title: Effect of LPS treatment on the viability and chemokine synthesis by epithelial cells and gingival fibroblasts.
abstract: OBJECTIVE
  Several local factors can affect the wound-healing process, delaying its progression and postponing tissue homeostasis. It is known that local inflammation is related to wound healing; however, the maintenance of the inflammatory reaction can impair the proliferation and migration of oral mucosal cells. The aim of this study was to evaluate the viability and chemokine expression of epithelial cells and gingival fibroblasts exposed to long-term lipopolysaccharide (LPS) treatment.
  
  
  METHODS
  Epithelial cells (HaCaT, Cell Lines Service, 300493) and human gingival fibroblasts (HGFs) were seeded (1×10(5) cells/well) in 24-well plates and incubated for 24h. To simulate the responses of cells to a local chronic oral mucosal inflammation, we added LPS of Escherichia coli (10 μg/ml) to Dulbecco's modified Eagle's medium (DMEM), kept in contact with fibroblasts and epithelial cells for 24, 48, and 72h. Then the cells were assessed for viability (alamarBlue assay), number (trypan blue assay), and expression of CCL2 and CCL5 inflammatory chemokines (enzyme-linked immunosorbent assay (ELISA)). Data were statistically analyzed by nonparametric Kruskal-Wallis and Mann-Whitney tests at a significance level of 5%.
  
  
  RESULTS
  Cell treatment with LPS caused significant decrease in viability for both cell lines. No time-dependent effect was observed for epithelial cells. However, reduction in fibroblast viability was greater at 48 and 72 h. CCL2 and CCL5 synthesis was significantly increased for both LPS-treated cells, and this expression decreased with time.
  
  
  CONCLUSIONS
  The maintenance of an inflammatory cell stimulus by LPS decreases the number and viability of cultured oral mucosal cells, which may be related to delayed wound healing.
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: F G
authors.initials: FG
authors.lastname: Basso
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: D G
authors.initials: DG
authors.lastname: Soares
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: T N
authors.initials: TN
authors.lastname: Pansani
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: A P S
authors.initials: AP
authors.lastname: Turrioni
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: D L
authors.initials: DL
authors.lastname: Scheffel
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil.
authors.forename: C A
authors.initials: CA
authors.lastname: de Souza Costa
authors.affiliation: UNESP - University Estadual Paulista, Araraquara, São Paulo, Brazil. Electronic address: jhebling@foar.unesp.br.
authors.forename: J
authors.initials: J
authors.lastname: Hebling
mesh_terms: D002452:Cell Count; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D005881:Gingiva / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*
chemical_list: D018925:Chemokines; D008070:Lipopolysaccharides
keywords: Epithelial cells; Fibroblasts; Inflammation; Wound healing