IADR Abstract Archives
Structure and Function of a 3D Gingivae-biofilm Interface Model
Objectives: Oral diseases such as gingivitis and periodontitis arise through a dysregulated immune response to microbial challenge. Here we combined a 3D model of gingival epithelium with a complex biofilm, to represent the periodontal pocket to study the bacterial-host cell interactions.
Methods: Human gingival epithelium was cultivated on an inert polycarbonate filter, and formed 6-8 layers, which histologically demonstrated characteristics of epithelium. In vitro grown ‘health-associated’, gingivitis-associated’ and ‘periodontitis-associated’ bacterial biofilms ± toothpaste treatment were co-cultured with the epithelium to investigate the host response to different biofilms. Following overnight exposure to oral biofilms, epithelial cell gene expression, protein release and viability was assessed. The morphology and integrity of the epithelium was evaluated by immunohistochemistry; the associated biofilms were assessed by confocal laser scanning microscope (CLSM) imaging.
Results: The ‘health-associated’ biofilm had minimal impact on the epithelium. Following co-culture with ‘gingivitis-associated’ biofilm, interleukin-8 (IL-8) release and IL8 gene expression of the tissue were significantly increased. The epithelial cell response to the ‘periodontitis-associated’ biofilm was intermediate between that of the ‘health-associated’ and ‘gingivitis-associated’ biofilms. All epithelial cells maintained good viability. Haematoxylin & Eosin stained sections of the epithelium revealed a greater level of degranulation in the prickle cell layer of the epithelium following incubation with the disease-associated biofilms. CLSM images of live/dead cells showed treatment specific effects on the biofilm.
Conclusions: Our results show 3D epithelial models may offer a substantial advantage for in vitro study of host-pathogen interactions. These tissue models remained viable following biofilm exposure and demonstrated responses that recapitulate those observed in inflammation at the oral mucosa. Such a platform allows in vitro dissection of the impact of biofilm and/or host treatment interventions in these systems, and therefore findings from this 3D model may contribute to our understanding of the pathogenesis and treatment of oral diseases in vivo.
Methods: Human gingival epithelium was cultivated on an inert polycarbonate filter, and formed 6-8 layers, which histologically demonstrated characteristics of epithelium. In vitro grown ‘health-associated’, gingivitis-associated’ and ‘periodontitis-associated’ bacterial biofilms ± toothpaste treatment were co-cultured with the epithelium to investigate the host response to different biofilms. Following overnight exposure to oral biofilms, epithelial cell gene expression, protein release and viability was assessed. The morphology and integrity of the epithelium was evaluated by immunohistochemistry; the associated biofilms were assessed by confocal laser scanning microscope (CLSM) imaging.
Results: The ‘health-associated’ biofilm had minimal impact on the epithelium. Following co-culture with ‘gingivitis-associated’ biofilm, interleukin-8 (IL-8) release and IL8 gene expression of the tissue were significantly increased. The epithelial cell response to the ‘periodontitis-associated’ biofilm was intermediate between that of the ‘health-associated’ and ‘gingivitis-associated’ biofilms. All epithelial cells maintained good viability. Haematoxylin & Eosin stained sections of the epithelium revealed a greater level of degranulation in the prickle cell layer of the epithelium following incubation with the disease-associated biofilms. CLSM images of live/dead cells showed treatment specific effects on the biofilm.
Conclusions: Our results show 3D epithelial models may offer a substantial advantage for in vitro study of host-pathogen interactions. These tissue models remained viable following biofilm exposure and demonstrated responses that recapitulate those observed in inflammation at the oral mucosa. Such a platform allows in vitro dissection of the impact of biofilm and/or host treatment interventions in these systems, and therefore findings from this 3D model may contribute to our understanding of the pathogenesis and treatment of oral diseases in vivo.