Abstract
A discrimination of the highly specialised annulus fibrosus (AF) and nucleus pulposus (NP) cells in the mature human intervertebral disc (IVD) is thus far still not possible in a reliable way. The aim of this study was to identify molecular markers that distinguish AF and NP cells in human disc tissue using microarray analysis as a screening tool. AF and NP samples were obtained from 28 cervical discs. First, all samples underwent quality sorting using two novel scoring systems for small-sized disc tissue samples including macroscopic, haptic and histological evaluation. Subsequently, samples with clear disc characteristics of either AF or NP that were free from impurities of foreign tissue (IVD score) and with low signs of disc degeneration on cellular level (DD score) were selected for GeneChip analysis (HGU1332P). The 11 AF and 9 NP samples showed distinctly different genome-wide transcriptomes. The majority of differentially expressed genes (DEGs) could be specifically assigned to the AF, whereas no DEG was exclusively expressed in the NP. Nevertheless, we identified 11 novel marker genes that clearly distinguished AF and NP, as confirmed by quantitative gene expression analysis. The novel established scoring systems and molecular markers showed the identity of AF and NP in disc starting material and are thus of great importance in the quality assurance of cell-based therapeutics in regenerative treatment of disc degeneration.
Highlights
Back pain is a major problem in today’s society
All samples included in this study first underwent a novel quality assessment system developed by us: the intervertebral disc (IVD) score and the disc degeneration (DD) score
The specific gene expression of the 5 annulus fibrosus (AF) and 6 nucleus pulposus (NP) markers proved the correct classification of AF and NP tissue samples
Summary
Back pain is a major problem in today’s society. One cause is the degeneration of the intervertebral disc, which is a naturally occurring phenomenon in aging humans. The disc is a cartilage-like tissue between the vertebrae that enables mobility of the spinal unit due to its specific structural composition and matrix organisation. The cells in the disc are highly specialised cells that produce and thereby maintain the disc matrix. The synthesis and degradation of extracellular matrix (ECM) molecules becomes imbalanced with time. This results in a loss of disc matrix and a decrease in cell number [1,2], causing the disc degenerative disease (DDD). Recent developments have used regenerative therapies to treat
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