Co-cultivation of mesenchymal stem cells and skin keratinocytes on electrospun scaffolds

Abstract

Available treatments in skin regeneration are insufficient for promoting healing. This study has aimed to produce a cutaneous substitute combining mesenchymal stem cells (MSCs), keratinocytes and a PDLLA biomaterial, constructed by electrospinning, into 3 groups: 1) PDLLA, 2) PDLLA/NaOH, scaffolds with hydrolyzed surfaces and 3) PDLLA/Lam, hydrolyzed and with laminin binding. The MSCs were seeded at the bottom of the scaffolds. After 24 hours, skin keratinocytes were seeded at the top. The scaffolds were characterized by morphology, fiber diameter, cell adhesion on the day of the seeding and viability on days 7, 14 and 21. As a result, the scaffolds presented randomly distributed, well formed fibers. The fiber diameter for all the groups was 4.58 mmfor the largest fibers and 574 nm for the smallest. The pore sizes of the scaffolds obtained were approximately 27.5 mm and 3.44 mm, for the largest and smallest fibers, respectively. In biological analysis, cell adhesion was greater in the PDLLA/Lam scaffolds with absorbance of 2.268 – 0.494, in comparison with 1.264 – 0.473 for the PDLLA and 1.159 – 0.120 for the PDLLA/NaOH scaffolds. There was an improvement in the number of viable cells from day 7 to day 21 of analysis. In general, the PDLLA/Lam group showed the best results for cell adhesion and viability tests. In conclusion, the PDLLA scaffolds, mainly the PDLLA/Lam groups, showed good results for the co-cultivation of the cells, with good cell adhesion and the presence of viable cells. Therefore, these scaffolds show promise to be suitable biomaterials for use in tissue engineering.