Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro.

This research aims to unearth the potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) to differentiate into bone cells within coralline hydroxyapatite scaffolds under laboratory conditions. The findings indicate that these cells can sustain their viability and demonstrate osteogenic potential when cultured in vitro in these scaffolds, which could be beneficial for bone-gap repair in horses.

Objective and Methodology

The objective of this study was to explore the potential of horse umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) for osteogenic differentiation within coralline hydroxyapatite scaffolds under lab healing conditions. This was carried out by:

• Culturing the scaffolds seeded with equine CB-MSC in cell expansion culture medium (control) or osteogenic induction medium (treatment).
• Testing cell viability and distribution using an MTT cell viability assay and DAPI nuclear fluorescence staining, respectively.
• Evaluating osteogenic differentiation after 10 days through reverse transcription polymerase chain reaction, alkaline phosphatase activity, and secreted osteocalcin concentration.
• Assessing cell morphology and matrix deposition using scanning electron microscopy (SEM) after 14 days in culture.

Results

The findings established that:

• The cells remained viable and evenly distributed throughout the scaffold.
• Osteogenic differentiation was successful within the scaffolds as indicated by increased expression of osteogenic markers like Runx2, osteopontin, osteonectin, collagen IA.
• There was an increase in alkaline phosphatase activity and osteocalcin protein secretion.
• Bone-like matrix presence was noticed in the scaffold pores upon SEM evaluation.

Conclusion

Following this study, it was concluded that equine CB-MSC can sustain their viability and showcase a promising potential for osteogenic differentiation in coralline hydroxyapatite scaffolds when cultured in a laboratory setting. Given these results, further investigation of Equine CB-MSC scaffold constructs is encouraged, particularly for its potential use as biologically active fillers in enhancing bone-gap repair in horses.