title: Interaction of rat alveolar macrophages with dental composite dust.
abstract: Dental composites have become the standard filling material to restore teeth, but during the placement of these restorations, high amounts of respirable composite dust (<5 μm) including many nano-sized particles may be released in the breathing zone of the patient and dental operator. Here we tested the respirable fraction of several composite particles for their cytotoxic effect using an alveolar macrophage model system. ​METHODS: Composite dust was generated following a clinical protocol, and the dust particles were collected under sterile circumstances. Dust was dispersed in fluid, and 5-μm-filtered to enrich the respirable fractions. Quartz DQ12 and corundum were used as positive and negative control, respectively. Four concentrations (22.5 μg/ml, 45 μg/ml, 90 μg/ml and 180 μg/ml) were applied to NR8383 alveolar macrophages. Light and electron microscopy were used for subcellular localization of particles. Culture supernatants were tested for release of lactate dehydrogenase, glucuronidase, TNF-α, and H2O2.
  
  
  
  Characterization of the suspended particles revealed numerous nano-sized particles but also many high volume particles, most of which could be removed by filtering. Even at the highest concentration (180 μg/ml), cells completely cleared settled particles from the bottom of the culture vessel. Accordingly, a mixture of nano- and micron-scaled particles was observed inside cells where they were confined to phagolysosomes. The filtered particle fractions elicited largely uniform dose-dependent responses, which were elevated compared to the control only at the highest concentration, which equaled a mean cellular dose of 120 pg/cell. A low inflammatory potential was identified due to dose-dependent release of H2O2and TNF-α. However, compared to the positive control, the released levels of H2O2and TNF-α were still moderate, but their release profiles depended on the type of composite.
  
  
  
  Alveolar macrophages are able to phagocytize respirable composite dust particle inclusive nanoparticles. Since NR8383 cells tolerate a comparatively high cell burden (60 pg/cell) of each of the five materials with minimal signs of cytotoxicity or inflammation, the toxic potential of respirable composite dust seems to be low. These results are reassuring for dental personnel, but more research is needed to characterize the actual exposure and uptake especially of the pure nano fraction.
authors.affiliation: KU Leuven BIOMAT, Department of Oral Health Sciences, University of Leuven & Dentistry University Hospitals Leuven, Kapucijnenvoer 7, Leuven, 3000, Belgium. kirsten.vanlanduyt@kuleuven.be.
authors.forename: K L
authors.initials: KL
authors.lastname: Van Landuyt
authors.affiliation: KU Leuven BIOMAT, Department of Oral Health Sciences, University of Leuven & Dentistry University Hospitals Leuven, Kapucijnenvoer 7, Leuven, 3000, Belgium.
authors.forename: S M
authors.initials: SM
authors.lastname: Cokic
authors.affiliation: Institute of Energy and Environmental Technology (IUTA) e.V, BliersheimerStraße 58-60, Duisburg, 47229, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Asbach
authors.affiliation: Research Unit Experimental Toxicology, Center for Environmental and Health Research, Department of Public Health and Primary Care, University of Leuven, Herestraat 49, Leuven, 3001, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Hoet
authors.affiliation: Research Unit Experimental Toxicology, Center for Environmental and Health Research, Department of Public Health and Primary Care, University of Leuven, Herestraat 49, Leuven, 3001, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Godderis
authors.affiliation: Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-University of Munich, Nussbaumstraße 26, Munich, 80336, Germany.
authors.forename: F X
authors.initials: FX
authors.lastname: Reichl
authors.affiliation: KU Leuven BIOMAT, Department of Oral Health Sciences, University of Leuven & Dentistry University Hospitals Leuven, Kapucijnenvoer 7, Leuven, 3000, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Van Meerbeek
authors.affiliation: IBE, IBE R&D gGmbH, Mendelstraße 11, Münster, 48149, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Vennemann
authors.affiliation: IBE, IBE R&D gGmbH, Mendelstraße 11, Münster, 48149, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Wiemann
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003188:Composite Resins; D004391:Dust; D016676:Macrophages, Alveolar / Q000378:metabolism*; D018384:Oxidative Stress; D051381:Rats
chemical_list: D003188:Composite Resins; D004391:Dust
keywords: Alveolar macrophage; Biotest; Dental composite; Dentist; Dust; Filler; Inhalable; NR8383; Nano-dust; Nanoparticle, Biocompatibility; Occupational; Resin