Elsevier

Differentiation

Volume 107, May–June 2019, Pages 24-34
Differentiation

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

https://doi.org/10.1016/j.diff.2019.05.002Get rights and content
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open access

Highlights

  • We evaluated the osteogenic potential of an aragonite-based scaffold using BM-MSCs.

  • The scaffold supports osteogenic differentiation and enhances proliferation of MSCs.

  • Osteogenesis involves scaffold surface modification with calcium phosphate deposits.

  • The Agili-C scaffold seems to be suitable for incorporation into bone voids.

  • The coral-based scaffold may promote rapid formation of high quality bone.

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

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

Abbreviations

ALP
alkaline phosphatase
AR
Alizarin red
BGLAP
bone gamma-carboxyglutamate
BM-MSC
bone marrow-derived mesenchymal stem cell
CC
control culture
DCP
dicalcium phosphate anhydrous
ECM
extracellular matrix
EDS
energy dispersive X-ray spectroscopy
GAPDH
glyceraldehyde 3-phosphate dehydrogenase
H&E
haematoxylin and eosin
HMDS
hexamethyldisilazane
hMSC
human mesenchymal stem cell
HPRT1
hypoxanthine phosphoribosyl-transferase 1
MSCGM
Mesenchymal Stem Cells Growth Medium
PBS
phosphate buffered saline
PCNA
proliferating cell nuclear antigen
PPIA
peptidyl-prolyl isomerase-A (cyclophilin-A)
RUNX2
runt-related transcription factor 2
SEC
scaffold-exposed culture
SEM
scanning electron microscopy
SPARC
secreted protein acidic rich in cysteine (osteonectin)
SPP1
osteopontin
TGF
transforming growth factor

Cited by (0)

1

These two authors equally contributed to the work.