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Survival and function of hepatocytes on a novel three-dimensional synthetic biodegradable polymer scaffold with an intrinsic network of channels.

机译：在具有固有通道网络的新型三维合成可生物降解聚合物支架上肝细胞的存活和功能。

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OBJECTIVE: To evaluate the survival and function of hepatocytes (HCs) on a novel three-dimensional (3D) synthetic biodegradable polymer scaffold with an intrinsic network of interconnected channels under continuous flow conditions. SUMMARY BACKGROUND DATA: The authors' laboratory has investigated HC transplantation using 3D biodegradable polymers as scaffolding as an alternative approach to treatment of end-stage liver disease. Previous studies have demonstrated survival of HCs transplanted on polymer discs in peripheral tissue sites and partial correction of single enzyme liver defects. One of the major limitations has been the insufficient survival of an adequate mass of transplanted cells; this is thought to be caused by inadequate oxygen diffusion. METHODS: HCs and nonparenchymal liver cells from Lewis rats were seeded onto 3D biodegradable polymer scaffolds. Microporous 3D polymers were created using 3D printing on copolymers of polylactide-coglycolide. The cell/polymer constructs were placed in static culture or continuous flow conditions. The devices were retrieved after 2 days and examined by scanning electron microscopy and histology. Culture medium was analyzed for albumin by enzyme-linked immunosorbent assay (ELISA). Differences in culture parameters including pH, PCO2, PO2, glucose, lactate, and HCO3 were examined. RESULTS: Scanning electron microscopy revealed successful attachment of HCs on the 3D polymer in both static and flow conditions. Histology demonstrated viable HCs in both conditions. ELISA demonstrated a significantly higher mean concentration of albumin in flow conditions than in static conditions. Culture parameter analysis revealed a significantly higher PO2 and glucose level, and a more physiologic pH in flow conditions than in static conditions. CONCLUSIONS: HCs cocultured with nonparenchymal cells can attach to and survive on the 3D polymer scaffolds in both static and flow conditions in the size and configuration used in this study. Flow conditions may provide a more conducive environment for HC metabolism and albumin synthesis than static conditions. The authors hypothesize that flow through directed channels will be necessary for the transfer of large masses of cells when implantation studies are initiated.
机译：目的：评估在连续流动条件下具有互连通道固有网络的新型三维（3D）合成生物可降解聚合物支架上肝细胞（HCs）的存活和功能。摘要背景数据：作者的实验室研究了使用3D可生物降解聚合物作为支架的HC移植，作为治疗终末期肝病的替代方法。先前的研究表明，在周围组织部位的聚合物圆盘上移植的HC的存活率和单酶肝缺陷的部分纠正。主要限制之一是足够数量的移植细胞存活不足。认为这是由于氧扩散不充分引起的。方法：将来自Lewis大鼠的HCs和非实质肝细胞接种到3D可生物降解的聚合物支架上。使用3D打印在聚丙交酯-乙交酯共聚物上创建微孔3D聚合物。将细胞/聚合物构建体置于静态培养或连续流动条件下。 2天后取回装置并通过扫描电子显微镜和组织学检查。通过酶联免疫吸附测定（ELISA）分析培养基中的白蛋白。检查了包括pH，PCO2，PO2，葡萄糖，乳酸盐和HCO3在内的培养参数的差异。结果：扫描电子显微镜显示HCs在静态和流动条件下均成功附着在3D聚合物上。组织学证明在两种情况下均存在可行的HC。 ELISA显示流动条件下的白蛋白平均浓度明显高于静态条件下的白蛋白浓度。培养参数分析显示，与静态相比，流动条件下的PO2和葡萄糖水平显着更高，并且生理pH更高。结论：在本研究中使用的大小和构型，与非实质细胞共培养的HCs可以在静态和流动条件下附着于3D聚合物支架并在其上存活。与静态条件相比，流动条件可能为HC代谢和白蛋白合成提供更有利的环境。作者假设，当着手进行植入研究时，流经定向通道对于大量细胞的转移将是必要的。

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