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Marine-Inspired Enzymatic Mineralization of Dairy-Derived Whey Protein Isolate (WPI) Hydrogels for Bone Tissue Regeneration

Engineering Department, Lancaster University, Lancaster LA1 4YW, UK
Department of Biomaterials and Composites, Faculty of Materials Science and Ceramics, AGH University of Science and Technology, 30-962 Kraków, Poland
INSERM U1008-Controlled Drug Delivery Systems and Biomaterials, Université de Lille, 59006 Lille, France
Chemistry Department, Lancaster University, Lancaster LA1 4YW, UK
Department of Biotechnology, Ghent University, B-9000 Gent, Belgium
Nanotechnology Department, Saratov State University, Saratov 410012, Russia
Laboratory of Biomaterials and Regenerative Medicine for Advanced Therapies, Indian Council of Medical Research-Regional Medical Research Center, Bhubaneswar, Odisha 751023, India
Materials Science Institute (MSI), Lancaster University, Lancaster LA1 4YW, UK
Author to whom correspondence should be addressed.
Mar. Drugs 2020, 18(6), 294;
Received: 27 April 2020 / Revised: 25 May 2020 / Accepted: 28 May 2020 / Published: 2 June 2020
Whey protein isolate (WPI) is a by-product from the production of cheese and Greek yoghurt comprising β-lactoglobulin (β-lg) (75%). Hydrogels can be produced from WPI solutions through heating; hydrogels can be sterilized by autoclaving. WPI hydrogels have shown cytocompatibility and ability to enhance proliferation and osteogenic differentiation of bone-forming cells. Hence, they have promise in the area of bone tissue regeneration. In contrast to commonly used ceramic minerals for bone regeneration, a major advantage of hydrogels is the ease of their modification by incorporating biologically active substances such as enzymes. Calcium carbonate (CaCO3) is the main inorganic component of the exoskeletons of marine invertebrates. Two polymorphs of CaCO3, calcite and aragonite, have shown the ability to promote bone regeneration. Other authors have reported that the addition of magnesium to inorganic phases has a beneficial effect on bone-forming cell growth. In this study, we employed a biomimetic, marine-inspired approach to mineralize WPI hydrogels with an inorganic phase consisting of CaCO3 (mainly calcite) and CaCO3 enriched with magnesium using the calcifying enzyme urease. The novelty of this study lies in both the enzymatic mineralization of WPI hydrogels and enrichment of the mineral with magnesium. Calcium was incorporated into the mineral formed to a greater extent than magnesium. Increasing the concentration of magnesium in the mineralization medium led to a reduction in the amount and crystallinity of the mineral formed. Biological studies revealed that mineralized and unmineralized hydrogels were not cytotoxic and promoted cell viability to comparable extents (approximately 74% of standard tissue culture polystyrene). The presence of magnesium in the mineral formed had no adverse effect on cell viability. In short, WPI hydrogels, both unmineralized and mineralized with CaCO3 and magnesium-enriched CaCO3, show potential as biomaterials for bone regeneration. View Full-Text
Keywords: hydrogel; composite; mineralization; enzyme; bioinspired; whey protein isolate hydrogel; composite; mineralization; enzyme; bioinspired; whey protein isolate
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Norris, K.; Kocot, M.; Tryba, A.M.; Chai, F.; Talari, A.; Ashton, L.; Parakhonskiy, B.V.; Samal, S.K.; Blanchemain, N.; Pamuła, E.; Douglas, T.E.L. Marine-Inspired Enzymatic Mineralization of Dairy-Derived Whey Protein Isolate (WPI) Hydrogels for Bone Tissue Regeneration. Mar. Drugs 2020, 18, 294.

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