April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Glial Cell Line-Derived Neurotrophic Factor (GDNF) Enriched Poly Lactic-co-Glycolic Acid (PLGA) Microspheres Suppress Apoptosis In Vitro
Author Affiliations & Notes
  • Sami Kamjoo
    Ophthalmology, Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • Vishal R. Sharma
    Ophthalmology, Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • Jing Yang
    Ophthalmology, Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • Patricia Checa
    Pharm & Pharmaceutical Tech Sch, Complutense University, Madrid, Spain
  • Irene Molina-Martinez
    Pharm & Pharmaceutical Tech Sch, Complutense University, Madrid, Spain
  • Rocio Herrero-Vanrell
    Pharm & Pharmaceutical Tech Sch, Complutense University, Madrid, Spain
  • G Astrid Limb
    Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, London, United Kingdom
  • Henry Klassen
    Ophthalmology, Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • M. C. Kenney
    Ophthalmology, Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • Baruch D. Kuppermann
    Ophthalmology, Gavin Herbert Eye Institute, UC Irvine, Irvine, California
  • Footnotes
    Commercial Relationships  Sami Kamjoo, None; Vishal R. Sharma, None; Jing Yang, None; Patricia Checa, None; Irene Molina-Martinez, None; Rocio Herrero-Vanrell, None; G Astrid Limb, None; Henry Klassen, None; M. C. Kenney, None; Baruch D. Kuppermann, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4443. doi:
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      Sami Kamjoo, Vishal R. Sharma, Jing Yang, Patricia Checa, Irene Molina-Martinez, Rocio Herrero-Vanrell, G Astrid Limb, Henry Klassen, M. C. Kenney, Baruch D. Kuppermann; Glial Cell Line-Derived Neurotrophic Factor (GDNF) Enriched Poly Lactic-co-Glycolic Acid (PLGA) Microspheres Suppress Apoptosis In Vitro. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4443.

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

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Abstract

Purpose: : To study effects of GDNF enriched PLGA microspheres on apoptosis induced by Benzo-e-Pyrene (BeP, a constituent of cigarette smoke) in human ARPE-19 and MIO-M1 retinal cells in vitro.

Methods: : ARPE-19 and MIO-M1 cells were pre-treated with 0.5mg/ml GDNF microspheres containing 7ng/ml of GDNF for 6 hours then incubated with 400µM BeP for 24 hours. Apoptosis was studied using the JC-1 assay which measures mitochondrial membrane potential (ΔΨm), and also the YO-PRO-1 Apoptosis kit which measures YO-PRO dye fluorescence in apoptotic cells. Controls included 0.5mg/ml blank microspheres and untreated cells.

Results: : ARPE-19 cells treated with BeP alone showed reduced ΔΨm levels (1.49±0.08 versus 3.68±0.14 for untreated controls, p<0.0001). GDNF microspheres plus BeP showed increased ΔΨm levels (2.01± 0.04, p=0.002) compared to BeP alone. MIO-M1 cell ΔΨm values were 3.24±0.22 for BeP versus 5.15±0.23 for GDNF-microspheres plus BeP (p=0.001) and 7.71±0.19 for untreated controls (p=0.0001 versus BeP alone). ARPE-19 cell apoptosis expressed as % fluorescence was 908.1±42.5 for BeP versus 98.25±0.47 for untreated controls (p<0.0001), 137.0±57 for GDNF microspheres plus BeP ( p<0.0001 vs BeP alone), and 669.3±20.83 for blank microspheres plus BeP (p<0.002 versus BeP alone). MIO-M1 apoptosis % fluorescence was 248.5±5.6 for BeP versus 98.25±0.47 for untreated controls (p<0.001), 156.5±15 for GDNF microspheres plus BeP (p=0.0016 vs BeP alone), and 181.9±11 for blank microspheres (p<0.001 vs BeP alone).

Conclusions: : GDNF microspheres partially protect ARPE-19 and MIO-M1 cells from the induction of apoptosis caused by BeP as measured by both JC-1 and YO-PRO assays. Blank microspheres also showed reduced apoptosis though to a lesser degree than GDNF microspheres. This reduced apoptosis by blank microspheres may be due to sequestering of BeP into the empty microspheres rather than any direct inhibitory effect of the microspheres.

Keywords: retinal pigment epithelium • apoptosis/cell death • retinal culture 
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