title: In vitro regulation of CCL3 and CXCL12 by bacterial by-products is dependent on site of origin of human oral fibroblasts.
abstract: BACKGROUND
  Production of chemokines by tissue resident cells is one of the main mechanisms involved in the inflammatory infiltrate formation during inflammation. The specific ability of fibroblasts from different oral tissues such as gingiva, periodontal ligament, and dental pulp from permanent and deciduous teeth in producing the chemokines CCL3 and CXCL12 under stimulation by bacterial products commonly found in endodontic infections was investigated.
  
  
  METHODS
  Cultures of fibroblasts from gingiva and periodontal ligament as well as from dental pulp from permanent and deciduous teeth were established by using an explant technique and stimulated with increasing concentrations of Escherichia coli lipopolysaccharide (EcLPS) and Enterococcus faecalis lipoteichoic acid (EfLTA) for 1, 6, and 24 hours. Supernatants were tested for CCL3 and CXCL12 by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  In general, CCL3 production was induced by EcLPS in the 4 fibroblast subtypes and by EfLTA in fibroblasts from gingiva and periodontal ligament. Constitutive CXCL12 synthesis decreased in all fibroblast subtypes especially under stimulation with EcLPS. Fibroblast from permanent deciduous teeth was the cell type presenting the most expressive reduction in CXCL12 release by both stimuli. On the basis of computational matching of CXCL12 mRNA with the microRNAs miR-141 and miR-200a, their expression was also investigated. Although detected in the fibroblasts, these molecules remained unaltered by bacterial by-product stimulation.
  
  
  CONCLUSIONS
  EcLPS and EfLTA induced the production of CCL3 and unbalanced the synthesis of CXCL12 in a manner dependent on the specific tissue origin of fibroblasts.
authors: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
authors: Carla Renata
authors: CR
authors: Sipert
authors: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
authors: Ana Carolina
authors: AC
authors: Morandini
authors: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
authors: Thiago José
authors: TJ
authors: Dionísio
authors: Department of Pediatric Dentistry, Orthodontics and Community Health, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
authors: Maria Aparecida Andrade Moreira
authors: MA
authors: Machado
authors: Department of Basic Sciences, Araçatuba School of Dentistry, Universidade Estadual Paulista, Araçatuba, São Paulo, Brazil.
authors: Sandra Helena Penha
authors: SH
authors: Oliveira
authors: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
authors: Ana Paula
authors: AP
authors: Campanelli
authors: Harvard Clinical and Translational Science Center, Laboratory for Innovative Translational Technologies, Harvard Medical School and Department of Developmental Biology, Harvard School of Dental Medicine, Boston, Massachusetts.
authors: Winston Patrick
authors: WP
authors: Kuo
authors: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil. Electronic address: cebola@usp.br.
authors: Carlos Ferreira
authors: CF
authors: Santos
mesh_terms: D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D054405:Chemokine CCL3 / Q000187:drug effects*; D054377:Chemokine CXCL12 / Q000187:drug effects*; D002648:Child; D004396:Coloring Agents; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects; D013293:Enterococcus faecalis; D004926:Escherichia coli; D005347:Fibroblasts / Q000187:drug effects*; D005881:Gingiva / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D035683:MicroRNAs / Q000032:analysis; D010513:Periodontal Ligament / Q000166:cytology* / Q000187:drug effects; D010641:Phenotype; D013682:Teichoic Acids / Q000494:pharmacology*; D013778:Tetrazolium Salts; D013844:Thiazoles; D014094:Tooth, Deciduous / Q000166:cytology; D055815:Young Adult
chemical_list: C515429:CCL3 protein, human; C515255:CXCL12 protein, human; D054405:Chemokine CCL3; D054377:Chemokine CXCL12; D004396:Coloring Agents; D008070:Lipopolysaccharides; C533895:MIRN141 microRNA, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D013682:Teichoic Acids; D013778:Tetrazolium Salts; D013844:Thiazoles; C009900:lipoteichoic acid; C022616:thiazolyl blue
keywords: CCL3; CXCL12; chemokines; dental pulp diseases; fibroblasts; microRNAs; periapical diseases