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Abstract
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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 (ɛ-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.
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