Can Extracorporeal Shockwave Promote Osteogenesis of Equine Bone Marrow-Derived Mesenchymal Stem Cells In Vitro?

The research article explores the effects of extracorporeal shockwave therapy (ESW) on the osteogenic property of equine bone marrow-derived mesenchymal stem cells (BMDMSCs). The results show that while ESW causes an initial increase in alkaline phosphatase (ALPL) production – a marker of osteogenic differentiation – it does not produce a lasting osteogenic effect.

Research Methodology

• The study began with the extraction of BMDMSCs from six horses which went through the process of culture expansion to passage 3, dissociation and placement in conical tubes.
• The treatment cells, termed “shocked” in the text, were exposed to 500 pulses of 0.16 mJ/mm energy.
• These cells were then reseeded and cultivated in either growth medium or osteogenic medium (a substrate conducible to bone growth).
• The researchers observed cellular proliferation and the cells’ trilineage potential (which examines its capacity to differentiate into three different cell types: adipocytes, osteoblasts, and chondrocytes).
• Cellular morphology was scored and cells were collected at five intervals: 1, 3, 7, 14, and 21 days. These cells were then analyzed for gene expression of osteogenic markers through rtPCR (reverse transcription Polymerase Chain Reaction) gene expression profiling.
• The researchers also studied media supernatants, the liquid part of the medium left after cells and cell debris have been removed, for secretion of Bone Morphogenetic Protein 2 (BMP-2), Vascular Endothelial Growth Factor (VEGF), Transforming Growth Factor Beta (TGFβ), and Prostaglandin Endoperoxidase synthase (PGE).
• Finally, the cellular lysates, or dissolved cell contents, were evaluated for ALPL production.

Research Findings

• There was no significant difference in the proliferative ability of the shocked cells versus the unshocked cells in either the growth or osteogenic mediums.
• In the shocked cells maintained in osteogenic medium, ALPL production was significantly higher than in the unshocked cells in the same medium at the third day of observation.
• ESW exposure led to a decrease in VEGF and TGFβ production at day 3, regardless of the type of medium used.
• No significant changes in gene expression were observed as confirmed by rtPCR analysis.

Conclusion

The study concluded that the exposure of BMDMSCs to ESW does not lead to any negative effects. While an initial significant increase in ALPL was detected, the treatment did not consistently boost the osteogenic effect, as observed in the cell expansion stage of the experiment, highlighting that ESW should not be relied upon to enhance the osteogenic potential of BMDMSCs in the treatment of bone fractures.