Evaluation of dermal growth of keratinocytes derived from foreskin in co-culture condition with mesenchymal stem cells on polyurethane/gelatin/amnion scaffold

An appropriate scaffold made of both synthetic and natural polymers can simultaneously supply desirable mechanical and biological characteristics such as tensile strength, elasticity, biocompatibility and bioactivity. Here, we investigated the synergistic effect of electrospun polyurethane (PU), gelatin and human amniotic polymers in the presence of mesenchymal stem cells (MSCs) and human keratinocyte (H-keratino) as cell sources for the regeneration of skin lesions. The in vitro biocompatibility was examined by the assay of (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and the epithelial differentiation was verified by real-time PCR after co-culture of MSCs and H-keratino on the proposed nanofibrous scaffold. A significant improvement in cell proliferation was found after cell culture on the electrospun scaffold containing the human amniotic membrane. In addition, gene expression after 14 days of co-culture process confirmed that both PU/gelatin and PU/gelatin-amnion electrospun scaffolds promote epithelial proliferation as well as dermal differentiation compared to H-keratino single-cell culture with regard to various markers such as Cytokeratin 10, Cytokeratin 14 and Involucrin. Also, the in vivo examination in mice indicated the considerable influence of the scaffold on the co-culturing of both cell types. Taking into account the results, the electrospun PU/gelatin-amnion nanofibrous scaffold in conjugation with the H-keratino/MSC co-culture could be considered as an exceptional nanofibrous substrate for applications in skin tissue engineering.

Automated summary

This study investigated the synergistic effect of an electrospun scaffold made of polyurethane, gelatin, and human amniotic polymers in the co-culture of mesenchymal stem cells and human keratinocytes. The results showed that this scaffold promoted cell proliferation and epithelial differentiation, and had a significant influence in mice experiments.