title: Engulfment of ceramic particles by fibroblasts does not alter cell behavior.
abstract: Despite many studies, the impact of ceramic particles on cell behavior remains unclear. The aim of the present study was to investigate the effects of nano-sized ceramic particles on fibroblastic cells. Fibroblasts (dermal fibroblasts freshly isolated from skin samples and WI26 fibroblastic cells) were cultured in a monolayer in the presence of alumina or cerium-zirconia particles (≈50 nm diameter) at two concentrations (100 or 500 μg ml-1). Fluorescent alumina particles were also used. The following properties were analyzed: cell morphology, cytoplasmic ceramic incorporation (using confocal and transmission electron microscopy) and migration (using a silicon insert). Sedimentation field-flow fractionation (SdFFF) was also used to evaluate the rate of incorporation of ceramic particles into the cells. Finally, after treatment with various concentrations of ceramic particles, fibroblasts were also included in a collagen type I lattice constituting a dermal equivalent (DE), and the collagen lattice retraction and cell proliferation were evaluated. In monolayer conditions, the presence of both alumina and cerium-zirconia ceramic particles did not cause any deleterious effects on cultured cells (dermal fibroblast and WI26 cells) and cell fate was not affected in any way by the presence of ceramic particles in the cytoplasm. Confocal (using fluorescent alumina particles) and electron microscopy (using both alumina and cerium-zirconia particles) showed that ceramic particles were internalized in the WI26 cells. Using fluorescent membrane labeling and fluorescent alumina particles, a membrane was observed around the particle-containing vesicles present in the cytoplasm. Electron microscopy on WI26 cells showed the presence of a classical bilayer membrane around the ceramic particles. Interestingly, SdFFF confirmed that some dermal fibroblasts contained many alumina ceramic particles while others contained very few; in WI26 cells, the uptake of alumina ceramic was more homogeneous. In DE, collagen lattice retraction and cell proliferation were unchanged when WI26 fibroblastic cells contained alumina or cerium-zirconia ceramic particles. Our data suggest that ceramic particles are internalized in the cells by endocytosis. The presence of ceramic particles in the cytoplasm has no affect on cell behavior, confirming the excellent biocompatibility of this material and anticipating a minimal harmful effect of potential wear debris.
authors.affiliation: University of Limoges, EA 6309 'Myeline maintenance and peripheral neuropathies', Limoges, F-87000 France.
authors.forename: Pierre-Antoine
authors.initials: PA
authors.lastname: Faye
authors.forename: Olivier
authors.initials: O
authors.lastname: Roualdes
authors.forename: Fabrice
authors.initials: F
authors.lastname: Rossignol
authors.forename: Daniel Jean
authors.initials: DJ
authors.lastname: Hartmann
authors.forename: Alexis
authors.initials: A
authors.lastname: Desmoulière
mesh_terms: D000537:Aluminum Oxide / Q000737:chemistry / Q000633:toxicity; D001672:Biocompatible Materials / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002516:Ceramics / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D002563:Cerium / Q000737:chemistry / Q000633:toxicity; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D008422:Materials Testing; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D015040:Zirconium / Q000737:chemistry / Q000633:toxicity
chemical_list: D001672:Biocompatible Materials; D002563:Cerium; D015040:Zirconium; D000537:Aluminum Oxide; C028541:zirconium oxide