Supporting Cell-Based Tendon Therapy: Effect of PDGF-BB and Ascorbic Acid on Rabbit Achilles Tenocytes in Vitro.

Cell-based tendon therapies with tenocytes as a cell source need effective tenocyte in vitro expansion before application for tendinopathies and tendon injuries. Supplementation of tenocyte culture with biomolecules that can boost proliferation and matrix synthesis is one viable option for supporting cell expansion. In this in vitro study, the impacts of ascorbic acid or PDGF-BB supplementation on rabbit Achilles tenocyte culture were studied. Namely, cell proliferation, changes in gene expression of several ECM and tendon markers (collagen I, collagen III, fibronectin, aggrecan, biglycan, decorin, ki67, tenascin-C, tenomodulin, Mohawk, α-SMA, MMP-2, MMP-9, TIMP1, and TIMP2) and ECM deposition (collagen I and fibronectin) were assessed. Ascorbic acid and PDGF-BB enhanced tenocyte proliferation, while ascorbic acid significantly accelerated the deposition of collagen I. Both biomolecules led to different changes in the gene expression profile of the cultured tenocytes, where upregulation of collagen I, Mohawk, decorin, MMP-2, and TIMP-2 was observed with ascorbic acid, while these markers were downregulated by PDGF-BB supplementation. Vice versa, there was an upregulation of fibronectin, biglycan and tenascin-C by PDGF-BB supplementation, while ascorbic acid led to a downregulation of these markers. However, both biomolecules are promising candidates for improving and accelerating the in vitro expansion of tenocytes, which is vital for various tendon tissue engineering approaches or cell-based tendon therapy.

Impact of UV sterilization and short term storage on the in vitro release kinetics and bioactivity of biomolecules from electrospun scaffolds.

To effectively translate bioactive scaffolds into a preclinical setting, proper sterilization techniques and storage conditions need to be carefully considered, as the chosen sterilization technique and storage condition might affect the structural and mechanical properties of the scaffolds, as well as the bioactivity and release kinetics of the incorporated biomolecules. Since rarely tested or quantified, we show here in a proof-of-concept study how these parameters are affected by UV sterilization and one week storage at different temperatures using bioactive electrospun DegraPol scaffolds that were specifically designed for application in the field of tendon rupture repair. Even though UV sterilization and the different storage conditions did not impact the morphology or the physicochemical properties of the bioactive scaffolds, UV sterilization caused significant attenuation of the growth factor release kinetics, here platelet derived growth factor (PDGF-BB) release (by approx. 85%) and slight decrease in ascorbic acid release (by approx. 20%). In contrast, 4 °C and -20 °C storage did not have a major effect on the release kinetics of PDGF-BB, while storage at room temperature caused increase in PDGF-BB released. All storage conditions had little effect on ascorbic acid release. Equally important, neither UV sterilization nor storage affected the bioactivity of the released PDGF-BB, suggesting stability of the bioactive scaffolds for at least one week and showing potential for bioactive DegraPol scaffolds to be translated into an off-the-shelf available product. These parameters are expected to be scaffold and protein-dependent.