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Stimulatory effect of Aggregatibacter actinomycetemcomitans DNA on proinflammatory cytokine expression by human gingival fibroblasts
Affiliation:1. Doctorado en Ciencias Biomédicas Básicas, Universidad Autónoma de San Luis Potosí, San Luis Potosi, 78290, Mexico;2. Doctorado Institucional de Ingeniería y Ciencia de los Materiales, Universidad Autónoma de San Luis Potosi, San Luis Potosi, 78290, Mexico;3. Facultad de Odontología, Universidad Autónoma de Chihuahua, Chihuahua, 31000, Mexico;4. Facultad de Ciencias, Universidad Autónoma de San Luis Potosi, San Luis Potosi, 78290, Mexico;1. Unit of Marine and Environmental Toxicology, UR 09-03, Sfax University, IPEIS, BP 1172, 3018 Sfax, Tunisia;2. Normandie Univ, UNICAEN, CMABio, SFR ICORE, 14000 Caen, France;3. Normandie Univ, UNICAEN, INSERM, COMETE, 14000 Caen, France;4. Normandie Univ, UNICAEN, ABTE, 14000 Caen, France;5. Centre François Baclesse, Avenue Général Harris, BP5026, F-14076 Caen Cedex-05, France;1. Department of Restorative Dentistry and Periodontology, Faculty of Dentistry, Chiang Mai University, Chiang Mai 50200, Thailand;2. Center of Excellence in Oral and Maxillofacial Biology, Department of Oral Biology and Diagnostic Sciences, Faculty of Dentistry, Chiang Mai University, Chiang Mai 50200, Thailand;3. Mineralized Tissue Research Unit, Faculty of Dentistry, Chulalongkorn University, Bangkok 10330, Thailand
Abstract:ObjectiveWhile different virulence factors have been reported of Aggregatibacter actinomycetemcomitans (Aa), there is little information about the stimulatory effect of its DNA. The main purpose of this study was to assess the inflammatory response of human gingival fibroblasts (HGFs) stimulated with A. actinomycetemcomitans DNA.DesignCytokine levels of IL-6, IL-1α and TNF-α were measured on the supernatant of HGFs activated with 10, 25, 50 and 100 μg/ml DNA of Aa during 24 h. Primary cultures of HGFs were infected with Aa and its DNA at different times and concentrations to compare its cytotoxic effect. Cell damage and adhesion of Aa to HGFs were evaluated under light microscopy and Scanning electron microscopy respectively.ResultsThere was a statistical difference (p < 0.05) in cytokine expression in HGFs activated by bacterial DNA with a dose dependent on IL-6 expression and a significantly elevated expression of IL-1α and TNF-α compared to Human DNA negative control. Substantial morphological alterations were observed after infection of A. actinomycetemcomitans in HGFs but not with bDNA exposure. Aggregatibacter actinomycetemcomitans showed a high rate of adhesion and cell damage to HGFs after 30 min.ConclusionsGenomic DNA of A. actinomycetemcomitans could be a factor in the pathogenesis of periodontitis that might play a major role in the inflammatory response.
Keywords:Bacterial DNA  Cytokines  Human gingival fibroblasts  Periodontitis
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