Appl Microbiol Biotechnol DOI 10.1007/s00253-014-5841-1
METHODS AND PROTOCOLS
Development of an oral mucosa model to study host-microbiome interactions during wound healing Tine De Ryck & Charlotte Grootaert & Laura Jaspaert & Frederiek-Maarten Kerckhof & Mireille Van Gele & Joachim De Schrijver & Pieter Van den Abbeele & Simon Swift & Marc Bracke & Tom Van de Wiele & Barbara Vanhoecke
Received: 30 December 2013 / Revised: 14 May 2014 / Accepted: 16 May 2014 # Springer-Verlag Berlin Heidelberg 2014
Abstract Crosstalk between the human host and its microbiota is reported to influence various diseases such as mucositis. Fundamental research in this area is however complicated by the time frame restrictions during which host-microbe interactions can be studied in vitro. The model proposed in this paper, consisting of an oral epithelium and biofilm, can be used to study microbe-host crosstalk in vitro in non-infectious conditions up to 72 h. Microbiota derived from oral swabs were cultured on an agar/mucin layer and challenged with monolayers of keratinocytes grown on plastic or collagen type I layers embedded with fibroblasts. The overall microbial biofilm composition in terms of diversity remained
representative for the oral microbiome, whilst the epithelial cell morphology and viability were unaffected. Applying the model to investigate wound healing revealed a reduced healing of 30 % in the presence of microbiota, which was not caused by a reduction of the proliferation index (52.1– 61.5) or a significantly increased number of apoptotic (1– 1.13) or necrotic (32–30.5 %) cells. Since the model allows the separate study of the microbial and cellular exometabolome, the biofilm and epithelial characteristics after co-culturing, it is applicable for investigations within fundamental research and for the discovery and development of agents that promote wound healing.
Electronic supplementary material The online version of this article (doi:10.1007/s00253-014-5841-1) contains supplementary material, which is available to authorized users.
Keywords Biofilm . Epithelium . Co-culture . In vitro model . Oral mucosa . Wound healing
T. De Ryck (*) : M. Bracke : B. Vanhoecke Laboratory of Experimental Cancer Research (LECR), Ghent University, De Pintelaan 185 1P7, B-9000 Ghent, Belgium e-mail: [email protected] C. Grootaert : L. Jaspaert : F.
METHODS AND PROTOCOLS
Development of an oral mucosa model to study host-microbiome interactions during wound healing Tine De Ryck & Charlotte Grootaert & Laura Jaspaert & Frederiek-Maarten Kerckhof & Mireille Van Gele & Joachim De Schrijver & Pieter Van den Abbeele & Simon Swift & Marc Bracke & Tom Van de Wiele & Barbara Vanhoecke
Received: 30 December 2013 / Revised: 14 May 2014 / Accepted: 16 May 2014 # Springer-Verlag Berlin Heidelberg 2014
Abstract Crosstalk between the human host and its microbiota is reported to influence various diseases such as mucositis. Fundamental research in this area is however complicated by the time frame restrictions during which host-microbe interactions can be studied in vitro. The model proposed in this paper, consisting of an oral epithelium and biofilm, can be used to study microbe-host crosstalk in vitro in non-infectious conditions up to 72 h. Microbiota derived from oral swabs were cultured on an agar/mucin layer and challenged with monolayers of keratinocytes grown on plastic or collagen type I layers embedded with fibroblasts. The overall microbial biofilm composition in terms of diversity remained
representative for the oral microbiome, whilst the epithelial cell morphology and viability were unaffected. Applying the model to investigate wound healing revealed a reduced healing of 30 % in the presence of microbiota, which was not caused by a reduction of the proliferation index (52.1– 61.5) or a significantly increased number of apoptotic (1– 1.13) or necrotic (32–30.5 %) cells. Since the model allows the separate study of the microbial and cellular exometabolome, the biofilm and epithelial characteristics after co-culturing, it is applicable for investigations within fundamental research and for the discovery and development of agents that promote wound healing.
Electronic supplementary material The online version of this article (doi:10.1007/s00253-014-5841-1) contains supplementary material, which is available to authorized users.
Keywords Biofilm . Epithelium . Co-culture . In vitro model . Oral mucosa . Wound healing
T. De Ryck (*) : M. Bracke : B. Vanhoecke Laboratory of Experimental Cancer Research (LECR), Ghent University, De Pintelaan 185 1P7, B-9000 Ghent, Belgium e-mail: [email protected] C. Grootaert : L. Jaspaert : F.
