Multiscale-Engineered Muscle Constructs: PEG Hydrogel Micro-Patterning on an Electrospun PCL Mat Functionalized with Gold Nanoparticles.

Megane Beldjilali-Labro,Augustin Lerebours,Claire Stewart,Alexander David Brown,Rachid Jellali,Murielle Dufresne,Alejandro Garcia Garcia,Jean-François Grosset,Cécile Legallais,Fahmi Bedoui,Erwann Guenin

doi:10.3390/ijms23010260

Abstract

The development of new, viable, and functional engineered tissue is a complex and challenging task. Skeletal muscle constructs have specific requirements as cells are sensitive to the stiffness, geometry of the materials, and biological micro-environment. The aim of this study was thus to design and characterize a multi-scale scaffold and to evaluate it regarding the differentiation process of C2C12 skeletal myoblasts. The significance of the work lies in the microfabrication of lines of polyethylene glycol, on poly(ε-caprolactone) nanofiber sheets obtained using the electrospinning process, coated or not with gold nanoparticles to act as a potential substrate for electrical stimulation. The differentiation of C2C12 cells was studied over a period of seven days and quantified through both expression of specific genes, and analysis of the myotubes’ alignment and length using confocal microscopy. We demonstrated that our multiscale bio-construct presented tunable mechanical properties and supported the different stages skeletal muscle, as well as improving the parallel orientation of the myotubes with a variation of less than 15°. These scaffolds showed the ability of sustained myogenic differentiation by enhancing the organization of reconstructed skeletal muscle. Moreover, they may be suitable for applications in mechanical and electrical stimulation to mimic the muscle’s physiological functions.

Highlights

Reconstructing lost function or mass of skeletal muscle, caused by chronic diseases and traumatic injuries, is often difficult to achieve despite the highly regenerative nature of this tissue [1,2]
This manufacturing process led to four different types of scaffold: (I) electrospun poly-ε-caprolactone (PCL), (II) the same coated with gold nanoparticles (PCL-Au), (III) electrospun PCL with patterning of polyethylene glycol (PEG) hydrogel lines (PCL-PEG), and (IV) an electrospun PCL coated with gold nanoparticles and patterned with PEG hydrogel lines (PCL-Au-PEG)
A mediumrange drum rotation speed of approximately 1000 rpm was applied during the electrospinning process

Summary

Introduction

Reconstructing lost function or mass of skeletal muscle, caused by chronic diseases and traumatic injuries, is often difficult to achieve despite the highly regenerative nature of this tissue [1,2]. Synthetic, or biohybrid biomaterials have been developed to replace or repair the structure of skeletal muscle. These constructs can represent suitable “native-like” tissue models for in vitro investigations, for better understanding of the muscle regeneration process, drug screening, the evaluation of physical protocols for the treatment of muscular diseases or injuries. The highly organized structure of skeletal muscle in long parallel conductive fibers is undoubtedly a challenge for its reconstruction [3,4]. Some studies focused on the impact of the size (nano or micro) and topograhical design on myoblast alignment, and of fusion on the biomaterial. [8,9]

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Conclusion