管状支架体外静态培养组织工程气管软骨  

Static culture of tissue-engineered tracheal cartilage with scaffolds in vitro

作　　者：杨林 武延格 纪玲 王正 

机构地区：[1]暨南大学第二临床医学院深圳市人民医院胸外科,广东省深圳市518020 [2]暨南大学第二临床医学院深圳市人民医院检验医学部,广东省深圳市518020

出　　处：《中国组织工程研究与临床康复》2008年第19期3601-3604,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30500499)~~

摘　　要：前期实验已证实人气管软骨细胞在DegraPol片状支架上保持了正常的形态和活力，同时分泌软骨特有基质成分。目的：以大鼠剑突软骨细胞为种子细胞种植于DegraPol管状泡沫支架，观察新形成的工程化组织在体外静态培养条件下的组织和力学特点。设计、时间及地点：单一样本观察，于2006-12／2008-02在深圳市人民医院医学研究中心完成。材料：DegraPol管状材料为瑞士联邦理工大学聚合材料研究所提供；实验动物为2周龄体质量50-60g的SPF级SD幼鼠。方法：收集第3代幼鼠剑突软骨细胞接种于DegraPol管状支架，形成软骨细胞-支架复合物，体外静态培养6周。主要观察指标：①AO／PI荧光染色观察软骨细胞在DegraPol管状泡沫支架中的存活情况，于培养3，6周取出软骨细胞-DegraPol支架复合体，MTT法测定细胞增殖情况。②培养3，6周，应力-应变机械力学方法测定最大应变和应力的变化。③培养3，6周，扫描电镜观察软骨细胞在DegraPol支架中培养后的超微结构。结果：①AO／PI荧光染色显示软骨细胞在DegraPol支架内保持良好的活性，MTT法结果显示培养6周组A值高于培养3周组（P〈0．05）。②应力-应变机械力学测定结果显示，培养3周组应力值和应变值高于培养6周组（P〈0．05）。③扫描电镜结果显示DegraPol支架与大鼠剑突软骨细胞有良好的生物相容性，培养6周后获得更多的软骨细胞增殖和更多的细胞外基质。结论：DegraPol管状泡沫支架有良好的生物相容性并且软骨细胞在支架上保持正常活性，但由于工程化复合物的生物力学强度较差，培养条件和培养方式有待进一步改善。BACKGROUND： The human cartilage tracheal chondrocytes can retain their normal morphons scaffold, and simultaneously excrete cartilage specific matrix. OBJECTIVE： Rat xiphoid chondrocytes were seeded and cultivated on DegraPol scaffolds, to aim directly at the histology and mechanical characteristic of tissue-engineered tracheal cartilage during the static culture in vitro. DESIGN, TIME AND SETTING： A single sample observation was carried out in the Medical Science Research Center of Shenzhen People＇s Hospital （Shenzhen, Guangdong, China） from December 2006 to February 2008. MATERIALS： DegraPol scaffolds were provided by Swiss Federal Institute of Technology （Zurich）. SD rats of SPF grade and aged 2 weeks, were provided by the Experimental Animal Center of Guangdong Province （China）. METHODS： The third passage of rat xiphoid chondrocytes were seeded onto DegraPol scaffolds to develop the chondrocyte-scaffold composites, which were cultured for 6 weeks in vitro. MAIN OUTCOME MEASURES： The survival status of chondrocytes was examined by AO/PI fluorescent stain, and MTT test was used to determine viable cells at weeks 3 and 6 when the chondrocyte-scaffold composites were took ont. The stress-strain mechanical force analysis of maximal stress and maximal strain, together with the ultrastracture of the chondrocytes on DegraPol scaffold was observed under scanning electron microscope. RESULTS： AO/PI stain showed chondrocytes possess favorable activity on DegraPol scaffolds. MTr assay showed the A values at week 3 of the culture was lower than that at week 6 （P 〈 0.05）. The maximal strain load strength test was （1.43＋0.58） turn/ram at week 3, （0.48±0.07） mm/mm at week 6. The stress-strain mechanical force analysis showed that, the maximal stress and the maximal strain were higher at week 3 of the culture compared with week 6 （P 〈 0.05）. Scanning electron microscopy showed more cell proliferation and more extracellular matrix were found at week 6, which indicated the good

关 键 词：组织工程 剑突软骨细胞 管状支架 生物相容性 生物材料 

分 类 号：R318[医药卫生—生物医学工程]