摘要
目的本研究通过类似于体内生理环境的关节软骨器官培养模型,评估利多卡因对关节软骨细胞活性的影响。方法实验材料取自女性骨性关节炎患者行膝关节置换术中切除的残余正常股骨髁,用环钻获取直径4mm的圆形新鲜全层厚度正常软骨,将其放置在培养液中1周,检测关节软骨器官培养模型的完整性。然后,通过不同临床相关浓度(0%、1%、2%)以及作用时间(30、60、120 min)的生理盐水和利多卡因溶液处理关节软骨,通过组织学HE染色和MTT方法定量测定关节软骨的组织学和细胞活性的变化,评估利多卡因对关节软骨的细胞毒性作用。结果新鲜关节软骨在培养液中经体外器官培养1周,软骨细胞的组织结构及细胞活性仅发生轻微的改变。相比较生理盐水组的软骨细胞活性改变为90%而言,时间效应组中1%利多卡因作用30、60、120 min后软骨细胞活性分别下降为79%、70%、56%。浓度效应组中当药物作用时间固定为1h,0.9%生理盐水、1%和2%利多卡因作用下的软骨细胞活性分别为92%、78%和70%。结论人关节软骨经体外器官培养1周,可维持组织学和细胞活性的完整性。仿生理状态下生物学特性和结构均完整的关节软骨经利多卡因作用后,随着作用时间延长和浓度增加,软骨细胞的活性明显降低,因此,临床相关浓度的利多卡因对人关节软骨有毒样作用。
Objective Lidocaine is a local anesthetic, which is commonly used for the treatment of osteoarthritis and perioperative analgesia. Cytotoxic effects of lidocaine on the articular cartilage in the cell culture in vitro have been reported. To evaluate the effects of lidocaine on cell viability using an articular cartilage organ culture model that provides a physiological environment similar to that in vivo. Methods The residual normal parts of the femoral condyle resected in knee arthroplasty for female patients with osteoarthritis were used as test materials. Normal cartilages with full thickness in the diameter of 4 mm were harvested by round drill. The articular cartilage was cultured in the media for 1 week, and the integrity of the organ culture model was tested. The articular cartilage was exposed to the saline and lidocaine of clinically relevant concentrations （0%, 1% and 2%） for different durations （30, 60 and 120 minutes）. The cytotoxic effects of lidocaine on the articular cartilage were evaluated by the changes of histological features and cell viability which were tested quantificationally by histological hematoxylin and eosin （HE） staining and 3- （4, 5） -dimethylthiahiazo （-z-y1） -3, 5-di- phenytetrazoliumromide （MTT） assay. Results Fresh articular cartilage showed little changes in histologic structure and cell viability when cultured in media in vitro for 1 week. Cell viability was decreased to 79%, 70% and 56% respectively when the articular cartilage was cultured in 1% lidocaine for 30 mins, 60 mins and 120 mins, and while the cell viability was changed to 90% when cultured in the saline. When exposed to 1% and 2% lidocaine for 1 hour, the articular cartilage would achieve about 78% and 70% cell viability while 0.9% saline exposure over the same amount of time resulted in 92% cell viability. Conclusions Human articular cartilage maintains the integrity of histology and cell viability when cultured in vivo for 1 week. Exposure of this biologically and structurally intact articular cartilage to lidocaine, which closely approximates the physiological environment in vivo, decreases cell viability in a dose- and time-dependent manner. This study demonstrates lidocaine at clinically relevant doses is toxic to human articular cartilage.
出处
《中国骨与关节杂志》
CAS
2013年第3期167-170,共4页
Chinese Journal of Bone and Joint
基金
北京市海淀区科学技术委员会基金资助项目(K20110111L)
关键词
关节软骨
器官培养
细胞活性
利多卡因
Articular cartilage
Organ culture
Cell viability
Lidocaine
