The effect of chitosan-gelatin scaffold pore size on amniotic epithelial cell attachment for use in tissue engineering

The amniotic membrane has gained much attention in regenerative medicine as a precious cell source. Recently, reparation of three dimensional matrices (scaffold) with appropriate specificity for cell culture, which depends on cell type, has been the subject of many studies. This study aimed to design optimal three-dimensional matrices in order to utilize amniotic epithelial cells for tissue engineering studies. In this experimental study, chitosan and chitosan-gelatin scaffolds were formed by freeze-drying technique at different pre-freezing temperatures (-20/-80/-196°C). Sample pore size was analyzed by scanning electron microscope (SEM) and porosity was measured by liquid displacement method. Amniotic epithelial cells isolated from amniotic membrane, were cultured on scaffolds with different pore sizes and the attachment of these cells was evaluated using MTT-Formazan assay and SEM images. Electron microscope results showed larger pore size at higher temperature (-20) and longer pre-freezing time. The porosity of 73% was desirable for cell culture. Furthermore, MTT findings confirmed the importance of pore size (100 µm) in cell adhesion to the scaffold. It seems that chitosan and chitosan-gelatin scaffolds with larger pore size increase the chance of amniotic epithelial cells attachment. Thus, these natural scaffolds can be used as suitable matrices for tissue engineering purposes.