The purpose of this study was to investigate the ultrastructure of the membrana limitans interna (internal limiting membrane, ILM) and to evaluate alterations to the retinal cell layers after membrane peeling with vital dyes. Twenty-five patients (25 eyes) who underwent macular hole surgery were included, whereby 12 indocyanine green (ICG)- and 13 brilliant blue G (BBG)-stained ILM were analyzed using light, transmission electron and scanning electron microscopy. Retinal cell fragments on the ILM were identified in both groups using immunohistochemistry. Comparing ICG- and BBG-stained membranes, larger cellular fragments were observed at a higher frequency in the BBG group. Thereby, the findings indicate that ICG permits an enhanced separation of the ILM from the underlying retina with less mechanical destruction. A possible explanation might be seen in the known photosensitivity of ICG, which induces a stiffening and shrinkage of the ILM but also generates retinal toxic metabolites.

Keywords:
Macular hole surgery, Pars plana vitrectomy, Membrana limitans interna, Ultrastructure, Brilliant blue G, Indocyanine green, Membrane peeling
1.
Lundkvist A, Reichenbach A, Betsholtz C, Carmeliet P, Wolburg H, Pekny M: Under stress, the absence of intermediate filaments from Muller cells in the retina has structural and functional consequences. J Cell Sci 2004;117:3481–3488.
2.
Bringmann A, Pannicke T, Biedermann B, Francke M, Iandiev I, Grosche J, Wiedemann P, Albrecht J, Reichenbach A: Role of retinal glial cells in neurotransmitter uptake and metabolism. Neurochem Int 2009;54:143–160.
3.
Ponsioen TL, van Luyn MJ, van der Worp RJ, van Meurs JC, Hooymans JM, Los LI: Collagen distribution in the human vitreoretinal interface. Invest Ophthalmol Vis Sci 2008;49:4089–4095.
4.
Kohno T, Sorgente N, Ishibashi T, Goodnight R, Ryan S: Immunofluorescent studies of fibronectin and laminin in the human eye. Invest Ophthalmol Vis Sci 1987;28:506–514.
5.
Russell S, Shepherd J, Hageman G: Distribution of glycoconjugates in the human retinal internal limiting membrane. Invest Ophthalmol Vis Sci 1991;32:1986–1995.
6.
Matsumoto B, Blanks J, Ryan S: Topographic variations in the rabbit and primate internal limiting membrane. Invest Ophthalmol Vis Sci 1984;25:71–82.
7.
Bainbridge J, Herbert E, Gregor Z: Macular holes: vitreoretinal relationships and surgical approaches. Eye 2008;22:1301–1309.
8.
Korda V, Dusova D, Studnicka J, Rencova E, Hejcmanova D: The surgical management of macular hole (in Czech). Cesk Slov Oftalmol 2005;61:316–320.
9.
Oie Y, Emi K, Takaoka G, Ikeda T: Effect of indocyanine green staining in peeling of internal limiting membrane for retinal detachment resulting from macular hole in myopic eyes. Ophthalmology 2007;114:303–306.
10.
Gandorfer A, Haritoglou C, Kampik A: Toxicity of indocyanine green in vitreoretinal surgery. Dev Ophthalmol 2008;42:69–81.
11.
Enaida H, Sakamoto T, Hisatomi T, Goto Y, Ishibashi T: Morphological and functional damage of the retina caused by intravitreous indocyanine green in rat eyes. Graefes Arch Clin Exp Ophthalmol 2002;240:209–213.
12.
Rodrigues E, Meyer C, Mennel S, Farah M: Mechanisms of intravitreal toxicity of indocyanine green dye: Implications for chromovitrectomy. Retina 2007;27:958–970.
13.
Iriyama A, Uchida S, Yanagi Y, Tamaki Y, Inoue Y, Matsuura K, Kadonosono K, Araie M: Effects of indocyanine green on retinal ganglion cells. Invest Ophthalmol Vis Sci 2004;45:943–947.
14.
Schmid-Kubista K, Lamar P, Schenk A, Stolba U, Binder S: Comparison of macular function and visual fields after membrane blue or infracyanine green staining in vitreoretinal surgery. Graefes Arch Clin Exp Ophthalmol 2010;248:381–388.
15.
Gass J: Idiopathic senile macular hole: its early stages and pathogenesis. Arch Ophthalmol 1988;106:629–639.
16.
Richardson K, Jarett L, Finke E: Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol 1960;35:313–323.
17.
Lois N, Burr JM, Norrie J, Vale L, Cook JA, McDonald A, Boachie C, Ternent L, McPherson G: Internal limiting membrane peeling versus no peeling for idiopathic full thickness macular hole: a pragmatic randomised controlled trial. Invest Ophthalmol Vis Sci 2011;52:1586–1592.
18.
Kwok AK, Lai TY, Man-Chan W, Woo DC: Indocyanine green assisted retinal internal limiting membrane removal in stage 3 or 4 macular hole surgery. Br J Ophthalmol 2003;87:71–74.
19.
Remy M, Thaler S, Schumann R, May C, Fiedorowicz M, Schuettauf F, Grüterich M, Priglinger S, Nentwich M, Kampik A, Haritoglou C: An in vivo evaluation of brilliant blue G in animals and humans. Br J Ophthalmol 2008;92:1142–1147.
20.
Engel E, Schraml R, Maisch T, Kobuch K, Konig B, Szeimies RM, Hillenkamp J, Baumler W, Vasold R: Light-induced decomposition of indocyanine green. Invest Ophthalmol Vis Sci 2008;49:1777–1783.
21.
Uemura A, Kanda S, Sakamoto Y, Kita H: Visual field defects after uneventful vitrectomy for epiretinal membrane with indocyanine green-assisted internal limiting membrane peeling. Am J Ophthalmol 2003;136:252–257.
22.
Yamashita T, Uemura A, Kita H, Nakao K, Sakamoto T: Long-term outcomes of visual field defects after indocyanine green-assisted macular hole surgery. Retina 2008;28:1228–1233.
23.
Yamashita T, Uemura A, Kita H, Sakamoto T: Analysis of the retinal nerve fiber layer after indocyanine green-assisted vitrectomy for idiopathic macular holes. Ophthalmology 2006;113:280–284.
24.
Ueno A, Hisatomi T, Enaida H, Kagimoto T, Mochizuki Y, Goto Y, Kubota T, Hata Y, Ishibashi T: Biocompatibility of brilliant blue G in a rat model of subretinal injection. Retina 2007;27:499–504.
25.
Enaida H, Ishibashi T: Brilliant blue in vitreoretinal surgery. Dev Ophthalmol 2008;42:115–125.
26.
Yuen D, Gonder J, Proulx A, Liu H, Hutnik C: Comparison of the in vitro safety of intraocular dyes using two retinal cell lines: a focus on brilliant blue G and indocyanine green. Am J Ophthalmol 2009;147:251–259.e252.
27.
Morales M, Freire V, Asumendi A, Araiz J, Herrera I, Castiella G, Corcóstegui I, Corcóstegui G: Comparative effects of six intraocular vital dyes on retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 2010;51:6018–6029.
28.
Biedermann B, Bringmann A, Franze K, Faude F, Wiedemann P, Reichenbach A: Gaba(a) receptors in Muller glial cells of the human retina. Glia 2004;46:302–310.
29.
Sebag J: Age-related differences in the human vitreoretinal interface. Arch Ophthalmol 1991;109:966–971.
30.
Gandorfer A, Haritoglou C, Kampik A: Toxicity of indocyanine green in vitreoretinal surgery. Dev Ophthalmol 2008;42:69–81.
31.
Wollensak G: Biomechanical changes of the internal limiting membrane after indocyanine green staining. Dev Ophthalmol 2008;42:82–90.
32.
Wollensak G, Wilsch M, Spoerl E, Seiler T: Collagen fiber diameter in the rabbit cornea after collagen crosslinking by riboflavin/UVA. Cornea 2004;23:503–507.
33.
Tokuda K, Zorumski C, Izumi Y: Involvement of illumination in indocyanine green toxicity after its washout in the ex vivo rat retina. Retina 2009;29:371–379.
34.
Da Mata AP, Burk SE, Foster RE, Riemann CD, Petersen MR, Nehemy MB, Augsburger JJ: Long-term follow-up of indocyanine green-assisted peeling of the retinal internal limiting membrane during vitrectomy surgery for idiopathic macular hole repair. Ophthalmology 2004;111:2246–2253.
© 2011 S. Karger AG, Basel
2011
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.