A Study On The Preparation And Characterization Of A Porous Scaffold For Three-dimensional Culture Of Airway Smooth Muscle Cell

Pore architecture in 3D polymeric scaffolds is known to play a critical role in tissue engineering as it provides the vital framework for the seeded cells to organize into a functioning tissue. The research fabricated chitosan-gelatin 3D scaffolds using freeze-drying technique (phase separation). This research used the ice crystals as porogen and got the porous scaffolds of different porosities and pore sizes through different freezing temperature regimes and different concentrations. From the scanning electron microscopic images, when the scaffold fabricated by lower concentration (1wt%) and higher pre-freezing temperature (-20℃), the scaffold formed the largest pore size (60-100μm). But highly interconnected pores with 93% porosity were observed when solution (1wt%) were frozen at -196℃.The research also investigated the density, water absorption and in vitro degradation of chitosan-gelatin scaffolds. The study found that the density of scaffolds became larger with the growth of the concentration of the chitosan-gelatin mixture; the water absorption ability advanced with the growth of pre-freezing temperature and the reduction of concentration of scaffolds; the time for the degradation of scaffolds decreased with the reduction of concentration of scaffolds and pre-freezing temperature.The paper built the method of isolating and culturing rat's airway smooth muscle cells. The research achieved obtaining robust rat's airway smooth muscle cells by attaching tissue pieces to the cell culture flasks. And the cells within seven generation can be used in this study.The research used the fourth generation of airway smooth muscle cell to seed into the scaffold of chitosan-gelatin. And by using MTT assay, we found the cells were in good condition. The experiment found that the porosity of the scaffold affected cell proliferation significantly. In other words, the cell proliferation advanced with the growth of the porosity of the scaffolds. Also we did scanning electron microscopy and fluorescent staining to find that cells were growing well in the scaffolds.This study indicates the freeze-drying technique as an alternative method to fabricate highly interconnected porous scaffolds for developing functional 3D chitosan-gelatin matrices for potential tissue engineering, biomedical and biotechnological applications.