Bone Morphogenetic Protein-2, But Not Mesenchymal Stromal Cells, Exert Regenerative Effects on Canine and Human Nucleus Pulposus Cells.
Chronic back pain is related to intervertebral disc (IVD) degeneration and dogs are employed as animal models to develop growth factor- and cell-based regenerative treatments. In this respect, the differential effects of transforming growth factor beta-1 (TGF-β1) and bone morphogenetic protein-2 (BMP2) on canine and human chondrocyte-like cells (CLCs) derived from the nucleus pulposus of degenerated IVDs were studied. Human and canine CLCs were cultured in 3D microaggregates in basal culture medium supplemented with/without TGF-β1(10 ng/mL) or BMP2 (100 or 250 ng/mL). Both TGF-β1and BMP2 increased proliferation and glycosaminoglycan (GAG) deposition of human and canine CLCs. TGF-β1induced collagen type I deposition and fibrotic (re)differentiation, whereas BMP2 induced more collagen type II deposition. In dogs, TGF-β1induced Smad1 and Smad2 signaling, whereas in humans, it only tended to induce Smad2 signaling. BMP2 supplementation increased Smad1 signaling in both species. This altogether indicates that Smad1 signaling was associated with collagen type II production, whereas Smad2 signaling was associated with fibrotic CLC (re)differentiation. As a step toward preclinical translation, treatment with BMP2 alone and combined with mesenchymal stromal cells (MSCs) was further investigated. Canine male CLCs were seeded in albumin-based hydrogels with/without female bone marrow-derived MSCs (50:50) in basal or 250 ng/mL BMP2-supplemented culture medium. Although the results indicate that a sufficient amount of MSCs survived the culture period, total GAG production was not increased and GAG production per cell was even decreased by the addition of MSCs, implying that MSCs did not exert additive regenerative effects on the CLCs.
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Frances C
FC
Bach
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Alberto
A
Miranda-Bedate
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Ferdi W M
FW
van Heel
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Frank M
FM
Riemers
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Margot C M E
MC
Müller
2 Department of Orthopaedics, University Medical Center Utrecht , Utrecht, The Netherlands .
Laura B
LB
Creemers
2 Department of Orthopaedics, University Medical Center Utrecht , Utrecht, The Netherlands .|3 Department of Biomedical Engineering, Eindhoven University of Technology , Eindhoven, The Netherlands .
Keita
K
Ito
4 TETEC AG , Reutlingen, Germany .
Karin
K
Benz
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Björn P
BP
Meij
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University , Utrecht, The Netherlands .
Marianna A
MA
Tryfonidou
D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000493:pharmacokinetics*; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D020219:Chondrogenesis / Q000187:drug effects*; D004285:Dogs; D005260:Female; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology / Q000502:physiology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D012038:Regeneration / Q000187:drug effects*; D051898:Smad1 Protein / Q000378:metabolism; D051899:Smad2 Protein / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; C497399:SMAD1 protein, human; C497782:SMAD2 protein, human; D051898:Smad1 Protein; D051899:Smad2 Protein; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1
bone morphogenetic protein-2; intervertebral disc; mesenchymal stromal cells; transforming growth factor beta-1