May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Targeted Mouse Vascular Endothelial Cell Killing Using Gold Nanospheres
Author Affiliations & Notes
  • K. L. Masselam
    New York Univ Medical Center, New York, New York
    Ophthalmology,
    Ophthalmology, Manhattan Eye, Ear and Throat Hospital, New York, New York
  • J. A. Young
    New York Univ Medical Center, New York, New York
    Ophthalmology,
  • J. Karen
    New York Univ Medical Center, New York, New York
    Dermatology,
  • C. Hunzeker
    New York Univ Medical Center, New York, New York
    Dermatology,
  • P.-J. Yu
    New York Univ Medical Center, New York, New York
    Cardiovascular Surgery,
  • P. Mignatti
    New York Univ Medical Center, New York, New York
    Cardiovascular Surgery,
  • Footnotes
    Commercial Relationships K.L. Masselam, None; J.A. Young, None; J. Karen, None; C. Hunzeker, None; P. Yu, None; P. Mignatti, None.
  • Footnotes
    Support None.
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 1744. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      K. L. Masselam, J. A. Young, J. Karen, C. Hunzeker, P.-J. Yu, P. Mignatti; Targeted Mouse Vascular Endothelial Cell Killing Using Gold Nanospheres. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1744.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose:: To employ surface plasmon resonance to target and kill endothelial cells by means of gold nanospheres conjugated to IgG.

Methods:: Mouse vascular endothelial cells were seeded at confluency in eight L-lysine coated LabTek wells. PCAM antibody was diluted to 0.1 mg/ml in 20 mM Hepes (pH 7.4) for a final concentration of 4 µg/ml antibody (80 µL used). Ten times the volume of gold (800 µL) was added to a 10:1 ratio of gold:antibody. BSA was added to a final concentration of 0.1 % and polyethylene glycol was added to a final concentration of 0.2 mg/ml in all wells. The suspension was then centrifuged at 5000 rpm in 4° C for 1 hour. The conjugate was then resuspended in PBS and 0.1% BSA. Endothelial monolayers were covered with 500 µl of media per well and 20 µL of conjugate was added to wells #1-4 (no conjugate was added to wells #5-8). The cells were incubated for 1 hour. The media was then replaced with PBS. Wells #1-3 and 5-7 were irradiated with 50 pulses per well of a 532-nm, 0.5 J/cm2, Q-switched Nd:YAG laser (6 mm spot). Media was removed and Trypan blue exclusion testing was performed to assay cell viability. Viable and non-viable cells were counted within the 6 mm irradiated spot for wells #1-4 and within a corresponding area for wells #5-8 by using an Axiovert 25 microscope (Carl Zeiss, Inc., Thornwood, NY) with a CWHK grid ocular (Olympus America Inc., Melville, NY) (ob 20×).

Results:: Irradiated wells with IgG-conjugated gold nanospheres (#1-3) demonstrated 45-65% cell death. Irradiated wells with unconjugated gold nanospheres (#5-7) showed 23-29% cell death. The conjugated (#4) and unconjugated (#8) non-irradiated wells yielded 24% and 22% cell death, respectively.

Conclusions:: Wells containing IgG-conjugated gold nanospheres demonstrated a higher yield cell death compared to wells containing unconjugated gold nanospheres. Cell death in the non-irradiated wells was likely explained by cell preparation. Conjugated gold nanospheres could potentially be used in ophthalmology to treat neovascularization as well as in other areas of medicine for targeted cell killing.

Keywords: neovascularization 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×