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H.S. Dua, R.S. McIntosh, P.J. Tighe, D.K. James, A. Hopkinson; Amniotic Membrane for Ocular Surface Reconstruction: Donor Variations and Handling Affect Membrane Constituents . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2718.
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© ARVO (1962-2015); The Authors (2016-present)
Despite widespread use of the amniotic membrane (AM) in ocular surgery, its characteristics, at the time of clinical application, are far from standardised. The effects of the methods of processing, storage and preoperative preparation on the membrane are not fully elucidated. Very little is known of the inter– and intra–donor variability within the membrane. The purpose of this study was to determine the effect of ‘handling’ on TGF–ß1 within fresh, processed and stored, and transplant ready AM (TRAM).
17 human AM, both fresh and handled, were analyzed for TGF–ß1 by immunohistochemistry, real time polymerase chain reaction, SDS PAGE electrophoresis and western blotting.
TGF–ß1 was the highest normalised expressed isoform of TGF–ß in all samples but varied between membranes from different donors and also at different sites within the same membrane. The highest concentration was noted in the spongy layer. Removal of the spongy layer successfully removed the bulk of TGF–ß1 from transplant ready membrane. Both latency associated protein (LAP) and a latent TGF–ß binding protein (LTBP) were also detected.
TGF–ß1 is present in various regulatory forms in the AM. There is a degree of inter– and intra–membrane variation, which is modified by handling. Key factors, which are critical in achieving the desired clinical effects may be inadvertently lost or retained depending on the handling procedure. Unless a standardised protocol is adopted that delivers a membrane with consistent constituents, clinical outcomes can vary substantially and comparisons may be invalid.
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