Accelerated Wound Closure - Differently Organized Nanofibers Affect Cell Migration and Hence the Closure of Artificial Wounds in a Cell Based In Vitro Model.

Maximilian Ottosson,Albin Jakobsson,Fredrik Johansson,Christophe Egles

doi:10.1371/journal.pone.0169419

Abstract

Nanofiber meshes holds great promise in wound healing applications by mimicking the topography of extracellular matrix, hence providing guidance for crucial cells involved in the regenerative processes. Here we explored the influence of nanofiber alignment on fibroblast behavior in a novel in vitro wound model. The model included electrospun poly-ε-caprolactone scaffolds with different nanofiber orientation. Fibroblasts were cultured to confluency for 24h before custom-made inserts were removed, creating cell-free zones serving as artificial wounds. Cell migration into these wounds was evaluated at 0-, 48- and 96h. Cell morphological analysis was performed using nuclei- and cytoskeleton stainings. Cell viability was assessed using a biochemical assay. This study demonstrates a novel in vitro wound assay, for exploring of the impact of nanofibers on wound healing. Additionally we show that it’s possible to affect the process of wound closure in a spatial manner using nanotopographies, resulting in faster closure on aligned fiber substrates.

Highlights

Increased understanding and advances in molecular- and cellular biology in recent years have greatly improved our understanding of the wound healing process [1]
N = 600 fibers/scaffold type were included for fiber diameter analysis, n = 60 pores/scaffold type for pore measurements and n = 3 scaffolds of each scaffold type were included for sample thickness and porosity measurement
The experimental results gathered throughout this project supports the hypothesis that an oriented scaffold would increase directed cell mobility as well as favor cell polarization and elongated cell morphologies compared to random fiber scaffolds and flat control surfaces, and accelerate the wound closure process. This is an important effect for several medical applications where orientation/polarization is of essence

Summary

Introduction

Increased understanding and advances in molecular- and cellular biology in recent years have greatly improved our understanding of the wound healing process [1]. This in turn has helped researchers and clinicians in their development of new improved methods for wound care, along with treatments to enhance the healing of wounds [2, 3]. The healing/regeneration of the skin incorporates a re-establishment of the dermis structure, i.e. vascularized granulation tissue, before a proper closure of the epidermis (epithelial cells) is meaningful. Triggering fibroblast migration into the wound area may speed up the remodeling of the dermis, and pave the way for faster closure of the epithelial layer. In order to accelerate these events, an artificial topographical guidance cue, resembling the native extracellular matrix (ECM) structure can be used

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Conclusion