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Differentiation. 2019 May - Jun;107:24-34. doi: 10.1016/j.diff.2019.05.002. Epub 2019 May 22.

Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells is enhanced by an aragonite scaffold.

Author information

1
Department of Anatomy, Histology and Embryology, Faculty of Medicine, University of Debrecen, Nagyerdei krt 98, Debrecen, 4032, Hungary. Electronic address: matta.csaba@med.unideb.hu.
2
Department of Anatomy, Histology and Embryology, Faculty of Medicine, University of Debrecen, Nagyerdei krt 98, Debrecen, 4032, Hungary. Electronic address: somogyics@anat.med.unideb.hu.
3
Department of Biomedical Sciences, Humanitas University, Via Rita Levi Montalcini 4, Rozzano, Milan, 20090, Italy; Humanitas Clinical and Research Center, Via Alessandro Manzoni 56, Rozzano, Milan, 20089, Italy. Electronic address: Elizaveta.kon@humanitas.it.
4
Orthopaedic Research & Foot and Ankle Unit, Rabin Medical Center, 39 Jabotinski St, Petah Tikva, 49100, Israel. Electronic address: dror@cartiheal.com.
5
CartiHeal 2009 Ltd, Atir Yeda 17, Kfar Saba, 4464313, Israel. Electronic address: tova@cartiheal.com.
6
CartiHeal 2009 Ltd, Atir Yeda 17, Kfar Saba, 4464313, Israel. Electronic address: nir@cartiheal.com.
7
Orthopaedic and Trauma Department, Uzsoki Hospital, Uzsoki ut 29, Budapest, 1145, Hungary. Electronic address: bertaagnes@hotmail.com.
8
Orthopaedic and Trauma Department, Uzsoki Hospital, Uzsoki ut 29, Budapest, 1145, Hungary. Electronic address: hangody.laszlo@hangody.hu.
9
Regenerative Medicine and Cellular Pharmacology Research Laboratory, Department of Dermatology and Allergology, Faculty of Medicine, University of Szeged, Koranyi fasor 6, Szeged, 6720, Hungary. Electronic address: vereb.zoltan@med.u-szeged.hu.
10
Department of Anatomy, Histology and Embryology, Faculty of Medicine, University of Debrecen, Nagyerdei krt 98, Debrecen, 4032, Hungary. Electronic address: roza@anat.med.unideb.hu.

Abstract

Bone graft substitutes and bone void fillers are predominantly used to treat bone defects and bone fusion in orthopaedic surgery. Some aragonite-based scaffolds of coralline exoskeleton origin exhibit osteoconductive properties and are described as useful bone repair scaffolds. The purpose of this study was to evaluate the in vitro osteogenic potential of the bone phase of a novel aragonite-based bi-phasic osteochondral scaffold (Agili-C™, CartiHeal Ltd.) using adult human bone marrow-derived mesenchymal stem cells (MSCs). Analyses were performed at several time intervals: 3, 7, 14, 21, 28 and 42 days post-seeding. Osteogenic differentiation was assessed by morphological characterisation using light microscopy after Alizarin red and von Kossa staining, and scanning electron microscopy. The transcript levels of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), bone gamma-carboxyglutamate (BGLAP), osteonectin (SPARC) and osteopontin (SPP1) were determined by quantitative PCR. Proliferation was assessed by a thymidine incorporation assay and proliferating cell nuclear antigen (PCNA) immunocytochemistry. Our results demonstrate that the bone phase of the bi-phasic aragonite-based scaffold supports osteogenic differentiation and enhanced proliferation of bone marrow-derived MSCs at both the molecular and histological levels. The scaffold was colonized by differentiating MSCs, suggesting its suitability for incorporation into bone voids to accelerate bone healing, remodelling and regeneration. The mechanism of osteogenic differentiation involves scaffold surface modification with de novo production of calcium phosphate deposits, as revealed by energy dispersive spectroscopy (EDS) analyses. This novel coral-based scaffold may promote the rapid formation of high quality bone during the repair of osteochondral lesions.

KEYWORDS:

Agili-C; Aragonite-based implant; Bone marrow-derived mesenchymal stem cell; Coralline scaffold; Energy dispersive x-ray spectroscopy; Osteogenesis

PMID:
31152959
DOI:
10.1016/j.diff.2019.05.002
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