Abstract

BackgroundThe main issue in cutaneous regeneration is to develop engineered scaffolds based on natural extracellular matrix to promote dynamics of skin progenitor cells and accelerate differentiation into mature keratinocytes.MethodsIn this study, nanofibrous scaffolds composed of a blend poly (ɛ-caprolactone) (PCL), silk fibroin (SF), soluble eggshell membrane (SESM), and Aloe vera (AV) gel were developed by electrospinning method and human basal cells were used to examine differentiation capacity toward keratinocyte-like cells. For this propose, cells were allocated to four distinct groups; control, PCL/SF, PCL/SF/SESM, and PCL/SF/SESM/AV. In all groups, cells were incubated with differentiation medium. Morphology, composition, hydrophilicity and mechanical features of PCL/SF, PCL/SF/SESM and PCL/SF/SESM/AV nanofibers were studied by scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FT-IR), water contact angle and tensile tests. To examine the orientation of basal cells to mature keratinocytes, we performed immunofluorescence analysis by monitoring cytokeratin-19. The expression of genes such as involucrin, keratin-14 and -5 was monitored by real-time PCR assay.ResultsPCL/SF, PCL/SF/SESM, and PCL/SF/SESM/AV had suitable physic chemical indices and biological activities to be applied as biomimetic scaffolds for the restoration cutaneous tissue. Compared to control, we found an increased basal cell proliferation at 7 and 14 days after plating on scaffolds and reach maximum levels in group PCL/SF/SESM/AV on day 14 (p < 0.05). Electron microscopy showed cell flattening, morphological adaptation. An integrated cell-to-cell connection was generated after cell seeding on scaffolds in all groups. Immunofluorescence imaging showed the ability of basal cells to synthesize cytokeratin-19 in PCL/SF, PCL/SF/SESM, and positive control cells after exposure to differentiation medium. However, these values were less in PCL/SF/SESM/AV compared to other groups. Real-time PCR analysis showed the potency of all scaffolds to induce the transcription of involucrin, keratin-14 and -5, especially involucrin in PCL/SF/SESM/AV group compared to the negative control.ConclusionModulation of scaffolds with natural biopolymers could enable us to synthesize structures appropriate for cutaneous regeneration.

Highlights

  • Cutaneous tissue is the largest organ that protects the body against infectious agents, mechanical, chemical and thermal stresses while prohibits the evaporation of biofluids [1, 2]

  • We aimed to investigate the regenerative potential of PCL/silk fibroin (SF), PCL/SF/soluble eggshell membrane (SESM), and PCL/SF/SESM/Aloe vera (AV) scaffold as natural biomaterials on the differentiation of human basal cells to keratinocytes over a period of 14 days

  • Because of the toxicity and high cost, the application of these solvents was limited, Only the acetic acid/formic acid solvent system seems applicable in PCL/SF, PCL/SF/SESM and PCL/SF/SESM/AV electrospinning due to the superior conductivity of formic acid which makes possible to the production of highquality nanofibers [43]

Read more

Summary

Introduction

Cutaneous tissue is the largest organ that protects the body against infectious agents, mechanical, chemical and thermal stresses while prohibits the evaporation of biofluids [1, 2]. Immune surveillance, sensory detection, and self-healing correlate with the normal activity of the skin [3]. In most circumstances, this organ has the potential to heal without any external manipulations. De novo modalities, such as tissue engineering approaches, are needed to afford these limitations [9]. In this regard, the fundamental criteria of an appropriate scaffold are to maintain cell functional activity such as adhesion, proliferation, migration, differentiation, and morphology [7, 10]. The main issue in cutaneous regeneration is to develop engineered scaffolds based on natural extracellular matrix to promote dynamics of skin progenitor cells and accelerate differentiation into mature keratinocytes

Objectives
Methods
Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call