Beyzadeoğlu Tahsin, Torun Köse Gamze, Ekinci Işın D, Bekler Halil, Yilmaz Cemil
Department of Orthopedics and Traumatology, Faculty of Medicine, Yeditepe University, İstanbul, Turkey.
Acta Orthop Traumatol Turc. 2012;46(3):201-7. doi: 10.3944/aott.2012.2606.
The aim of this study was to evaluate the effects of both in vivo and in vitro bupivacaine, levobupivacaine and tramadol on articular cartilage and chondrocytes in experimental rat models.
Thirty mature Sprague Dawley rats weighing 230-300 g were randomized into 3 groups. Bupivacaine (Group 1), levobupivacaine (Group 2) and tramadol (Group 3) were injected into the right knee joints and a physiological 0.9% saline into the left. From each group, 5 rats were executed 48 hours following drug administration after 5 and 10 days. The specimens were fixed, decalcified and stained with hematoxylin & eosin and toluidine blue. All samples were histopathologically evaluated according to the recommendation of ICRS' osteoarthritis and cartilage histopathology grading and staging system. Articular cartilage cells of the rats were cultured and seeded into cell culture flasks. Cartilage cell seeded samples (104 cells/ml) were incubated in three different anesthetic agents (0.5%); bupivacaine, levobupivacaine, and tramadol, respectively. CellTiter 96(®) Non-Radioactive Cell Proliferation (MTS) assay was used to determine the cell density on the samples.
Statistically significant higher OARSI grades and OA stage and scores were detected when comparing the group injected with levobupivacaine and executed after 10 days with the levobupivacaine injected group killed after 48 hours (p<0.01 [p=0.008]). Although, statistical analysis could not be done due to insufficient number of samples in the in vitro part of the experiment, it can be concluded that tramadol is cytotoxic to rat chondrocyte in vitro after 30 min of exposure. Additionally, cell numbers in both the bupivacaine and levobupivacaine treated wells showed decrease throughout 15, 30 and 60 minute exposures.
Although chondrotoxicity of bupivacaine was less harmful than levobupivacaine and tramadol, these findings suggest that local anesthetics may negatively affect articular cartilage and chondrocytes.
本研究旨在评估在实验大鼠模型中，布比卡因、左旋布比卡因和曲马多在体内和体外对关节软骨和软骨细胞的影响。
将30只体重230 - 300克的成年Sprague Dawley大鼠随机分为3组。将布比卡因（第1组）、左旋布比卡因（第2组）和曲马多（第3组）注入右膝关节，左膝关节注入生理0.9%盐水。给药后5天和10天，每组分别于给药48小时后处死5只大鼠。标本固定、脱钙，并用苏木精和伊红以及甲苯胺蓝染色。所有样本均根据国际软骨修复学会（ICRS）骨关节炎和软骨组织病理学分级及分期系统的建议进行组织病理学评估。培养大鼠的关节软骨细胞并接种到细胞培养瓶中。将接种软骨细胞的样本（104个细胞/毫升）分别在三种不同的麻醉剂（0.5%）中孵育；布比卡因、左旋布比卡因和曲马多。使用CellTiter 96(®)非放射性细胞增殖（MTS）测定法来确定样本上的细胞密度。
当比较注射左旋布比卡因并在10天后处死的组与注射左旋布比卡因并在48小时后处死的组时，检测到骨关节炎研究学会国际分会（OARSI）分级、骨关节炎（OA）分期和评分在统计学上有显著更高的值（p<0.01 [p = 0.008]）。尽管由于实验体外部分样本数量不足无法进行统计分析，但可以得出结论，曲马多在体外暴露30分钟后对大鼠软骨细胞具有细胞毒性。此外，在整个15、30和60分钟的暴露过程中，布比卡因和左旋布比卡因处理孔中的细胞数量均呈下降趋势。
尽管布比卡因的软骨毒性比左旋布比卡因和曲马多小，但这些发现表明局部麻醉剂可能会对关节软骨和软骨细胞产生负面影响。
