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通过视网膜下注射脂质过氧化氢并连续进行Tookad初步光动力治疗对色素兔实验性诱导脉络膜新生血管形成的临床评估

Clinical evaluation of experimentally induced choroidal neovascularizations in pigmented rabbits by subretinal injection of lipid hydroperoxide and consecutive preliminary photodynamic treatment with Tookad.

作者信息

Framme Carsten, Sachs Helmut G, Kobuch Karin, Flucke Barbara, Birngruber Reginald

机构信息

Medical Laser Center Lübeck, Lübeck, Germany.

出版信息

Ophthalmologica. 2008;222(4):254-64. doi: 10.1159/000136903. Epub 2008 Jun 5.

Abstract

PURPOSE

Up to date several approaches have been undertaken to achieve an 'easy-to-handle' animal model of choroidal neovascularizations (CNVs) in rabbits; however, so far in none of the studies could healthy retinal tissue be maintained, which is mandatory to further investigate the effects of photodynamic therapy (PDT) or anti-vascular-endothelial-growth-factor treatments. It was our aim to reevaluate and verify the method of inducing experimental CNVs in rabbits using subretinally injected linoleic acid hydroperoxide (LHP) as proposed by Tamai et al. and to use it for experimental PDT.

MATERIAL AND METHODS

In 33 eyes of Chinchilla breed rabbits LHP of two different concentrations (25 and 100 microg/50 microl) was injected into the subretinal space via a transvitreal approach under guidance of an operation microscope. Ophthalmoscopic and angiographic examinations were performed on days 3, 7, 14 and 28 after surgery. Preliminary PDT with different experimental parameter sets was performed in 3 eyes using the new photosensitizer Tookad.

RESULTS

Using LHP in the higher concentration, an angiographically determined CNV induction was observed in 27% of all injection sites (n = 34) on days 14 and 28 revealing early well-demarcated and progressive leakage. No CNV was detected at the lower LHP concentration (60 injection sites). Subretinal CNV was verified histologically revealing vessel formation above the retinal pigment epithelium level. Herein, a significant damage to the outer retinal layers was always observed; however, the general structure of the choriocapillary layer was maintained. Tookad PDT was clinically able to completely stop leakage in 1 case and reduce leakage in 2 cases. Histologically the choriocapillary layer was occluded.

CONCLUSION

Subretinal injection of LHP induces angiographically well-demarcated classic CNVs in rabbits; however, the CNV rate was low, and histology revealed severe damage of the outer retinal layers but not of the choriocapillary layer, which is important for studying PDT interactions. Preliminary experimental PDT could clinically stop or reduce leakage from angiographic CNV. Due to the small CNV rate and the significant collateral retinal tissue damage, this model seems to be only of partial suitability for investigating new treatment modalities in CNV.

摘要

目的

目前已经采取了几种方法来建立一种“易于操作”的兔脉络膜新生血管(CNV)动物模型;然而,到目前为止,在所有研究中均未能维持健康的视网膜组织,而这对于进一步研究光动力疗法(PDT)或抗血管内皮生长因子治疗的效果是必不可少的。我们的目的是重新评估并验证Tamai等人提出的通过视网膜下注射氢过氧化亚油酸(LHP)诱导兔实验性CNV的方法,并将其用于实验性PDT。

材料与方法

在33只青紫蓝兔的眼中,在手术显微镜引导下,通过经玻璃体途径将两种不同浓度(25和100μg/50μl)的LHP注射到视网膜下间隙。术后第3、7、14和28天进行眼底镜和血管造影检查。使用新型光敏剂Tookad对3只眼进行了不同实验参数设置的初步PDT。

结果

使用较高浓度的LHP时,在第14天和28天,在所有注射部位(n = 34)的27%观察到血管造影确定的CNV诱导,显示早期界限清楚且逐渐进展的渗漏。在较低LHP浓度下未检测到CNV(60个注射部位)。视网膜下CNV经组织学证实,显示在视网膜色素上皮层上方有血管形成。在此,总是观察到视网膜外层有明显损伤;然而,脉络膜毛细血管层的总体结构得以维持。Tookad PDT在临床上能够使1例渗漏完全停止,2例渗漏减少。组织学上脉络膜毛细血管层闭塞。

结论

视网膜下注射LHP可在兔中诱导血管造影界限清楚的典型CNV;然而,CNV发生率较低,并且组织学显示视网膜外层有严重损伤,但脉络膜毛细血管层未受损伤,这对于研究PDT相互作用很重要。初步实验性PDT在临床上能够阻止或减少血管造影CNV的渗漏。由于CNV发生率低且视网膜组织有明显的附带损伤,该模型似乎仅部分适用于研究CNV的新治疗方式。

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