In vitro evaluation of a multi-layer radial-flow bioreactor based on galactosylated chitosan nanofiber scaffolds.
In vitro evaluation of a multi-layer radial-flow bioreactor based on galactosylated chitosan nanofiber scaffolds.
复制标题
DOI：
10.1016/j.biomaterials.2009.05.020
复制
发表时间：
2009-09
期刊：
影响因子：
14
通讯作者：
X. Chu;Xiao-lei Shi;Zhangqi Feng;Jin-yang Gu;Hai-Yun Xu;Yue Zhang;Z. Gu;Yi-tao Ding
中科院分区：
文献类型：
--
作者：
X. Chu;Xiao-lei Shi;Zhangqi Feng;Jin-yang Gu;Hai-Yun Xu;Yue Zhang;Z. Gu;Yi-tao Ding
生物人工肝的临床应用很大程度上依赖于生物反应器的发展。本研究以半乳糖基化壳聚糖纳米纤维为支架材料，构建了多层径向流生物反应器，并对其体外效果进行了评价。该生物反应器包含65层堆叠的平板，纳米纤维支架被静电纺丝固定和聚集肝细胞。含猪红细胞的培养基由中心向外灌注，使交换材料足够供氧。我们测定了肝细胞特异性功能和一般代谢参数，并测量了耗氧量（OCR）。显微镜和扫描电镜观察显示细胞与支架之间黏附紧密。与对照组（无纳米纤维支架的生物反应器）相比，我们的生物反应器中粘附的细胞数量增加了1.59倍；肝细胞蛋白质合成能力为1.73倍，尿素合成能力为2.86倍。有红细胞的生物反应器的OCR约为无红细胞生物反应器的1.91倍。将半乳糖化壳聚糖纳米纤维支架引入到新型生物反应器中，大大增强了细胞的粘附性和功能，并且添加到培养基中的红细胞能够为肝细胞提供足够的氧气。重要的是，我们的新生物反应器显示出令人兴奋的效率，它可能为肝功能衰竭患者提供短期支持。
Clinical use of bioartificial livers (BAL) strongly relies on the development of bioreactors. In this study, we developed a multi-layer radial-flow bioreactor based on galactosylated chitosan nanofiber scaffolds and evaluated its efficacy in vitro. The bioreactor contains 65 layers of stacked flat plates, on which the nanofiber scaffolds were electrospinned for hepatocyte immobilization and aggregation. Culture medium containing pig red blood cells (RBCs) was perfused from the center to periphery, so that exchange materials are sufficient to afford enough oxygen. We determined the parameters for hepatocyte-specific function and general metabolism and also measured the oxygen consumption rate (OCR). Microscope and scanned electron microscopy observation showed a tight adhesion between cells and scaffolds. Compared with the control (bioreactors without nanofiber scaffolds), the number of adhered cells in our bioreactor was 1.59-fold; the protein-synthesis capacity of hepatocytes was 1.73-fold and urea was 2.86-fold. Moreover, the OCR of bioreactors with RBCs was about 1.91-fold that of bioreactors without RBCs. The galactosylated chitosan nanofiber scaffolds introduced into our new bioreactor greatly enhanced cell adhesion and function, and the RBCs added into the culture medium were able to afford enough oxygen for hepatocytes. Importantly, our new bioreactor showed an exciting efficiency, and it may afford the short-term support of patients with hepatic failure.