The Bilayer Skin Substitute Based on Human Adipose-Derived Mesenchymal Stem Cells and Neonate Keratinocytes on the 3D Nanofibrous Pcl-Platelet Gel Scaffold Publisher



Ranjbarvan P^{1, 2} ; Golchin A^{1, 3} ; Azari A⁴ ; Niknam Z⁵ Show Affiliations
Source: Polymer Bulletin Published:2022

Abstract

Abstract: In the present study, neonate keratinocytes were cocultured with human adipose-derived mesenchymal stem cells (hAMSCs) on the electrospun polycaprolactone-platelet gel (PCL–PG) scaffold. To evaluate its potential of wound healing and skin tissue engineering, neonate keratinocytes must be differentiated. The PCL scaffold prepared by the electrospinning technique was fabricated and coated by PG. Then scaffolds were fully characterized by SEM, contact angle, FTIR–ATR, and tensile test. Following seeding hAMSCs as a feeder layer, neonate keratinocytes were cocultured directly on PCL–PG scaffolds and neat PCL. The hAMSCs and neonate keratinocytes' viability was measured using MTT assay for up to 10 days (1st, 3rd, 5th, 7th, and 10th days). To examine the epidermal maturation, cytokeratin 10 and loricrin determinants detection was used through immunocytochemistry (ICC). In addition, real-time PCR was used to examine keratinocyte marker genes in keratin 10, keratin 14, and Involucrin. The MTT assay results showed higher cell viability and the proliferation of neo-keratinocytes on PCL–PG fibrillar scaffolds than the PCL scaffold. On the PCL–PG scaffolds in the presence of hAMSCs as a feeder layer, the PCR and ICC analysis had a higher cell differentiation compared to neat PCL scaffolds. Moreover, SEM images showed that the keratinocytes cocultured with hAMSCs demonstrated a better proliferation and adhesion on PCL–PG nanofiber scaffolds. Based on results, PG increased the nanofibrous PCL scaffold's biological properties, including hydrophilicity, cell attachment, cell viability, and expression of keratinocyte markers in the neat keratinocytes and hAMSCs coculture system. The findings support the potential of this engineered construct for engineering skin tissue and wound dressing. Graphic abstract: [Figure not available: see fulltext.]. © 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.