Tissue Engineering for Orthopedics & Mechanobiology (TOM), Bone & Joint Program, Department for BioMedical Research (DBMR), Faculty of Medicine, University of Bern, CH-3008 Bern, Switzerland.
Graduate School for Cellular and Biomedical Sciences (GCB), University of Bern, CH-3012 Bern, Switzerland.
Int J Mol Sci. 2024 Nov 2;25(21):11803. doi: 10.3390/ijms252111803.
Low back pain (LBP) is a significant global health issue, contributing to disability and socioeconomic burdens worldwide. The degeneration of the human intervertebral disc (IVD) is a critical factor in the pathogenesis of LBP. Recent studies have emphasized the significance of a specific set of genes and extracellular matrix (ECM) in IVD health. In particular, Noggin has emerged as a critical gene due to its high expression levels in healthy nucleus pulposus cells (NPCs) observed in our previous research. In this study, it was hypothesized that decreased Noggin expression in NPCs is associated with IVD degeneration and contributes to LBP development. A lentivirus-mediated RNAi was applied to knock down Noggin expression in primary NPCs from six human donors. The NPCs after transduction were evaluated through cell viability analysis, XTT assay, and cell apoptosis analyses. After two weeks, a colony formation assay was used to examine the anchor-independent growth ability of transduced cells. At the transcript level, anabolic and catabolic markers were quantified using RT-qPCR. The results demonstrated that lentivirus-mediated downregulation of Noggin significantly inhibited cell proliferation, reduced cell viability, and suppressed colony formation, while inducing apoptosis in human NPCs in vitro. Notably, it disrupted cellular anabolic processes and promoted catabolic activity in human NPCs post-transduction. Our findings indicated that the degeneration of human IVD is possibly related to decreased Noggin expression in NPCs. This research provides valuable insights into the role of Noggin in IVD homeostasis and its implications in LBP treatment.
下背痛（LBP）是一个重大的全球健康问题，导致全球残疾和社会经济负担。人类椎间盘（IVD）的退化是 LBP 发病机制的一个关键因素。最近的研究强调了一组特定基因和细胞外基质（ECM）在 IVD 健康中的重要性。特别是，Noggin 因其在我们之前的研究中观察到的健康核髓核细胞（NPC）中的高表达水平而成为一个关键基因。在这项研究中，假设 NPC 中 Noggin 表达的降低与 IVD 退化有关，并导致 LBP 的发展。应用慢病毒介导的 RNAi 敲低来自六位人类供体的原代 NPC 中的 Noggin 表达。转导后的 NPC 通过细胞活力分析、XTT 测定和细胞凋亡分析进行评估。两周后，使用集落形成测定法检查转导细胞的无锚定生长能力。在转录水平上，使用 RT-qPCR 定量分析合成代谢和分解代谢标记物。结果表明，慢病毒介导的 Noggin 下调显著抑制细胞增殖，降低细胞活力，并抑制 NPC 体外集落形成，同时诱导细胞凋亡。值得注意的是，它破坏了 NPC 中的细胞合成代谢过程并促进了分解代谢活性。我们的研究结果表明，人类 IVD 的退化可能与 NPC 中 Noggin 表达的降低有关。这项研究为 Noggin 在 IVD 动态平衡中的作用及其在 LBP 治疗中的意义提供了有价值的见解。
