Repetitive strikes loading organ culture model to investigate the biological and biomechanical responses of the intervertebral disc

IF 3.4 3区医学 Q1 ORTHOPEDICS JOR Spine Pub Date : 2024-01-19 DOI:10.1002/jsp2.1314

Jiaxiang Zhou, Jianmin Wang, Jianfeng Li, Zhengya Zhu, Zhongyuan He, Junhong Li, Tao Tang, Hongkun Chen, Yukun Du, Zhen Li, Manman Gao, Zhiyu Zhou, Yongming Xi

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Abstract

Background

Disc degeneration is associated with repetitive violent injuries. This study aims to explore the impact of repetitive strikes loading on the biology and biomechanics of intervertebral discs (IVDs) using an organ culture model.

Methods

IVDs from the bovine tail were isolated and cultured in a bioreactor, with exposure to various loading conditions. The control group was subjected to physiological loading, while the model group was exposed to either one strike loading (compression at 38% of IVD height) or repetitive one strike loading (compression at 38% of IVD height). Disc height and dynamic compressive stiffness were measured after overnight swelling and loading. Furthermore, histological morphology, cell viability, and gene expression were analyzed on Day 32. Glycosaminoglycan (GAG) and nitric oxide (NO) release in conditioned medium were also analyzed.

Results

The repetitive one strike group exhibited early disc degeneration, characterized by decreased dynamic compression stiffness, the presence of annulus fibrosus clefts, and degradation of the extracellular matrix. Additionally, this group demonstrated significantly higher levels of cell death (p < 0.05) and glycosaminoglycan (GAG) release (p < 0.05) compared to the control group. Furthermore, upregulation of MMP1, MMP13, and ADAMTS5 was observed in both nucleus pulposus (NP) and annulus fibrosus (AF) tissues of the repetitive one strike group (p < 0.05). The one strike group exhibited annulus fibrosus clefts but showed no gene expression changes compared to the control group.

Conclusions

This study shows that repetitive violent injuries lead to the degeneration of a healthy bovine IVDs, thereby providing new insights into early-stage disc degeneration.

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研究椎间盘的生物和生物力学反应的重复冲击加载器官培养模型

背景椎间盘退化与重复性暴力损伤有关。本研究旨在利用器官培养模型探讨重复性打击负荷对椎间盘（IVD）的生物学和生物力学的影响。方法在生物反应器中分离和培养牛尾椎间盘，并将其置于各种加载条件下。对照组承受生理负荷，而模型组则承受单击负荷（以 IVD 高度的 38% 压缩）或重复单击负荷（以 IVD 高度的 38% 压缩）。经过一夜的膨胀和加载后，对椎间盘高度和动态压缩刚度进行了测量。此外，第 32 天还对组织形态、细胞活力和基因表达进行了分析。此外，还分析了条件培养基中糖胺聚糖（GAG）和一氧化氮（NO）的释放情况。结果重复一击组表现出早期椎间盘退变，其特征是动态压缩硬度下降、出现纤维环裂隙和细胞外基质降解。此外，与对照组相比，该组的细胞死亡水平（p < 0.05）和糖胺聚糖（GAG）释放水平（p < 0.05）明显更高。此外，在重复一击组的髓核（NP）和环状纤维（AF）组织中都观察到了 MMP1、MMP13 和 ADAMTS5 的上调（p < 0.05）。与对照组相比，一击组出现环状纤维裂隙，但基因表达无变化。结论本研究表明，重复性暴力损伤会导致健康牛 IVD 退化，从而为早期椎间盘退化提供了新的见解。

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