Different combinations of growth factors for the tenogenic differentiation of bone marrow mesenchymal stem cells in monolayer culture and in fibrin-based three-dimensional constructs.
Tendon injuries are severe burdens in clinics. The poor tendon healing is related to an ineffective response of resident cells and inadequate vascularization. Thanks to the high proliferation and multi-lineage differentiation capability, bone marrow-derived mesenchymal stem cells (BMSCs) are a promising cell source to support the tendon repair. To date, the association of various growth factors to induce the in vitro tenogenic differentiation of multipotent progenitor cells is poorly investigated. This study aimed to investigate the tenogenic differentiation of rabbit BMSCs by testing the combination of bone morphogenetic proteins (BMP-12 and 14) with transforming growth factor beta (TGF-β) and vascular endothelial growth factor (VEGF) both in 2D and 3D cultures within fibrin-based constructs. After 7 and 14 days, the tenogenic differentiation was assessed by analyzing cell metabolism and collagen content, the gene expression of tenogenic markers and the histological cell distribution and collagen deposition within 3D constructs. Our results demonstrated that the association of BMP-14 with TGF-β3 and VEGF enhanced the BMSC tenogenic differentiation both in 2D and 3D cultures. This study supports the use of fibrin as hydrogel-based matrix to generate spheroids loaded with tenogenic differentiated BMSCs that could be used to treat tendon lesions in the future.
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161 Milan, Italy.
Marta
M
Bottagisio
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161 Milan, Italy.
Silvia
S
Lopa
Department of Veterinary Medicine (DiMeVet), University of Milan, Via Celoria 10, 20133 Milan, Italy.
Valentina
V
Granata
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161 Milan, Italy.
Giuseppe
G
Talò
Department of Veterinary Medicine (DiMeVet), University of Milan, Via Celoria 10, 20133 Milan, Italy.
Chiara
C
Bazzocchi
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161 Milan, Italy; Regenerative Medicine Technologies Lab, Ente Ospedaliero Cantonale (EOC), Lugano, Switzerland; Swiss Institute of Regenerative Medicine (SIRM), Lugano, Switzerland; Fondazione Cardiocentro Ticino, Lugano, Switzerland.
Matteo
M
Moretti
Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Via R. Galeazzi 4, 20161 Milan, Italy. Electronic address: arianna.lovati@grupposandonato.it.
Arianna Barbara
AB
Lovati
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002454:Cell Differentiation; D002478:Cells, Cultured; D005337:Fibrin / Q000494:pharmacology; D008233:Lymphotoxin-alpha / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D011817:Rabbits; D013710:Tendons / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D019485:Bone Morphogenetic Proteins; D008233:Lymphotoxin-alpha; D042461:Vascular Endothelial Growth Factor A; C086559:growth differentiation factor 7; D005337:Fibrin
Bone marrow mesenchymal stem cells; Fibrin glue; Growth factors; Rabbit; Tenogenic differentiation