Culture of human mesenchymal stem cells using electrosprayed porous chitosan microbeads

The aim of this study was to fabricate porous chitosan microbeads using an electrospraying method into liquid nitrogen, then thawing and refreezing. The microbeads were then used to evaluate their potential for tissue engineering of human mesenchymal stem cells (hMSCs). Scanning electron microscopy (SEM) and a mercury porosimeter were used to show the morphology of the scaffolds formed and to determine their pore size and porosity. As the chitosan concentration increased (0.5, 1, 1.5, and 2 wt %), the diameter of the porous microbead increased from 350 to 890 microm, and the average pore size and the porosity decreased from 65 to 21 microm and 95 to 38%, respectively. The hMSCs were cultured onto the porous microbeads in a spinner flask. SEM images and methyl tetrazolium salt assays at 3, 7, 14, and 21 days of culture revealed that hMSCs had successfully attached and proliferated inside the porous microbeads. This study demonstrated that electrosprayed porous chitosan microbeads can be used as three-dimensional scaffolds for tissue engineering.