Preparation and characterization of hepatocyte-specific macroporous microcarrier

doi:10.3969/j.issn.2095-4344.2013.16.021

Abstract

Abstract:

BACKGROUND: Hepatocyte culture is the key to the hepatocyte transplantation and bioartificial liver study. It has turned into the first problem how to get adequate number of better activity and well-functioning hepatocytes.
OBJECTIVE: To prepare a high-performance silk fibroin/galactosylated chitosan macroporous microcarrier with hepatocyte-specific characteristics, for hepatocyte adhesion culture in the three-dimensional environment in vitro.
METHODS: Silk fibroin, chitosan, and lactose acid served as raw materials to prepare a hepatocyte-specific silk fibroin/galactosylated chitosan macroporous microcarrier using comprehensive application of emulsified-chemical crosslinking, polarity solution and freeze-drying. Optical microscope, scanning electron microscope, proton nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, and cell biocompatibility test were used to characterize its morphology, physical and chemical properties.
RESULTS AND CONCLUSION: The microcarrier was successfully prepared in the project. Furthermore, the pores were occupied inside and outside of the microcarrier and interconnected, which presented hepatocyte-specific characteristics. Under the scanning electron microscope, the macroporous microcarrier exhibited a trumpet-shaped porous structure with the pore size of 40-80 µm and even pore distribution, providing an appropriate environment for high-density culture of hepatocytes. The galactosyl was successfully compounded into the scaffold material which was confirmed by the proton nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy. Cell biocompatibility test showed a hepatocyte-specific affinity to the silk fibroin/galactosylated chitosan macroporous microcarrier, and the cells grew and aggregated around the microcarrier exhibiting a good activity. These findings indicate that combination of emulsified-chemical crosslinking, polarity solution and freeze-drying can prepare a silk fibroin/galactosylated chitosan macroporous microcarrier with hepatocyte-specific characteristics.

Key words: biomaterials, material mechanics and surface modification, hepatocyte-specific characteristics, macroporous microcarrier, silk fibroin, cell culture, chitosan, galactosyl, freeze-drying, bioartificial liver, National “863&rdquo, Project of China

CLC Number:

R318

Guo Wei, Gong Du-hui, Hu Zhi-wei, Cheng Yuan, Gao Yi, Pan Ming-xin. Preparation and characterization of hepatocyte-specific macroporous microcarrier[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(16): 2994-3001.

Figures/Tables 7

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