生物羊膜冷冻干燥工艺优化研究  

Study of Process Optimization on Freeze Drying of Human Amniotic Membrane

作　　者：高健[1] 刘玲玲[1] 刘文[1] 宋晶[1] 李坤[1] 洪运佳[1] 鲁乐水[1] 吕虎[1] 

机构地区：[1]江西师范大学生命科学学院,南昌330022

出　　处：《生物医学工程学杂志》2012年第4期705-709,共5页Journal of Biomedical Engineering

基　　金：国家中小企业创新基金资助项目(06C26213601339)

摘　　要：为探索羊膜冷冻干燥保存的最佳工艺条件,取健康剖宫产产妇胎盘,钝性剥离羊膜。以保存羊膜组织形态变化、胶原蛋白酶降解速度、生物力学特征、细胞因子含量为考察指标,对羊膜冷冻干燥工艺中的关键因素进行优化。结果显示,改良冷冻干燥羊膜上皮细胞、纤维基质层形态结构与新鲜羊膜基本相同,但纤维基质厚度略有增加,上皮细胞表面微绒毛略有减少;Ⅳ型胶原酶在溶液中降解速度有所加快;生物力学特征与新鲜羊膜无明显差别;6种细胞因子含量明显低于新鲜羊膜。结合前期工作,与常规冷冻干燥法比较,改良冷冻干燥工艺对保存羊膜组织结构和生物学活性因子影响较小,并能使保存羊膜具有更好的生物力学特性。The study was aimed to investigate the optimum conditions of freeze drying preservation of amniotic mem- brane （AM）. The AM from the health puerperal woman was preserved by freeze drying at optimized way. The key factors of freeze drying process, including abstersion aqua, conservation liquor, the curve of freezing temperature,and the ingredient of protective agent, were optimized. All their morphologic structure was observed by light microscope and scanning electron microscope. The degradation rates by collagenase IV and the characterization of biomechanies were analyzed. The radio-immunologic method was used to investigate the cytokines quantity. All properties of freeze dried AM were compared with those of fresh AM. Light micrographs showed that the five structure-layers ex- ist both in the fresh AM and in those preserved by freeze drying,while the fibro-material was tight-structured in the fresh AM,but loose slightly; the thickness of fibro-material was larger slightly in freeze dried AM. Scanning electron mierographs show that the micro-hairs of epithelial cells in fresh AM were decreased slightly in optimized drying AM, the collagen fibre of fresh AM and of optimized drying AM were well in morphological structures and arranged tightly. The degradation rate by collagenase IV was faster in optimized drying AM,compared with that of the fresh AM. There were insignificant diversity in biomechanical characters （tensile strength, elongation at break and elastic modulus）of the optimized drying AM compared with fresh AM. The cytokines quantity in optimized drying AM de- creased significantly compared with fresh AM. The improved freeze drying process has better advantage in keeping the morphological structure, preferable biomechanics and biological vitality of AM,compared with the early research.

关 键 词：羊膜 冷冻干燥 组织形态 细胞因子 生物力学 

分 类 号：R779.6[医药卫生—眼科]