Lassota Nathan, Kiilgaard Jens Folke, la Cour Morten, Scherfig Erik, Prause Jan Ulrik
Department of Neuroscience and Pharmacology, Eye Pathology Section, University of Copenhagen, Copenhagen, Denmark.
Acta Ophthalmol. 2008 Aug;86(5):495-503. doi: 10.1111/j.1600-0420.2007.01127.x.
To study an expanded time course of surgically induced choroidal neovascularization (CNV) in a porcine model applying fluorescence angiography and immunohistology.
Twenty-two porcine eyes underwent vitrectomy, a retinal bleb was raised and the detached retina perforated using endodiathermy. Bruch's membrane was perforated with a retinal perforator at a site where the overlying neuroretina was normal. Eyes were enucleated in a time interval between 30 min and 42 days after the perforation, and the pigs were subsequently killed. Immediately prior to enucleation, fundus photographs and fluorescein angiograms were obtained. Sections of paraffin-embedded eyes were immunohistochemically stained.
On fluorescein angiography, membranes aged 14 days or less exhibited leakage in 10/11 cases while the remaining showed persistent staining. The propensity to leak diminished with time and only 1/3 of the oldest membranes leaked. In eyes enucleated immediately after surgery, neuroretinas overlying the induced lesions were intact without apparent atrophy of cells. At day 3, macrophages and myofibroblasts formed membrane-like structures in the subretinal space. At day 7, the outer surface of the membrane was covered by retinal pigment epithelium (RPE) cells and the neuroretinas had suffered damage in the form of outer segment loss. In the time period 14-42 days, the CNV membrane became completely enveloped by RPE cells. The degree of membrane vascularization increased with time and was at its maximum after 42 days. Intact outer segments were identified over the oldest membranes.
The formation of surgical CNV membranes followed the normal reparatory pathway and the degree of vascularization of CNV membranes continued to increase during the 42 days. However, propensity to leak diminished with time. We believe that this was because of the fact that RPE cells completely enveloped older membrane and thus prevented leakage from the newly formed vessels. Photoreceptor outer segments, which had atrophied after 7 days, were able to regenerate over CNV membranes and could be identified again after 42 days.
应用荧光血管造影术和免疫组织学方法,研究猪模型中手术诱导脉络膜新生血管(CNV)的扩展时间进程。
对22只猪眼进行玻璃体切除术,形成视网膜下液泡,并用透热疗法使脱离的视网膜穿孔。在神经视网膜正常的部位,用视网膜穿孔器穿透 Bruch 膜。在穿孔后30分钟至42天的时间间隔内摘除眼球,随后处死猪。在摘除眼球前立即拍摄眼底照片和荧光素血管造影照片。对石蜡包埋的眼球切片进行免疫组织化学染色。
在荧光素血管造影中,14天及以内的新生血管膜在11例中有10例出现渗漏,其余表现为持续染色。渗漏倾向随时间降低,最老的新生血管膜只有1/3发生渗漏。术后立即摘除的眼球中,诱导病变上方的神经视网膜完整,细胞无明显萎缩。第3天,巨噬细胞和成肌纤维细胞在视网膜下间隙形成膜样结构。第7天,新生血管膜外表面被视网膜色素上皮(RPE)细胞覆盖,神经视网膜出现外节缺失形式的损伤。在14 - 42天期间,CNV膜完全被RPE细胞包裹。新生血管膜的血管化程度随时间增加,42天后达到最大值。在最老的新生血管膜上方发现完整的外节。
手术诱导的CNV膜形成遵循正常的修复途径,CNV膜的血管化程度在42天内持续增加。然而,渗漏倾向随时间降低。我们认为这是因为RPE细胞完全包裹了较老的新生血管膜从而阻止了新形成血管的渗漏。7天后萎缩的光感受器外节能够在CNV膜上再生,并在42天后再次被识别。