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The Incorporation of Marine Coral Microparticles into Collagen-Based Scaffolds Promotes Osteogenesis of Human Mesenchymal Stromal Cells via Calcium Ion Signalling.

ABSTRACT: Composite biomaterial scaffolds consisting of natural polymers and bioceramics may offer an alternative to autologous grafts for applications such as bone repair. Herein, we sought to investigate the possibility of incorporating marine coral microparticles into a collagen-based scaffold, a process which we hypothesised would enhance the mechanical properties of the scaffold as well its capacity to promote osteogenesis of human mesenchymal stromal cells. Cryomilling and sieving were utilised to achieve coral microparticles of mean diameters 14 µm and 64 µm which were separately incorporated into collagen-based slurries and freeze-dried to form porous scaffolds. X-ray diffraction and Fourier transform infrared spectroscopy determined the coral microparticles to be comprised of calcium carbonate whereas collagen/coral composite scaffolds were shown to have a crystalline calcium ethanoate structure. Crosslinked collagen/coral scaffolds demonstrated enhanced compressive properties when compared to collagen only scaffolds and also promoted more robust osteogenic differentiation of mesenchymal stromal cells, as indicated by increased expression of bone morphogenetic protein 2 at the gene level, and enhanced alkaline phosphatase activity and calcium accumulation at the protein level. Only subtle differences were observed when comparing the effect of coral microparticles of different sizes, with improved osteogenesis occurring as a result of calcium ion signalling delivered from collagen/coral composite scaffolds. These scaffolds, fabricated from entirely natural sources, therefore show promise as novel biomaterials for tissue engineering applications such as bone regeneration.

SUBMITTER: Sheehy EJ

PROVIDER: S-EPMC7073845 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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The Incorporation of Marine Coral Microparticles into Collagen-Based Scaffolds Promotes Osteogenesis of Human Mesenchymal Stromal Cells via Calcium Ion Signalling.

Sheehy Eamon J EJ Lemoine Mark M Clarke Declan D Gonzalez Vazquez Arlyng A O'Brien Fergal J FJ

Marine drugs 20200123 2

Composite biomaterial scaffolds consisting of natural polymers and bioceramics may offer an alternative to autologous grafts for applications such as bone repair. Herein, we sought to investigate the possibility of incorporating marine coral microparticles into a collagen-based scaffold, a process which we hypothesised would enhance the mechanical properties of the scaffold as well its capacity to promote osteogenesis of human mesenchymal stromal cells. Cryomilling and sieving were utilised to a ...[more]

PMID: 31979233

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Project description:Osteogenic cell signaling pathway disruption varies among bone diseases. This investigation was designed to identify adipose-derived multipotent stromal cell (ASC) and bone graft scaffold combinations for local, targeted restoration of gene expression and extracellular matrix (ECM) deposition. Human ASC osteogenesis on bone graft materials was quantified following culture in stromal (S), osteogenic (O), or osteogenic for 48 h followed by stromal medium (OS) to test the two-part hypothesis: (1) identical ASC isolates on distinct bone graft scaffolds demonstrate unique viability, differentiation, ECM production, and gene expression in the same culture conditions; (2) identical ASC-bone graft scaffold combinations have different cell viability, differentiation, ECM production, and gene expression when cultured in S, O, or OS medium. Three commercially available bone graft scaffold materials, type I bovine collagen (C), hydroxyapatite + β-tricalcium phosphate + type I bovine collagen (HT), and β-tricalcium phosphate + type I bovine collagen (CT) were evaluated. Passage 3 ASCs were loaded onto scaffold blocks with a spinner flask bioreactor, and constructs were cultured up to 28 days. Cell viability, gene expression (alkaline phosphatase [ALPL], osteoprotegerin [TNFRSF11B], osteocalcin [BGLAP], cannabinoid receptors type I [CNR1] and II [CNR2], receptor activator of nuclear factor kappa β ligand [TNFSF11]), as well as ECM DNA, collagen, sulfated glycosaminoglycan, and protein content were quantified. Matrix organization was evaluated with scanning electron microscopy. Effects of scaffold, medium, or culture duration on cell viability were minimal. Significantly higher initial ALPL expression decreased with time, while BGLAP expression increased in HT constructs in O medium, and the constructs had the most abundant ECM components and ultrastructural organization. There was a similar, although delayed, pattern of gene expression and greater ECM collagen with less organization in C constructs in O medium. Higher CNR1 expression in C versus higher TNFRSF11B/TNFSF11 expression in HT constructs throughout the study support stimulation of unique osteogenic signaling pathways by identical cell isolates. These results suggest that bone scaffold composition may be used to selectively target specific osteogenic cell signaling pathways in ASC constructs to stimulate ECM deposition based on therapeutic needs.

Dataset Information

The Incorporation of Marine Coral Microparticles into Collagen-Based Scaffolds Promotes Osteogenesis of Human Mesenchymal Stromal Cells via Calcium Ion Signalling.

Publications

The Incorporation of Marine Coral Microparticles into Collagen-Based Scaffolds Promotes Osteogenesis of Human Mesenchymal Stromal Cells via Calcium Ion Signalling.

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