课题背景：课题系重庆医科大学附属第一医院承担的重庆市科技计划项目“新型生物医学材料—羊膜的制备及临床前研究”的系列子课题之一。根据国家对医用生物材料/制品的相关规定，对羊膜进行规范化的制备、保存、并进行全面的临床前研究；研发出可供临床及科研使用的具有良好的生物安全性、稳定性和有效性的羊膜制品，符合国家对医用生物材料和三类医疗器械的要求。

应用要点：文章观察不同活性羊膜移植对兔眼角膜碱烧伤的治疗效果，并分析其生物活性成分作用。羊膜尤其是新鲜羊膜移植可以减轻眼表碱烧伤急性期的炎症反应，加速眼表的稳定和修复，但即使新鲜羊膜的移植效果优于现有的保存羊膜也不能完全取代保存羊膜。甘油保存羊膜组和真空干燥羊膜组保存了大部分羊膜中生物活性物质，能够取得较为满意的临床效果，乙醇保存羊膜由于所含生物活性物质被固定丧失了其生物学作用，移植后临床效果差。

同行评价：近年来羊膜在医学中的应用发展迅速，在眼科针对的都是常见病多发病，基层医院亦可开展。但羊膜的作用机制至今仍不清楚，该实验选用不同活性羊膜分组，建立动物模型，比较不同活性羊膜移植治疗角膜碱烧伤的效果，从一个方面反映了羊膜的作用机制。配合其母项目，评估不同活性羊膜制备的有效性，也成为鉴定羊膜制备的一种新的有效的方法。

摘要
目的: 近年来羊膜移植成为治疗角膜碱烧伤较为有效的手段之一，但其生物学基础尚不完全清楚。观察不同活性羊膜移植对兔眼角膜碱烧伤的治疗效果，并分析其生物活性成分作用。
方法：实验于2004-01/07于重庆医科大学动物实验中心实验室完成，动物实验方法符合动物伦理学要求。①实验材料及分组：健康产妇剖宫产胎盘由重庆医科大学附属第一医院产科提供。选用新西兰大白兔45只，按随机数字表法分为5组（n =9）：新鲜羊膜组、甘油保存羊膜组、乙醇保存羊膜组、真空干燥羊膜组及对照组。②实验方法：于新鲜羊膜制备后24 h内，保存羊膜保存1个月后行羊膜移植。③实验评估：术后2~21 d每日、21 d后每3日观察眼表角膜透明度及新生血管生长情况；术后14，30，60 d行荧光素染色，观察角膜上皮的修复情况；术后2周、1个月、2个月行组织病理和透射电镜观察。
结果：白兔43只进入结果分析。①在角膜透明性及角膜上皮修复速度上新鲜羊膜组、甘油保存羊膜组和真空干燥羊膜组无明显差异，均明显优于乙醇保存羊膜组和对照组，乙醇保存羊膜组与对照组无明显差异。②在抑制新生血管方面，新鲜羊膜组明显优于甘油保存羊膜组及真空干燥羊膜组。
结论：真空干燥羊膜、甘油保存羊膜和新鲜羊膜移植后均能减轻眼表碱烧伤的炎症反应，促进角膜上皮修复。其中新鲜羊膜效果最好，可能与其较其他羊膜含有更多的生物活性物质有关。
关键词：羊膜/移植；烧伤，化学；保存，生物学

胡柯，赵敏，张琪，李鸿.新鲜羊膜及几种保存羊膜移植治疗角膜碱烧伤[J].中国组织工程研究与临床康复，2008，12(5):857-860
[www.zglckf.com/zglckf/ejournal/upfiles/08-5/5k-857(ps).pdf]

Transplantation of fresh amnion and preserved amnion for corneal alkali burn

Abstract

AIM：Recently, amnion transplantation has been one of effective methods for curing corneal alkali burn, but its biomechanism is unclear. This study was to discuss the efficiency of different kinds of amnion on rabbit corneal alkali burn and to find the bioactive functions of the amnion.
METHODS: The experiment was carried out in the Central Laboratory of Experimental Animals at Chongqing University of Medical Sciences between January and July in 2004. All the disposals were in accordance with the ethical standard of animals.①Placenta from healthy puerperants underwent uterine-incision delivery were offered by Department of Obstetrics in the First Affiliated Hospital of Chongqing University of Medical Sciences. A total of 45 New Zealand rabbits were used for corneal alkali burn models and divided into fresh amniotic membrane (FAM), glycerin preserved amniotic membrane (GPAM), vacuum xeransis amniotic membrane (VXAM), alcolhol preserved amniotic membrane (APAM) and control groups, every group had 9 rabbits.②The FAM was prepared within 24 hours, and the preserved amnion were transplanted after 1 month.③The corneal opacity and corneal neovascularization were observed everyday during days 2–21 after operation and once every three days after days 21; corneal fluorescein staining was conducted to observe the repair of corneal epithelium on days 14, 30, 60 after operation; 2 weeks, 1 month and 2 months after operation, we did histopathological examination and scanning electron microscope observation.
RESULTS: Forty-three rabbits were involved in the result analysis.①There were no significant differences in the corneal opacity and repair velocity of corneal epithelium among FAM, GPAM and VXAM, which were remarkably superior to APAM and controls. APAM group and control group had not distinct difference.②FAM had better effects on inhibiting corneal neovascularization than GPAM and VXAM.
CONCLUSION: After transplantation, VXAM, GPAM and FAM can all accelerate corneal epithelial healing and lighten inflammation. FAM containing more bioactive materials, plays best.

Hu K, Zhao M, Zhang Q, Li H.Transplantation of fresh amnion and preserved amnion for corneal alkali burn.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(5):857-860(China)
[www.zglckf.com/zglckf/ejournal/upfiles/08-5/5k-857(ps).pdf]