title: Toxicity of formaldehyde to human oral fibroblasts and epithelial cells: influences of culture conditions and role of thiol status.
abstract: The toxicity of formaldehyde, a monomer released from certain polymeric dental materials, was studied in cultured human oral fibroblasts and epithelial cells. The influences of growth conditions were evaluated for both cell types, as well as the role of the internal and external thiol states. A one-hour exposure to formaldehyde decreased the colony-forming efficiency (CFE) of both cell types in a concentration-dependent manner, although the toxicity varied up to 100-fold with the conditions. Clearly, the presence of serum and the thiol cysteine counteracted the toxicity in fibroblasts. Similarly, pituitary extract and cysteine, or a mixture of amino acids and ethanolamines, counteracted the formaldehyde toxicity in serum-free cultures of epithelial cells. In contrast, a growth-promoting surface matrix of fibronectin and collagen did not influence the formaldehyde toxicity, as shown by both the CFE assay and a dye reduction assay. Further, a short-term change to the various growth media per se with or without the supplements serum or cysteine did not significantly alter the CFE. Analysis of the thiol state demonstrated significant differences between epithelial cells and fibroblasts, i.e., comparatively lower cellular levels of the free low-molecular-weight thiols glutathione and cysteine in fibroblasts. This result correlated to significantly higher formaldehyde toxicity in the fibroblasts than in the epithelial cells. Taken together, the results indicated the cytoprotective function of both intracellular and extracellular thiols toward formaldehyde, as well as the usefulness of thiol-free and chemically defined conditions for toxicity assessments in oral epithelial cells and fibroblasts. We conclude that the combined use of a controlled external milieu and the presumed target cell type may be advantageous in evaluations of oral toxicity mechanisms or the toxic potency of dental materials, particularly those which, like formaldehyde, may react with thiols or amines.
authors.affiliation: Division of Experimental Carcinogenesis, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: J A
authors.initials: JA
authors.lastname: Nilsson
authors.forename: X
authors.initials: X
authors.lastname: Zheng
authors.forename: K
authors.initials: K
authors.lastname: Sundqvist
authors.forename: Y
authors.initials: Y
authors.lastname: Liu
authors.forename: L
authors.initials: L
authors.lastname: Atzori
authors.forename: A
authors.initials: A
authors.lastname: Elfwing
authors.forename: K
authors.initials: K
authors.lastname: Arvidson
authors.forename: R C
authors.initials: RC
authors.lastname: Grafström
mesh_terms: D001681:Biological Assay / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods; D003470:Culture Media; D003764:Dental Materials / Q000633:toxicity*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005557:Formaldehyde / Q000633:toxicity*; D006801:Humans; D007202:Indicators and Reagents; D009061:Mouth Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013778:Tetrazolium Salts
chemical_list: D003470:Culture Media; D003764:Dental Materials; D007202:Indicators and Reagents; D013438:Sulfhydryl Compounds; D013778:Tetrazolium Salts; D005557:Formaldehyde