Low back pain is experienced by a large number of people in western countries and may be caused and influenced by many different pathologies and psychosocial factors including disc degeneration. Disc degeneration involves the increased expression of proinflammatory cytokines and matrix metalloproteinases (MMPs) in the disc environment, which leads to the loss of extracellular matrix (ECM) and the viability of the native disc cells (DCs). Treatment approaches using growth factors and cell therapy have been proposed due to the compelling results that growth factors and mesenchymal stem cells (MSCs) can influence the degenerated discs. The aim of this study was to investigate the effects of conditioned media (CM) from human MSCs (hMSCs) and connective tissue growth factor (CTGF) and TGF-β on disc cells, and hMSCs isolated from patients with degenerative discs and severe low back pain. The aim was also to examine the constituents of CM in order to study the peptides that could bring about intervertebral disc (IVD) regeneration. DCs and hMSC pellets (approx.. 200,000 cells) were cultured and stimulated with hMSC-derived CM or CTGF and TGF-β over 28 days. The effects of CM and CTGF on DCs and hMSCs were assessed via cell viability, proteoglycan production, the expression of ECM proteins, and chondrogenesis in 3D pellet culture. To identify the constituents of CM, CM was analyzed with tandem mass spectrometry. The findings indicate that CM enhanced the cellular viability and ECM production of DCs while CTGF and the control exhibited nonsignificant differences. The same was observed in the hMSC group. Mass spectrometry analysis of CM identified >700 peptides, 129 of which showed a relative abundance of ≥2 (CTGF among them). The results suggest that CM holds potential to counter the progression of disc degeneration, likely resulting from the combination of all the substances released by the hMSCs. The soluble factors released belong to different peptide families. The precise mechanism underlying the regenerative effect needs to be investigated further, prior to incorporating peptides in the development of new treatment strategies for low back pain that is potentially caused by IVD degeneration.

中文翻译：

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间充质干细胞分泌因子对退化椎间盘细胞影响的研究

腰痛在西方国家很多人都经历过，可能由许多不同的病理和社会心理因素（包括椎间盘退变）引起和影响。椎间盘退变涉及椎间盘环境中促炎细胞因子和基质金属蛋白酶 (MMP) 的表达增加，这导致细胞外基质 (ECM) 的丢失和天然椎间盘细胞 (DC) 的活力。由于生长因子和间充质干细胞 (MSC) 可以影响退化椎间盘的令人信服的结果，已经提出了使用生长因子和细胞疗法的治疗方法。本研究的目的是研究来自人 MSC (hMSC) 的条件培养基 (CM) 和结缔组织生长因子 (CTGF) 和 TGF-β 对椎间盘细胞的影响，从患有退行性椎间盘和严重腰痛的患者中分离出 hMSCs。目的还在于检查 CM 的成分，以研究可导致椎间盘 (IVD) 再生的肽。DCs 和 hMSC 颗粒（大约 200,000 个细胞）被培养并用 hMSC 衍生的 CM 或 CTGF 和 TGF-β 刺激超过 28 天。CM 和 CTGF 对 DCs 和 hMSCs 的影响通过细胞活力、蛋白多糖产生、ECM 蛋白的表达和 3D 颗粒培养中的软骨形成进行评估。为了鉴定 CM 的成分，用串联质谱法分析了 CM。研究结果表明，CM 增强了 DCs 的细胞活力和 ECM 产生，而 CTGF 和对照表现出不显着的差异。在 hMSC 组中也观察到了同样的情况。CM 的质谱分析鉴定 > 700 个肽段，其中 129 个肽段的相对丰度≥2（其中 CTGF）。结果表明，CM 具有对抗椎间盘退变进展的潜力，这可能是由于 hMSC 释放的所有物质的组合所致。释放的可溶性因子属于不同的肽家族。在将肽用于开发可能由 IVD 变性引起的腰痛的新治疗策略之前，需要进一步研究再生效应的确切机制。释放的可溶性因子属于不同的肽家族。在将肽用于开发可能由 IVD 变性引起的腰痛的新治疗策略之前，需要进一步研究再生效应的确切机制。释放的可溶性因子属于不同的肽家族。在将肽用于开发可能由 IVD 变性引起的腰痛的新治疗策略之前，需要进一步研究再生效应的确切机制。