title: Regulation of hypoxia-induced cell death in human tenocytes.
abstract: Degenerate shoulder tendons display evidence of hypoxia. However tendons are relatively avascular and not considered to have high oxygen requirements and the vulnerability of tendon cells to hypoxia is unclear. Cultured human tenocytes were exposed to hypoxia and the cellular response detected using QPCR, Western blotting, viability, and ELISA assays. We find that tenocytes respond to hypoxia in vitro by activating classical HIF-1α-driven pathways. Total hypoxia caused significant tenocyte apoptosis. Transcription factors typically involved in hypoxic response, HIF-1α and FOXO3A, were upregulated. Hypoxia caused sustained upregulation of several proapoptotic proteins known to mediate hypoxia-induced apoptosis, such as Bnip3 and Nix, but others were unchanged although they were reportedly hypoxia-sensitive in other cell types. Antiapoptotic proteins Bcl2 and Bcl-xL were unchanged by hypoxia. Normal human tenocytes expressed all isoforms of the hypoxia-induced vascular growth factor VEGF except VEGF-D. Hypoxia markedly upregulated VEGF-A mRNA, followed by increased VEGF protein secretion. However treatment with VEGF did not improve tenocyte survival. As a protective strategy for tenocytes at risk of hypoxic death we added prosurvival growth factors insulin or platelet rich plasma (PRP). Both agents strongly protected tenocytes from hypoxia-induced death over 48 h, suggesting possible efficacy in the acute postrupture tendon or integrating graft.
authors.affiliation: Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford OX3 7LD, UK ; Department of Periodontology, Guanghua School of Stomatology and Hospital of Stomatology, Sun Yat-sen University, 56 Lingyuan Road West, Guangzhou 510055, China.
authors.forename: Min
authors.initials: M
authors.lastname: Liang
authors.forename: Hannah R
authors.initials: HR
authors.lastname: Cornell
authors.forename: Nasim
authors.initials: N
authors.lastname: Zargar Baboldashti
authors.forename: Mark S
authors.initials: MS
authors.lastname: Thompson
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Carr
authors.forename: Philippa A
authors.initials: PA
authors.lastname: Hulley