Castillo Romero, YormarisLafaurie, Gloria InesAlfonso Torres, Jennifer TatianaLaverde González, Paula Andrea2020-04-282020-04-282020https://hdl.handle.net/20.500.12495/2431Dental plaque is one of the most complex bio-film systems in nature and the main agent of oral infections such as caries and periodontal disease; hindering the progress of a symbiotic oral bio-film to a dysbiotic state is one of the main research goals. To standardize a periodontal multi-species bio-film model in vitro in order of colonization. Bacterial inocula with strains of Streptococcus sanguinis ATCC 10556, Streptococcus oralis ATCC 35037, Streptococcus mutans ATCC 25175, Actinomyces isrraelii ATCC 12012 Fusobacterium nucleatum ATCC 25586 and Porphyromonas gingivalis ATCC 33277 were adjusted in order to reach a concentration of >1x108UFC/ml. The biofilms were formed in colonization order on layers of silicate glass lacquer. Bacterial growth was confirmed by means of culture on Brucella agar, bacterial viability with the LIVE/DEAD kit BacLight™ and evaluated with epifluorescence microscopy. This was done in order to identify live cells from dead ones calculating the viability percentage with pixel intensity analysis using ImageJ software. Additionally, the formation of biofilm was determined using the violet crystal method. Tri-dimensional conformation of bio-film was confirmed with scanning electron microscopy. All experiments were carried out in triplicate and data presented as descriptive statistics. Bio-films in a pure and viable state were obtained after seven days with micro-organisms initially inoculated into the film. The bio-films formed over layers without saliva presented higher proportion of dead cells (56%) compared with those with live cells (44%) and conformation was more disperse. Bio-films formed over layers conditioned with saliva present greater proportion of live cells and a cluster formation, with less area spread but better film conformation. Oral periodontal multi-species bio-films were formed with six bacteria on layers of glass and lacquer with a viability percentage greater than 50%. Conditioning of lacquer and glass did not favour important changes on the bio-film’s viability.application/pdfspaAttribution-NonCommercial-ShareAlike 4.0 InternationalTesis de doctoradoEnfermedades periodontalesinstname:Universidad El Bosquereponame:Repositorio Institucional Universidad El Bosquerepourl:https://repositorio.unbosque.edu.coAcceso abiertoinfo:eu-repo/semantics/openAccesshttps://purl.org/coar/access_right/c_abf22019