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

Olivera Evrova,Maurizio Calcagni,Johanna Buschmann,Damian Kellenberger,Viola Vogel

doi:10.3390/ijms21020458

Olivera Evrova, Maurizio Calcagni + Show 3 more

Open Access

https://doi.org/10.3390/ijms21020458

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Abstract

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.

Highlights

Different cell-based therapies, including biological and decellularized tissues [1,2], cell-seeded grafts from natural and synthetic biomaterials [3,4] or stem cell-based approaches [5] have been explored for the treatment of tendinopathies and tendon injuries
Ascorbic acid and platelet-derived growth factor-BB (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, matrix metalloproteases (MMPs)-2, and TIMP-2 was observed with ascorbic acid, while these markers were downregulated by PDGF-BB supplementation
In order to assess the effect of both biomolecules on cell culture proliferation, tenocytes cultured in both serum+ and serum-free medium were supplemented with different concentrations of PDGF-BB and ascorbic acid (AA)

Summary

Introduction

Different cell-based therapies, including biological and decellularized tissues [1,2], cell-seeded grafts from natural and synthetic biomaterials [3,4] or stem cell-based approaches [5] have been explored for the treatment of tendinopathies and tendon injuries. While the application of stem cells and cues for tenogenic differentiation have been reported extensively [16,17,18,19], the use of tenocytes to treat tendon diseases is currently not investigated widely. This may be due to two facts: first, tenocytes occur in relatively low numbers in tendon tissue and have to be expanded in vitro prior to application; second, tenocytes may undergo a phenotypic drift in culture, losing typical tendon markers during passaging, and providing a limited range of low passages convenient for cellular therapy [20]. Tenocytes present an attractive cell source for tendon tissue engineering applications

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