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Invest Ophthalmol Vis Sci 2003;44: E-Abstract 481.
© 2003 ARVO


481—B456

Carbon Nanotube Bucky Paper as an Artificial Support Membrane and Bruch's Membrane Patch in Subretinal RPE and IPE Transplantation

T. Leng1, P. Huie1, K.V. Bilbao1, M.S. Blumenkranz1, D.J. Loftus2 and H.A. Fishman2

1 Ophthalmology, Stanford University, Stanford, CA, United States
2 NASA Ames Research Center, Moffett Field, CA, United States

Commercial Relationships: T. Leng, None; P. Huie, None; K.V. Bilbao, None; M.S. Blumenkranz, None; D.J. Loftus, None; H.A. Fishman, None.

Grant Identification: Stanford Bio-X Interdisciplinary Initiatives Program and Visx, Inc.

Abstract

Purpose: To explore the use of porous carbon nanotube bucky paper as an artificial support membrane for the subretinal transplantation of retinal pigment epithelial (RPE) and iris pigment epithelial (IPE) cells in an effort to provide an autologous replacement for damaged RPE and Bruch's membrane in patients with age-related macular degeneration (AMD).
Methods: A suspension of multiwalled carbon nanotubes (MWCNTs) was cleansed of impurities through washes with sulfuric and nitric acid. The end result was an aqueous MWCNT suspension that was filtered through Whatman filter paper on a Buchner funnel. The filtered MWCNTs were allowed to air dry in an oven at 70° C for 1 hr to finalize the bucky paper. Human RPE cells (ARPE-19) and IPE cells harvested from New Zealand White (NZW) rabbits via papain-based enzymatic dissection were cultured on sterilized bucky paper (UV-irradiated) with serum-supplemented media at 37°C and 6.5% CO2. Analysis of the cultured cells was accomplished through fixation of the samples for light and electron microscopy. Furthermore, bucky paper was implanted into 5 NZW rabbits via standard 3-port pars plana vitrectomy and the creation of a retinal bleb with infusion of balanced salt solution through a 42-G needle. A 1 mm retinotomy was created and a 1 x 1 mm piece of bucky paper was inserted into the subretinal space. The retina was reattached by air-fluid exchange. The eyes were enucleated 2 weeks and 1 month after implantation and fixed for histological analysis.
Results: We found that carbon nanotube bucky paper provided a suitable substrate for in vitro culture of RPE cells, allowing the cells to form a uniform monolayer. IPE cells, however, tended to spread out when cultured on bucky paper and did not establishe a uniform monolayer. In addition, bucky paper proved to have good handling qualities during surgery; it was sufficiently rigid to slide flat into the subretinal space. Lastly, bucky paper was well tolerated in the subretinal space.
Conclusions: Carbon nanotube bucky paper is a strong, flexible, and porous material that has the potential to serve as an artificial substrate for RPE/IPE transplantation or as a patch for damaged Bruch's membrane in the treatment of AMD. To date, it has proven to be biocompatible enough to allow for in vitro culture of RPE and IPE cells and to be well tolerated in the subretinal space.

Keywords: retinal pigment epithelium • age-related macular degeneration • transplantation

 © 2003, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any part of this abstract, contact the ARVO Office at arvo{at}arvo.org.





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