Effect of fibrin on osteogenic differentiation and VEGF expression of bone marrow stromal cells in mineralised scaffolds: a three-dimensional analysis

Abstract

The aim of the present study was to test the hypothesis that a fibrin matrix enhances the osteogenic differentiation and expression of vascular endothelial growth factor (VEGF) by human bone marrow stromal cells (hBMSCs) seeded into mineralised scaffolds. Porous calcium carbonate scaffolds were droplet seeded with hBMSCs using a matrix containing 3 % fibrinogen and cultured for 3 weeks. Seeded scaffolds without the fibrin matrix served as controls. The scaffolds were evaluated, using undecalcified thick sections, for fluorescence staining for nuclei, osteocalcin (OC) and VEGF. The sections were systematically scanned using optical sectioning and three dimensional distributions of cells and positive staining indicating expression of OC and VEGF were reconstructed from the z-stacks. The fibrin matrix maintained a significantly higher level of cell numbers after 2 d and 1 week and delayed the onset of osteogenic differentiation while sustaining a significantly higher level of OC and VEGF expression after 2 and 3 weeks, starting from the periphery of the scaffolds. There was a decrease in cell density from the periphery to the centre of the scaffolds in both groups The percentage of cells expressing OC and VEGF was significantly different between the centre and the periphery of the scaffolds in the fibrin(+) group but not in the controls. It is concluded that the fibrin matrix used appears to be a useful adjunct for supporting and sustaining osteogenic and angiogenic activity of hBMSCs in tissue engineered constructs. This could help to improve their performance in a clinical setting.

Platelet-Rich Plasma Favors Proliferation of Canine Adipose-Derived Mesenchymal Stem Cells in Methacrylate-Endcapped Caprolactone Porous Scaffold Niches

“…This bioactive factor/scaffold has disadvantages when used as scaffolds, such as low rigidity that can produce shrinkage of scaffols and rapid degradation that would not permit proper bone regeneration. Therefore, other proposals suggest the use of fibrin as an adjunt element for sustaining osteogenic capacity of stem cells in mineralized scaffolds [26]. PRGF has also been considered as a natural scaffold [19] with advantages such as flexibility, non toxic, no immune reaction, complete resorption [27,28].…”

A review of fibrin and fibrin composites for bone tissue engineering

“…However, the maximum level of expression of OC and VEGF in hBMSCs did not increase through the use of fibrin. 111 Kneser et al 42 immobilized osteogenic cells in fibrin and seeded them into a processed bovine cancellous bone (PBCB) matrix. Later, scaffolds with and without cells were implanted into a rat calvarial defect.…”

The effects of scaffold architecture and fibrin gel addition on tendon cell phenotype

“…The gel degradation added variability to protein expression data, and highlighted the improved stability of the construct with a scaffold. This has been noted in the literature, where it has been demonstrated that scaffolds and fibrin gels stimulate the greatest healing response when used together [11,12,37].…”

“…With the addition of fibrin gels to scaffolds, tissue engineering outcomes, such as in vitro blood vessel formation and osteogenic differentiation, have been shown to improve [11,12]. While the advantage of adding fibrin to tissue engineering constructs is clear, the effects of combining fibrin gel and architectural cues has, to our knowledge, never been examined.…”