A NOVEL DUAL-CHAMBER CULTURE SYSTEM WITH BIOMIMETIC TRIPHASIC SCAFFOLD FOR OSTEOCHONDRAL TISSUE ENGINEERING
A NOVEL DUAL-CHAMBER CULTURE SYSTEM WITH BIOMIMETIC TRIPHASIC SCAFFOLD FOR OSTEOCHONDRAL TISSUE ENGINEERING
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用于骨软骨组织工程的具有仿生三相支架的新型双室培养系统
DOI：
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发表时间：
2017-03
影响因子：
0.1
通讯作者：
Yang, Liu
中科院分区：
文献类型：
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作者：
Liu, Jun-Li;Liu, Qing-Yu;Tang, Hong;Yang, Liu
在这项研究中，一个仿生三相支架构建模仿天然软骨软骨下骨组织结构。该支架包括软骨层、软骨钙化带和软骨下骨层。软骨层为II型胶原海绵，CZC和软骨下骨层为脱细胞猪膝关节。为了构建软骨层和软骨下骨层分离的微环境，采用计算机辅助设计方法设计了双腔生物反应器，以聚乳酸为材料，以CZC作为双腔生物反应器的屏障，利用3D打印机制作了双腔生物反应器。这两个腔室中的培养基通过蠕动泵分别循环。将羊膜间充质干细胞接种于该支架上，荧光标记法进行细胞追踪，总DNA含量分析法检测细胞增殖情况，诱导培养基用于指导干细胞分化。培养7 d后，细胞在支架上呈规则分布，细胞的粘附和增殖不受影响。未发生细胞跨CZC迁移。总DNA含量分析显示，支架中的细胞以时间依赖的方式增加。成软骨和成骨培养液可分别诱导这两个腔室中的干细胞分化为软骨细胞和骨细胞。初步研究表明，仿生三相支架双室培养系统是可行的，因此，该系统将进一步改进和体内测试。
In this study, a biomimetic triphasic scaffold was constructed to mimic the native cartilage-subchondral bone tissue structure. This scaffold contained chondral layer, calcified zone of cartilage (CZC) and subchondral bone layer. The chondral layer was type II collagen sponge, the CZC and the subchondral bone layer were derived from normal pig knee by decellularization. In order to build separate microenvironment for chondral layer and subchondral bone layer, a dual-chamber bioreactor was designed by computer aided design, manufactured by 3D printer using Poly Lactic Acid, with CZC as the barrier of these two chambers. Culture medium in these two chambers was circulated separately by peristaltic pumps. Amniotic mesenchymal stem cells were seeded in this scaffold, fluorescence labeling was used for cell tracking, total DNA content analysis was used to indicate cell proliferation, and inducing medium was used to direct stem cells differentiation. After 7 days culture, the cells regularly distributed in the scaffold, cell adhesion and proliferation was not affected. No cell migration across CZC occurred. Total DNA content analysis showed that cells in scaffold increased in a time-dependent manner. Chondrogenic and osteogenic medium could induce stem cells in these two chambers to differentiate into chondrocytes and osteocytes, respectively. Our pilot study showed that the dual-chamber culture system with biomimetic triphasic scaffold was feasible, therefore this system will be further modified and tested in vivo.