猪膀胱无细胞基质生物支架材料中转化生长因子β1的含量测定  

Contents of transforming growth factor beta 1 in pig bladder acellular matrix as biologic scaffold materials

作　　者：袁铭[1] 李汉忠[2] 张玉石[2] 石炳毅[1] 

机构地区：[1]解放军总医院第二附属医院全军器官移植中心,北京市100091 [2]中国医学科学院中国协和医科大学北京协和医院泌尿外科,北京市100730

出　　处：《中国组织工程研究与临床康复》2008年第27期5305-5308,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:天然生物支架材料可能含有一定量的生长因子,在组织工程膀胱构建中存有其潜在的生物学行为。目的:观察去细胞处理后猪膀胱无细胞基质支架材料中转化生长因子β1含量的变化。设计、时间及地点:细胞组织工程学实验,于2006-07/2007-03在中国医学科学院北京协和医院中心实验室完成。材料:取材0.5h内的猪新鲜膀胱标本12个,采集自北京市资源肉联厂,随机分为对照组、去细胞组,6个/组。方法:去细胞组采用低渗-去污剂洗涤-核酸酶消化法对新鲜猪膀胱进行脱细胞处理,制备膀胱无细胞基质,对照组不做任何处理。主要观察指标:苏木精-伊红染色观察膀胱去细胞效果,采用二辛可宁酸法测定可溶性蛋白含量,酶联免疫吸附试验测定转化生长因子β1含量。结果:对照组新鲜膀胱组织细胞结构完整,去细胞组膀胱组织细胞成分已基本去除,未见有核成分残留。去细胞组可溶性蛋白含量为(0.143±0.053)%,转化生长因子β1含量为(113.31±43.02)ng/kg,均明显低于对照组(t=15.56,P<0.001;t=24.63,P<0.001)。结论:采用低渗-去污剂洗涤-核酸酶消化法可有效去除猪膀胱细胞成分,但仍有一定量的转化生长因子β1残留,可能为种子细胞生长提供有利因素。BACKGROUND： Natural biological scaffold material maybe contains growth factor, and has potential biological behavior in tissue-engineered bladder. OBJECTIVE： To observe the contents of the transforming growth factor beta 1 （TGF- β 1） in porcine bladder acellular matrix as biologic scaffold materials for tissue engineering. DESIGN, TIME AND SETTING： The cell tissue engineering experiment was performed at the Central Laboratory of Peking Union Hospital of Chinese Academy of Medical Sciences from July 2006 to March 2007. MATERIALS： Twelve fresh pig bladder samples within half an hour were obtained from Beijing Ziyuan Factory, and randomly divided into a control group and an experimental group, 6 in each group. METHODS： A hypotension and detergent and nucleic acid enzymatic digestion extraction process had been developed to remove all the cellular components from the porcine bladder to prepare the bladder acelhilar matrix （BAM）. Samples in the control group were left intact. MAIN OUTCOME MEASURES： The hematoxylin and eosin （HE） staining was routinely assayed to observe bladder acelhilar effect. The dissolvable protein and TGF- β 1 were undergone by bicinchoninic acid （BCA） and enzyme linked immunosorbent assay （ELISA） respectively. RESULTS： The fresh bladder tissue had a good cellular structure in the control group, while the decellularized BAM had not any cellular and nucleus components, The contents of the dissolvable protein and TGF- 13 1 with the fresh bladder and BAM in the experimental group （0.143±0.53）% and （113.31±43.02） ng/kg were significantly lower than in the control group （t=15.56, P 〈 0.001; t =24.63, P 〈 0.001）. CONCLUSION： This method of the hypotension and detergent and nucleic acid enzymatic digestion extraction shows a good effect to remove all the bladder cellular components, while still some TGF- β 1 remained in BAM, which maybe indicated an important function in reconstructing the tissue-engineered bladder.

关 键 词：膀胱无细胞基质 组织工程 转化生长因子Β1 酶联免疫吸附试验 

分 类 号：R318[医药卫生—生物医学工程]