IADR Abstract Archives
An in Vitro co-Culture Model System for Investigating Host-Pathogen Interactions
Objectives: The aim of this study was to investigate the effects of disease-state biofilm models on host cells following therapeutic interventions using in vitro organotypic co-culture systems.
Methods: A complex multi-species model representative of gingival inflammation (e.g., gingivitis) was used for the biofilm studies. Once mature, these biofilms were treated bi-daily with toothpaste alongside mechanical brushing, to mimic our recommended oral healthcare regimes. Following treatment, biofilm cells were exposed to oral keratinocyte cell lines and levels of inflammation were assessed using gene and protein expression profiling with qPCR and ELISAs, respectively. Host cells were also imaged with immunofluorescence.
Results: Bi-daily toothpaste treatment with mechanical debridement of the biofilm model systems reduced bioburden and cellular viability of the microbial consortia, compared to untreated controls. Consequently, biofilm treatment influenced the host response: repeat interventions had greater impact on reducing levels of inflammation in the oral keratinocytes, at a transcriptional and protein level. Gene expression and protein release of interleukin-8, a pro-inflammatory biomarker of the oral cavity, was increased following exposure of the cells to untreated biofilm cells. Such an inflammatory effect was alleviated with appropriate treatment regimes.
Conclusions: The use of in vitro models for investigating host-pathogen interactions are essential for testing of novel and conventional therapeutics in oral healthcare. Here we have demonstrated the use of a co-culture organotypic system for simulating bi-daily toothpaste and brushing interventions on a complex biofilm model.
Methods: A complex multi-species model representative of gingival inflammation (e.g., gingivitis) was used for the biofilm studies. Once mature, these biofilms were treated bi-daily with toothpaste alongside mechanical brushing, to mimic our recommended oral healthcare regimes. Following treatment, biofilm cells were exposed to oral keratinocyte cell lines and levels of inflammation were assessed using gene and protein expression profiling with qPCR and ELISAs, respectively. Host cells were also imaged with immunofluorescence.
Results: Bi-daily toothpaste treatment with mechanical debridement of the biofilm model systems reduced bioburden and cellular viability of the microbial consortia, compared to untreated controls. Consequently, biofilm treatment influenced the host response: repeat interventions had greater impact on reducing levels of inflammation in the oral keratinocytes, at a transcriptional and protein level. Gene expression and protein release of interleukin-8, a pro-inflammatory biomarker of the oral cavity, was increased following exposure of the cells to untreated biofilm cells. Such an inflammatory effect was alleviated with appropriate treatment regimes.
Conclusions: The use of in vitro models for investigating host-pathogen interactions are essential for testing of novel and conventional therapeutics in oral healthcare. Here we have demonstrated the use of a co-culture organotypic system for simulating bi-daily toothpaste and brushing interventions on a complex biofilm model.