title: Preparation of three-dimensional macroporous chitosan-gelatin B microspheres and HepG2-cell culture.
abstract: 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.
authors.affiliation: Department of General Surgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Huang
authors.affiliation: Department of General Surgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Long
authors.initials: L
authors.lastname: Cui
authors.affiliation: Department of General Surgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Cheng-Hong
authors.initials: CH
authors.lastname: Peng
authors.affiliation: Department of General Surgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.|Department of General Surgery, Central Hospital, Minghang District, Shanghai, China.
authors.forename: Xu-Bo
authors.initials: XB
authors.lastname: Wu
authors.affiliation: Department of General Surgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.|Department of General Surgery, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Bao-San
authors.initials: BS
authors.lastname: Han
authors.affiliation: Department of General Surgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Ya-Dong
authors.initials: YD
authors.lastname: Dong
mesh_terms: D002465:Cell Movement; D060833:Cellular Microenvironment; D048271:Chitosan / Q000737:chemistry*; D005780:Gelatin / Q000737:chemistry*; D056945:Hep G2 Cells; D006801:Humans; D008863:Microspheres; D016062:Porosity
chemical_list: D005780:Gelatin; D048271:Chitosan
keywords: 3D culture; chitosan; gelatin; microsphere; poly-HEMA; porosity; scaffold; tissue engineering