自制染色干燥细胞外基质胶羊膜生物学特性及修补结膜缺损的实验研究  

Biological characteristics and its conjunctivarepair effects of self-made stained drying extracellular matrix amniotic membrane

作　　者：江威[1,2] 邹晶[1] 邵毅[1] 高桂平[1] 李云燕[1] 邓军萍 杨璐[1] 

机构地区：[1]南昌大学第一附属医院眼科,江西省南昌市330006 [2]高安市人民医院眼科,江西省高安市330800

出　　处：《眼科新进展》2015年第8期704-707,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81160118;81300729;81400372);全国临床医药研究专项基金(编号:L2012052);江西省远航工程(编号:2014022);江西省学位与研究生教育教学改革研究项目(编号:2015);江西省科技平台建设项目(编号:2013116);江西省青年科学基金(编号:20151BAB215016);江西省科技支撑计划项目(编号:20151BBG70223);江西省教育厅科技计划项目(编号:GJJ14170);江西省卫计委科技计划项目(编号:20155154);赣江学者计划(编号:2015)~~

摘　　要：目的研究自制染色干燥细胞外基质胶羊膜(extracellular matrix amniotic membrane,ECM-AM)的生物活性因子表达情况、生物力学特征以及在兔结膜修补术中的应用效果。方法使用光镜和HE染色对自制染色干燥ECM-AM进行形态学观察,并通过免疫荧光染色对比其与单纯冻干羊膜中不同生物活性因子Laminin 5、β-catenin和CollagenⅣ的表达情况。利用负荷传感器对比自制染色干燥ECM-AM与单纯冻干羊膜的最大承受拉伸力、弹性模量和拉伸长度,且把保存12个月的自制染色干燥ECM-AM应用于兔羊膜-结膜修补术,并在术后2周观察兔结膜愈合情况。结果自制染色干燥ECMAM可以分辨羊膜的折叠处、皱褶及边缘,HE染色表明自制染色干燥ECMAM基底膜的完整度和厚度均优于保存相应天数(12个月)的冻干羊膜。保存12个月的自制染色干燥ECM-AM中Laminin 5、β-catenin和CollagenⅣ均比单纯冻干羊膜要高。自制染色干燥ECM-AM的最大承受拉伸力以及弹性模量均明显高于于单纯冻干羊膜(均为P<0.05),但二者平均拉伸长度上差异无统计学意义(P>0.05)。自制染色干燥ECM-AM常温保存12个月后应用于兔羊膜-结膜修补术,术后2周羊膜上有兔结膜上皮长入且K19分布于整个结膜上皮细胞,并无明显的排斥反应和炎症反应。结论自制染色干燥ECM-AM含有丰富的生物活性因子,具有优越的生物力学特性,为眼表重建提供了更加安全有效的材料。Objective To study the bioactive factor, biomechani- cai characteristics of self-made stained drying extracellular matrix amni- otic membrane （ECM-AM）, and observe its effects in rabbit conjunctiva- repair surgery. Methods The morphology of self-made stained drying ECM-AM were observed by HE staining with microscope, and the biome- chanical characteristics including maximum tolerance, elasticity modulus and tensile elongation, and expression of bioactive factor such as Laminin 5, β-catenin and Collagen IV in self-made stained drying ECM-AM and single drying amniotic membrane were compared by immunofluores- cence. Besides,the ECM-AM which had been saved 12 months was used in rabbit conjunctiva-repair surgery, and the conjunctival healing was ob- served at postoperative 12 months. Results After saving 12 months, the self-made stained drying ECM-AM could distinguish the edge and creases easily. The HE staining showed that the self-made stained drying ECM-AM was more integrity and thicker than single drying amniotic membrane. The expression of bioactive factor such as Laminin 5, β-cate- nin and Collagen IV in self-made stained drying ECM-AM were more than those in single drying amniotic membrane. The biomechanical characteris- tics of self-made stained drying ECM-AM including maximum tolerance, elasticity modulus were better than that of single drying amniotic mem- brane（ all P 〈 0.05 ）, but there was no difference in tensile elongation （P 〉 0. 05 ）. Two weeks after conjunctiva-repair surgery with self-made stained drying ECM-AM in rabbits, the conjtmctivai epithelial of rabbits grew on the surface of amniotic membrane and K19 distributed through- out the conjunctival epithelial cells with no obvious rejection and inflam- mation. Conclusion Self-made stained drying ECM-AM has abundant bioactive factors and superior biomechanical characteristics, and provides more safe and effective material for reconstruction of ocular surface.

关 键 词：细胞外基质胶羊膜 单纯冻干羊膜 生物活性因子 结膜修补术 

分 类 号：R361.2[医药卫生—病理学]