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Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

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Abstract

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.

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Acknowledgments

We thank Prof. Anthea Rowlerson (King’s College London, UK) for manuscript language revision. This work was supported by a grant from the University of Padova, Italy (prot. n. CPDA138242, PRAT 2013).

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Authors and Affiliations

  1. Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, Legnaro, 35020, Padova, Italy

    Chiara Gomiero, Giulia Bertolutti, Tiziana Martinello & Marco Patruno

  2. Global Stem Cell Technology-ANACURA Group, Noorwegenstraat 4, 9940, Evergem, Belgium

    Nathalie Van Bruaene, Sarah Y. Broeckx & Jan H. Spaas

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  1. Chiara Gomiero
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  2. Giulia Bertolutti
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  3. Tiziana Martinello
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  4. Nathalie Van Bruaene
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  5. Sarah Y. Broeckx
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  6. Marco Patruno
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  7. Jan H. Spaas
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Correspondence to Marco Patruno.

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JS and SB are employers of GST and inventors of several patents owned by the company.

None of the other authors of this paper has a financial or personal relationship with other people or organisations that could inappropriately influence or bias the content of the paper.

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Marco Patruno and Jan Spaas Contributed equally.

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Supplementary data associated with this article can be found, in the online version of the Vet Res Comm journal.

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Population dubling time of PB-MSC: the kinetics shows the means of four horses values and error bars indicate ± SD. (GIF 7 kb)

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Gomiero, C., Bertolutti, G., Martinello, T. et al. Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology. Vet Res Commun 40, 39–48 (2016). https://doi.org/10.1007/s11259-016-9652-y

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