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Hypoxic preconditioning of myoblasts implanted in a tissue engineering chamber significantly increases local angiogenesis via upregulation of myoblast vascular endothelial growth factor-A expression and downregulation of miRNA-1, miRNA-206 and angiopoietin-1.
- Source :
-
Journal of tissue engineering and regenerative medicine [J Tissue Eng Regen Med] 2018 Jan; Vol. 12 (1), pp. e408-e421. Date of Electronic Publication: 2017 Jul 25. - Publication Year :
- 2018
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Abstract
- Vascularization is a major hurdle for growing three-dimensional tissue engineered constructs. This study investigated the mechanisms involved in hypoxic preconditioning of primary rat myoblasts in vitro and their influence on local angiogenesis postimplantation. Primary rat myoblast cultures were exposed to 90 min hypoxia at <1% oxygen followed by normoxia for 24 h. Real time (RT) polymerase chain reaction evaluation indicated that 90 min hypoxia resulted in significant downregulation of miR-1 and miR-206 (p < 0.05) and angiopoietin-1 (p < 0.05) with upregulation of vascular endothelial growth factor-A (VEGF-A; p < 0.05). The miR-1 and angiopoietin-1 responses remained significantly downregulated after a 24 h rest phase. In addition, direct inhibition of miR-206 in L6 myoblasts caused a significant increase in VEGF-A expression (p < 0.05), further establishing that changes in VEGF-A expression are influenced by miR-206. Of the myogenic genes examined, MyoD was significantly upregulated, only after 24 h rest (p < 0.05). Preconditioned or control myoblasts were implanted with Matrigelâ„¢ into isolated bilateral tissue engineering chambers incorporating a flow-through epigastric vascular pedicle in severe combined immunodeficiency mice and the chamber tissue harvested 14 days later. Chambers implanted with preconditioned myoblasts had a significantly increased percentage volume of blood vessels (p = 0.0325) compared with chambers implanted with control myoblasts. Hypoxic preconditioned myoblasts promote vascularization of constructs via VEGF upregulation and downregulation of angiopoietin-1, miR-1 and miR-206. The relatively simple strategy of hypoxic preconditioning of implanted cells - including non-stem cell types - has broad, future applications in tissue engineering of skeletal muscle and other tissues, as a technique to significantly increase implant site angiogenesis.<br /> (Copyright © 2017 John Wiley & Sons, Ltd.)
- Subjects :
- Animals
Biomarkers metabolism
Cell Hypoxia genetics
Cells, Cultured
Desmin metabolism
Male
Mice, SCID
MicroRNAs metabolism
Muscle Development genetics
Myoblasts metabolism
Platelet Endothelial Cell Adhesion Molecule-1 metabolism
RNA, Messenger genetics
RNA, Messenger metabolism
Rats, Sprague-Dawley
Tissue Scaffolds chemistry
Vascular Endothelial Growth Factor A metabolism
Down-Regulation genetics
Implants, Experimental
MicroRNAs genetics
Myoblasts pathology
Neovascularization, Physiologic
Tissue Engineering instrumentation
Up-Regulation genetics
Vascular Endothelial Growth Factor A genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1932-7005
- Volume :
- 12
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Journal of tissue engineering and regenerative medicine
- Publication Type :
- Academic Journal
- Accession number :
- 28477583
- Full Text :
- https://doi.org/10.1002/term.2440