Preparation of three-dimensional macroporous chitosan-gelatin B microspheres and HepG2-cell culture
Preparation of three-dimensional macroporous chitosan-gelatin B microspheres and HepG2-cell culture
复制标题
三维大孔壳聚糖明胶B微球的制备及HepG2细胞培养
DOI：
10.1002/term.1888
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发表时间：
2016-12-01
影响因子：
3.3
通讯作者：
Dong, Ya-Dong
中科院分区：
文献类型：
--
作者：
Huang, Fang;Cui, Long;Dong, Ya-Dong
采用冷冻干燥、静电交联和离子交联相结合的三步法制备了具有开放、互连、大孔（100-200 μ m）结构的壳聚糖-明胶B微球。为了防止壳聚糖-明胶B微球多孔结构的破坏和提高微球的生物稳定性，选用饱和三聚磷酸乙醇溶液（85%乙醇）作为交联剂，N-（3-二甲氨基丙基）-N-乙基-碳二亚胺/N-羟基琥珀酰亚胺作为第二交联剂，与明胶A反应，固定壳聚糖-明胶B微球，以提高微球的生物相容性。三维大孔壳聚糖-明胶B微球（3D-P-CGMs）的吸水率为12.84，孔隙率为85.45%。在体外溶菌酶降解1，3，5，7，10，14，和21天后，在3D-P-CGMs的生物降解性提高。在3D-P-CGMs上培养的人肝癌细胞系（HepG 2细胞）的形态是球形的，不同于在传统的二维条件下培养的细胞。扫描电子显微镜和石蜡切片用于确认3D-P-CGMs的多孔结构。HepG 2细胞能够通过孔迁移到内部。细胞增殖、白蛋白和乳酸脱氢酶水平表明，3D-P-CGMs可提供更大的比表面积和合适的细胞生长和存活的微环境。因此，3D-P-CGMs非常适合作为组织工程和细胞载体研究中细胞培养的大孔支架。版权所有（c）2014约翰威利父子有限公司
Chitosan-gelatin B microspheres with an open, interconnected, highly macroporous (100-200 mu m) structure were prepared via a three-step protocol combining freeze-drying with an electrostatic and ionic cross-linking method. Saturated tripolyphosphate ethanol solution (85% ethanol) was chosen as the crosslinking agent to prevent destruction of the porous structure and to improve the biostability of the chitosan-gelatin B microspheres, with N-(3-dimethylaminopropyl)-N-ethyl-carbodiimide/N-hydroxysuccinimide as a second crosslinking agent to react with gelatin A and fixed chitosan-gelatin B microspheres to attain improved biocompatibility. Water absorption of the three-dimensional macroporous chitosan-gelatin B microspheres (3D-P-CGMs) was 12.84, with a porosity of 85.45%. In vitro lysozyme degradation after 1, 3, 5, 7, 10, 14, and 21days showed improved biodegradation in the 3D-P-CGMs. The morphology of human hepatoma cell lines (HepG2 cells) cultured on the 3D-P-CGMs was spherical, unlike that of cells cultured under traditional two-dimensional conditions. Scanning electron microscopy and paraffin sections were used to confirm the porous structure of the 3D-P-CGMs. HepG2 cells were able to migrate inside through the pore. Cell proliferation and levels of albumin and lactate dehydrogenase suggested that the 3D-P-CGMs could provide a larger specific surface area and an appropriate microenvironment for cell growth and survival. Hence, the 3D-P-CGMs are eminently suitable as macroporous scaffolds for cell cultures in tissue engineering and cell carrier studies. Copyright (c) 2014 John Wiley & Sons, Ltd.