Bone-mimetic porous hydroxyapatite/whitlockite scaffolds: preparation, characterization and interactions with human mese
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Bone-mimetic porous hydroxyapatite/whitlockite scaffolds: preparation, characterization and interactions with human mesenchymal stem cells Leonard Bauer1,* , Maja Antunovic´1, Anamarija Rogina1, Marica Ivankovic´1, and Hrvoje Ivankovic´1 1
Faculty of Chemical Engineering and Technology, University of Zagreb, Marulic´ev trg 19, HR-10000 Zagreb, Croatia
Received: 8 June 2020
ABSTRACT
Accepted: 21 October 2020
Bone-mimetic highly porous Mg-substituted calcium phosphate scaffolds, composed of hydroxyapatite (HAP) and whitlockite (WH), were synthesized by hydrothermal method at 200 °C, using calcium carbonate skeletons of cuttlefish bone, ammonium dihydrogenphosphate (NH4H2PO4) and magnesium chloride hexahydrate (MgCl2 9 6H2O) or magnesium perchlorate (Mg(ClO4)2) as reagents. The effect of Mg content on the compositional and morphological properties of scaffolds was studied by means of X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis and scanning electron microscopy (SEM) with energy-dispersive X-ray analysis. Structural refinements performed by Rietveld method indicated that Mg2? ions were preferentially incorporated into the WH phase. SEM images of all prepared scaffolds showed that the interconnected structure of the cuttlefish bone was completely maintained after the hydrothermal synthesis. Results of compression tests showed a positive impact of the whitlockite phase on the mechanical properties of scaffolds. Human mesenchymal stem cells (hMSCs) were cultured on scaffolds in osteogenic medium for 21 days. Immunohistochemical staining showed that Mg-CaP scaffolds with the HAP:WH wt ratio of 90:10 and 70:30 exhibited higher expression of collagen type I and osteocalcin than pure HAP scaffold. Calcium deposition was confirmed by Alizarin Red staining. Positive effect of Mg2? ions on the differentiation of hMSCs on porous 3D scaffolds was also confirmed by reverse transcription-quantitative polymerase chain reaction analysis.
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Springer Science+Business
Media, LLC, part of Springer Nature 2020
Handling Editor: Christopher Blanford.
Address correspondence to E-mail: [email protected]
https://doi.org/10.1007/s10853-020-05489-3
J Mater Sci
GRAPHICAL ABSTRACT
Introduction Over the last three decades, calcium phosphate (CaP) ceramics have been extensively studied and widely used as bone substitute materials [1–4]. In addition to the most researched hydroxyapatite (HAP, Ca10(PO4)6(OH)2), tricalcium phosphate (b-TCP, Ca3(PO4)2) and the biphasic mixture of HAP and b-TCP (BCPs) have also attracted considerable attention. In order to better mimic natural bone mineral, ionic substitutions within the apatite lattice have been widely employed. Several reviews have been published on the preparation of ion substituted calcium phosphate materials [5–8]. Mg2? ion plays an important role in bone metabolism [9], which is the reason why it has attracted such remarkable interest. Magnesium deficiency can result in decreased bone mass, reduced bone growth and osteoporosis. To incorporate Mg2
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