March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Absorption Of Spio Nanoparticles Using Different Media On Arpe-19 And Hcec Cell Cultures
Author Affiliations & Notes
  • Gustavo T. Grottone
    Ophthalmology, UNIFESP/Santa Casa de Santos, Santos, Brazil
  • Renata R. Loureiro
    Ophthalmology, UNIFESP, Santos, Brazil
  • Joyce Couvre
    Ophthalmology, UNIFESP, Santos, Brazil
  • Lionel Gamarra
    Oncology, Instituto Israelita de Pesquisas Albert Einstein, São Paulo, Brazil
  • Priscila Cristovam
    Ophthalmology, UNIFESP/Santa Casa de Santos, Santos, Brazil
  • José Álvaro P. Gomes
    Ophthalmology, UNIFESP, Santos, Brazil
  • Footnotes
    Commercial Relationships  Gustavo T. Grottone, None; Renata R. Loureiro, None; Joyce Couvre, None; Lionel Gamarra, None; Priscila Cristovam, None; José Álvaro P. Gomes, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 6551. doi:
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      Gustavo T. Grottone, Renata R. Loureiro, Joyce Couvre, Lionel Gamarra, Priscila Cristovam, José Álvaro P. Gomes; Absorption Of Spio Nanoparticles Using Different Media On Arpe-19 And Hcec Cell Cultures. Invest. Ophthalmol. Vis. Sci. 2012;53(14):6551.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Locate and evaluate the absorption of two commercial available super paramagnetic iron oxide nanoparticles incubated in different media on HCEC and ARPE-19 using light microscopy and electron microscopy.

Methods: : We used two different commercial nano particles with different particles sizes (100 nm and 300 nm) and different coating(dextran-coated and silicon-coated). After an incubation period of 12 hours, using a variety of incubation media(RPMI, RPMI + FBS 5%, RPMI + FBS 10%, DMEM, DMEM + FBS 5%, + DMEM + FBS 10%), ARPE-19 cultures and Human Corneal Endothelial Cells were observed on light microscope using prussian blue staining. The evaluation was qualitative on the density of blue pigment(iron-oxide) at a 10x field. Futhermore, we analyzed the pigment location(surface of the cell or inside the cell). To corroborate the data observed at light microscope we did scanning electron microscopy and transmission electron microscopy to find the correct location of the superparamagnetic nano particles.

Results: : The groups which we used RPMI-only or DMEM-only incubation media had a poor absorption of 100nm nano particles which increased with the addition of FBS 5% or FBS 10%. This result was maximum at RPMI + FBS 10% group either in HCEC or ARPE-19 cells. In the other hand, there was no variation on 300nm nano particles using any of the incubation media. At light and electron microscopy, the granules of 100 nm nano particles were on cytoplasm of the cells. Regarding the 300nm nano particles they were found at intercellular spaces and adherent to cell surface.

Conclusions: : Our study showed an evidence that adding FBS at incubation media when using SPIO nanoparticles, provide a better absorption on 100nm groups which seems to have a linear correlation. In the other hand, as the 300nm showed no cell absorption, there was no difference on using FBS or not. RPMI had the best results considering 100nm nano particles cell absorption when compared to DMEM.

Keywords: cornea: endothelium • retinal pigment epithelium • microscopy: light/fluorescence/immunohistochemistry 
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