Toxicity Assessment of Intravitreal Triamcinolone and Bevacizumab in a Retinal Explant Mouse Model Using Two-Photon Microscopy

被引:14
作者
Schlichtenbrede, Frank C. [1 ]
Mittmann, Wolfgang [2 ]
Rensch, Florian [1 ]
vom Hagen, Franziska [1 ]
Jonas, Jost B. [1 ]
Euler, Thomas [2 ]
机构
[1] Heidelberg Univ, Dept Ophthalmol, Med Fac Mannheim, Heidelberg, Germany
[2] Max Planck Inst Med Res, Dept Biomed Opt, Heidelberg, Germany
关键词
STARBURST AMACRINE CELLS; CALCIUM SIGNALS; GANGLION-CELLS; IN-VIVO; ACETONIDE; RABBIT;
D O I
10.1167/iovs.08-3078
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
PURPOSE. Intravitreal drug administration leads to high intraocular concentrations with potentially toxic effects on ocular tissues. This study was an assessment of the toxicity of triamcinolone and bevacizumab in living retinal explants using two-photon (2P) microscopy. METHODS. Wild-type mice received intravitreal injections of triamcinolone, bevacizumab, or vehicle. Ten and 45 days after injection, wholemounted retinal explants were incubated with the fluorescent dye sulforhodamine 101 (SR101) to analyze morphology and tissue damage with 2P microscopy ex vivo. Retinas that received the same treatment were stained for apoptosis (TUNEL) and glial activation (GFAP). An intravitreal injection of NMDA (N-methyl-D-aspartate) was used as a positive control to ensure the fidelity of detection of retinal damage with ex vivo 2P microscopy. RESULTS. Overall retinal morphology was undisturbed after all procedures and time points. NMDA injection resulted in a strong increase in the number of SR101-labeled cells and increased apoptosis and glial activation when compared with sham-injected eyes. This result was in contrast to exposure to bevacizumab, which caused no appreciable damage. After triamcinolone treatment, marked damage in the inner retina was observed. However, damaged cells were restricted to sharply demarcated areas, and only mild changes in TUNEL-positive cells and GFAP activation was observed when compared to sham-injected eyes. CONCLUSIONS. 2P microscopy in combination with SR101 staining allows fast morphologic assessment of living retinal explants and can be used to evaluate adverse effects on retinal viability of test substances. Bevacizumab treatment did not cause any detectable retinal damage, whereas triamcinolone was associated with substantial, although spatially restricted, damage. (Invest Ophthalmol Vis Sci. 2009;50:5880-5887) DOI: 10.1167/iovs.08-3078
引用
收藏
页码:5880 / 5887
页数:8
相关论文
共 32 条
[1]   Anti-vascular endothelial growth factor therapy for ocular neovascular disease [J].
Andreoli, Christopher M. ;
Miller, Joan W. .
CURRENT OPINION IN OPHTHALMOLOGY, 2007, 18 (06) :502-508
[2]   Optical recording of light-evoked calcium signals in the functionally intact retina [J].
Denk, W ;
Detwiler, PB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (12) :7035-7040
[3]   2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY [J].
DENK, W ;
STRICKLER, JH ;
WEBB, WW .
SCIENCE, 1990, 248 (4951) :73-76
[4]   Photon upmanship: Why multiphoton imaging is more than a gimmick [J].
Denk, W ;
Svoboda, K .
NEURON, 1997, 18 (03) :351-357
[5]  
Denk Winfried, 1995, P445
[6]   Directionally selective calcium signals in dendrites of starburst amacrine cells [J].
Euler, T ;
Detwiler, PB ;
Denk, W .
NATURE, 2002, 418 (6900) :845-852
[7]   Eyecup scope-optical recordings of light stimulus-evoked fluorescence signals in the retina [J].
Euler, Thomas ;
Hausselt, Susanne E. ;
Margolis, David J. ;
Breuninger, Tobias ;
Castell, Xavier ;
Detwiler, Peter B. ;
Denk, Winfried .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 2009, 457 (06) :1393-1414
[8]   Comparison of different techniques for purification of triamcinolone acetonide suspension for intravitreal use [J].
García-Arumí, J ;
Boixadera, A ;
Giralt, J ;
Martinez-Castillo, V ;
Gomez-Ulla, F ;
Corcostegui, B ;
García-Arumí, E .
BRITISH JOURNAL OF OPHTHALMOLOGY, 2005, 89 (09) :1112-1114
[9]   A dendrite-autonomous mechanism for direction selectivity in retinal starburst amacrine cells [J].
Hausselt, Susanne E. ;
Euler, Thomas ;
Detwiler, Peter B. ;
Denk, Winfried .
PLOS BIOLOGY, 2007, 5 (07) :1474-1493
[10]  
Jeon CJ, 1998, J NEUROSCI, V18, P8936