Preparation of three-dimensional macroporous chitosan–gelatin B microspheres and HepG2-cell culture
Article first published online: 14 APR 2014
Copyright © 2014 John Wiley & Sons, Ltd.
Journal of Tissue Engineering and Regenerative Medicine
How to Cite
Huang, F., Cui, L., Peng, C.-H., Wu, X.-B., Han, B.-S. and Dong, Y.-D. (2014), Preparation of three-dimensional macroporous chitosan–gelatin B microspheres and HepG2-cell culture. J Tissue Eng Regen Med. doi: 10.1002/term.1888
- Article first published online: 14 APR 2014
- Manuscript Accepted: 24 FEB 2014
- Manuscript Revised: 1 DEC 2013
- Manuscript Received: 9 FEB 2013
- 3D culture;
- tissue engineering
Chitosan–gelatin B microspheres with an open, interconnected, highly macroporous (100–200 µm) 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 21 days 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 © 2014 John Wiley & Sons, Ltd.