Electrospun gelatin/methylcellulose hybrid nanofibers promoted the maturation of human cutaneous tissue progenitor cells toward keratinocyte-like cells

Until the present time, the development and fabrication of ideal engineered grafts for cutaneous regeneration is the subject of debate. Previously, we showed regenerative properties of electrospun silk fibroin (SF), poly (e-caprolactone) (PCL), and soluble eggshell membrane (SESM) nanofibers on human skin progenitor cells in vitro. Here, we aimed to assess the potency of electrospun PCL/SF/SESM/gelatin (Gel) and PCL/SF/SESM/methylcellulose (MC) hybrid scaffolds on skin progenitor cells in vitro. To this end, PCL/SF/SESM electrospun nanofibers containing Gel and MC were used to measure the differentiation of skin basal cells (BCCs) into keratinocyte-like cells. Physicochemical characterization of prepared nanofibers was studied using SEM, FT-IR, tensile, and water contact angle assays. BCCs viability, attachment, survival, and differentiation capacity were monitored using MTT, immunofluorescence (IF), and real-time PCR analyses. Data indicated the addition of Gel and MC into electrospun PCL/SF/SESM nanofibers increased mean diameter size and surface wettability. In contrast to MC, the addition of Gel caused a slight decrease in tensile strength value. In vitro data exhibited improved cell viability coincided with cell-to-cell connection (p < 0.05). IF showed the increase of keratinocyte specific marker, cytokeratin-19, in BCCs plated on PCL/SF/SESM/Gel and PCL/SF/SESM/MC scaffolds related to control cells cultured on the plastic surface. Real-time PCR analysis of the up-regulation of involucrin, keratin-14, and -5 in PCL/SF/SESM/Gel and PCL/SF/SESM/MC groups compared to matched control groups. Based on our data, these effects were significantly more in PCL/SF/SESM/MC groups compared to other groups (p < 0.0001). It is suggested that PCL/SF/SESM scaffold blended with Gel and MC can be used as supporting substrates for the regeneration of injured cutaneous tissue.

Automated summary

This study evaluated the effects of PCL/SF/SESM/Gel and PCL/SF/SESM/MC hybrid scaffolds on skin progenitor cells. The results showed that the addition of Gel and MC increased the diameter size and surface wettability of the nanofibers, improved cell viability, and promoted the differentiation of skin cells into keratinocyte-like cells.