Eva Veiseth-Kent

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.

This chapter outlines a method for sample preparation for bottom-up proteomics by LC-MSMS analysis of in vitro proliferated bovine primary skeletal muscle cells. The protocol describes the isolation of bovine primary skeletal muscle cells, extraction of proteins, proteolytic digestion of proteins, and desalting of the final peptide samples. The final peptide samples can be analyzed using various LC-MSMS systems after reconstitution in a suitable elution buffer.

"Proteomics as a tool to better understand beef tenderness" published on 28 Nov 2014 by Wageningen Academic.

"Protein changes in muscles with varying meat tenderness" published on 01 Nov 2014 by Wageningen Academic.

"Changes in protein abundance in the vertebral columnof Atlantic salmon (Salmosalar) fed variable dietary P levels" published on 01 Nov 2013 by Wageningen Academic.