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Optimisation of polymer scaffolds for ocular cell transplantation

Optimisation of polymer scaffolds for ocular cell transplantation
Optimisation of polymer scaffolds for ocular cell transplantation
Purpose: the retinal pigment epithelium would be the primary target for cellular replacement in degenerative ocular diseases such as age related macular degeneration. Transplantation of cells into the eye as a suspension has a number of disadvantages including cell death and incorrectly localised or disorganised grafts. Recent studies have demonstrated that growth of RPE cells on polymer scaffolds can facilitate the transplantation of cells as an intact monolayer which may reduce cell death by up to 10 fold (1). However there have only been limited investigations to determine the optimal polymer composition for such scaffolds.

Methods: using a technique described by (2). We manufactured and evaluated five different blends of poly(L-lactic acid) (PLLA) with poly(D, L-lactic-glycolic acid) (PLGA). Scanning electron microscopy (SEM) and 13C Nuclear magnetic resonance spectroscopy were used to characterise the polymer surface and verify the ratios produced. Polymer surface chemistry was modified using the extracellular matrix protein laminin. The polymer blends were then seeded with ARPE-19 cells and maintained in culture for up to 4 weeks. Cell adherence and proliferation were assessed using SEM and immunofluorescence. Cell survival was quantified by measurement of apoptosis and cell membrane integrity. Maintenance of phenotypic characteristics was also investigated using immunocytochemistry.

Results and Conclusion: highly porous polymers with an average thickness of 180µm were produced. The ARPE-19 cell line proliferated, remained viable and retained phenotypic characteristics most efficiently on the 25% PLLA:75% PLGA blend ratio. Further in vivo studies are now required to confirm the functionality of cells on such polymer scaffolds.

transplantation, retinal pigment epithelium, cell survival
6230
Thomson, Heather Anne Jane
7db0d6dd-4e3c-47b0-a281-9a04c0b70252
Treharne, Andrew John
a158bb09-9d41-4b3a-ae02-ccbd76dd9417
Walker, Paul
9235b363-f0a5-46ff-99c6-22de755d7ed5
Grossel, Martin Christopher
403bf3ff-6364-44e9-ab46-52d84c6f0d56
Lotery, Andrew John
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514
Thomson, Heather Anne Jane
7db0d6dd-4e3c-47b0-a281-9a04c0b70252
Treharne, Andrew John
a158bb09-9d41-4b3a-ae02-ccbd76dd9417
Walker, Paul
9235b363-f0a5-46ff-99c6-22de755d7ed5
Grossel, Martin Christopher
403bf3ff-6364-44e9-ab46-52d84c6f0d56
Lotery, Andrew John
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514

Thomson, Heather Anne Jane, Treharne, Andrew John, Walker, Paul, Grossel, Martin Christopher and Lotery, Andrew John (2009) Optimisation of polymer scaffolds for ocular cell transplantation. ARVO Meeting Abstracts, 50 (5), 6230.

Record type: Article

Abstract

Purpose: the retinal pigment epithelium would be the primary target for cellular replacement in degenerative ocular diseases such as age related macular degeneration. Transplantation of cells into the eye as a suspension has a number of disadvantages including cell death and incorrectly localised or disorganised grafts. Recent studies have demonstrated that growth of RPE cells on polymer scaffolds can facilitate the transplantation of cells as an intact monolayer which may reduce cell death by up to 10 fold (1). However there have only been limited investigations to determine the optimal polymer composition for such scaffolds.

Methods: using a technique described by (2). We manufactured and evaluated five different blends of poly(L-lactic acid) (PLLA) with poly(D, L-lactic-glycolic acid) (PLGA). Scanning electron microscopy (SEM) and 13C Nuclear magnetic resonance spectroscopy were used to characterise the polymer surface and verify the ratios produced. Polymer surface chemistry was modified using the extracellular matrix protein laminin. The polymer blends were then seeded with ARPE-19 cells and maintained in culture for up to 4 weeks. Cell adherence and proliferation were assessed using SEM and immunofluorescence. Cell survival was quantified by measurement of apoptosis and cell membrane integrity. Maintenance of phenotypic characteristics was also investigated using immunocytochemistry.

Results and Conclusion: highly porous polymers with an average thickness of 180µm were produced. The ARPE-19 cell line proliferated, remained viable and retained phenotypic characteristics most efficiently on the 25% PLLA:75% PLGA blend ratio. Further in vivo studies are now required to confirm the functionality of cells on such polymer scaffolds.

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More information

Published date: May 2009
Venue - Dates: The Association for Research in Vision and Ophthalmology Conference, Fort Lauderdale, United States, 2009-04-30
Keywords: transplantation, retinal pigment epithelium, cell survival

Identifiers

Local EPrints ID: 181039
URI: http://eprints.soton.ac.uk/id/eprint/181039
PURE UUID: 185a5f61-3c39-48a7-af26-3922abd9a598
ORCID for Martin Christopher Grossel: ORCID iD orcid.org/0000-0001-7469-6854
ORCID for Andrew John Lotery: ORCID iD orcid.org/0000-0001-5541-4305

Catalogue record

Date deposited: 15 Apr 2011 08:24
Last modified: 11 Dec 2021 03:51

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Contributors

Author: Heather Anne Jane Thomson
Author: Andrew John Treharne
Author: Paul Walker

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