Fluoride at non-toxic dose affects odontoblast gene expression in vitro.
Elevated fluoride intake may lead to local tissue disturbances, known as fluorosis. Towards an understanding of this effect, fluoride-induced molecular responses were analyzed in MO6-G3 cultured odontoblasts cells. NaF at 1mM changed expression of genes implicated in tissue formation and growth, without affecting cell proliferation or inducing stress factor RNAs. Up to 1mM NaF, DNA accumulation was not inhibited, whereas at 3mM, cells detached from their support and did not proliferate. Intracellular structures, characterized by EM, were normal up to 1mM, but at 3mM, necrotic features were evident. No sign of apoptotic transformation appeared at any NaF concentration. Fluoride-sensitive genes were identified by microarray analysis; expression levels of selected RNAs were determined by conventional and real-time RT-PCR. At 1mM fluoride, RNAs encoding the extracellular matrix proteins asporin and fibromodulin, and the cell membrane associated proteins periostin and IMT2A were 10-fold reduced. RNA coding for signaling factor TNF-receptor 9 was diminished to one-third, whereas that for the chemokine Scya-5 was enhanced 2.5-fold. These RNAs are present in vivo in tooth forming cells. This was demonstrated by in situ hybridization and RT-PCR on RNA from dissected tissue samples; for the presence and functioning of fibromodulin in dentin matrix, a more comprehensive study has earlier been performed by others [Goldberg, M., Septier, D., Oldberg, A., Young, M.F., Ameye, L.G., 2006. Fibromodulin deficient mice display impaired collagen fibrillogenesis in predentin as well as altered dentin mineralization and enamel formation. J. Histochem. Cytochem. 54, 525-537]. Expression of most other RNA species, in particular of stress factor coding RNAs, was not altered. It was concluded that fluoride could influence the transcription pattern without inducing cell stress or apoptosis. In odontoblasts in vivo, aberrant expression of these fluoride-sensitive genes may impair the formation of the extracellular matrix and influence cell communication, with the possible consequence of fluorotic patterns of normal and deviant dentin.
Centre de Recherche des Cordeliers, UMRS 872 - Equipe 5, Biologie Oro-faciale et Pathologie, Escalier E, 2e étage,15 rue de l'Ecole de Médecine, 75 270 Paris Cedex 06, France. Tilmann.Wurtz@crc.jussieu.fr <Tilmann.Wurtz@crc.jussieu.fr>
Tilmann
T
Wurtz
Sophia
S
Houari
Nicole
N
Mauro
Mary
M
MacDougall
Heiko
H
Peters
Ariane
A
Berdal
D000818:Animals; D000831:Animals, Newborn; D002461:Cell Line, Transformed; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D022162:Cytoplasmic Vesicles / Q000187:drug effects / Q000648:ultrastructure; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D017403:In Situ Hybridization; D051379:Mice; D009336:Necrosis / Q000139:chemically induced / Q000473:pathology; D009804:Odontoblasts / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D012969:Sodium Fluoride / Q000633:toxicity*
D016326:Extracellular Matrix Proteins; D012333:RNA, Messenger; D012969:Sodium Fluoride