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  • Development of a New Bone-Mimetic Surface Treatment Platform: Nanoneedle Hydroxyapatite (nnHA) Coating.

Development of a New Bone-Mimetic Surface Treatment Platform: Nanoneedle Hydroxyapatite (nnHA) Coating.

Advanced healthcare materials (2020-10-29)
Kian F Eichholz, Stanislas Von Euw, Ross Burdis, Daniel J Kelly, David A Hoey
ABSTRACT

The hierarchical structure of bone plays pivotal roles in driving cell behavior and tissue regeneration and must be considered when designing materials for orthopedic applications. Herein, it is aimed to recapitulate the native bone environment by using melt electrowriting to fabricate fibrous microarchitectures which are modified with plate-shaped (pHA) or novel nanoneedle-shaped (nnHA) crystals. Nuclear magnetic resonance spectroscopy, scanning electron microscopy, transmission electron microscopy, and X-ray diffraction demonstrate that these coatings replicate the nanostructure and composition of native bone. Human mesenchymal stem/stromal cell (MSC) mineralization is significantly increased fivefold with pHA scaffolds and 14-fold with nnHA scaffolds. Given the protein stabilizing properties of mineral, these materials are further functionalized with bone morphogenetic protein 2 (BMP2). nnHA treatment facilitates controlled release of BMP2 which further enhance MSC mineral deposition. Finally, the versatility of this nnHA treatment method, which may be used to coat different architectures/materials including fused deposition modeling (FDM) scaffolds and Ti6Al4V titanium, is demonstrated. This study thus outlines a method for fabricating scaffolds with precise fibrous microarchitectures and bone-mimetic nnHA extrafibrillar coatings which significantly enhance MSC osteogenesis and therapeutic protein delivery, and leverages these results to show how this surface treatment method may be applied to a wider field for multiple orthopedic applications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Polycaprolactone, average Mn 80,000
Sigma-Aldrich
Hydroxyapatite, nanopowder, <200 nm particle size (BET), ≥97%, synthetic
Sigma-Aldrich
Direct Red 80, Dye content 25 %
Sigma-Aldrich
Sodium phosphate tribasic dodecahydrate, BioXtra, ≥98.0% (titration)
Sigma-Aldrich
SIGMAFAST p-Nitrophenyl phosphate Tablets, tablet, To prepare 5 mL
Sigma-Aldrich
Calcium chloride dihydrate, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
Human BMP2 ELISA Kit, for serum, plasma, cell culture supernatant and urine
Sigma-Aldrich
Phosphatase, Alkaline from bovine intestinal mucosa, buffered aqueous solution, ≥2,000 DEA units/mg protein
Sigma-Aldrich
p-Nitrophenyl Phosphate Liquid Substrate System, liquid
Sigma-Aldrich
Alizarin Red S, certified by the Biological Stain Commission