大鼠角膜移植后新生血管演变观察(英文)

目的:观察大鼠同种异体穿透性角膜移植后无干预下角膜新生血管的生长演变。方法:SPF级雌性SD大鼠34眼行同种异体穿透性角膜移植,术后4,7,15和30d在显微镜下观察角膜新生血管的生长情况,通过公式C/12×3.14×[r2-(r-I)2]计算角膜新生血管的面积。结果:在移植的34眼中出现新生血管的29眼(85%)。新生血管最初环角膜缘呈毛刷状生长,后向角膜中央蔓延,为扭曲粗大的血管,末端呈树枝状分支。高峰时期交叉密布成网状,以后逐渐萎缩。角膜移植后4,7,15和30d角膜新生血管平均面积分别为11.8±3.5mm2,18.5±4.0mm2,14.4±4.3mm2和6.0±1.8mm2,总体平均面积为12.7±1.9mm2;新生血管面积占角膜面积百分比值分别为30.8%±8.7%,65.3%±12.8%, 59.4%±14.5%和36.2%±10.9%,总体为48.7%±6.4%。结论:大鼠同种异体穿透性角膜移植后角膜新生血管在第4d时出现,7d时面积达最大值;以后逐渐下降,30d时约降至7d时面积的1/2。     

大鼠角膜移植后新生血管的演变

目的 现察大鼠同种异体穿透性角膜移植后无干预下角膜新生血管的生长演变.方法 SPF级雌性SD大鼠34只(34眼)行同种异体穿透性角膜移植,术后不进行任何治疗,分别于术后第4天、第7天、第15天和第30天在裂隙灯显微镜下观察角膜新生血管的生长情况,利用公式C/12×3.14×[r2-(r-I)2]计算角膜新生血管的面积,并比较不同时间点角膜新生血管面积占全角膜面积的百分比.结果 移植后的32眼(剔除1只死亡鼠及1只丢失鼠)中出现新生血管29眼,占90.6%.新生血管最初环角膜缘呈毛刷状生长,后向角膜中央蔓延.高峰时期交叉密布成网状,以后逐渐萎缩.角膜移植后第4天、第7天、第15天和第30天角膜新生血管平均面积分别为(11.83±3.35)mm2、(18.52±3.95)mm2、(14.38±4.28)mm2和(6.00±1.84)mm2,总体平均面积为(12.68mm±1.87)mm2;新生血管面积占角膜面积百分比值均数分别为(30.76±8.72)%、(65.25±12.78)%、(59.35±14.48)%和(36.15±10.90)%,总体为(48.67±6.38)%.结论 大鼠同种异体穿透性角膜移植后无干预下角膜新生血管发生率极高,且新生血管来源于结膜角膜缘的毛细血管丛.术后血管新生立即开始,因此应尽早开始抗炎治疗,防止新生血管出现.     

人Kringle1-5蛋白滴眼液抑制兔角膜新生血管的研究

目的:观察人Kringle1-5(K1-5)蛋白滴眼液来观察其抑制角膜新生血管的效果。方法:随机将缝线法诱导单侧角膜新生血管的28只新西兰白兔分为两组。K1-5组给予人K1-5蛋白滴眼液点眼,PBS组给予PBS液点眼。观察两组角膜新生血管形成时间、最长新生血管、新生血管面积并进行统计学分析。随机选择处死实验兔,取下角膜组织进行病理检查。结果:K1-5组最早于3d,平均3.50±0.52d可见新生血管芽进入角膜缘,9～15d新生血管生长最为旺盛;20d时最长新生血管为4.38±0.27mm,新生血管面积27.51±4.19mm2。而对照组最早于2d,平均2.28±0.47d可见新生血管芽进入角膜缘,4～10d时新生血管生长最为旺盛;至20d时新生血管最长为7.33±0.46mm,新生血管面积为38.96±4.11mm2。两组角膜新生血管出现时间、最长新生血管、新生血管面积方面均具有统计学差异(P<0.01)。在不同时段里K1-5组角膜基质层缝线周围新生血管较少,局部有少量浆细胞及成纤维细胞;PBS组新生血管较多,局部有较多浆细胞、淋巴细胞、成纤维细胞。K1-5组在不同时段里角膜组织中VEGF121阳性表达少于PBS组。结论:人K1-5蛋白点眼能够抑制缝线法诱导的角膜新生血管形成和生长。     

FTY720抑制S1P诱导的角膜新生血管的研究

目的：：探讨芬戈莫德(fingolimod,FTY720)对大鼠角膜新生血管( CNV)的抑制作用。方法：分别采用MTT法和划痕法观察不同浓度FTY720对人脐静脉内皮细胞( HUVECs )的增生和S1 P诱导下的细胞迁移的影响。应用大鼠角膜微囊袋模型,检测FTY720对S1 P诱导的CNV的作用。将30只SD大鼠按随机数字表法分成A、B和C组,每组10只,在各组角膜基质层内植入S1 P的同时依次植入0,5,20μg FTY720缓释颗粒。术后对CNV生长情况观察,并在12 d行组织病理学检查。实验结果采用单因素方差分析。结果：10,102,103,104 nmol/L FTY720与HUVECs共孵育72h可抑制细胞增生(P<0.01),10,100nmol/L FTY720作用24 h后,可抑制由S1 P诱导的细胞迁移,随FTY720浓度增加其抑制细胞增生和迁移的作用均增强,A、B、C组大鼠角膜微囊袋缓释微粒体植入后12d,CNV面积分别为10.05±1.19,6.59±0.95,2.70±0.68mm2(F=145.155, P<0.01),A与B组、B与C组间均有统计学差异(P<0.01)。     

螺内酯抑制大鼠角膜碱烧伤新生血管形成的作用

目的:观察螺内酯(spironolactone,SL)对大鼠角膜碱烧伤后角膜新生血管(corneal neovascularization,CRNV)形成的抑制作用。方法:健康SD大鼠42只随机分为3组,6只6眼为正常组,其余36只36眼建立碱烧伤诱导的大鼠CRNV模型后随机分为实验组和对照组,每组18只。实验组给予SL(100mg/kg)灌胃,对照组给予等量生理盐水灌胃,1次/d。术后第4,7,14d在裂隙灯显微镜下观察CRNV的面积。结果:对照组在碱烧伤后4d,大鼠角巩膜缘处有新生血管芽成刷状生长,生长速度迅速,至14d新生血管交织成网状,几乎布满整个角膜。实验组大鼠CRNV管径细、分布生长速度缓慢。在碱烧伤后4,7和14d实验组CRNV面积均明显小于对照组(P<0.01)。     

角膜缝线诱导建立大鼠角膜新生血管模型

目的探讨在大鼠角膜基质缝线诱发角膜新生血管(CNV)模型的制作技巧及特点。方法20只SD大鼠,在手术显微镜下用10-0号尼龙线铲针在角膜基质层间置3针缝线(11点、12点、1点3个钟点位置),自距离角膜缘略小于2.0mm向瞳孔中心方向进针。采用裂隙灯显微数字图像处理系统动态观察角膜新生血管的生长情况,并观察CNV模型的角膜组织病理学变化。结果①角膜缝线后4d可见明显从角膜缘伸入角膜的毛刷状小血管、垂直角膜缘切线方向,角膜缝线处水肿;随着角膜水肿继续加重,8d新生血管延伸到达或超过缝线位置,分支密集并互相吻合形成袢状血管网、生长范围不超过50%圆周;②HE染色显示在正常角膜切片基质层仅见成纤维细胞和纤维母细胞,而在CNV模型角膜切片则见角膜上皮层水肿增厚、浅层基质内密集大量新生血管、管腔大小不等,新生血管周围有大量分叶核炎性细胞浸润。结论大鼠角膜显微缝线诱导的新生血管生长稳定、适于定量研究。     

大鼠角膜碱烧伤后CD105在新生血管化角膜中的表达

目的:研究大鼠角膜碱烧伤后CD105在新生血管化角膜组织中的表达。方法:采用角膜碱烧伤法对60只Wistar大鼠制作角膜新生血管模型。裂隙灯显微镜下观察大鼠角膜碱烧伤后角膜新生血管生长情况,免疫组化法检测碱烧伤后不同时间角膜组织中CD105的表达。结果:大鼠角膜碱烧伤后2d角膜新生血管开始形成,3～7d生长旺盛,新生血管面积达到最大,10～14d角膜新生血管生长停滞。在新生血管化角膜组织中,CD105于3～7d表达最明显,14d后表达微弱。结论:大鼠角膜碱烧伤后CD105在新生血管化角膜组织中有明显表达。     

恒温下兔角膜热烧伤后继发新生血管建模初步研究

目的：初步研究恒温条件下兔角膜热烧伤后继发的角膜新生血管（ CNV）快速制模的合适条件。 方法：新西兰大白兔45只,随机分为（ A,B,C,D,E）五组恒温烧灼器诱导实验性CNV模型,除E组为5只,其他组均为10只。 A组：100℃, B组：200℃, C组：300℃, D组：400℃,E组：空白对照。以左眼为实验眼,比较建模后第4,7,14,30 d在裂隙灯显微镜下观察各组角膜新生血管的生长情况并计算角膜新生血管生长面积。采用SPSS 19.0统计软件包对数据进行分析,计算资料用x-±s表示,各时间点各自新生血管面积的比较采用重复测量数据的方差分析;显著性标准为α=0.05。 结果：制模后第4,7,14,30d,A组角膜热烧伤后无明显新生血管生长,仅少量留有角膜云翳（n=2）。 B组新生血管面积分别为：9.16±1.45,37.73±5.49,62.44±7.54,40.28±7.39mm2;C组新生血管面积分别为：11.45±1.04,44.51±4.64,66.13±4.13,43.04±2.33mm2;D组新生血管面积分别为：13.23±0.86,47.26±4.59,67.57±4.56,45.59±4.44mm2;D组部分角膜局部出现焦化（n=4）,3d内出现穿孔（ n=6）。 E组未见新生血管生长。各时间点的CNV面积比较,差异有显著性（P〈0.05）,B,C,D分组之间有统计学差异（P〈0.05）。 结论：角膜热烧伤4~7d时,制模温度越高,新生血管生长越快,面积越大;14~30 d 新生血管面积无明显差别,但400℃角膜制模失败率高,200℃~300℃制作的兔角膜新生血管模型均一性及重复性高。     

bFGF缓释微囊颗粒诱导角膜新生血管形成治疗兔大泡性角膜病变

目的:观察bFGF诱导角膜新生血管形成,对兔大泡性角膜病变的治疗作用。方法:采用0.5g/L苯扎溴铵溶液(BAKB)前房灌注复制兔大泡性角膜病变模型。然后将bFGF缓释微囊颗粒植入病变兔眼的角膜基质层间,诱导角膜新生血管生长,观察随角膜新生血管的形成,病变角膜水肿及大泡的改变。结果:在微囊颗粒植入后4wk病变角膜中央厚度2.61±0.25(CT),角膜水肿度评分0分,角膜新生血管评分0分,病变大泡呈吸收性改变。结论:大泡性角膜病变动物模型角膜基质层间植入bFGF缓释微囊颗粒,能够较为迅速的诱导角膜新生血管形成。同时,伴随CNV的生长,病变角膜的临床症状逐渐缓解。因此,该实验方法可能为大泡性角膜病变的治疗提供了另一种思路和方法。     

瘦素和血管内皮生长因子在碱烧伤诱导的大鼠角膜新生血管模型中的表达

目的 观察碱烧伤后不同时期角膜的组织病理学变化,以及瘦素和血管内皮生长因子(vascular endothelial growth factor,VFGF)的表达,探讨它们与角膜新生血管(corneal neovascularization,CNV)的关系.方法 使用碱烧伤诱导大鼠角膜新生血管模型,采用浸润1 mol·L-1 NaOH溶液、直径为3 mm的单片滤纸,准确置于大鼠右眼角膜中央30 s,制作CNV模型.25只SD大鼠右眼为实验眼,随机分成5组,每组5眼,左眼为正常对照眼.每天用裂隙灯显微镜观察角膜新生血管,分别计算新生血管长度和面积.并行HE染色和免疫组织化学检测.结果 碱烧伤后4 d、7 d、14 d、21 d,实验组CNV面积分别为:(2.55±0.15)m2、(4.15±0.36)mm2、(10.21±0.45)mm2、(8.56±0.06)mm2.免疫组织化学检测显示角膜碱烧伤后1 d,实验组瘦素、VEGF的表达开始增高,碱烧伤后14 d达高峰,14 d后瘦素、VEGF的表达下降,21 d后显著下降;其表达部位主要分布在形成的新生血管区域和上皮层;正常对照组瘦素可微弱表达于角膜上皮层和基质层,VEGF没有表达或仅在上皮基底膜有微弱表达.实验组角膜瘦素、VEGF阳性表达率较对照组明显增加(χ2=29.07,P=0.001;χ2=35.51,P=0.001),瘦素与VEGF的表达具有正相关性(r=0.95,P=0.001).结论 瘦素和VEGF表达水平与角膜新生血管的生成具有相关性,瘦素可能成为治疗CNV的靶点.     

Angiogenesis induced by micropocket assay on rat cornea

AIM: To explore the skills and characteristics of corneal neovascular model in rat induced by micropocket assay. METHODS: Nine eyes of nine Sprague-Dawley rats were studied. Pellets made of vascular endothelial growth factor (VEGF), poly-2-hydroxylethylmethacrylate and sucralfate were implanted into the corneal stroma no closer than 1mm from the limbus. Biomicroscopic features of corneal neovascular were observed on 1, 3, 5, 7th day after the implantation. RESULTS: On day 1 after operation, the limbal vessels were dilated, with no angiogenesis appeared On day 3, angiogenesis began to invade pericornea with a brush shape, the area of CNV was (2.23±0.59)mm². On day 5, new vessels reached the lower margin of pellet densely, and the area of CNV was (6.81±1.35)mm². On day 7, new vessels continued to elongate, parts of them extended as loops toward the pellet, and the area of CNV was (8.92 ±1.79)mm². Neither hyphema or other complications occurred. CONCLUSION: Corneal neovascularization induced by micropocket assay in rat grows steadily without complication, and is suitable for quantitative researches.     

Quantification of angiogenesis due to basic fibroblast growth factor in a modified rabbit corneal model.

The rabbit corneal angiogenesis assay was modified to allow quantification of the neovascular response induced by growth factors. Basic Fibroblast Growth Factor (bFGF) was used for this experiment. bFGF was diluted in phosphate buffer solution (PBS) in such a way that 2 microliter of solution contained concentrations ranging from 17 to 1,200 ng. Each drop was absorbed by a 1 x 2 mm particle of a dried 70% hydratable hydrogel and implanted into a mid-stroma corneal pocket, 2 mm from the limbus. Iodinated bFGF was used to measure growth factor diffusion in the cornea, which was found to be isotropic. Autoradiography showed that bFGF was stored in the cornea at both epithelial and endothelial level. Corneal neovascularization occurred on the second day after implantation and was maximal on the 7th day. At this time the neovascular surface was measured by planimetry on corneal photographs and compared with controls. This method allows precise definition of the dose of any substance to be implanted into the cornea and induces a rapid neovascular response, thus allowing quantitative evaluation of neovascularization within one week. A neovascular response was detectable for doses as low as 35 ng of bFGF and increased proportionally to the dose of bFGF implanted.     

A method to quantify neovascularization in the mouse cornea.

A method for microscopical observation and quantification of corneal neovascularization in the living mouse is described. Measurements of the corneal vasculature were made on still-mode video-pictures of the cornea. The validity of the method was shown by the high correlation between the morphometric measurements and computerized values on the same vessel structures. The method was applied to describe the neovascular response of heat-killed tumour fragments implanted into corneal pockets. The neovascular response that was observed in some corneas was dependent on the distance from the edge of the pocket with heat-killed material to the nearest vessel loop. The pocket area was vascularized only if this distance (measured the 3rd day after implantation) was less than 0.70 mm. Resorption of heat-killed tissue occurred in corneas without any neovascular response and it was not altered by penetration of vessel into the cornea. The described method proved to be suitable for studying the neovascularization induced by corneal pocket implants.     

Local suppression of IL-1 by receptor antagonist in the rat model of corneal alkali injury

Proinflammatory cytokines, including interleukin-1 (IL-1) have been implicated in the inflammation that follows corneal alkali injury. The purpose of this series of experiments was to test whether topically applied interleukin-1 receptor antagonist (IL-1ra) could suppress corneal inflammation and promote transparency after alkali injury. Alkali injury was induced on day 0 by application of 1N NaOH to both eyes of Wistar rats (n = 28). Immediately thereafter, eyes received either topical IL-1ra (20 mg ml(-1)) in 0.2% sodium hyaluronate or vehicle alone three times daily during days 0-14. Biomicroscopic features including corneal opacity and neovascularization were assessed using a standard grading scheme. Inflammation was quantified histologically. Corneas excised at day 3 and 7 (randomly selected six eyes in each group per time point studied) were homogenized, and levels of IL-1alpha, IL-1beta, IL-6, IL-8, IL-10, and RANTES were quantified by enzyme-linked immunosorbent assay. Epithelial wound healing was examined by computed analysis of fluorescein stained corneal photographs taken daily until day 14. According to these evaluations, eyes treated with IL-1ra maintained corneal transparency with minimal neovascular invasion. Additionally, corneal damage and cell infiltration were reduced on day 7 (infiltration cells were almost 40% decreased). All cytokine/chemokine levels in IL-1ra treated eyes were significantly lower at day 3, and IL-6 and IL-10 remained significantly lower at day 7 compared to vehicle-treated eyes. IL-1ra treatment retarded epithelial wound healing in the early stage (day 1-4); however, subsequently IL-1ra treated eyes had enhanced healing with full epithelial closure at nearly the same time point as vehicle-treated eyes (day 10). We conclude that local antagonism of IL-1 after alkali injury can significantly decrease corneal inflammation and lead to enhanced corneal transparency.     

Effect of bevacizumab on corneal neovascularization in experimental rabbit model

Purpose:  To investigate the effect of bevacizumab in an experimental rabbit model of corneal neovascularization.
Methods:  The right eyes of 24 white New Zealand rabbits were included in a corneal neovascularization model using alkaline burn. They were divided into four groups. Topical bevacizumab was installed three times daily in group 1, 5 mg bevacizumab subconjunctivally every 2 days in group 2, 10 mg bevacizumab subconjunctivally every 2 days in group 3 and 0.2 cc of normal saline in the same way in group 4 (control group). All eyes were treated for 7 days. Then the animals were killed and corneal specimens sent for histopathological analysis. Tear film and aqueous humour samples were obtained to assess vascular endothelial growth factor (VEGF) levels.
Results:  Seven days after topical bevacizumab treatment the neovascular index in group 1 was lower than that in the control group ( P  = 0.028). In groups 2 and 3 the neovascular index was lower 2 days after subconjunctival bevacizumab treatment than that in control group ( P  = 0.009 and P  = 0.009, respectively). In the control group the VEGF level in aqueous humour increased by 66% from day 7 to 14. In groups 1–3 it decreased by 49.80%, 70.20% and 76.44%, respectively ( P  = 0.043). The VEGF level in tear film of the control group increased by 35.23% from day 7 to 14, which was not significant ( P  = 0.893), while in groups 1–3 it decreased by 57.26%, 34.59% and 67.97%, respectively, which was only significant in groups 1 and 3 ( P  = 0.043).
Conclusions:  Subconjunctival 5 mg/mL bevacizumab is effective in reducing corneal neovascularization in animal models and in reducing VEGF levels. Further research is needed to assess the potential side effects and minimal effective dose.     

Central corneal thickness in patients with neovascular age-related macular degeneration

PURPOSE: To compare the central corneal thickness (CCT) measurements of patients with neovascular age-related macular degeneration (AMD) and control subjects. METHODS: The CCT value (measured with ultrasound corneal pachymetry) of 130 eyes (130 patients, 1 eye from each patient) with neovascular AMD (AMD group) and 98 eyes (98 patients, 1 eye from each patient) of similar age, sex, and eye's axial length healthy control subjects (normal group) was compared. RESULTS: The mean age (AMD group: 69.1 years vs. control group: 69.5 years, P = 0.81), sex (AMD group: 77 women, 59% vs. control group: 59 women, 60%, P = 0.77), and eye's axial length (AMD group: 25.05-mm vs. control group: 24.61-mm, P = 0.38) of patients with neovascular AMD and healthy control subjects were comparable. There were no statistically significant differences in the mean CCT measurements in the neovascular AMD group in comparison with the control group (549.44 vs. 544.35 microm, P = 0.11). CONCLUSIONS: CCT measurements do not differ in patients with neovascular AMD compared with healthy control subjects.     

雷帕霉素抑制大鼠角膜移植免疫排斥反应的实验研究

目的探讨雷帕霉素(RAPA)对大鼠角膜移植排斥反应的防治效果。方法以SD大鼠为供体,Wistar大鼠为受体建立角膜移植实验模型。采用完全随机分组设计方案,将68只Wistar鼠(68只眼)随机分为4组,A组为Wistar鼠自体原位角膜移植,B、C、D组为SDWistar鼠之间进行同种异体角膜移植。术后灌胃给药,A组和B组给予空白液,C组和D组分别给予RAPA(3mg·kg-1·d-1)和环孢霉素A(CsA)(10mg·kg-1·d-1),连续用药12d。根据Holland排斥反应评分系统,判断术后植片排斥情况。比较各组角膜植片的平均存活时间和植片存活率。采用广义估计方程对角膜的新生血管评分进行统计学分析。术后14d,对各组角膜进行组织学检查。结果RAPA组发生排斥反应的时间明显延迟,同种异体移植对照组植片平均存活时间(MST)为(11.0±1.5)d,RAPA组MST为(36.1±14.9)d,两组比较差异有统计学意义(P<0.05)。RAPA组角膜植片存活情况较CsA组好,但两组植片存活率比较差异无统计学意义(P>0.05)。RAPA组术后角膜新生血管的生长明显低于异体移植对照组(P<0.05)和CsA组(P<0.05)。组织学检查证实,术后14d,RAPA组角膜植片未见明显淋巴细胞浸润。结论口服RAPA能够延缓大鼠角膜移植排斥反应的发生,并能够抑制术后新生血管的生成。     

^99Tc-MDP对碱烧伤性大鼠角膜新生血管生成的抑制作用

目的：探讨^99Tc-MDP对碱烧伤性大鼠角膜新生血管生成的抑制作用。方法：采用碱烧伤法制备角膜新生血管模型,24只健康SD大鼠随机分成两组,实验组予以腹腔注射^99Tc-MDP,每天1次,至模型建立后14d。对照组以等体积的生理盐水腹腔注射。观察新生血管形成时间、第21d时角膜新生血管长度和新生血管面积。结果：实验组新生血管长入的时间为6．25±1．93d,对照组长入时间为5．83±1．86d,差异无统计学意义（P=0．30）。新生血管长度实验组为1．7±0．33mm,对照组为2．82±0．25mm;新生血管的面积实验组为17．3±3．26mm^2,对照组为24．8±5．10mm^2,两者之间差异均有统计学意义（P〈0．01）。结论：使用^99Tc-MDP可以有效抑制碱烧伤引起的角膜新生血管的生成。     

Bevacizumab inhibits corneal neovascularization in an alkali burn induced model of corneal angiogenesis

BACKGROUND: New and uncontrolled blood vessel development in the cornea is a pivotal process in the pathogenesis of several corneal diseases. These corneal diseases may finally cause blindness and managing them therapeutically is problematic. The data supporting a causal role for vascular endothelial growth factor in corneal neovascularization are extensive. This study aimed to evaluate the effect of subconjunctival bevacizumab (Avastin) on experimental corneal neovascularization in rabbits. METHODS: Chemical cauterization of the cornea was performed by touching central cornea with a 5-mm-diameter NaOH-soaked cotton applicator for 10 s in 20 eyes of 20 White New Zealand rabbits. The rabbits were then divided randomly into two equal groups. Bevacizumab (2.5 mg) was administered to 10 eyes (group 1) by a subconjunctival injection immediately after chemical cauterization of corneal surface. As a control, 10 eyes (group 2) received an injection of distilled water. Rabbits were examined daily for detection of the first signs of neovascularization. Three weeks later, the extent of corneal neovascularization was evaluated by direct examination and photograph analyses. Total corneal neovascularization area, degree of circumference involved and longest neovascular pedicle length were assessed. RESULTS: Bevacizumab significantly decreased the total neovascularization area (P < 0.009), the circumference involved (P < 0.011) and the longest neovascular pedicle length (P < 0.023). CONCLUSION: Local injection of bevacizumab has a significant effect on inhibition of alkali burn-induced corneal neovascularization. This shows the potential value of bevacizumab in the treatment of corneal neovascularization.     

Expression of cell adhesion molecules on limbal and neovascular endothelium in corneal inflammatory neovascularization.

PURPOSE: To investigate the expression of cell-adhesion molecules on corneolimbal and neovascular endothelium and the associated leukocyte infiltration in an experimental model of inflammatory corneal neovascularization (NV). METHODS: Corneal NV was induced in BALB/c mice by placement of nylon sutures. Interleukin-1 receptor antagonist (IL-1ra) was used topically to determine whether suppression of IL-1 could affect adhesion molecule expression and leukocytic infiltration. At set time points, corneal samples were analyzed immunohistochemically for expression of P-selectin, E-selectin, intercellular adhesion molecule (ICAM)-1, vascular adhesion molecule (VCAM)-1, and platelet- endothelial adhesion molecule (PECAM)-1. Leukocytic infiltration at different time points was quantified histologically. In companion experiments mice deficient in ICAM-1 were investigated to determine the functional relevance of this molecule in corneal leukocyte infiltration. RESULTS: Significant enhanced expression of ICAM-1 was detected on the corneolimbal vascular endothelium as early as 8 hours and on the newly formed corneal NV by day 3, and treatment with IL-1ra led to significant suppression of this expression. IL-1ra-induced suppression of ICAM-1 expression was accompanied by a profound decrease in corneal leukocytic infiltration by 44.6% at day 1 (P < 0.003), 71.8% at day 3 (P < 0.001), 60.1% at day 7 (P < 0.001), and 63.8% at day 14 (P < 0.001), compared with control corneas. Similarly, in ICAM-1 knockout mice, the corneal leukocytic infiltration was 50.3%, 52.9%, and 36.4%, compared with wild-type control animals on day 1 (P < 0.001), day 7 (P < 0.005), and day 14 (P < 0.001), respectively. Expression of PECAM-1 was constitutively present on perilimbal vascular endothelium and had no response to IL-1ra treatment. No significant expression of P-selectin, E-selectin, or VCAM-1 was detected in this experimental model. CONCLUSIONS: These results suggest that leukocytic infiltration in this model of inflammatory corneal NV is closely associated with ICAM-1 expression, and that topical IL-1ra displays corneal anti-inflammatory effects, largely by suppressing ICAM-1 expression on vascular endothelial cells.