Poly-lactic acid nanoparticles (PLA-NP) promote physiological modifications in lung epithelial cells and are internalized by clathrin-coated pits and lipid rafts

Journal of Nanobiotechnology

Research

Aniela Sommer¹  Ilse Foissner¹  Albert Duschl²  Martin Himly²  Matthew Samuel Powys Boyles³  Camila Macedo da Luz⁴  Nathalia Balthazar Martins⁴  Priscila Falagan-Lotsch⁴  Henrique Rudolf Tutumi⁴  Paulo Emílio Corrêa Leite⁵  José Mauro Granjeiro⁶  Eidy de Oliveira Santos⁷  Mariana Rodrigues Pereira⁸ 

[1] Department of Cell Biology, University of Salzburg, Salzburg, Austria;Department of Molecular Biology, University of Salzburg, Salzburg, Austria;Department of Molecular Biology, University of Salzburg, Salzburg, Austria;Heriot-Watt University, Edinburg, UK;Laboratory of Bioengineering and in Vitro Toxicology, Directory of Metrology Applied to Life Sciences (Dimav), National Institute of Metrology Quality and Technology (INMETRO), Duque De Caxias, RJ, Brazil;Laboratory of Bioengineering and in Vitro Toxicology, Directory of Metrology Applied to Life Sciences (Dimav), National Institute of Metrology Quality and Technology (INMETRO), Duque De Caxias, RJ, Brazil;Av. Nossa Senhora das Gracas 50, LABET - Dimav, Predio 27, Duque de Caxias, Xerem, 25250-020, Rio de Janeiro, Brazil;Laboratory of Bioengineering and in Vitro Toxicology, Directory of Metrology Applied to Life Sciences (Dimav), National Institute of Metrology Quality and Technology (INMETRO), Duque De Caxias, RJ, Brazil;Dental School, Fluminense Federal University, Niteroi, RJ, Brazil;Laboratory of Bioengineering and in Vitro Toxicology, Directory of Metrology Applied to Life Sciences (Dimav), National Institute of Metrology Quality and Technology (INMETRO), Duque De Caxias, RJ, Brazil;Laboratory of Biochemistry, State University Center of West Zone (UEZO), Rio de Janeiro, RJ, Brazil;Laboratory of Chemical Signaling in Nervous System, Biology Institute, Fluminense Federal University, Niteroi, RJ, Brazil;

关键词: Nanoparticles;    Drug delivery;    Endocytosis;    Lipid rafts;    Clathrin-coated pits;   

DOI  :  10.1186/s12951-016-0238-1

 received in 2016-09-08, accepted in 2016-12-03,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundPoly-lactic acid nanoparticles (PLA-NP) are a type of polymeric NP, frequently used as nanomedicines, which have advantages over metallic NP such as the ability to maintain therapeutic drug levels for sustained periods of time. Despite PLA-NP being considered biocompatible, data concerning alterations in cellular physiology are scarce.MethodsWe conducted an extensive evaluation of PLA-NP biocompatibility in human lung epithelial A549 cells using high throughput screening and more complex methodologies. These included measurements of cytotoxicity, cell viability, immunomodulatory potential, and effects upon the cells’ proteome. We used non- and green-fluorescent PLA-NP with 63 and 66 nm diameters, respectively. Cells were exposed with concentrations of 2, 20, 100 and 200 µg/mL, for 24, 48 and 72 h, in most experiments. Moreover, possible endocytic mechanisms of internalization of PLA-NP were investigated, such as those involving caveolae, lipid rafts, macropinocytosis and clathrin-coated pits.ResultsCell viability and proliferation were not altered in response to PLA-NP. Multiplex analysis of secreted mediators revealed a low-level reduction of IL-12p70 and vascular epidermal growth factor (VEGF) in response to PLA-NP, while all other mediators assessed were unaffected. However, changes to the cells’ proteome were observed in response to PLA-NP, and, additionally, the cellular stress marker miR155 was found to reduce. In dual exposures of staurosporine (STS) with PLA-NP, PLA-NP enhanced susceptibility to STS-induced cell death. Finally, PLA-NP were rapidly internalized in association with clathrin-coated pits, and, to a lesser extent, with lipid rafts.ConclusionsThese data demonstrate that PLA-NP are internalized and, in general, tolerated by A549 cells, with no cytotoxicity and no secretion of pro-inflammatory mediators. However, PLA-NP exposure may induce modification of biological functions of A549 cells, which should be considered when designing drug delivery systems. Moreover, the pathways of PLA-NP internalization we detected could contribute to the improvement of selective uptake strategies.

【 授权许可】

CC BY   
© The Author(s) 2017