2017 ISAKOS Biennial Congress ePoster #317

 

Platelet-Rich Plasma And Tgf-Beta Antagonist Act Sinergisticaly In Treatment Of Muscle Injuries

Robi Kelc, MD, PhD, Maribor SLOVENIA
Matjaz Vogrin, MD, PhD, Limbus SLOVENIA

Dep. of Orthopaedic Surgery, University Medical Centre Maribor, Slovenia, Maribor, , SLOVENIA

FDA Status Not Applicable

Summary

Antifibrotic therapy for muscle repair after injury

Abstract

Background

Regeneration of skeletal muscle after injury is limited by scar formation, slow healing time and a high recurrence rate. Concerns have been raised that platelet-rich plasma (PRP) derived TGF-ß could contribute to fibrotic remodelling after injury.
Materials & Methods
Using human myoblast cell culture we measured metabolic activity of the cells and their proliferation rate, TGF- ß and myostatin expression and myogenic commitment through MyoD/myogenin and desmin positive cells.

Results

The metabolic activity of cells determined by MTT assay was increased by more than 400% (p<0,001) after PRP treatment when compared with control while decorin lead to a 140% (p<0,005) increase. Decorin did not affect cell proliferation; however, PrP treatment lead to 5-fold (p<0,001) increase in viable cell count when compared with control.
The concentration of main inhibitory cytokine in muscle regeneration process TGF-ß was significantly down regulated by its inhibitor decorin (18%, p<0,001) but to even greater extent by PRP (30%, p<0,001) although it contains this cytokine in a concentrated quantity.
Immunocytochemistry qualitatively revealed the shift towards mature myocites in a myogenetic differentiation process. In PRP-treated group a 32,5% increase in myogenin positive cells was detected when compared with control and 24% decrease in MyoD positive cells. While decorin lead to only a slight further shift, it has shown to have a synergistic effect when added to PRP as further 11,5% increase in cells positive for myogenin together with a 44% decrease in those positive for MyoD were detected when treating the cultured cells with a combination of PRP and decorin. Further, the expression of desmin, an intermediate filament and a marker of late stage myogenic differentiation was up-regulated when treated with PRP alone in combination with decorin.

Discussion

Our results clearly showed that PRP treated myoblasts have a significant increase in the mitochondrial activity and in the cell proliferation rate as compared to those treated with decorin and control cells. At the same time lower expression of TGF-ß and MSTN was evident in PRP treated myoblasts, although PRP itself contains some amount of TGF-ß.
Despite concerns about promoting fibrosis developement, PRP inhibits the TGF-ß and MSTN activity to even greater extent than decorin. We can conclude that PRP can be a highly potential therapeutic agent for skeletal muscle regeneration and repair, especially if in combination with TGF-ß antagonist.
This work offers first scientific data on the molecular effects of PrP, not only in skeletal muscle, but any musculoskeletal tissue. It answers the major questions regarding its use in sports medicine and it also shows how its effects can be enhanced for the use in skeletal muscle therapy.