Phenotypic Study of Human Gingival Fibroblasts Labeled With Superparamagnetic Anionic Nanoparticles

• Autores: Antoine Lafont, Bruno Gogly, Adrien Naveau, Pierre Smirnov, Christine Ménager, Florence Gazeau, Olivier Clément
• Localización: Journal of periodontology, ISSN 0022-3492, Vol. 77, Nº. 2, 2006, págs. 238-247
• Idioma: inglés
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• Resumen

  • Phenotypic Study of Human Gingival Fibroblasts Labeled With Superparamagnetic Anionic Nanoparticles Adrien Naveau,* Pierre Smirnov,†‡ Christine Ménager,§ Florence Gazeau,‡ Olivier Clément,† Antoine Lafont,* and Bruno Gogly* *Laboratory of Arterial Development Research, Paris Descartes Medicine University, National Institute of Health and Medical Research Combined Team 0016, Paris, France.

    †Laboratory of Imagery Research, Paris Descartes Medicine University.

    ‡Laboratory of Complex Matters and Systems, Denis Diderot University, National Center for Scientific Research Joint Research Unit 7057, Paris, France.

    §Laboratory of Ionic Liquids and Loaded Interfaces, Pierre and Marie Curie University, National Center for Scientific Research Joint Research Unit 7612, Paris, France.

    Correspondence: Dr. Adrien Naveau, Laboratory of Arterial Development Research, Paris Descartes Medicine University, INSERM EMI 0016, 156 rue de Vaugirard, 75015 Paris, France. Fax: 33-01-49-81-31-46; e-mail: adrien.naveau@laposte.net.

    Background: A specific labeler of the human gingival fibroblast (HGF) does not exist. Anionic maghemite nanoparticles allow labeling of a wide cell variety and their recognition in cellular, organotypical, and animal models.

    Methods: We studied internalization effects of nanoparticles on an HGF phenotype in vitro, evaluating transcription and secretion of connective tissue remodeling molecules, i.e., matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and cytokines controlling their activation/inhibition, i.e., transforming growth factor-β (TGF-β1), tumor necrosis factor-α (TNF-α), and interleukins 1β and 4 (IL-1β and IL-4). After proliferation kinetics, cellular uptake was studied by Perls coloration and magnetophoresis on labeled culture. Dot blotting, Western blotting, and zymography were used to detect MMP-1, -2, and -3 and TIMP-1 and -2 secretions in culture supernatants, and reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the mRNA expression of these molecules. Enzyme-linked immunosorbent assay (ELISA) tests were used to determine TGF-β1, TNF-α, IL-1β, and IL-4 levels.

    Results: Our data indicated high (15.3 ± 5.8 pg/cell) but heterogeneous distribution of nanoparticles in HGF. Twenty-four hours after labeling, MMP-1, -2, and -3 and TIMP-2 secretion increased (P <0.001) with RT-PCR confirmation at 12 hours, whereas TIMP-1 did not. IL-1β increased at day 1 (D1) (P <0.001) and IL-4 at D3 (P <0.01), but not TGF-β1 or TNF-α.

    Conclusions: After labeling with these maghemite nanoparticles, HGF increased secretion of IL-1β at D1, probably inducing the increase of MMP-1, -2, and -3 and TIMP-2. The increase of IL-4 secretion began with the decreased synthesis of MMPs and TIMPs at D3. Despite this transitory inflammatory reaction at 3 days following internalization, maghemite nanoparticles did not affect HGF phenotype, thereby authorizing their use as labelers.

    KEYWORDS: Cytokines, matrix metalloproteinases, nanoparticles Cited by Lavanya Venkataraman, Balakrishnan Sivaraman, Pratik Vaidya and Anand Ramamurthi. (2014) Nanoparticulate delivery of agents for induced elastogenesis in three-dimensional collagenous matrices. Journal of Tissue Engineering and Regenerative Medicine, n/a-n/a.

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    CrossRef Bruno Gogly, Benjamin Fournier, Ludovic Couty, Adrien Naveau, Camille Brasselet, Eric Durand, Bernard Coulomb and Antoine Lafont. (2009) Gingival fibroblast inhibits MMP-7: Evaluation in an ex vivo aorta model. Journal of Molecular and Cellular Cardiology 47, 296-303.

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