Cai Xian-Yi, Xiong Li-Ming, Yang Shu-Hua, Shao Zeng-Wu, Xie Mao, Gao Fei, Ding Fan
Department of Orthopaedic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 430022, Wuhan, China.
Department of Orthopaedic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 430022, Wuhan, China.
Spine J. 2014 Mar 1;14(3):483-90. doi: 10.1016/j.spinee.2013.06.041. Epub 2013 Aug 23.
It has been shown that bupivacaine, the most commonly used local anesthetic to relieve or control pain in interventional spine procedures, is cytotoxic to intervertebral disc (IVD) cells in vitro. However, some other common local anesthetics, such as ropivacaine and lidocaine, are also frequently used in the treatment of spine-related pain, and the potential effects of these agents remain unclear.
The purpose of this study was to evaluate the effect of various local anesthetics on rabbit IVD cells in vitro and further compare the cytotoxicity of ropivacaine, bupivacaine, lidocaine, and saline solution control.
Controlled laboratory study.
Rabbit annulus fibrosus (AF) and nucleus pulposus (NP) cells were isolated from Japanese white rabbits.
Both AF and NP cells at the second generation maintained in monolayer were exposed to various concentrations of local anesthetics (eg, bupivacaine) or different durations of exposure and evaluated for cell viability by use of cell counting kit-8 (CCK-8). In addition, to compare the cytotoxicity of ropivacaine, bupivacaine, lidocaine, and saline solution control in commercial concentration, the viability was analyzed by flow cytometry after 60-minute exposure, and the morphologic changes were observed by the phase-contrast microscopy. Apoptosis and necrosis of IVD cells were confirmed by using fluorescence microscopy with double staining of Hoechst 33342 and propidium iodide.
Rabbit IVD cell death demonstrated a time and dose dependence in response to bupivacaine and lidocaine. However, ropivacaine only exerted a significant time-dependent effect on IVD cells. There was no significant difference in IVD viability after treatment with different doses of ropivacaine. In addition, the results showed that lidocaine was the most toxic of the three local anesthetics and that ropivacaine presented less cytotoxicity than lidocaine and bupivacaine. Fluorescence microscopy also confirmed that the short-term toxic effect of local anesthetics on both AF and NP cells was mainly caused by necrosis rather than apoptosis.
Results show that bupivacaine and lidocaine decrease cell viability in rabbit IVD cells in a dose- and time-dependent manner. All local anesthetics should be avoided if at all possible. Ropivacaine may be a choice if necessary, but it is also toxic. The increase in cell death is more related with cell necrosis rather than cell apoptosis. If these results can be corroborated in tissue explant models or animal studies, caution regarding diagnosing, treating, and controlling spine-related pain with local anesthetics is prompted.
已有研究表明，布比卡因是介入性脊柱手术中最常用的用于缓解或控制疼痛的局部麻醉剂，在体外对椎间盘（IVD）细胞具有细胞毒性。然而，其他一些常用的局部麻醉剂，如罗哌卡因和利多卡因，也常用于治疗脊柱相关疼痛，这些药物的潜在影响仍不明确。
本研究旨在评估各种局部麻醉剂对兔IVD细胞的体外作用，并进一步比较罗哌卡因、布比卡因、利多卡因和生理盐水对照的细胞毒性。
对照实验室研究。
从日本白兔中分离出兔纤维环（AF）和髓核（NP）细胞。
将第二代单层培养的AF和NP细胞暴露于不同浓度的局部麻醉剂（如布比卡因）或不同的暴露时间，使用细胞计数试剂盒-8（CCK-8）评估细胞活力。此外，为了比较罗哌卡因、布比卡因、利多卡因和生理盐水对照在商业浓度下的细胞毒性，在暴露60分钟后通过流式细胞术分析细胞活力，并通过相差显微镜观察形态学变化。使用Hoechst 33342和碘化丙啶双重染色的荧光显微镜确认IVD细胞的凋亡和坏死。
兔IVD细胞死亡对布比卡因和利多卡因表现出时间和剂量依赖性。然而，罗哌卡因仅对IVD细胞产生显著的时间依赖性作用。用不同剂量的罗哌卡因处理后，IVD活力没有显著差异。此外，结果表明利多卡因是三种局部麻醉剂中毒性最大的，罗哌卡因的细胞毒性低于利多卡因和布比卡因。荧光显微镜也证实局部麻醉剂对AF和NP细胞的短期毒性作用主要由坏死而非凋亡引起。
结果表明布比卡因和利多卡因以剂量和时间依赖性方式降低兔IVD细胞的活力。应尽可能避免使用所有局部麻醉剂。如有必要，罗哌卡因可能是一种选择，但它也有毒性。细胞死亡的增加与细胞坏死而非细胞凋亡更相关。如果这些结果能在组织外植体模型或动物研究中得到证实，则提示在使用局部麻醉剂诊断、治疗和控制脊柱相关疼痛时需谨慎。
