Processed Eggshell Membrane Powder Is a Promising Biomaterial for Use in Tissue Engineering.

Sissel B Rønning,Paul Barham,Ralf Schmidt,Ragnhild S Berg,Mona E Pedersen,Henri-Pierre Suso,Eva Veiseth-Kent,Eirik Haugen,Vibeke Høst,Christian R Wilhelmsen

doi:10.3390/ijms21218130

Sissel B Rønning, Paul Barham + Show 8 more

Open Access

https://doi.org/10.3390/ijms21218130

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Abstract

The purpose of this study was to investigate the tissue regenerating and biomechanical properties of processed eggshell membrane powder (PEP) for use in 3D-scaffolds. PEP is a low-cost, natural biomaterial with beneficial bioactive properties. Most importantly, this material is available as a by-product of the chicken egg processing (breaking) industry on a large scale, and it could have potential as a low-cost ingredient for therapeutic scaffolds. Scaffolds consisting of collagen alone and collagen combined with PEP were produced and analyzed for their mechanical properties and the growth of primary fibroblasts and skeletal muscle cells. Mechanical testing revealed that a PEP/collagen-based scaffold increased the mechanical hardness of the scaffold compared with a pure collagen scaffold. Scanning electron microscopy (SEM) demonstrated an interconnected porous structure for both scaffolds, and that the PEP was evenly distributed in dense clusters within the scaffold. Fibroblast and skeletal muscle cells attached, were viable and able to proliferate for 1 and 2 weeks in both scaffolds. The cell types retained their phenotypic properties expressing phenotype markers of fibroblasts (TE7, alpha-smooth muscle actin) and skeletal muscle (CD56) visualized by immunostaining. mRNA expression of the skeletal muscle markers myoD, myogenin, and fibroblasts marker (SMA) together with extracellular matrix components supported viable phenotypes and matrix-producing cells in both types of scaffolds. In conclusion, PEP is a promising low-cost, natural biomaterial for use in combination with collagen as a scaffold for 3D-tissue engineering to improve the mechanical properties and promote cellular adhesion and growth of regenerating cells.

Highlights

Tissue regeneration and healing is an intricate process where the skin, skeletal muscle or other body tissues repair themselves after injury [1,2]
We have previously demonstrated that industrially processed eggshell membrane powder (PEP) is biocompatible and has wound healing and regeneration properties in vitro and in vivo [7,8]
We demonstrated that PEP has immune-modulating properties in vitro, by increasing anti-inflammatory cytokine production IL-10 and increased expression of heme-detoxification enzyme heme oxygenase-1 (HO-1), a highly expressed protein in the anti-inflammatory M2 macrophages

Summary

Introduction

Tissue regeneration and healing is an intricate process where the skin, skeletal muscle or other body tissues repair themselves after injury [1,2]. The extracellular matrix (ECM), a complex network of fibrous proteins, carbohydrates and glycoproteins surrounding the cells, is the main regulator in tissue regeneration. It works as the structural support and scaffold for cell growth and differentiation, the supplier and regulator of essential growth factors and cytokines, and cell enzyme activities (for review [3]). Tissue engineering is a promising therapeutic treatment to repair and replace destroyed tissue, where biodegradable scaffolds mimick the ECM support the regeneration process [4]. Several different materials have been proposed, as a possible replacement for the ECM, with the majority being either biological or synthetic polymers such as a collagen or polyester [5]. One promising material in this respect is using eggshell membrane

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