Alterations of bovine nucleus pulposus cells with aging

Author:

Molinos Maria123,Fiordalisi Morena F.123,Caldeira Joana12,Almeida Catarina R.124,Barbosa Mário A.123,Gonçalves Raquel M.123ORCID

Affiliation:

1. i3S – Instituto de Investigação e Inovação em Saúde Universidade do Porto Porto Portugal

2. INEB – Instituto de Engenharia Biomédica Universidade do Porto Porto Portugal

3. ICBAS – Instituto de Ciências Biomédicas Abel Salazar Universidade do Porto Porto Portugal

4. iBiMED – Institute of Biomedicine, Department of Medical Sciences University of Aveiro Aveiro Portugal

Abstract

AbstractAging is one of the major etiological factors driving intervertebral disc (IVD) degeneration, the main cause of low back pain. The nucleus pulposus (NP) includes a heterogeneous cell population, which is still poorly characterized. Here, we aimed to uncover main alterations in NP cells with aging. For that, bovine coccygeal discs from young (12 months) and old (10–16 years old) animals were dissected and primary NP cells were isolated. Gene expression and proteomics of fresh NP cells were performed. NP cells were labelled with propidium iodide and analysed by flow cytometry for the expression of CD29, CD44, CD45, CD146, GD2, Tie2, CD34 and Stro‐1. Morphological cell features were also dissected by imaging flow cytometry. Elder NP cells (up‐regulated bIL‐6 and bMMP1 gene expression) presented lower percentages of CD29+, CD44+, CD45+ and Tie2+ cells compared with young NP cells (upregulated bIL‐8, bCOL2A1 and bACAN gene expression), while GD2, CD146, Stro‐1 and CD34 expression were maintained with age. NP cellulome showed an upregulation of proteins related to endoplasmic reticulum (ER) and melanosome independently of age, whereas proteins upregulated in elder NP cells were also associated with glycosylation and disulfide bonds. Flow cytometry analysis of NP cells disclosed the existence of 4 subpopulations with distinct auto‐fluorescence and size with different dynamics along aging. Regarding cell morphology, aging increases NP cell area, diameter and vesicles. These results contribute to a better understanding of NP cells aging and highlighting potential anti‐aging targets that can help to mitigate age‐related disc disease.

Funder

Fundação para a Ciência e a Tecnologia

EUROSPINE

Publisher

Wiley

Subject

Cell Biology,Aging

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