Alfredo Franco-Obregón1,2, Elena Cambria3, Helen Greutert3, Timon Wernas4, Wolfgang Hitzl5,6, Marcel Egli4, Miho Sekiguchi7, Norbert Boos8, Oliver Hausmann9, Stephen J Ferguson3, Hiroshi Kobayashi3,7, Karin Wuertz-Kozak10,11,12,13. 1. Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore. 2. BioIonic Currents Electromagnetic Pulsing Systems Laboratory, BICEPS, National University of Singapore, Singapore, Singapore. 3. Institute for Biomechanics, D-HEST, ETH Zurich, Hönggerbergring 64, 8093, Zurich, Switzerland. 4. School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, Lucerne, Switzerland. 5. Research Office (Biostatistics), Paracelsus Private Medical University, Salzburg, Austria. 6. Department of Ophthalmology and Optometry, Paracelsus Medical University Salzburg, Salzburg, Austria. 7. Department of Orthopaedic Surgery, School of Medicine, Fukushima Medical University, Fukushima, Japan. 8. Prodorso Center for Spinal Medicine, Zurich, Switzerland. 9. Neuro- and Spine Center, Hirslanden Klinik St. Anna, Lucerne, Switzerland. 10. Institute for Biomechanics, D-HEST, ETH Zurich, Hönggerbergring 64, 8093, Zurich, Switzerland. kwuertz@ethz.ch. 11. Spine Center, Schön Klinik München Harlaching, 81547, Munich, Germany. kwuertz@ethz.ch. 12. Academic Teaching Hospital and Spine Research Institute, Paracelsus Private Medical University, Salzburg, Austria. kwuertz@ethz.ch. 13. Department of Health Science, University of Potsdam, Potsdam, Germany. kwuertz@ethz.ch.
Abstract
PURPOSE: Prolonged bed rest and microgravity in space cause intervertebral disc (IVD) degeneration. However, the underlying molecular mechanisms are not completely understood. Transient receptor potential canonical (TRPC) channels are implicated in mechanosensing of several tissues, but are poorly explored in IVDs. METHODS: Primary human IVD cells from surgical biopsies composed of both annulus fibrosus and nucleus pulposus (passage 1-2) were exposed to simulated microgravity and to the TRPC channel inhibitor SKF-96365 (SKF) for up to 5 days. Proliferative capacity, cell cycle distribution, senescence and TRPC channel expression were analyzed. RESULTS: Both simulated microgravity and TRPC channel antagonism reduced the proliferative capacity of IVD cells and induced senescence. While significant changes in cell cycle distributions (reduction in G1 and accumulation in G2/M) were observed upon SKF treatment, the effect was small upon 3 days of simulated microgravity. Finally, downregulation of TRPC6 was shown under simulated microgravity. CONCLUSIONS: Simulated microgravity and TRPC channel inhibition both led to reduced proliferation and increased senescence. Furthermore, simulated microgravity reduced TRPC6 expression. IVD cell senescence and mechanotransduction may hence potentially be regulated by TRPC6 expression. This study thus reveals promising targets for future studies. These slides can be retrieved under Electronic Supplementary Material.
PURPOSE: Prolonged bed rest and microgravity in space cause intervertebral disc (IVD) degeneration. However, the underlying molecular mechanisms are not completely understood. Transient receptor potential canonical (TRPC) channels are implicated in mechanosensing of several tissues, but are poorly explored in IVDs. METHODS: Primary humanIVD cells from surgical biopsies composed of both annulus fibrosus and nucleus pulposus (passage 1-2) were exposed to simulated microgravity and to the TRPC channel inhibitor SKF-96365 (SKF) for up to 5 days. Proliferative capacity, cell cycle distribution, senescence and TRPC channel expression were analyzed. RESULTS: Both simulated microgravity and TRPC channel antagonism reduced the proliferative capacity of IVD cells and induced senescence. While significant changes in cell cycle distributions (reduction in G1 and accumulation in G2/M) were observed upon SKF treatment, the effect was small upon 3 days of simulated microgravity. Finally, downregulation of TRPC6 was shown under simulated microgravity. CONCLUSIONS: Simulated microgravity and TRPC channel inhibition both led to reduced proliferation and increased senescence. Furthermore, simulated microgravity reduced TRPC6 expression. IVD cell senescence and mechanotransduction may hence potentially be regulated by TRPC6 expression. This study thus reveals promising targets for future studies. These slides can be retrieved under Electronic Supplementary Material.
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