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Advances in allograft processing have opened new horizons forclinical adaptation of flexor tendon allografts as delivery scaffolds for antifibrotictherapeutics. Recombinant adeno-associated-virus (rAAV) gene delivery of the growth anddifferentiation factor 5 (GDF-5) has been previously associated with antifibroticeffects in a mouse model of flexor tendoplasty. In this study, we compared the effectsof loading freeze-dried allografts with different doses of GDF-5 protein or rAAV-Gdf5 onflexor tendon healing and adhesions. We first optimized the protein and viral loadingparameters using reverse transcription polymerase chain reaction (RT-PCR), enzyme-linkedimmunosorbent assay (ELISA), and in vivo bioluminescent imaging. We then reconstructedflexor digitorum longus (FDL) tendons of the mouse hindlimb with allografts loaded withlow and high doses of recombinant GDF-5 protein and rAAV-Gdf5 and evaluated jointflexion and biomechanical properties of the reconstructed tendon. In vitro optimizationstudies determined that both the loading time and concentration of the growth factor andviral vector had dose-dependent effects on their retention on the freeze-driedallograft. In vivo data suggest that protein and gene delivery of GDF-5 had equivalenteffects on improving joint flexion function, in the range of doses used. Within thedoses tested, the lower doses of GDF-5 had more potent effects on suppressing adhesionswithout adversely affecting the strength of the repair. These findings indicateequivalent antifibrotic effects of Gdf5 gene and protein delivery, but suggest thatlocalized delivery of this potent factor should also carefully consider the dosage usedto eliminate untoward effects, regardless of the delivery mode

Freeze-dried allograft-mediated gene or protein delivery of growth and differentiation factor 5 reduces reconstructed murine flexor tendon adhesions