Department of Anesthesiology, University Medical Center Regensburg, Regensburg, Germany.
Arthroscopy. 2013 Oct;29(10):1676-84. doi: 10.1016/j.arthro.2013.06.018. Epub 2013 Aug 29.
The purpose of this study was to investigate the cytotoxic potency of local anesthetics on human mesenchymal stem cells (MSCs) before and after chondrogenic differentiation.
MSCs were exposed to equal and equipotent concentrations of bupivacaine, ropivacaine, and mepivacaine for 1 hour. Cell viability, apoptosis, and necrosis were determined using flow cytometry and live/dead staining. After chondrogenic differentiation, MSC viability was determined in aggregates exposed to equipotent concentrations of the named agents, applying fluorescence microscopy.
All local anesthetics showed detrimental cytotoxic effects on MSC monolayer cultures in a concentration- and time-specific manner. Minimum viability rates were found 96 hours after a 1-hour exposure. Bupivacaine 0.5% caused a reduction of vital MSCs to 5% ± 1%. Sixteen percent ± 2% viable cells were detected after treatment with 0.75% ropivacaine. Exposure to 2% mepivacaine decreased vitality rates to 1% ± 0%. Ropivacaine was significantly less cytotoxic than were bupivacaine and mepivacaine. Immediate cell death was mainly caused by necrosis followed by apoptosis afterward. Viability rates of MSCs embedded in cartilaginous tissue after chondrogenic differentiation were not reduced by local anesthetic treatment.
Local anesthetics are cytotoxic to MSCs in a concentration-, time-, and agent-dependent manner in monolayer cultures but not in whole-tissue probes.
MSCs are applied for treatment of cartilage defects. Intra-articular application of local anesthesia is a common procedure in pain management and has shown chondrotoxic effects. Therefore, it is crucial to evaluate the impact of local anesthetics on human MSCs and regenerative cartilage tissue engineering.
本研究旨在探讨局部麻醉剂在软骨分化前后对人骨髓间充质干细胞（MSCs）的细胞毒性作用。
将 MSC 暴露于等浓度和等效浓度的布比卡因、罗哌卡因和甲哌卡因中 1 小时。采用流式细胞术和活/死染色法测定细胞活力、凋亡和坏死。在软骨分化后，将含有命名剂的等效力浓度的聚集物暴露于 MSC 中，应用荧光显微镜测定 MSC 的活力。
所有局部麻醉剂均以浓度和时间依赖的方式对 MSC 单层培养物产生有害的细胞毒性作用。在 1 小时暴露后 96 小时发现最小存活率。0.5%布比卡因导致活 MSC 减少到 5%±1%。用 0.75%罗哌卡因处理后，检测到 16%±2%的活细胞。暴露于 2%甲哌卡因可将活力率降低至 1%±0%。罗哌卡因的细胞毒性明显低于布比卡因和甲哌卡因。即刻细胞死亡主要由坏死引起，随后是凋亡。软骨分化后嵌入软骨组织中的 MSC 的存活率未因局部麻醉剂处理而降低。
局部麻醉剂在单层培养物中以浓度、时间和药物依赖性方式对 MSCs 具有细胞毒性，但在整个组织探针中则不然。
MSCs 用于治疗软骨缺陷。关节内局部麻醉的应用是疼痛管理中的常见程序，已显示出软骨毒性作用。因此，评估局部麻醉剂对人 MSCs 和再生软骨组织工程的影响至关重要。
