Oh Eunha G, Xiao Li, Xu Zhiwen, Xing Yuan, Zhang Yi, Anbaei Parastoo, Chi Jialun A, Jin Li, Pompano Rebecca R, Li Xudong
Department of Orthopaedic Surgery, University of Virginia, Charlottesville, VA 22908, USA.
Department of Spine Surgery, Spinal Deformity Center, The Second Xiangya Hospital of Central South University, Changsha 410011, China.
Cells. 2025 Aug 8;14(16):1230. doi: 10.3390/cells14161230.
Intervertebral disc degeneration is a leading cause of back and leg pain and a major contributor to disability worldwide. Despite its prevalence, treatments remain limited due to incomplete understanding of its pathology. In vivo models pose challenges for controlled conditions, while in vitro cell cultures lack key cell-cell and cell-matrix interactions. To address these limitations, we developed a novel tissue slice culture model of mouse discs, in which intact mouse discs were sliced down to 300 μm thickness with a vibratome and cultured ex vivo at various time points. The cell viability, matrix components, structure integrity, inflammatory responses, and macrophage interactions were evaluated with biochemistry, gene expression, histology, and 3D imaging analyses. Disc slices maintained structural integrity and cell viability, with preserved extracellular matrix in the annulus fibrosus (AF) and mild degeneration in nucleus pulposus (NP) by day 5. Interleukin-1 (IL-1) induced disc degeneration manifested by increased glycosaminoglycan release in media and reduced and mRNA levels in disc cells. Cultured disc slices promoted macrophages towards pro-inflammatory phenotype with elevated mRNA levels of , , and . Macrophage overlay and 3D imaging demonstrated macrophage infiltration into the NP and AF tissues up to ~100 µm in depth. The disc tissue slice model captures key features of intervertebral discs and can be used for investigating mechanisms of disc degeneration and therapeutic evaluation.
椎间盘退变是导致腰腿痛的主要原因，也是全球范围内导致残疾的主要因素。尽管其发病率很高，但由于对其病理机制的认识不完整，治疗方法仍然有限。体内模型在控制条件方面存在挑战，而体外细胞培养缺乏关键的细胞间和细胞与基质的相互作用。为了解决这些局限性，我们开发了一种新型的小鼠椎间盘组织切片培养模型，其中完整的小鼠椎间盘用振动切片机切成300μm厚，并在不同时间点进行离体培养。通过生物化学、基因表达、组织学和三维成像分析评估细胞活力、基质成分、结构完整性、炎症反应和巨噬细胞相互作用。椎间盘切片在第5天时保持结构完整性和细胞活力，纤维环（AF）中的细胞外基质得以保留，髓核（NP）出现轻度退变。白细胞介素-1（IL-1）诱导的椎间盘退变表现为培养基中糖胺聚糖释放增加，椎间盘细胞中 和 信使核糖核酸水平降低。培养的椎间盘切片促使巨噬细胞向促炎表型转变， 、 和 的信使核糖核酸水平升高。巨噬细胞覆盖和三维成像显示巨噬细胞浸润到NP和AF组织中，深度可达约100μm。椎间盘组织切片模型捕捉到了椎间盘的关键特征，可用于研究椎间盘退变机制和治疗评估。
