Lin Chen-Huan, Hsu Shan-hui, Huang Chi-En, Cheng Wen-Tung, Su Jang-Ming
Department of Chemical Engineering, National Chung Hsing University, Taichung, Taiwan, ROC.
Biomaterials. 2009 Sep;30(25):4117-26. doi: 10.1016/j.biomaterials.2009.04.028. Epub 2009 May 17.
A scaffold-bioreactor system was developed for growing tissue-engineered trachea and the effect of fluid flow on producing trachea-like neotissue was investigated. Chondrocytes were seeded in the poly(epsilon-caprolactone)-type II collagen scaffold and grown in the bioreactor operated under continuous flow at a rotational speed from 5 to 20 rpm. Flow analysis showed that the maximal and minimal shear stress in the bioreactor was 0.189-0.752 dyne/cm(2) and 30.3x10(-5)-104x10(-5) dyne/cm(2), respectively. After 4 and 8 weeks, the constructs were harvested from the bioreactor and analyzed. The application of rotation increased cell proliferation, GAG and collagen content in the constructs. Especially at 15 rpm, a two-fold increase in cell number, 170% increase in GAG, and 240% increase in collagen were found compared to static culture at 8 weeks. H&E staining showed the formation of neocartilage and the alignment of chondrocytes along the flow direction. The constructs grown under 15 rpm was selected for implantation into tracheal defects of rabbits. The mean survival of six animals was 52 days. The re-epithelialization of respiratory epithelium from the anastomotic sites was observed, with granulation tissue overgrowth. This successful initial step would allow us to make further improvement in applying tissue-engineering techniques to regenerate tracheas for practical use.
开发了一种用于培养组织工程气管的支架 - 生物反应器系统，并研究了流体流动对产生气管样新组织的影响。将软骨细胞接种到聚（ε - 己内酯） - II型胶原支架中，并在以5至20 rpm的转速连续流动操作的生物反应器中培养。流动分析表明，生物反应器中的最大和最小剪应力分别为0.189 - 0.752达因/平方厘米和30.3×10⁻⁵ - 104×10⁻⁵达因/平方厘米。4周和8周后，从生物反应器中取出构建体并进行分析。旋转的应用增加了构建体中的细胞增殖、糖胺聚糖（GAG）和胶原蛋白含量。特别是在15 rpm时，与8周的静态培养相比，细胞数量增加了两倍，GAG增加了170%，胶原蛋白增加了240%。苏木精 - 伊红（H&E）染色显示形成了新软骨，并且软骨细胞沿流动方向排列。选择在15 rpm下生长的构建体植入兔的气管缺损处。六只动物的平均存活时间为52天。观察到吻合部位的呼吸道上皮重新上皮化，伴有肉芽组织过度生长。这一成功的初步步骤将使我们能够在应用组织工程技术再生气管以供实际使用方面进一步改进。
