Peng Gongze, Li Saina, Peng Qing, Li Yang, Weng Jun, Jia Zhidong, Kang Jiyao, Lei Xiongxin, Zhang Guifeng, Gao Yi
1 Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center of Artificial Organ and Tissue Engineering, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
2 National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Beijing, China.
J Biomater Appl. 2017 Jul;32(1):93-103. doi: 10.1177/0885328217709607. Epub 2017 May 15.
Background/aims The critical part of a bio-artificial liver device is establishment of a bioreactor filled with liver cells. However, it is still unclear how to maintain benign cell function while achieving the sufficient cell quantity. In the current study, we aim to establish a novel carrier for the culture of HepG2 cells, a liver cell line, by modifying polypropylene nonwoven fabrics with native type I collagen. Methods "Piranha" solution, KH-550 and glutaraldehyde subsequently were used to bridge native type I collagen and polypropylene nonwoven fabrics. The type I collagen-coupled polypropylene nonwoven fabric was characterized by XPS, SEM, ATR-FTIR and water contact angle measurement. Furthermore, the biocompatibility between HepG2 cells and fiber film is evaluated by the ability of cell proliferation, albumin secretion, as well as urea synthesis. Results The coating of collagen onto polypropylene fabrics was more efficient using the chemical covalent binding method than direct immersion, which was validated by the presence of collagen-related elements and chemical bond. The adding of collagen in polypropylene fabrics promoted hydrophilicity and HepG2 cell adherence. Additionally, enhanced cell proliferation, increased albumin secretion and urea synthesis were observed in HepG2 cells growing on collagen-coated polypropylene fabrics. Conclusions The collagen coated polypropylene nonwoven fabrics, acting as a feasible substrate for HepG2 cell culture, may be used as a promising liver cell carrier for artificial liver reactor.
背景/目的 生物人工肝装置的关键部分是建立一个充满肝细胞的生物反应器。然而,目前尚不清楚如何在实现足够细胞数量的同时维持细胞的良性功能。在本研究中,我们旨在通过用天然I型胶原修饰聚丙烯无纺布来建立一种用于培养肝癌细胞系HepG2的新型载体。方法 依次使用“食人鱼”溶液、KH-550和戊二醛将天然I型胶原与聚丙烯无纺布连接起来。通过X射线光电子能谱(XPS)、扫描电子显微镜(SEM)、衰减全反射傅里叶变换红外光谱(ATR-FTIR)和水接触角测量对I型胶原偶联的聚丙烯无纺布进行表征。此外,通过细胞增殖能力、白蛋白分泌能力以及尿素合成能力来评估HepG2细胞与纤维膜之间的生物相容性。结果 采用化学共价结合法将胶原包被在聚丙烯织物上比直接浸泡更有效,这通过胶原相关元素和化学键的存在得到了验证。聚丙烯织物中添加胶原可促进亲水性和HepG2细胞黏附。此外,在胶原包被的聚丙烯织物上生长的HepG2细胞中观察到细胞增殖增强、白蛋白分泌增加和尿素合成增加。结论 胶原包被的聚丙烯无纺布作为HepG2细胞培养的可行基质,有望成为人工肝反应器的肝细胞载体。