Influence of triamcinolone intravitreal injection on retinochoroidal healing processes
Influence of triamcinolone intravitreal injection on retinochoroidal healing processes
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DOI：
10.1016/j.exer.2007.01.024
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发表时间：
2007-06-01
影响因子：
3.4
通讯作者：
Jeanny, J. C.
中科院分区：
文献类型：
--
作者：
Dot, C.;Behar-Cohen, F.;Jeanny, J. C.
为了分析玻璃体内注射曲安奈德对氩激光视网膜光凝后伤口愈合过程的影响，8-12周龄的野生型C57 BL/6 J小鼠经历标准氩激光光凝方案。在戊巴比妥麻醉和瞳孔放大后，诱导氩激光损伤（50 μ m，400 mW，0.05 s）。两次光凝冲击在双眼中从视神经产生两个圆盘直径。将光凝小鼠分为四组：组I（n = 12），光凝对照，不接受任何玻璃体内注射。Ⅱ组（n = 12）玻璃体腔内注射1 μ l平衡盐溶液（BSS）。第三组（n = 12），玻璃体内注射含15 μ g曲安奈德（TAAC）的BSS溶液1 μ l。在光凝后1、3、7、14天以及2和4个月处死这三组中的每一组的两只小鼠。组IV（n = 10）接受1.5、3、7.5、15或30 μ g TAAC，并在第14天全部处死。对摘除的眼睛进行了系统的分析，细胞重塑过程中发生的激光损伤及其附近。为此，使用针对GFAP、von Willebrand因子、F4/80和K167的特异性抗体来检测星形胶质细胞、活化的Muller细胞、血管内皮细胞、浸润的炎性细胞和活跃增殖的细胞。沿着TUNEL反应和核DAPI染色。所产生的光凝损伤的时间和空间观察表明，氩激光束照射后24小时，在组I和组II的小鼠中观察到RPE细胞和脉络膜的局部和良好描绘的影响。这些小鼠眼睛中的视网膜内层被保留，而在视网膜外核层水平观察到TUNEL阳性（凋亡）细胞。在这个阶段，GFAP的强染色与整个激光路径中激活的视网膜星形胶质细胞和Muller细胞相关。从光凝后第3天开始，在激光损伤的边缘检测到扩张的新脉络膜毛细血管。这些过程伴随着炎性细胞的浸润和病变部位内增殖细胞的存在。用15 μ g/μ l的曲安西龙处理的组III中的小鼠显示浸润炎性细胞和增殖细胞的数量减少，这与未注射的激光处理的对照相比没有统计学显著性。在光凝后的前2个月内，激光损伤边缘的新脉络膜毛细血管的发育也受到抑制。然而，在第4个月，在这些用TAAC处理的小鼠中观察到新血管的生长。即使在小剂量下，第IV组小鼠也未显示任何新毛细血管的形成。氩激光光凝小鼠眼后，玻璃体内注射曲安奈德显着影响视网膜和脉络膜的重建和愈合过程。曲安西龙是该模型中新血管形成的强效抑制剂。然而，这种抑制是短暂的。这些观察结果应提供一个实用的见解的模式TAAC使用湿性AMD患者。(c)2007爱思唯尔有限公司保留所有权利。
To analyze the effects of triamcinolone intravitreal injection on the wound healing processes after argon laser retinal photocoagulation, wild type C57BL/6J mice, 8-12 weeks old underwent a standard argon laser photocoagulation protocol. After pentobarbital anesthesia and pupil dilatation, argon laser lesions were induced (50 mu m, 400 mW, 0.05 s). Two photocoagulation impacts created two disc diameters from the optic nerve in both eyes. The photocoagulated mice were divided into four groups: Group I (n = 12), photocoagulation controls, did not receive any intravitreous injection. Group II (n = 12), received an intravitreous injection of 1 mu l of balanced salt solution (BSS). Group III (n = 12), received an intravitreous injection of 1 mu l containing 15 mu g of triamcinolone acetonide (TAAC) in BSS. Two mice from each of these three groups were sacrificed at 1, 3, 7, 14 days and 2 and 4 months after photocoagulation. Group IV (n = 10) received 1.5, 3, 7.5, 15, or 30 mu g of TAAC and were all sacrificed on day 14. The enucleated eyes were subjected to systematic analysis of the cellular remodeling processes taking place within the laser lesion and its vicinity. To this purpose, specific antibodies against GFAP, von Willebrand factor, F4/80 and K167 were used for the detection of astrocytes, activated Muller cells, vascular endothelial cells, infiltrating inflammatory cells and actively proliferating cells. TUNEL reaction was also carried out along with nuclear DAPI staining. Temporal and spatial observations of the created photocoagulation lesions demonstrate that 24 h following the argon laser beam, a localized and well-delineated affection of the RPE cells and choroid is observed in mice in Groups I and II. The inner retinal layers in these mice eyes are preserved while TUNEL positive (apoptotic) cells are observed at the retinal outer nuclear layer level. At this stage, intense staining with GFAP is associated with activated retinal astrocytes and Muller cells throughout the laser path. From day 3 after photocoagulation, dilated new choroidal capillaries are detected on the edges of the laser lesion. These processes are accompanied by infiltration of inflammatory cells and the presence of proliferating cells within the lesion site. Mice in Group III treated with 15 mu g/mu l of triamcinolone showed a decreased number of infiltrating inflammatory cells and proliferating cells, which was not statistically significant compared to uninjected laser treated controls. The development of new choroidal capillaries on the edges of the laser lesion was also inhibited during the first 2 months after photocoagulation. However, on month 4 the growth of new vessels was observed in these mice treated with TAAC. Mice of Group IV did not show any development of new capillaries even with small doses. After argon laser photocoagulation of the mouse eye, intravitreal injection of triamcinolone markedly influenced the retina and choroid remodeling and healing processes. Triamcinolone is a powerful inhibitor of the formation of neovessels in this model. However, this inhibition is transient. These observations should provide a practical insight for the mode of TAAC use in patients with wet AMD. (c) 2007 Elsevier Ltd. All rights reserved.