贝伐单抗结膜下注射对青光眼滤过术后滤过泡瘢痕化的抑制作用

背景 青光眼滤过手术术后滤过道的瘢痕化是导致手术失败的主要原因.传统的抑制滤过道瘢痕化的方法是丝裂霉素C的应用,但存在较多的并发症.研究表明贝伐单抗具有抑制新生血管和纤维增生的作用,其对青光眼滤过术后滤过泡瘢痕化是否有抑制作用受到关注. 目的 观察贝伐单抗结膜下注射对兔眼小梁切除术后滤过泡纤维瘢痕形成的抑制效果. 方法 按随机数字表法将7～9周龄新西兰大白兔40只随机分为4个组,各组兔右眼均行常规小梁网切除术.贝伐单抗单次注射组兔眼术毕结膜下注射贝伐单抗0.05 ml(25 mg/ml),贝伐单抗多次注射组兔眼分别于术毕及术后3、7d注射贝伐单抗,每次均注射0.05 ml,丝裂霉素C组兔眼术毕局部涂用丝裂霉素C,生理盐水组兔眼术毕以同样的方法注射0.05 ml生理盐水.所有兔眼术后每隔1日用Schi(o)tz眼压计测量眼压,行裂隙灯显微镜检查以观察滤过泡形态及其表面的血管分布,并用卡尺测量和计算滤过泡面积.分别于术后14d和28 d摘取实验眼行滤过泡组织病理学检查,采用免疫组织化学法检测滤过道组织中血管内皮细胞标志物CD31的表达以计算微血管数目. 结果 各组兔眼术后眼压值的总体比较差异无统计学意义(F=0.88,P=0.47).与贝伐单抗单次注射组、丝裂霉素C组和生理盐水组兔眼滤过泡的形态比较,术后7d贝伐单抗多次注射组兔眼滤过泡高度隆起且弥散,表面血管稀疏.贝伐单抗多次注射组兔眼滤过泡生存时间为27 d,而贝伐单抗单次注射组、丝裂霉素C组均为19d,生理盐水组为13d.术后14d各组兔眼滤过道胶原纤维百分比分别为(49.18±1.54)％、(26.41±1.23)％、(50.68±1.87)％和(70.63±1.81)％,贝伐单抗单次注射组、贝伐单抗多次注射组、丝裂霉素C组滤过道胶原纤维百分比均低于生理盐水组,贝伐单抗多次注射组低于贝伐单抗单次注射组,差异均有统计学意义(P＜0.05);术后28 d贝伐单抗多次注射组胶原纤维百分比为(66.82±1.53)％,其他3个组出现瘢痕化.术后14d贝伐单抗多次注射组兔眼滤过道组织中微血管数目明显低于贝伐单抗单次注射组、丝裂霉素C组和生理盐水组,差异均有统计学意义(均P＜0.05).术后28 d贝伐单抗多次注射组兔眼滤过道组织中微血管数且为3.51±0.31,均高于贝伐单抗注射组、丝裂霉素组和生理盐水组,差异均有统计学意义(均P＜0.05). 结论 青光眼滤过术后结膜下注射贝伐单抗有助于维持功能滤过泡的形态,抑制滤过道瘢痕化,提高手术的成功率.     

结膜下注射贝伐单抗对兔角膜新生血管的抑制作用

目的 观察碱烧伤后不同时间结膜下注射贝伐单抗（Bevacizumab）角膜新生血管（CNV）的形成与转归.方法 新西兰白兔54只,制成单眼碱烧伤模型,随机分为3组,每组18只眼,A组碱烧伤后结膜下立即注射贝伐单抗2.5 mg（0.1 ml）,B组碱烧伤后3d结膜下注射贝伐单抗2.5 mg（0.1 ml）,C组结膜下注射生理盐水0.1ml,为对照组.共观察28 d.裂隙灯显微镜下观察角膜新生血管生长情况,行眼前段照相并计算其面积,伤后7、14、28 d各组随机取6例角膜行共焦显微镜检查,观察角膜组织炎性细胞浸润情况及角膜新生血管形态学变化.结果 A、B及C组角膜新生血管开始出现的时间分别为（5.9＋0.8）d、（3.5＋0.6）d及（3.4＋1.1）d,其中A组明显较C组延长（P＜0.05）,B组与C组差异无统计学意义（P =0.068）.伤后各时间点A、B组角膜新生血管的生长面积均明显较C组减少（P＜0.05）,A组与B组角膜新生血管面积比较,差异有统计学意义（P＜0.05）.共焦显微镜检查可见C组烧伤区大量炎性细胞浸润及新生血管形成,而A组角膜炎性细胞较少,烧伤区无新生血管形成,B组见少量新生血管侵入烧伤区.3组基质层均可见纤维及瘢痕组织增生,其中治疗组纤维增生程度与瘢痕组织均较对照组轻.结论 结膜下注射贝伐单抗可抑制角膜炎性细胞形成,改善损伤角膜基质,促进角膜愈合,从而减少碱烧伤引起的角膜新生血管的生长,在早期注射能取得更好的疗效.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

贝伐单抗在眼表新生血管相关性疾病中的应用

临床应用玻璃体腔注射贝伐单抗(bevacizumab)治疗虹膜、脉络膜新生血管性疾病已经取得了良好的短期疗效,而眼表新生血管相关性疾病的治疗也一直是眼科界的一大难题.近年来眼科学者开始应用贝伐单抗结膜下注射或者点眼治疗眼表新生血管的动物模型,同时部分学者也开始在临床尝试性应用贝伐单抗治疗眼表新生血管相关性疾病,如角膜炎症、翼状胬肉、Stevens-Johnson综合征以及与角膜移植手术相关的新生血管,取得了一定的疗效,也发现了一些问题.本文就近几年来在眼表疾病中应用贝伐单抗的安全性、给药途径、给药剂量和疗效以及相关问题加以综述.     

Inhibition of corneal neovascularization after alkali burn: comparison of different doses of bevacizumab in monotherapy or associated with dexamethasone

Background: To compare the effects of different doses of bevacizumab with both saline and dexamethasone on inflammatory angiogenesis in the rat cornea induced by small chemical lesions. Methods: Corneal chemical cauterization was performed on 24 rats. Animals were divided randomly into six groups and received a daily subconjunctival injection for 7 days of: balanced salt solution 0.1 mL or dexamethasone phosphate 4 mg/day or bevacizumab 2.5 mg/day, 3.75 mg/day, 5.0 mg/day or bevacizumab 5.0 mg/day + dexamethasone phosphate 4 mg/day. Clinical examination under slit lamp was performed daily for 7 days to evaluate corneal opacity and vessel size evolution. Computer‐assisted quantitative image analysis was used to measure the total corneal area covered by neovascularization. Results: At final examination, the dexamethasone, bevacizumab 5.0 mg/day and dexamethasone + bevacizumab groups showed a significant lowering in corneal opacity score as compared with control (P = 0.024, P = 0.006 and P = 0.013, respectively). Also, a significant reduction on new vessels size score was observed. Surface of corneal neovascularization was significantly reduced in dexamethasone, bevacizumab 5.0 mg/day and dexamethasone + bevacizumab groups compared with control (P = 0.045, P = 0.047 and P = 0.044, respectively). Conclusion: Our study demonstrates the ability of a 5.0 mg/day bevacizumab subconjunctival injection, in monotherapy or associated with dexamethasone, to cause a short‐term involution of corneal neovascularization after corneal alkali burn. Combination of both of these treatments may have advantages to monotherapy approaches.     

The inhibitory effect of different concentrations of topical bevacizumab on corneal neovascularization

Acta Ophthalmol. 2010: 88: 862–867

Abstract.

Purpose: This study aimed to evaluate the effects of different concentrations of topically administered bevacizumab (Avastin) on experimental corneal neovascularization (NV) in rats. Methods: Corneal NV was induced by chemical cauterization with silver nitrate sticks applied to the centre of the corneas of 37 Wistar rats. The rats were then randomized to four topical treatment groups: group 1 (n = 10) received 4 mg/ml bevacizumab; group 2 (n = 9) received 2 mg/ml bevacizumab; group 3 (n = 10) received 1 mg/ml bevacizumab, and group 4 (n = 8) represented a control group and received saline. All drops were initiated immediately after cauterization and applied twice per day for 7 days. Corneal NV was assessed 8 days after cauterization in a masked fashion, both qualitatively by clinical evaluation and quantitatively by blood vessel count in photographs of histological sections. Results: On clinical evaluation, groups 1 and 2 showed significantly less NV compared with the saline‐treated control group (p = 0.006 and p = 0.024, respectively). Histopathological evaluation showed that only group 1 differed significantly from controls (5% significance level) and normal corneal epithelium was seen in all groups. Conclusions: Topically administered bevacizumab at a concentration of 4 mg/ml significantly reduces corneal NV according to both clinical and histopathological evaluations; lower concentrations were less effective on both parameters. No corneal epitheliopathy was found using these concentrations.     

淋巴管形成与单纯疱疹病毒性角膜炎的关系

单纯疱疹病毒性角膜炎（herpes simplex keratitis,HSK）所致的角膜新生血管形成是全世界范围内各年龄段人群视力下降甚至致盲的主要原因。虽现在大量研究已了解到HSK除了可以诱导角膜新生血管生成外,还能诱导淋巴管扩展到角膜,并且淋巴管形成在HSK发病过程中起重要作用,但针对淋巴管形成的研究进程却远落后于角膜新生血管。本文对近年来淋巴管在HSK中作用的研究进展进行综述,力求进一步了解淋巴管形成与HSK的关系,为角膜疾病的治疗探索新的契机。     

结膜下注射AMD3100对小鼠角膜碱烧伤血管新生的治疗作用

目的探讨结膜下注射AMD3100对小鼠角膜碱烧伤血管新生的治疗作用及其机制。方法采用碱烧伤法诱导小鼠角膜血管新生（CNV）形成。在碱烧伤后当天,治疗组和对照组分别在球结膜下注射AMD31005μg（10μL）和生理盐水（10μL）,每日1次,连续用药7d。裂隙灯显微镜下观察不同时间点角膜炎症指数。病理组织学检查观察炎症细胞数量,并采用免疫组化检测角膜组织微血管密度。结果实验组在碱烧伤后不同时间点的炎症指数、炎症细胞数量和微血管密度均明显少于对照组（P〈0．05）。结论AMD3100可有效抑制CNV,可能与减轻碱烧伤后炎症反应有关。     

The Effect of Bevacizumab on Corneal Neovascularization in Rabbits

Purpose

To determine the efficacy of topical application and subconjunctival injection of bevacizumab in the treatment of corneal neovascularization.

Methods

Corneal neovascularization was induced with a silk suture of the corneal stroma in 12 rabbits (24 eyes). One week after suturing, four rabbits were treated with topical bevacizumab at 5 mg/mL (group A) and another four rabbits were treated with topical bevacizumab 10 mg/mL (group B) in the right eyes twice a day for two weeks. A subconjunctival injection of bevacizumab 1.25 mg/mL was done in the right eyes of four rabbits (group C). All of the left eyes (12 eyes) were used as controls. The area of corneal neovascularization was measured after one and two weeks, and the concentration of vascular endothelial growth factor (VEGF) in corneal tissue was measured after two weeks.

Results

The neovascularized area was smaller in all treated groups than in the control group (p<0.001). Upon analysis of the neovascularized area, there was no significant difference between groups A and B. However, the mean neovascularized area of group B was significantly smaller than that of group C after two weeks of treatment (p=0.043). The histologic examination revealed fewer new corneal vessels in all treated groups than the control group. The concentration of VEGF was significantly lower in all treated groups compared to the control group (p<0.01), but no difference was shown between treated groups.

Conclusions

Topical and subconjunctival bevacizumab application may be useful in the treatment of corneal neovascularization and further study is necessary.     

角膜新生血管对角膜损伤神经再生影响的研究

目的 探讨角膜新生血管对大鼠角膜损伤神经再生的影响。设计 实验研究。研究对象 SD大鼠。方法 采用随机数字表法将18只SD大鼠分为3组,每组6只。A组行缝线铲针角膜基质层间切开及缝线诱导新生血管术,术后0、3、7天给予结膜下注射贝伐单抗;B组行缝线铲针角膜基质层间切开及缝线诱导新生血管术;C组0、3、7天行结膜下注射贝伐单抗操作。分别在术后1天、1周、2周、4周,采用裂隙灯照相法观察记录角膜新生血管面积;角膜共聚焦显微镜记录神经长度。采用Cochet-Bonnet知觉仪测量缝线区的角膜知觉,采用Schirmer试验泪液线测量右眼的泪液分泌量。术后4周角膜全层铺片免疫荧光染色,记录上皮下神经密度。主要指标 角膜新生血管面积比、神经长度、上皮下神经密度、角膜知觉、泪液分泌量。结果 A、B组术后1、2周有角膜新生血管生长,术后4周消退闭锁,C组无角膜新生血管生长。A组术后1、2周新生血管面积比为（10.86±1.57）%和（1.87±0.69）%,分别小于B组的（25.42±2.65）%和（6.48±1.10）%（P均=0.000）。术后1天A、B组神经长度分别为（151.02±4.74）μm、（149.69±4.32）μm（P=0.306）;术后1、2、4周,A组神经长度均长于B组,分别为（193.84±2.25）μm与（155.73±2.98）μm、（217.15±2.08）μm与（166.21±2.41）μm、（220.70±1.41）μm与（203.76±1.74）μm（P均=0.000）。术后A、B组神经长度均有减少并有恢复趋势,C组无明显变化。术后4周A组损伤区上皮下神经密度（22.60%±2.02%）明显高于B组（9.41%±2.01%）（P=0.000）。A、B组上皮下神经短小稀疏、密度低,C组形态正常。A、B组术后1、2、4周时角膜知觉及泪液分泌量均无统计学差异（P均＞0.05）。A、B组均有下降并恢复趋势,C组无明显变化。结论 角膜新生血管可能抑制角膜损伤神经再生,抑制角膜新生血管有利于神经再生。（眼科, 2017, 26： 106-111）     

Subconjunctival bevacizumab injection for corneal neovascularization

PURPOSE: To report on the clinical use of subconjunctival bevacizumab in patients with corneal neovascularization. METHODS: The charts of 10 consecutive patients with corneal neovascularization who received subconjunctival injections of bevacizumab (2.5 mg/0.1 mL) were reviewed. Digital photographs of the cornea were graded by 2 masked observers for density, extent, and centricity of corneal vascularization. Image analysis was used to determine the area of cornea covered by neovascularization as a percentage of the total corneal area. RESULTS: No significant ocular or systemic adverse events were observed during 3.5 +/- 1.1 months of follow-up. Seven patients showed partial regression of vessels. The extent decreased from 6.0 +/- 1.2 (SD) clock hours before the injection to 4.6 +/- 1.0 clock hours after bevacizumab injection (P = 0.008). Density decreased from 2.7 +/- 0.2 to 1.9 +/- 0.3, respectively. (P = 0.007). No change was noticed in the centricity of corneal vessels. Corneal neovascularization covered, on average, 14.8% +/- 2.5% (SD) of the corneal surface before the injections, compared with 10.5% +/- 2.8% (P = 0.36, t test) after bevacizumab injection. Therefore, bevacizumab decreased corneal neovascularization by 29%. CONCLUSIONS: Short-term results suggest that subconjunctival bevacizumab is well tolerated and associated with a partial regression of corneal neovascularization.     

Inhibition of corneal neovascularization by subconjunctival bevacizumab in an animal model

PURPOSE: To evaluate the effect of subconjunctival injection of bevacizumab on experimentally induced corneal neovascularization. DESIGN: Experimental animal study. METHODS: Twelve New Zealand white rabbits were involved, divided equally into four groups. Only one eye per rabbit was used. Topical instillation of 10 microl 5% NaOH solution was used, under general anesthesia, to induce corneal neovascularization secondary to corneal alkali burn in groups 2, 3, and 4. A single dose of 3.75 mg (25 mg/ml) bevacizumab was injected subconjunctivally. Group 1 (control group 1) was neither cauterized nor treated. Group 2 (control group 2) received a sham injection of balanced salt solution on day 14. Group 3 was treated on day 14 (after corneal neovascularization had been established). Group 4 was treated on day 1. Digital photographs were obtained and analyzed during the entire 28-day procedure. The area of neovascularization and scarring were measured in terms of the percentage of corneal surface affected. RESULTS: On day 28, the difference of neovascularization between groups 2, 3, and 4 was found to be statistically significant at the .05 level (one-way analysis of variance [ANOVA]): group 4 (4.7%+/-3.1%).1, one-way ANOVA). No side effects were noted. CONCLUSIONS: Subconjunctival administration of bevacizumab inhibits corneal neovascularization effectively in the rabbit experimental model, especially if administered early.     

Measurement of central corneal thickness and pre-corneal tear film thickness of rabbits using the Scheimpflug system

AIM:To measure central corneal thickness (CCT) and pre-corneal tear film thickness using the Galilei dual-Scheimpflug analyzer (GSA) in New Zealand white rabbits.METHODS:Ten normal New Zealand white rabbits (20 eyes) were included in this study. With the assistance of 0.1% fluorescein, the pre-corneal tear film can be well visualized. Both eyes of each rabbit were scanned once with the GSA pre- and post-instillation of 1μL 0.1% fluorescein. The difference between the two measurements of CCT (4-mm diameter) was recorded as the pachymetric values of the central tear film.RESULTS:The CCT of pre- and post-instillation was 388.8±9.5μm and 407.0±10.5μm, respectively. After a paired t-test analysis, the central pre-corneal tear film thickness of 4mm diameter was 18.2±5.31μm with a 95% confidence interval of (15.7, 20.6)μm (P<0.001).CONCLUSION:GSA can be used to measure CCT and analyze central tear film thickness of rabbits with the help of fluorescein.