S
Whawell
Integrated Biosciences, University of Sheffield, School of Clinical Dentistry, 19 Claremont Crescent, Sheffield S10 2TA, UK. Electronic address: i.douglas@sheffield.ac.uk.
C W Ian
CW
Douglas
D000818:Animals; D016207:Cytokines / Q000378:metabolism; D050296:Microbial Viability; D008962:Models, Theoretical; D009061:Mouth Mucosa / Q000382:microbiology*; D009924:Organ Culture Techniques / Q000379:methods; D016966:Porphyromonas gingivalis / Q000254:growth & development / Q000276:immunology / Q000502:physiology*; D017208:Rats, Wistar
D016207:Cytokines
Chemokines; Epithelial cells; Innate immunity; Oral mucosa; Periodontitis; Porphyromonas gingivalis