September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Biocompatibility Assessment of End-Thiolated Hyaluronate Coated Gold Nanoparticles on Retinal Pigment Epithelial (ARPE-19) cells
Author Affiliations & Notes
  • Bedia Begum Karakocak
    Energy&Environmental and Chemical Engineering, Washington University, Saint Louis, Missouri, United States
    Department of Ophthalmology and Visual Sciences, Washington University, Saint Louis, Missouri, United States
  • Joshua Tyler Davis
    Department of Ophthalmology and Visual Sciences, Washington University, Saint Louis, Missouri, United States
    Veterans Affairs Medical Center , Saint Louis, Missouri, United States
  • Pratim Biswas
    Energy&Environmental and Chemical Engineering, Washington University, Saint Louis, Missouri, United States
  • Nathan Ravi
    Department of Ophthalmology and Visual Sciences, Washington University, Saint Louis, Missouri, United States
    Veterans Affairs Medical Center , Saint Louis, Missouri, United States
  • Footnotes
    Commercial Relationships   Bedia Karakocak, None; Joshua Davis, None; Pratim Biswas, None; Nathan Ravi, None
  • Footnotes
    Support  NIH Grant EY021620; Core Grant: P30EY02687; Research to Prevent Blindness
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 289. doi:
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    • Get Citation

      Bedia Begum Karakocak, Joshua Tyler Davis, Pratim Biswas, Nathan Ravi; Biocompatibility Assessment of End-Thiolated Hyaluronate Coated Gold Nanoparticles on Retinal Pigment Epithelial (ARPE-19) cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):289.

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

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Abstract

Purpose : Gold nanoparticles (Au NPs) are promising targeting agents in drug and gene delivery for various cell types, including ocular cells due to their unique optical, magnetic properties, and ease in manipulating the surface characteristics. However, nascent Au NPs have been reported to have significant toxicity. The goal of this study is to create biocompatible Au NPs, which can be used as delivery agents for ocular cells.

Methods : In this study, an established retinal pigmented epithelial (ARPE-19) cell line was used to assess toxicity. Au NPs were synthesized via citrate reduction method and further coated with end thiolized hyaluronate (HA). Total organic carbon (TOC) and thermal gravimetric analysis (TGA) measurements were performed to quantify the amount of HA coating on Au NPs. HA conjugated Au NPs were tracked inside the cell via confocal microscopy (Figure 1A). The cell proliferation behavior was monitored continuously via electrical cell-substrate impedance sensing (ECIS; Applied Biophysics, NY) for 96 hours (Figure 1B). The biocompatibility of resultant HA-Au NPs was also tested with MTT and Apo Tox-GloTM (Promega, CA) assays (Figure 2).

Results : The presence of hyaluronate coating on Au NPs was confirmed and quantified. ECIS, MTT and Apo Tox-GloTM results show that the ARPE-19 cells were able to proliferate and maintain a monolayer in the presence of the HA-Au NPs at concentrations where nascent Au NPs were toxic (Figures 1&2). Evidence is also provided to show that the HA Au NP’s crossed the cell membrane and were observed within the cells.

Conclusions : Our results indicate that hyaluronate coated Au NPs have potential as delivery agents for ocular cells as they are shown to have enhanced biocompatibility compared to the nascent Au NPs with ARPE-19 cells.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

Figure 1. (A) The 3-D confocal images of HA coated Au NPs within the ARPE-19 cells. (B) Comparison of cell attachment impedance measurements at 4000Hz as a function of time for nascent and Hyaluronate (HA) coated Gold (Au) NPs. The exposure concentrations used for comparison are the critical concentrations for nascent Au NPs (0.05 mg/ml (5nm), 0.09 mg/ml (10nm), and 0.15 mg/ml (20nm).

Figure 1. (A) The 3-D confocal images of HA coated Au NPs within the ARPE-19 cells. (B) Comparison of cell attachment impedance measurements at 4000Hz as a function of time for nascent and Hyaluronate (HA) coated Gold (Au) NPs. The exposure concentrations used for comparison are the critical concentrations for nascent Au NPs (0.05 mg/ml (5nm), 0.09 mg/ml (10nm), and 0.15 mg/ml (20nm).

 

Figure 2. Biocompatibility results for hyaluronate (HA) coated vs nascent Au NPs with (A) MTT (B); Apo Tox-GloTM assays.

Figure 2. Biocompatibility results for hyaluronate (HA) coated vs nascent Au NPs with (A) MTT (B); Apo Tox-GloTM assays.

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