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  • Human Mesenchymal Stem Cell Response to Lactoferrin-based Composite Coatings.

Human Mesenchymal Stem Cell Response to Lactoferrin-based Composite Coatings.

Materials (Basel, Switzerland) (2019-10-23)
Madalina Icriverzi, Anca Bonciu, Laurentiu Rusen, Livia Elena Sima, Simona Brajnicov, Anisoara Cimpean, Robert W Evans, Valentina Dinca, Anca Roseanu
ABSTRACT

The potential of mesenchymal stem cells (MSCs) for implantology and cell-based therapy represents one of the major ongoing research subjects within the last decades. In bone regeneration applications, the various environmental factors including bioactive compounds such as growth factors, chemicals and physical characteristics of biointerfaces are the key factors in controlling and regulating osteogenic differentiation from MSCs. In our study, we have investigated the influence of Lactoferrin (Lf) and Hydroxyapatite (HA) embedded within a biodegradable PEG-PCL copolymer on the osteogenic fate of MSCs, previous studies revealing an anti-inflammatory potential of the coating and osteogenic differentiation of murine pre-osteoblast cells. The copolymer matrix was obtained by the Matrix Assisted Pulsed Laser Evaporation technique (MAPLE) and the composite layers containing the bioactive compounds (Lf, HA, and Lf-HA) were characterised by Scanning Electron Microscopy and Atomic Force Microscopy. Energy-dispersive X-ray spectroscopy contact angle and surface energy of the analysed coatings were also measured. The characteristics of the composite surfaces were correlated with the viability, proliferation, and morphology of human MSCs (hMSCs) cultured on the developed coatings. All surfaces were found not to exhibit toxicity, as confirmed by the LIVE/DEAD assay. The Lf-HA composite exhibited an increase in osteogenic differentiation of hMSCs, results supported by alkaline phosphatase and mineralisation assays. This is the first report of the capacity of biodegradable composite layers containing Lf to induce osteogenic differentiation from hMSCs, a property revealing its potential for application in bone regeneration.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Edelfosine, ≥95% (HPLC)
Sigma-Aldrich
Hydroxyapatite, nanopowder, <200 nm particle size (BET), ≥97%, synthetic
Sigma-Aldrich
Poly(ethylene glycol)-block-poly(ε−caprolactone) methyl ether, PEG average Mn ~5,000, PCL average Mn ~5,000
Sigma-Aldrich
Lactoferrin from human milk, ≥85% (SDS-PAGE), lyophilized powder
Sigma-Aldrich
Quantitative Alkaline Phosphatase ES Characterization Kit, This Quantitative Alkaline Phosphatase Embryonic Stem (ES) Characterization Kit easily quantifies the amount of alkaline phosphatase present within their embryonic stem cell cultures using a convenient 96-well colorimetric enzymatic assay.