Journal
SPINE JOURNAL
Volume 14, Issue 3, Pages 455-468Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.spinee.2013.10.018
Keywords
Intervertebral disc cells; Mesenchymal stem cells; Chondrogenesis; Aromatase; Testosterone and three-dimensional cultures; Sex hormones
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Funding
- Swiss Paraplegic Foundation
- Swiss National Foundation [CR3I3_140717/1]
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BACKGROUND CONTEXT: Testosterone (T) is a hormone and regulator involved in the processes of development of the organism (ie, promoting development of bone and muscle mass). Although T effects on the mesenchyme-derived muscle, bone, and adipose tissues are well studied, T effects on intervertebral disc (IVD) have not been reported. PURPOSE: The aim was to test the following hypothesis: if a physiological concentration of T (similar to 30 nM) can improve in vitro chondrogenesis of human IVD cells and mesenchymal stem cells (MSCs). STUDY DESIGN/SETTING: Human IVD cells and MSCs were differentiated to chondrogenic lineage on gelatin scaffolds for 4 weeks, in the presence or absence of T. METHODS: Chondrogenesis was assessed by cell viability, by measuring gene expression with quantitative polymerase chain reaction and extracellular matrix (ECM) accumulation with immunoblotting, immunohistochemical, and biochemical methods. RESULTS: Supplementation of T to chondrogenic culture did not affect viability. In male IVD cells, T had a beneficial impact on chondrogenesis, particularly in nucleus pulposus cells, demonstrated by an increased expression of aggrecan, collagen type I, and especially collagen type II. Conversely, T had no effects on chondrogenesis of female IVD cells or MSCs from both genders. A gene expression array of transforming growth factor beta/bone morphogenetic protein signaling cascade showed that in male IVD cells, T promoted a stable general but nonsignificant increase in gene expression. Furthermore, aromatase inhibitor anastrazole repressed the effect of T on ECM expression by IVD cells. The results suggest that T increased ECM accumulation in male IVD cells in combination with its conversion to estradiol by the enzyme aromatase. CONCLUSIONS: We demonstrated that T effectively enhances in vitro chondrogenesis in male IVD cells, rising the interest in the possible role of sex hormones in IVD degeneration. Nevertheless, T does not affect chondrogenic differentiation of female IVD cells and MSCs from both genders. (C) 2014 Elsevier Inc. All rights reserved.
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