旋转生物反应器用于提高组织工程气管软骨力学强度的实验研究  

In vitro Rotating Bioreactor for Improving the Mechanic Characteristics of Tissue Engineered Tracheal Cartilage

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,深圳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,深圳518020

出　　处：《中国生物医学工程学报》2009年第1期103-107,共5页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30500499)

摘　　要：针对体外静态培养方法的缺点,采用旋转生物反应器培养组织工程气管软骨,探讨工程化软骨合适的体外培养条件。选用大鼠剑突软骨细胞种植到DegraPol管状支架上,然后分别于传统静态培养和生物反应器内培养。于体外培养3周和6周后,取出软骨细胞-DegraPol支架复合物,以噻唑蓝(MTT)法测定细胞增殖活性,GAG浓度测定细胞外基质分泌情况,应力-应变机械力学方法测定最大应变和应力的变化,并制备扫描电镜标本观察软骨细胞在DegraP01支架中培养后的超微结构。体外培养3周应用MTT法测定A值分别为:静态培养的细胞-支架复合物组0.12±0.01,生物反应器组0.17±0.05(每组n=6)。GAG浓度测定静态培养组为(0.14±0.03)μg/mg,生物反应器组为(0.22±O.03)μg/mg(每组n=6)。应力-应变机械力学测定结果为,体外培养3周应变值:生物反应器组为(3.53±0.91),静态培养组为(1.71±0.13)。应力值生物反应器组为(0.33±0.04)MPa,静态培养组为(0.26±0.01)MPa。体外培养6周应变值:生物反应器组为(0.57±0.10),静态培养组为(0.48±0.07),应力值生物反应器组为(0.16±0.02)MPa,静态培养组为(0.09±0.02)MPa(每组n=4)。扫描电镜观察显示生物反应器组获得更好的软骨样结构和更多的细胞外基质。应用旋转生物反应器能够提供适宜的机械应力刺激,可作为体外构建组织工程化气管软骨的可行的培养方法。This study is to aim directly at the shortcoming of static culture in vitro, optimize the cuture condition of tissue engineered tracheal cartilage by rotating bioreactor in vitro, to improve the mechanic characteristics of tissue engineered tracheal cartilage. Rat xiphoid chondrocytes were seeded onto DegraPol scaffolds and cultivated respectively by static condition and rotaing bioreactor in vitro. The proliferation of the constructs was examined by MTI＂ assay. Stress-strain mechanical force analysis was coducted. The uhrastructure of the constructs was observed under electron microscope at 3 weeks of culture. MTI＂ assay showed the cell viability were 0.12 ± 0.01 in static culture and 0.17 ± 0.05 in bioreactor culture （n = 6 for each group）. The maximal strain was （3.53 ± 0.91 ） in the bioreactor culture, （ 1.71 ± 0.13） in the static culture, and（0.32 ± 0.09） for the control. The tensile strength was（0.33 ± 0.04） MPa in the bioreactor culture, （0.26 ± 0.01） MPa in the static culture, and（0.09 ± 0.01） MPa for the control. Electron microscopyobservation indicated that cell proliferation and ECM synthesis were both heavier in the bio constructs. The rotating bioreactor can provide better cultural condition and could be used for the formation of mature tissue engineered cartilage.

关 键 词：组织工程 软骨细胞 生物反应器 

分 类 号：R318[医药卫生—生物医学工程]