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Development of a protocol of isolation of nanoparticles from patients' broncho-alveolar lavages for their in vitro toxicity assessment.
Bernal, Kévin; Kose, Ozge; Leclerc, Lara; Vergnon, Jean-Michel; Pourchez, Jérémie; Forest, Valérie.
Afiliação
  • Bernal K; Mines Saint-Etienne, Univ Jean Monnet, INSERM U 1059 Sainbiose, Centre CIS, CS 62362 42023, 158 cours Fauriel F-42023 Saint-Etienne Cedex 2 France vforest@emse.fr +33477499776.
  • Kose O; Mines Saint-Etienne, Univ Jean Monnet, INSERM U 1059 Sainbiose, Centre CIS, CS 62362 42023, 158 cours Fauriel F-42023 Saint-Etienne Cedex 2 France vforest@emse.fr +33477499776.
  • Leclerc L; Mines Saint-Etienne, Univ Jean Monnet, INSERM U 1059 Sainbiose, Centre CIS, CS 62362 42023, 158 cours Fauriel F-42023 Saint-Etienne Cedex 2 France vforest@emse.fr +33477499776.
  • Vergnon JM; Univ Lyon, Univ Jean Monnet, INSERM U 1059 Sainbiose F-42023 Saint-Etienne France.
  • Pourchez J; CHU Saint-Etienne, Service de Pneumologie F-42055 Saint-Etienne France.
  • Forest V; Mines Saint-Etienne, Univ Jean Monnet, INSERM U 1059 Sainbiose, Centre CIS, CS 62362 42023, 158 cours Fauriel F-42023 Saint-Etienne Cedex 2 France vforest@emse.fr +33477499776.
Nanoscale Adv ; 6(2): 458-466, 2024 Jan 16.
Article em En | MEDLINE | ID: mdl-38235099
ABSTRACT
To investigate potential correlations between human exposure to inhaled particles and pathological effects, the biological monitoring of nanoparticles in broncho-alveolar lavages (BAL) from patients has been proposed. To better understand the underlying mechanisms of toxicity, we propose to couple this biomonitoring of nanoparticles to their in vitro toxicity assessment. However, BAL obtained from regular clinical practice are conditioned with sodium hypochlorite solution (in a 50% v/v ratio), which is toxic to cells. The aim of this study was to develop a protocol to neutralize sodium hypochlorite, allowing to properly investigate the toxicity of the nanoparticles BAL contain. We first tried to neutralize chemically the sodium hypochlorite using H2O2, ascorbic acid or sodium ascorbate but this approach was unsuccessful. In addition, standard toxicology assays (MTT, LDH) could not be used because of interference with neutralizing solutions. We thus changed strategy and used ultracentrifugation to isolate nanoparticles from the sodium hypochlorite solution, with satisfactory extraction yields (88 to 100%). We then incubated the extracted nanoparticles with macrophages from the RAW264.7 cell line and assessed the cell viability and pro-inflammatory response. This study can be used as a proof-of-concept for further study of the biological impact of nanoparticles. This approach paves the way for studies aiming at a better understanding of the aetiology of some idiopathic diseases and underlying mechanisms.

Texto completo: 1 Base de dados: MEDLINE Tipo de estudo: Guideline Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Tipo de estudo: Guideline Idioma: En Ano de publicação: 2024 Tipo de documento: Article