Age-dependent changes in intervertebral disc cell mitochondria and bioenergetics
1 Department of Physical Medicine and Rehabilitation, University of Pittsburgh, 3471 5th Avenue, Pittsburgh, PA 15213, USA
2 University of Pittsburgh Medical Centre Enterprises, Pittsburgh, PA 15213, USA
3 Department of Orthopaedic Surgery, University of Pittsburgh, 200 Lothrop Street, E1641 Biomedical Science Tower, Pittsburgh, PA 15261, USA
4 Centre for Biologic Imaging, University of Pittsburgh, BST S224, 3500 Terrace St, Pittsburgh, PA 15261, USA
5 Institute on the Biology of Aging and Metabolism, Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 312 Church Street, Minneapolis, MN, USA
6 Department of Bioengineering, University of Pittsburgh, 3700 O’Hara Street, Pittsburgh, PA 15213, USA
7 Department of Pharmacology and Chemical Biology, University of Pittsburgh, 2.6e Hillman Cancer Centre, 5117 Centre Avenue, Pittsburgh, PA 15213, USA
Abstract
Robust cellular bioenergetics is vital in the energy-demanding process of maintaining matrix homeostasis in the intervertebral disc. Age-related decline in disc cellular bioenergetics is hypothesised to contribute to the matrix homeostatic perturbation observed in intervertebral disc degeneration. The present study aimed to measure how ageing impacted disc cell mitochondria and bioenergetics. Age-related changes measured included matrix content and cellularity in disc tissue, as well as matrix synthesis, cell proliferation and senescence markers in cell cultures derived from annulus fibrosus (AF) and nucleus pulposus (NP) isolated from the discs of young (6-9 months) and older (36-50 months) New Zealand White rabbits. Cellular bioenergetic parameters were measured using a Seahorse XFe96 Analyzer, in addition to quantitating mitochondrial morphological changes and membrane potential. Ageing reduced mitochondrial number and membrane potential in both cell types. Also, it significantly reduced glycolytic capacity, mitochondrial reserve capacity, maximum aerobic capacity and non-glucose-dependent respiration in NP. Moreover, NP cells exhibited age-related decline in matrix synthesis and reduced cellularity in older tissues. Despite a lack of changes in mitochondrial respiration with age, AF cells showed an increase in glycolysis and altered matrix production. While previous studies report age-related matrix degenerative changes in disc cells, the present study revealed, for the first time, that ageing affected mitochondrial number and function, particularly in NP cells. Consequently, age-related bioenergetic changes may contribute to the functional alterations in aged NP cells that underlie disc degeneration.
Graphical Abstract

Keywords
- Mitochondria
- bioenergetics
- ageing
- disc
- senescence
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