title: Heat-killed Enterococcus faecalis alters nitric oxide and CXCL12 production but not CXCL8 and CCL3 production by cultured human dental pulp fibroblasts.
abstract: BACKGROUND
  Fibroblasts are the most abundant cells in dental pulp. To investigate their capacity to produce the chemokines CCL3, CXCL8, and CXCL12 as well as nitric oxide (NO), we evaluated the production of these mediators in supernatants of cultured human dental pulp fibroblasts (HDPF) stimulated by heat-killed Enterococcus faecalis (HKEF).
  
  
  METHODS
  Primary cultures of HDPF were stimulated with medium alone or HKEF (1:1, 10:1, or 100:1 bacteria:fibroblast) for 1, 6, and 24 hours. Chemokines and NO were assessed through enzyme-linked immunosorbent assay and Griess reaction, respectively. Statistical analysis was performed by using analysis of variance and Tukey post test.
  
  
  RESULTS
  CCL3 was not detected, whereas constitutive CXCL8 was not affected. Production of CXCL12 was increased at 1 and 6 hours, and NO was increased at the concentration of 1:1 bacteria:fibroblast at 24 hours. Viability and proliferation assays did not reveal cell number differences.
  
  
  CONCLUSIONS
  These findings demonstrate that heat-killed E. faecalis is able to increase production of CXCL12 and NO by HDPF.
authors.affiliation: Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.
authors.forename: Carla R
authors.initials: CR
authors.lastname: Sipert
authors.forename: Ivaldo G
authors.initials: IG
authors.lastname: Moraes
authors.forename: Norberti
authors.initials: N
authors.lastname: Bernardinelli
authors.forename: Roberto B
authors.initials: RB
authors.lastname: Garcia
authors.forename: Clóvis M
authors.initials: CM
authors.lastname: Bramante
authors.forename: Thaís H
authors.initials: TH
authors.lastname: Gasparoto
authors.forename: Eduardo A
authors.initials: EA
authors.lastname: Figueira
authors.forename: Thiago J
authors.initials: TJ
authors.lastname: Dionísio
authors.forename: Ana P
authors.initials: AP
authors.lastname: Campanelli
authors.forename: Sandra H P
authors.initials: SH
authors.lastname: Oliveira
authors.forename: Fernando Q
authors.initials: FQ
authors.lastname: Cunha
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Santos
mesh_terms: D049109:Cell Proliferation; D002473:Cell Wall / Q000737:chemistry; D002478:Cells, Cultured; D054405:Chemokine CCL3 / Q000096:biosynthesis; D054377:Chemokine CXCL12 / Q000096:biosynthesis*; D017077:Culture Media, Conditioned; D003782:Dental Pulp / Q000166:cytology / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology*; D013293:Enterococcus faecalis / Q000737:chemistry* / Q000276:immunology; D005347:Fibroblasts / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D006358:Hot Temperature; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis; D009569:Nitric Oxide / Q000096:biosynthesis*; D015854:Up-Regulation; D037521:Virulence Factors
chemical_list: C515429:CCL3 protein, human; D054405:Chemokine CCL3; D054377:Chemokine CXCL12; D017077:Culture Media, Conditioned; D016209:Interleukin-8; D037521:Virulence Factors; D009569:Nitric Oxide