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

Abstract

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.

Graphical Abstract

Keywords:

• Human keratinocyte
• mesenchymal stem-cells
• amnion
• co-culture
• dermal tissue

Acknowledgment

We gratefully thank from the financial grant of Shahid Beheshti University of Medical Sciences by the ethical number of IR.SBMU.RETECH.REC.1397.875 and ID code of 14269. Also, it should be noted that none of the authors do not any conflict about the article publication.