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Effects of HEMA on type I collagen protein in human gingival fibroblasts

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Abstract

The cytotoxicity of dental composites has been attributed to the release of residual monomers from polymerized adhesive systems due to degradation processes or the incomplete polymerization of materials. 2-Hydroxyethyl methacrylate (HEMA) is one of the major components released from dental adhesives. Cytotoxic effects due to high concentrations of HEMA have already been investigated, but the influence of minor toxic concentrations on specific proteins such as type I collagen has not been studied in depth. The objective of this project was to study the effect of minor toxic concentrations of HEMA on human gingival fibroblasts (HGFs), investigating modification in cell morphology, cell viability, and the influence on type I collagen protein. Primary lines of human gingival fibroblasts were exposed to 3 mmol/L HEMA for different periods of time (24 h, 72 h, 96 h). The cell vitality was determined by MTT assay, and high-resolution scanning electron microscopy analysis was performed to evaluate differences in cell morphology before and after treatment. The presence and localization of type I collagen was determined by immunofluorescence in HGFs treated with HEMA for the same period of time. The vitality of the cells decreased after 72 h of exposure. The HGFs grown in monolayer and observed by field emission in-lens scanning electron microscopy demonstrated a preserved surface morphology after 24 h of treatment, while they showed an altered morphology after 96 h of treatment. Immunofluorescence demonstrated a reduction of type I collagen due to HEMA exposure after 96 h. From these results, we conclude that low concentrations of HEMA can significantly alter the morphology of human gingival fibroblasts and interfere with the presence of type I collagen protein.

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Abbreviations

HEMA:

2-Hydroxyethyl methacrylate

HGF:

human gingival fibroblast

FEISEM:

field emission in-lens scanning electron microscopy

TC50 :

(concentration responsible for 50% of cell death)

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Authors and Affiliations

  1. Dipartimento di Scienze Anatomiche Umane e Fisiopatologia dell’Apparato Locomotore, Università di Bologna, Italy

    M. Falconi, G. Teti, M. Zago & G. Mazzotti

  2. Dipartimento di Igiene e Sanità Pubblica, sezione Medicina Legale, Università di Bologna, Italy

    S. Pelotti

  3. Division of Dental Sciences and Biomaterials, Department of Biomedicine, University of Trieste, Trieste, Italy

    L. Breschi

  4. Dip. Scienze Anatomiche Umane e Fisiopatologia dell’Apparato Locomotore, Via Irnerio, 48, 40126, Bologna, Italy

    M. Falconi

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  1. M. Falconi
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  2. G. Teti
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  3. M. Zago
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  4. S. Pelotti
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  5. L. Breschi
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  6. G. Mazzotti
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Correspondence to M. Falconi.

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Falconi, M., Teti, G., Zago, M. et al. Effects of HEMA on type I collagen protein in human gingival fibroblasts. Cell Biol Toxicol 23, 313–322 (2007). https://doi.org/10.1007/s10565-006-0148-3

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  • DOI: https://doi.org/10.1007/s10565-006-0148-3

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