Skip to main content
Top
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology 9/2005

01-09-2005 | Laboratory Investigation

Toxic effects of indocyanine green, infracyanine green, and trypan blue on the human retinal pigmented epithelium

Authors: L. Kodjikian, T. Richter, M. Halberstadt, F. Beby, F. Flueckiger, M. Boehnke, J. G. Garweg

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 9/2005

Login to get access

Abstract

Background

Indocyanine green, infracyanine green, and trypan blue are frequently used as aids to visualize structures removed during vitreoretinal surgery. But they may have toxic effects on the retina. We therefore compared the acute and chronic toxicities of these stains on cultured human retinal pigmented epithelial (RPE) cells using clinically relevant concentrations and an identical experimental setup for each agent.

Methods

Monolayers of RPE cells were incubated with various concentrations of indocyanine green, infracyanine green (each at 0.005%, 0.05%, and 0.5%) or trypan blue (0.05%, 0.06%, 0.1%, 0.15%, and 0.5%) for 5 min (acute exposure) or 6 days (chronic exposure). Using the propidium iodide assay, acute cytotoxicity was monitored at 15-min intervals for up to 3 h. Chronic cytotoxicity was assessed by monitoring cell calcein esterase activity, cell proliferation, and cell morphology (viability) after 6 days of exposure.

Results

Indocyanine and infracyanine green induced acute and chronic toxicities at a concentration above 0.05%. Trypan blue evoked no acute toxicity, but it was chronically cytotoxic at all tested concentrations.

Conclusions

Despite thorough rinsing after application, significant amounts of the not sufficiently water soluble indocyanine and infracyanine green are retained after surgery by the eye. Trypan blue, being more water-soluble than ICG, is probably retained to the least degree. This circumstance is fortunate given that trypan blue exhibits a chronic cytotoxicity comparable to ICG at all clinically relevant concentrations. During vitrectomy, surgeons should aim to expose retinal tissue to only low concentrations of these stains and for as short a period as possible.
Literature
1.
go back to reference Ashikari M, Ozeki H, Tomida K, Sakurai E, Tamai K, Ogura Y (2003) Retention of dye after indocyanine green-assisted internal limiting membrane peeling. Am J Ophthalmol 136:172–174 Ashikari M, Ozeki H, Tomida K, Sakurai E, Tamai K, Ogura Y (2003) Retention of dye after indocyanine green-assisted internal limiting membrane peeling. Am J Ophthalmol 136:172–174
2.
go back to reference Ciardella AP, Schiff W, Barile G, Vidne O, Sparrow J, Langton K, Chang S (2003) Persistent indocyanine green fluorescence after vitrectomy for macular hole. Am J Ophthalmol 136:174–177 Ciardella AP, Schiff W, Barile G, Vidne O, Sparrow J, Langton K, Chang S (2003) Persistent indocyanine green fluorescence after vitrectomy for macular hole. Am J Ophthalmol 136:174–177
3.
go back to reference Da Mata AP, Burk SE, Riemann CD, Rosa RH Jr, Snyder ME, Petersen MR, Foster RE (2001) Indocyanine green-assisted peeling of the retinal limiting membrane during vitrectomy surgery for macular hole repair. Ophthalmology 108:1187–1192 Da Mata AP, Burk SE, Riemann CD, Rosa RH Jr, Snyder ME, Petersen MR, Foster RE (2001) Indocyanine green-assisted peeling of the retinal limiting membrane during vitrectomy surgery for macular hole repair. Ophthalmology 108:1187–1192
4.
go back to reference Dietz FB, Jaffe RA (2003) Indocyanine green. Evidence of neurotoxicity in spinal root axons. Anesthesiology 98:516–520 Dietz FB, Jaffe RA (2003) Indocyanine green. Evidence of neurotoxicity in spinal root axons. Anesthesiology 98:516–520
5.
go back to reference Eckardt C, Eckardt U, Groos S, Luciano L, Reale E (1997) Removal of the internal limiting membrane in macular holes: clinical and morphological findings. Ophthalmologe 94:545–551 Eckardt C, Eckardt U, Groos S, Luciano L, Reale E (1997) Removal of the internal limiting membrane in macular holes: clinical and morphological findings. Ophthalmologe 94:545–551
6.
go back to reference Enaida H, Sakamoto T, Hisatomi T, Goto Y, Ishibashi T (2002) Morphological and functional damage of the retina caused by intravitreous indocyanine green in rat eyes. Graefe Arch Clin Exp Ophthalmol 240:209–213 Enaida H, Sakamoto T, Hisatomi T, Goto Y, Ishibashi T (2002) Morphological and functional damage of the retina caused by intravitreous indocyanine green in rat eyes. Graefe Arch Clin Exp Ophthalmol 240:209–213
7.
go back to reference Engelbrecht NE, Freeman J, Sternberg P, Aaberg TM, Aaberg TM Jr, Martin DF, Sippy BD (2002) Retinal pigment epithelium after macular hole surgery with indocyanine green-assisted internal limiting membrane peeling. Am J Ophthalmol 133:89–94 Engelbrecht NE, Freeman J, Sternberg P, Aaberg TM, Aaberg TM Jr, Martin DF, Sippy BD (2002) Retinal pigment epithelium after macular hole surgery with indocyanine green-assisted internal limiting membrane peeling. Am J Ophthalmol 133:89–94
8.
go back to reference Feron EJ, Veckeneer M, Parys-Van Ginderdeuren R, Van Lommel A, Melles GRJ, Stalmans P (2002) Trypan blue staining of epiretinal membranes in proliferative vitreoretinopathy. Arch Ophthalmol 120:141–144 Feron EJ, Veckeneer M, Parys-Van Ginderdeuren R, Van Lommel A, Melles GRJ, Stalmans P (2002) Trypan blue staining of epiretinal membranes in proliferative vitreoretinopathy. Arch Ophthalmol 120:141–144
9.
go back to reference Gale JS, Proulx AA, Gonder JR, Mao AJ, Hutnik CM (2004) Comparison of the in vitro toxicity of indocyanine green to that of trypan blue retinal pigment epithelium cell cultures. Am J Ophthalmol 138:64–69 Gale JS, Proulx AA, Gonder JR, Mao AJ, Hutnik CM (2004) Comparison of the in vitro toxicity of indocyanine green to that of trypan blue retinal pigment epithelium cell cultures. Am J Ophthalmol 138:64–69
10.
go back to reference Gandorfer A, Messmer EM, Ulbig MW, Kampik A (2000) Resolution of diabetic macular edema after surgical removal of the posterior hyaloid and the inner limiting membrane. Retina 20:126–133 Gandorfer A, Messmer EM, Ulbig MW, Kampik A (2000) Resolution of diabetic macular edema after surgical removal of the posterior hyaloid and the inner limiting membrane. Retina 20:126–133
11.
go back to reference Gandorfer A, Haritoglou C, Gass CA, Ulbig MW, Kampik A (2001) Indocyanine green-assisted peeling of the internal limiting membrane may cause retinal damage. Am J Ophthalmol 132:431–433 Gandorfer A, Haritoglou C, Gass CA, Ulbig MW, Kampik A (2001) Indocyanine green-assisted peeling of the internal limiting membrane may cause retinal damage. Am J Ophthalmol 132:431–433
12.
go back to reference Gandorfer A, Messmer EM, Ulbig MW, Kampik A (2001) Indocyanine green selectively stains the internal limiting membrane. Am J Ophthalmol 131:387–388 Gandorfer A, Messmer EM, Ulbig MW, Kampik A (2001) Indocyanine green selectively stains the internal limiting membrane. Am J Ophthalmol 131:387–388
13.
go back to reference Gandorfer A, Haritoglou C, Gandorfer A, Kampik A (2003) Retinal damage from indocyanine green in experimental macular surgery. Invest Ophthalmol Vis Sci 44:316–323 Gandorfer A, Haritoglou C, Gandorfer A, Kampik A (2003) Retinal damage from indocyanine green in experimental macular surgery. Invest Ophthalmol Vis Sci 44:316–323
14.
go back to reference Grisanti S, Szurman P, Gelisken F, Aisenbrey S, Oficjalska-Mlynczak J, Bartz-Schmidt KU (2004) Histological findings in experimental macular surgery with indocyanine green. Invest Ophthalmol Vis Sci 45:282–286 Grisanti S, Szurman P, Gelisken F, Aisenbrey S, Oficjalska-Mlynczak J, Bartz-Schmidt KU (2004) Histological findings in experimental macular surgery with indocyanine green. Invest Ophthalmol Vis Sci 45:282–286
15.
go back to reference Haritoglou C, Gass CA, Schaumberger M, Ehrt O, Gandorfer A, Kampik A (2001) Macular changes after peeling of the internal limiting membrane in macular hole surgery. Am J Ophthalmol 132:363–368 Haritoglou C, Gass CA, Schaumberger M, Ehrt O, Gandorfer A, Kampik A (2001) Macular changes after peeling of the internal limiting membrane in macular hole surgery. Am J Ophthalmol 132:363–368
16.
go back to reference Haritoglou C, Gandorfer A, Gass CA, Schaumberger M, Ulbig MW, Kampik A (2002) Indocyanine green-assisted peeling of the internal limiting membrane in macular hole surgery affects visual outcome: a clinicopathologic correlation. Am J Ophthalmol 134:836–841 Haritoglou C, Gandorfer A, Gass CA, Schaumberger M, Ulbig MW, Kampik A (2002) Indocyanine green-assisted peeling of the internal limiting membrane in macular hole surgery affects visual outcome: a clinicopathologic correlation. Am J Ophthalmol 134:836–841
17.
go back to reference Haritoglou C, Gandorfer A, Gass CA, Schaumberger M, Ulbig MW, Kampik A (2003) The effect of indocyanine-green on functional outcome of macular pucker surgery. Am J Ophthalmol 135:328–337 Haritoglou C, Gandorfer A, Gass CA, Schaumberger M, Ulbig MW, Kampik A (2003) The effect of indocyanine-green on functional outcome of macular pucker surgery. Am J Ophthalmol 135:328–337
18.
go back to reference Haritoglou C, Gandorfer A, Gass CA, Kampik A (2004) Histology of the vitreoretinal interface after staining of the internal limiting membrane using glucose 5% diluted indocyanine and infracyanine green. Am J Ophthalmol 137:345–348 Haritoglou C, Gandorfer A, Gass CA, Kampik A (2004) Histology of the vitreoretinal interface after staining of the internal limiting membrane using glucose 5% diluted indocyanine and infracyanine green. Am J Ophthalmol 137:345–348
19.
go back to reference Haritoglou C, Eibl K, Schaumberger M, Mueller AJ, Priglinger S, Alge C, Kampik A (2004) Functional outcome after trypan blue-assisted vitrectomy for macular pucker: a prospective, randomized, comparative trial. Am J Ophthalmol 138:1–5 Haritoglou C, Eibl K, Schaumberger M, Mueller AJ, Priglinger S, Alge C, Kampik A (2004) Functional outcome after trypan blue-assisted vitrectomy for macular pucker: a prospective, randomized, comparative trial. Am J Ophthalmol 138:1–5
20.
go back to reference Hirata A, Inomata Y, Kawaji T, Tanihara H (2003) Persistent subretinal indocyanine green induces retinal pigment epithelium atrophy. Am J Ophthalmol 136:353–355 Hirata A, Inomata Y, Kawaji T, Tanihara H (2003) Persistent subretinal indocyanine green induces retinal pigment epithelium atrophy. Am J Ophthalmol 136:353–355
21.
go back to reference Ho JD, Tsai RJF, Chen SN, Chen HC (2003) Toxic effect of indocyanine green on retinal pigment epithelium related to osmotic effects of the solvent. Am J Ophthalmol 135:258 Ho JD, Tsai RJF, Chen SN, Chen HC (2003) Toxic effect of indocyanine green on retinal pigment epithelium related to osmotic effects of the solvent. Am J Ophthalmol 135:258
22.
go back to reference Iriyama A, Uchida S, Yanagi Y, Tamaki Y, Inoue Y, Matsuura K, Kadonosono K, Araie M (2004) Effects of indocyanine green on retinal ganglion cells. Invest Ophthalmol Vis Sci 45:943–947 Iriyama A, Uchida S, Yanagi Y, Tamaki Y, Inoue Y, Matsuura K, Kadonosono K, Araie M (2004) Effects of indocyanine green on retinal ganglion cells. Invest Ophthalmol Vis Sci 45:943–947
23.
go back to reference Jackson TL, Hillenkamp J, Knight BC, Zhang JJ, Thomas D, Stanford MR, Marshall J (2004) Safety testing of indocyanine green and trypan blue using retinal pigment epithelium and glial cell cultures. Invest Ophthalmol Vis Sci 45:2778–2785 Jackson TL, Hillenkamp J, Knight BC, Zhang JJ, Thomas D, Stanford MR, Marshall J (2004) Safety testing of indocyanine green and trypan blue using retinal pigment epithelium and glial cell cultures. Invest Ophthalmol Vis Sci 45:2778–2785
24.
go back to reference Kadonosono K, Itoh N, Uchio E, Nakamura S, Ohno S (2000) Staining of the internal limiting membrane in macular hole surgery. Arch Ophthalmol 118:1116–1118 Kadonosono K, Itoh N, Uchio E, Nakamura S, Ohno S (2000) Staining of the internal limiting membrane in macular hole surgery. Arch Ophthalmol 118:1116–1118
25.
go back to reference Kadonosono K, Takeuchi S, Yabuki K, Yamakawa T, Mekada A, Uchio E (2003) Absorption of short wavelengths of endoillumination in indocyanine green solution: implications for internal limiting membrane removal. Graefe Arch Clin Exp Ophthalmol 241:284–286 Kadonosono K, Takeuchi S, Yabuki K, Yamakawa T, Mekada A, Uchio E (2003) Absorption of short wavelengths of endoillumination in indocyanine green solution: implications for internal limiting membrane removal. Graefe Arch Clin Exp Ophthalmol 241:284–286
26.
go back to reference Kwok AKH, Li VWY, Pang CP, Lai TYY, Yam GHF, Chan NR, Lam DSC (2001) Indocyanine green staining and removal of internal limiting membrane in macular hole surgery: histology and outcome. Am J Ophthalmol 132:178–183 Kwok AKH, Li VWY, Pang CP, Lai TYY, Yam GHF, Chan NR, Lam DSC (2001) Indocyanine green staining and removal of internal limiting membrane in macular hole surgery: histology and outcome. Am J Ophthalmol 132:178–183
27.
go back to reference Kwok AK, Lai TY, Yuen KS, Tam BS, Wong VW (2003) Macular hole surgery with or without indocyanine green stained internal limiting membrane peeling. Clin Experiment Ophthalmol 31:470–475 Kwok AK, Lai TY, Yuen KS, Tam BS, Wong VW (2003) Macular hole surgery with or without indocyanine green stained internal limiting membrane peeling. Clin Experiment Ophthalmol 31:470–475
28.
go back to reference Kwok AK, Lai TY, Li WW, Yew DT, Wong VW (2004) Trypan blue- and indocyanine green-assisted epiretinal membrane surgery: clinical and histopathological studies. Eye 18:882–888 Kwok AK, Lai TY, Li WW, Yew DT, Wong VW (2004) Trypan blue- and indocyanine green-assisted epiretinal membrane surgery: clinical and histopathological studies. Eye 18:882–888
29.
go back to reference Li K, Wong D, Stanga P, Groenewald C, McGalliard J (2003) Trypan blue staining of internal limiting membrane and epiretinal membrane during vitrectomy: visual results and histopathological findings. Br J Ophthalmol 87:216–219 Li K, Wong D, Stanga P, Groenewald C, McGalliard J (2003) Trypan blue staining of internal limiting membrane and epiretinal membrane during vitrectomy: visual results and histopathological findings. Br J Ophthalmol 87:216–219
30.
go back to reference Meyer CH, Rodrigues EB, Kroll P (2004) Reduced concentration and incubation of intravitreal indocyanine green can improve the functional outcome in macular hole surgery. Am J Ophthalmol 137:386 Meyer CH, Rodrigues EB, Kroll P (2004) Reduced concentration and incubation of intravitreal indocyanine green can improve the functional outcome in macular hole surgery. Am J Ophthalmol 137:386
31.
go back to reference Norn MS (1980) Per operative trypan blue vital staining of corneal endothelium: eight years’ follow up. Acta Ophthalmol Scand 58:550–558 Norn MS (1980) Per operative trypan blue vital staining of corneal endothelium: eight years’ follow up. Acta Ophthalmol Scand 58:550–558
32.
go back to reference Sakamoto T, Itaya K, Noda Y, Ishibashi T (2002) Retinal pigment epithelial changes after indocyanine green-assisted vitrectomy. Retina 22:794–796 Sakamoto T, Itaya K, Noda Y, Ishibashi T (2002) Retinal pigment epithelial changes after indocyanine green-assisted vitrectomy. Retina 22:794–796
33.
go back to reference Schmidt JC, Rodrigues EB, Meyer CH, Hoerle S, Kroll P (2004) A modified technique to stain the internal limiting membrane with indocyanine green. Ophthalmologica 218:176–179 Schmidt JC, Rodrigues EB, Meyer CH, Hoerle S, Kroll P (2004) A modified technique to stain the internal limiting membrane with indocyanine green. Ophthalmologica 218:176–179
34.
go back to reference Sippy BD, Engelbrecht NE, Hubbard GB, Moriarty SE, Jiang S, Aaberg TM, Aaberg TM Jr, Grossniklaus HE, Sternberg P (2001) Indocyanine green effect on cultured human retinal pigment epithelial cells: implication for macular hole surgery. Am J Ophthalmol 132:433–435 Sippy BD, Engelbrecht NE, Hubbard GB, Moriarty SE, Jiang S, Aaberg TM, Aaberg TM Jr, Grossniklaus HE, Sternberg P (2001) Indocyanine green effect on cultured human retinal pigment epithelial cells: implication for macular hole surgery. Am J Ophthalmol 132:433–435
35.
go back to reference Spaide RF (2002) Persistent intraocular indocyanine green staining after macular hole surgery. Retina 22:637–639 Spaide RF (2002) Persistent intraocular indocyanine green staining after macular hole surgery. Retina 22:637–639
36.
go back to reference Stalmans P, Feron EJ, Parys-Van Ginderdeuren R, Van Lommel A, Melles GRJ, Veckeneer M (2003) Double vital staining using trypan blue and infracyanine green in macular pucker surgery. Br J Ophthalmol 87:713–716 Stalmans P, Feron EJ, Parys-Van Ginderdeuren R, Van Lommel A, Melles GRJ, Veckeneer M (2003) Double vital staining using trypan blue and infracyanine green in macular pucker surgery. Br J Ophthalmol 87:713–716
37.
go back to reference Stalmans P, Van Aken EH, Melles G, Veckeneer M, Feron EJ, Stalmans I (2003) Trypan blue not toxic for retinal pigment epithelium in vitro. Am J Ophthalmol 135:234–236 Stalmans P, Van Aken EH, Melles G, Veckeneer M, Feron EJ, Stalmans I (2003) Trypan blue not toxic for retinal pigment epithelium in vitro. Am J Ophthalmol 135:234–236
38.
go back to reference Stalmans P, Van Lommel A, Parys-Van Ginderdeuren R (2003) Indocyanine green-assisted peeling of the internal limiting membrane in macular hole surgery affects visual outcome: a clinicopathological correlation. Am J Ophthalmol 136:961–962 Stalmans P, Van Lommel A, Parys-Van Ginderdeuren R (2003) Indocyanine green-assisted peeling of the internal limiting membrane in macular hole surgery affects visual outcome: a clinicopathological correlation. Am J Ophthalmol 136:961–962
39.
go back to reference Tadayoni R, Paques M, Girmens JF, Massin P, Gaudric A (2003) Persistent of fundus fluorescence after use of indocyanine green for macular surgery. Ophthalmology 110:604–608 Tadayoni R, Paques M, Girmens JF, Massin P, Gaudric A (2003) Persistent of fundus fluorescence after use of indocyanine green for macular surgery. Ophthalmology 110:604–608
40.
go back to reference Veckeneer M, van Overdam K, Monzer J, Kobuch K, van Marle W, Spekreijse H, van Meurs J (2001) Ocular toxicity study of trypan blue injected into the vitreous cavity of rabbit eyes. Graefe Arch Clin Exp Ophthalmol 239:698–704 Veckeneer M, van Overdam K, Monzer J, Kobuch K, van Marle W, Spekreijse H, van Meurs J (2001) Ocular toxicity study of trypan blue injected into the vitreous cavity of rabbit eyes. Graefe Arch Clin Exp Ophthalmol 239:698–704
41.
go back to reference Ventura AC, Bohnke M (1999) Toxicity of pentoxifylline on monolayers of highly proliferative cells of epithelial origin. J Ocular Pharmacol Ther 15:525–535 Ventura AC, Bohnke M (1999) Toxicity of pentoxifylline on monolayers of highly proliferative cells of epithelial origin. J Ocular Pharmacol Ther 15:525–535
42.
go back to reference Vlckova A, Gasparic J, Horakova K (1971) Investigation of the cytotoxicity of different trypan blue commercial products. Bull Acad Pol Sci Biol 19:763–770 Vlckova A, Gasparic J, Horakova K (1971) Investigation of the cytotoxicity of different trypan blue commercial products. Bull Acad Pol Sci Biol 19:763–770
43.
go back to reference Weinberger AWA, Kirchhof B, Mazinani BE, Schrage NF (2001) Persistent indocyanine green (ICG) fluorescence 6 weeks after intraocular ICG administration for macular hole surgery. Graefe Arch Clin Exp Ophthalmol 239:388–390 Weinberger AWA, Kirchhof B, Mazinani BE, Schrage NF (2001) Persistent indocyanine green (ICG) fluorescence 6 weeks after intraocular ICG administration for macular hole surgery. Graefe Arch Clin Exp Ophthalmol 239:388–390
44.
go back to reference Weinberger AWA, Schlossmacher B, Dahlke C, Hermel M, Kirchhof B, Schrage NF (2002) Indocyanine green-assisted internal limiting membrane peeling in macular hole surgery—a follow-up study. Graefe Arch Clin Exp Ophthalmol 240:913–917 Weinberger AWA, Schlossmacher B, Dahlke C, Hermel M, Kirchhof B, Schrage NF (2002) Indocyanine green-assisted internal limiting membrane peeling in macular hole surgery—a follow-up study. Graefe Arch Clin Exp Ophthalmol 240:913–917
Metadata
Title
Toxic effects of indocyanine green, infracyanine green, and trypan blue on the human retinal pigmented epithelium
Authors
L. Kodjikian
T. Richter
M. Halberstadt
F. Beby
F. Flueckiger
M. Boehnke
J. G. Garweg
Publication date
01-09-2005
Publisher
Springer-Verlag
Published in
Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 9/2005
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI
https://doi.org/10.1007/s00417-004-1121-6

Other articles of this Issue 9/2005

Graefe's Archive for Clinical and Experimental Ophthalmology 9/2005 Go to the issue