title: Palatal mucosa derived fibroblasts present an adaptive behavior regarding cytokine secretion when grafted onto the gingival margin.
abstract: BACKGROUND
  Considering that grafted gingival tissue might have to be adapted to the receptor area and that fibroblasts have the ability to respond to bacterial stimuli through the release of various cytokines, this study investigated whether fibroblasts from the palatal mucosa behave differently when grafted onto the gingival margin regarding cytokine secretion.
  
  
  METHODS
  Biopsies from the palatal mucosa were collected at the time of free gingival graft surgery, and after four months re-collection was performed upon surgery for root coverage. Fibroblasts were isolated by the explant technique, cultured and stimulated with Porphyromonas gingivalis (Pg) and Escherichia coli (Ec) LPS for 24 or 48 h for comparative evaluation of the secretion of cytokines and chemokines, such as IL-6, IL-8/CXCL8, MIP-1α/CCL3, TGF-β, VEGF and CXCL16. Unstimulated cells were used as the control group. Cells were tested for viability through MTT assay, and secretion of cytokines and chemokines was evaluated in the cell supernatants by Enzyme-Linked Immunosorbent Assay (ELISA).
  
  
  RESULTS
  Fibroblasts from the palatal mucosa maintained the same secretion pattern of IL-6 when grafted onto the gingival margin. On the contrary, fibroblasts from the marginal gingival graft showed increased secretion of IL-8/CXCL8 even in the absence of stimulation. Interestingly, MIP-1α/CCL3 secretion by fibroblasts from the marginal gingival graft was significantly increased after 48 hours of stimulation with Pg LPS and after 24 h with Ec LPS. Only fibroblasts from the marginal gingival graft showed secretion of TGF-β. VEGF and CXCL16 secretion were not detected by both subsets of fibroblasts.
  
  
  CONCLUSIONS
  Fibroblasts from the palatal mucosa seem to be adapted to local conditions of the site microenvironment when grafted onto the gingival marginal area. This evidence supports the effective participation of fibroblasts in the homeostasis of the marginal periodontium through secretion modulation of important inflammatory mediators.
authors.forename: Fabíola Pontes
authors.initials: FP
authors.lastname: Azevedo
authors.forename: Ana Carolina Faria
authors.initials: AC
authors.lastname: Morandini
authors.forename: Carla Renata
authors.initials: CR
authors.lastname: Sipert
authors.forename: Thiago José
authors.initials: TJ
authors.lastname: Dionísio
authors.affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil. cebola@usp.br.
authors.forename: Carlos Ferreira
authors.initials: CF
authors.lastname: Santos
authors.forename: Carla Andreotti
authors.initials: CA
authors.lastname: Damante
authors.forename: Maria Lúcia Rubo
authors.initials: ML
authors.lastname: de Rezende
authors.forename: Adriana Campos Passanezi
authors.initials: AC
authors.lastname: Sant'ana
authors.forename: Sebastião Luiz Aguiar
authors.initials: SL
authors.lastname: Greghi
mesh_terms: D056704:Adaptive Immunity / Q000276:immunology*; D000328:Adult; D064592:Autografts / Q000637:transplantation; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000502:physiology; D054405:Chemokine CCL3 / Q000378:metabolism; D000075743:Chemokine CXCL16; D019743:Chemokines, CXC / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism*; D004926:Escherichia coli; D005260:Female; D005347:Fibroblasts / Q000276:immunology / Q000637:transplantation*; D005881:Gingiva / Q000166:cytology / Q000637:transplantation*; D005889:Gingival Recession / Q000601:surgery; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D008875:Middle Aged; D010159:Palate; D016966:Porphyromonas gingivalis; D051116:Receptors, Scavenger; D016212:Transforming Growth Factor beta / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: C515429:CCL3 protein, human; C423727:CXCL16 protein, human; C508606:CXCL8 protein, human; D054405:Chemokine CCL3; D000075743:Chemokine CXCL16; D019743:Chemokines, CXC; D016207:Cytokines; C508600:IL6 protein, human; D015850:Interleukin-6; D016209:Interleukin-8; D008070:Lipopolysaccharides; D051116:Receptors, Scavenger; D016212:Transforming Growth Factor beta; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A