Biomolecular Sciences Research Centre, Sheffield Hallam University, City Campus, Howard Street, Sheffield S1 1WB,
Eur Cell Mater. 2020 Jul 16;40:21-37. doi: 10.22203/eCM.v040a02.
Low-back pain affects 80 % of the world population at some point in their lives and 40 % of the cases are attributed to intervertebral disc (IVD) degeneration. Over the years, many animal models have been developed for the evaluation of prevention and treatment strategies for IVD degeneration. Ex vivo organ culture systems have also been developed to better control mechanical loading and biochemical conditions, but a reproducible ex vivo model that mimics moderate human disc degeneration is lacking. The present study described an ex vivo caprine IVD degeneration model that simulated the changes seen in the nucleus pulposus during moderate human disc degeneration. Following pre-load under diurnal, simulated physiological loading (SPL) conditions, lumbar caprine IVDs were degenerated enzymatically by injecting collagenase and chondroitinase ABC (cABC). After digestion, IVDs were subjected to SPL for 7 d. No intervention and phosphate-buffered saline injection were used as controls. Disc deformation was continuously monitored to assess disc height recovery. Histology and immunohistochemistry were performed to determine the histological grade of degeneration, matrix expression, degrading enzyme and catabolic cytokine expression. Injection of collagenase and cABC irreversibly affected the disc mechanical properties. A decrease in extracellular matrix components was found, along with a consistent increase in degradative enzymes and catabolic proteins [interleukin (IL)-1β, -8 and vascular endothelial growth factor (VEGF)]. The changes observed were commensurate with those seen in moderate human-IVD degeneration. This model should allow for controlled ex vivo testing of potential biological, cellular and biomaterial treatments of moderate human-IVD degeneration.
下背痛影响全球 80%的人口在其生命中的某个时刻，40%的病例归因于椎间盘（IVD）退变。多年来，已经开发了许多动物模型来评估预防和治疗 IVD 退变的策略。还开发了离体器官培养系统，以更好地控制机械加载和生化条件，但缺乏模拟中度人类椎间盘退变的可重复离体模型。本研究描述了一种模拟中度人类椎间盘退变过程中髓核变化的山羊离体椎间盘退变模型。在日间模拟生理加载（SPL）条件下预加载后，通过注射胶原酶和软骨素酶 ABC（cABC）对山羊腰椎 IVD 进行酶促退变。消化后，将 IVD 置于 SPL 下 7 天。未进行干预和磷酸盐缓冲盐水注射作为对照。连续监测椎间盘变形以评估椎间盘高度恢复。进行组织学和免疫组织化学检查以确定退变的组织学等级、基质表达、降解酶和分解代谢细胞因子表达。胶原酶和 cABC 的注射不可逆地影响了椎间盘的机械性能。发现细胞外基质成分减少，同时降解酶和分解代谢蛋白[白细胞介素（IL）-1β、-8 和血管内皮生长因子（VEGF）]持续增加。观察到的变化与中度人类-IVD 退变一致。该模型应允许对中度人类-IVD 退变的潜在生物、细胞和生物材料治疗进行体外受控测试。
