OBJECTIVE: We hypothesize that intervertebral disc (IVD) cells from distinct region respond differently to oxygen environment, and that IVD cells from patients with disc degeneration can benefit from hypoxia condition. Therefore, we aimed to determine the transcriptional response and extracellular matrix (ECM) production of nucleus pulposus (NP) and annulus fibrosus (AF) cells to different oxygen tension. METHOD: Human NP and AF from degenerated IVD were seeded in 3D scaffolds and subjected to varying oxygen tension (2% and 20%) for 3 weeks. Changes in ECM were evaluated using quantitative real-time reverse transcriptase polymerase chain reaction, histological and immunohistological analyses. RESULTS: Hypoxia significantly enhances NP cells phenotype, which resulted in greater production of sulfated glycosaminoglycan (GAG) and collagen type II within the constructs and the cells expressed higher levels of genes encoding NP ECM. A significantly stronger fluorescent signal for hypoxia-inducible factor (HIF-1α) as also found in the NP cells under the hypoxic than normoxic condition. However, there was little effect of hypoxia on the AF cells. CONCLUSIONS: The NP and AF cells respond differently to hypoxia condition on the 3D scaffold, and hypoxia could enhance NP phenotype. When used in concert with appropriate scaffold material, human NP cells from degenerated disc could be regenerated for tissue engineering application.
OBJECTIVE: We hypothesize that intervertebral disc (IVD) cells from distinct region respond differently to oxygen environment, and that IVD cells from patients with disc degeneration can benefit from hypoxia condition. Therefore, we aimed to determine the transcriptional response and extracellular matrix (ECM) production of nucleus pulposus (NP) and annulus fibrosus (AF) cells to different oxygen tension. METHOD:Human NP and AF from degenerated IVD were seeded in 3D scaffolds and subjected to varying oxygen tension (2% and 20%) for 3 weeks. Changes in ECM were evaluated using quantitative real-time reverse transcriptase polymerase chain reaction, histological and immunohistological analyses. RESULTS:Hypoxia significantly enhances NP cells phenotype, which resulted in greater production of sulfated glycosaminoglycan (GAG) and collagen type II within the constructs and the cells expressed higher levels of genes encoding NP ECM. A significantly stronger fluorescent signal for hypoxia-inducible factor (HIF-1α) as also found in the NP cells under the hypoxic than normoxic condition. However, there was little effect of hypoxia on the AF cells. CONCLUSIONS: The NP and AF cells respond differently to hypoxia condition on the 3D scaffold, and hypoxia could enhance NP phenotype. When used in concert with appropriate scaffold material, human NP cells from degenerated disc could be regenerated for tissue engineering application.
Authors: Thomas R Christiani; Katelynn Toomer; Joseph Sheehan; Angelika Nitzl; Amanda Branda; Elizabeth England; Pamela Graney; Cristina Iftode; Andrea J Vernengo Journal: J Vis Exp Date: 2016-10-26 Impact factor: 1.355
Authors: Deborah J Gorth; Katherine E Lothstein; Joseph A Chiaro; Megan J Farrell; George R Dodge; Dawn M Elliott; Neil R Malhotra; Robert L Mauck; Lachlan J Smith Journal: J Orthop Res Date: 2015-03-08 Impact factor: 3.494
Authors: Frances C Bach; Lisanne T Laagland; Michael P Grant; Laura B Creemers; Keita Ito; Björn P Meij; Fackson Mwale; Marianna A Tryfonidou Journal: PLoS One Date: 2017-11-08 Impact factor: 3.240