Recently, milk-derived proteins have attracted attention for applications in the biomedical field such as tissue regeneration. Whey protein isolate (WPI), especially its main component β-lactoglobulin, can modulate immunity and acts as an antioxidant, antitumor, antiviral, and antibacterial agent. There are very few reports of the application of WPI in tissue engineering, especially in bone tissue engineering. In this study, we tested the influence of different concentrations of WPI on behavior of human osteoblast-like Saos-2 cells, human adipose tissue-derived stem cells (ASC), and human neonatal dermal fibroblasts (FIB). The positive effect on growth was apparent for Saos-2 cells and FIB but not for ASC. However, the expression of markers characteristic for early osteogenic cell differentiation [type-I collagen (COL1) and alkaline phosphatase (ALP)] as well as ALP activity, increased dose-dependently in ASC. Importantly, Saos-2 cells were able to deposit calcium in the presence of WPI, even in a proliferation medium without other supplements that support osteogenic cell differentiation. The results indicate that, depending on the cell type, WPI can act as an enhancer of cell proliferation and osteogenic differentiation. Therefore, enrichment of biomaterials for bone regeneration with WPI seems a promising approach, especially due to the low cost of WPI.

Department of Biomaterials and Tissue Engineering Institute of Physiology of the Czech Academy of Sciences Videnska 1083 142 20 Prague the Czech Republic

Department of Food Technology Institute of Human Nutrition and Food Science Christian Albrechts Universität zu Kiel Heinrich Hecht Platz 10 24118 Kiel Germany

Department of Molecular Biotechnology Faculty of Bioscience Engineering Ghent University Coupure Links 653 9000 Gent Belgium; Centre for Nano and Biophotonics Ghent University 9000 Ghent Belgium

Department of Molecular Biotechnology Faculty of Bioscience Engineering Ghent University Coupure Links 653 9000 Gent Belgium; Engineering Department Gillow Avenue Lancaster University Lancaster LA1 4YW United Kingdom

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