title: Pulp fibroblasts synthesize functional complement proteins involved in initiating dentin-pulp regeneration.
abstract: The complement system is an efficient plasma immune surveillance system that controls tissue injury and infection. Although the liver constitutes the primary circulating complement protein synthesis site, extrahepatic synthesis is known to optimize local tissue inflammatory reaction. Because dentin-pulp regeneration is known to be regulated locally, we investigated activation of the local complement system within the dental pulp and its role in initiating the regeneration process. Membrane attack complex (C5b-9) formation and Gram's staining revealed that complement activation is correlated with the presence of Gram-positive bacteria in carious human teeth. RT-PCR analysis demonstrated that cultured human pulp fibroblasts stimulated with lipoteichoic acid produce all the proteins required for efficient complement activation. This was demonstrated in vitro by C5b-9 formation and C5a active fragment production in the absence of plasma proteins. Finally, the dynamic migration assays performed in μ-Slide chemotaxis chambers and use of a C5aR-specific antagonist (W54011) demonstrated that the activation of complement proteins synthesized by pulp fibroblasts and the subsequent release of C5a specifically induced pulp progenitor cell recruitment. Our study reveals human pulp fibroblasts as the first nonimmune cell type capable of synthesizing all complement proteins. These fibroblasts cells contribute significantly to tissue regeneration by recruiting pulp progenitors via complement activation, which suggests to a potential therapeutic strategy of targeting pulp fibroblasts in dentin-pulp regeneration.
authors.affiliation: CNRS, ISM UMR 7287, 13288, Aix Marseille Université, Marseille, France.
authors.forename: Fanny
authors.initials: F
authors.lastname: Chmilewsky
authors.affiliation: CNRS, ISM UMR 7287, 13288, Aix Marseille Université, Marseille, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Jeanneau
authors.affiliation: CNRS, ISM UMR 7287, 13288, Aix Marseille Université, Marseille, France; Odontology Service, APHM, Hôspital Timone, Marseille, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Laurent
authors.affiliation: CNRS, ISM UMR 7287, 13288, Aix Marseille Université, Marseille, France. Electronic address: imad.about@univ-amu.fr.
authors.forename: Imad
authors.initials: I
authors.lastname: About
mesh_terms: D054443:Cell Migration Assays; D002478:Cells, Cultured; D003167:Complement Activation; D015938:Complement Membrane Attack Complex / Q000378:metabolism*; D003470:Culture Media; D003731:Dental Caries / Q000378:metabolism; D003782:Dental Pulp / Q000166:cytology*; D003804:Dentin / Q000502:physiology*; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides; D012038:Regeneration; D013234:Stem Cells / Q000378:metabolism; D013682:Teichoic Acids
chemical_list: D015938:Complement Membrane Attack Complex; D003470:Culture Media; D008070:Lipopolysaccharides; D013682:Teichoic Acids; C009900:lipoteichoic acid