Electrospun triple‐layered PLLA/gelatin. PRGF/PLLA scaffold induces fibroblast migration

Abstract

The function of fibroblast cells in wounded areas results in reconstruction of the extra cellular matrix and consequently resolution of granulation tissue. It is suggested that the use of platelet-rich plasma can accelerate the healing process in nonhealing or slow-healing wounds. In this study, a simple and novel method has been used to fabricate an electrospun three-layered scaffold containing plasma rich in growth factor with the aim of increasing the proliferation and migration of fibroblast cells in vitro. First, plasma rich in growth factor was derived from platelet rich plasma, and then a three-layered scaffold was fabricated using PLLA nanofibers as the outer layers and plasma rich in growth factor-containing gelatin fibers as the internal layer. The growth morphology of cells seeded on this scaffold was compared to those seeded on one layered PLLA scaffold. The study of the cell growth rate on different substrates and the migration of cells in response to the drug release of multilayered scaffold was investigated by the cell quantification assay and a modified under agarose assay. Scanning electron microscopy and fluorescence images showed that cells seeded on multilayered scaffold were completely oriented 72 hours after seeding compared to those seeded on PLLA scaffold. The cell quantification assay also indicated significant increase in proliferation rate of cells seeded on three-layered scaffold compared to those seeded on PLLA scaffold and finally, monitoring cell migration proved that cells migrate significantly toward the three-layered scaffold up to 48 to 72 hours and afterwards start to show a diminished migration rate toward this scaffold. K E Y W O R D S electrospinning, fibroblast migration, multilayered scaffolds, platelet rich plasma, skin tissue engineering

Evaluation of in vitro fibroblast migration by electrospun triple-layered PU-CA/gelatin.PRGF/PU-CA scaffold using an AAVS1 targeted EGFP reporter cell line

“…Furthermore, the correlation between gene expression, release profile, and cell behavior displayed that the scaffold containing PRGF can be potentially used as a wound dresser. 5 MTT results showed an increase in the number of active cells on days 1, 3, and 7 in the TCPS group, while reducing viable cells was observed after day 7. In addition, the cells showed a proper proliferation with reasonable speed on these scaffolds until day 14.…”

“…The multi-layered scaffolds, similar to skin architecture, are employed as potential engineered skin substitutes applicable to wound care. 5 Efficient genome engineering tools and reliable reporter cell lines are invaluable tools to investigate the role and behavior of cells during the treatment processes. Sitespecific gene integration at pre-defined safe harbour loci facilitates rapid generation of stable cell lines with limited transgene silencing and few detrimental effects.…”

“… 3 Studies have indicated that PRGF contains several bioactive proteins and growth factors that facilitate the process of tissue regeneration and wound healing. 4 , 5 Therefore, the transport of growth factors and cells to the wound area is vital in the wound healing process. The application of PRGF in the scaffold along with fibrillar and cellular components is highly beneficial for the generation of elastic, dense, and haemostatic fibrin during the re-epithelialization phase of soft tissue repair.…”

Electrospun Nanofibers: Recent Applications in Drug Delivery and Cancer Therapy

“…With this type of gelatin-based scaffolds, in addition to drug transport and use for tissue engineering, the migration of certain cells can also be induced or favored. This is the case in the study of Piran et al [47] that synthesized electrospun three-layered scaffold with plasma enriched with growth factor to promote the migration and growth of fibroblast, which is a very important aspect in the regeneration of wounds.…”

Electrospinning process parameters optimization for biofunctional curcumin/gelatin nanofibers

“…Gelatin nanofibers (like collagen) have been used in biomedical applications such as cosmetics, wound dressing, tissue engineering, surgical treatments, etc [34]. Gelatin nanofibers (having sufficient mechanical properties to be used in these applications as nanofibrous mats) have been electrospun for ultrathin nanofibers [34][35][36][37][38][39][40][41][42][43][44]. These authors also reported that the properties of these nanofibers can be tailored as per requirements by optimizing input parameters such as voltage, viscosity, distance between the spinneret and rotating drum collector, and flow rate.…”