Seshadri Venkat, Coyle Christian H, Chu Constance R
Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
Arthroscopy. 2009 Apr;25(4):337-47. doi: 10.1016/j.arthro.2009.01.003. Epub 2009 Feb 13.
This study examined the viability of bovine articular chondrocytes after exposure to methylprednisolone, methylprednisolone with lidocaine, and methylprednisolone in a simulated inflammatory environment.
Bovine articular chondrocytes were suspended in alginate beads and cultured in Dulbecco's modified Eagle's medium/F-12 for 1 week before experimentation. Suspended chondrocytes were exposed to 0.9% saline solution (negative control), methylprednisolone (4, 8, and 16 mg/mL), methylprednisolone (8 mg/mL) with 1% lidocaine, or methylprednisolone (8 mg/mL) and saline solution in a simulated inflammatory environment (interleukin [IL] 1beta exposure, 10 ng/mL) for 15, 30, and 60 minutes. Flow cytometry was performed 1 day, 4 days, and 7 days after exposure by use of annexin V and propidium iodide to assess chondrocyte viability.
Chondrocyte viability decreased from 84% in saline solution to 62%, 38%, and 2.4% 1 day after 60 minutes of exposure to 4, 8, and 16 mg/mL of methylprednisolone, respectively (n = 7, P < .05). Chondrotoxicity increased with increasing time of exposure to methylprednisolone and with increasing time after exposure. In IL-1beta-activated chondrocytes, viability decreased from 76% in saline solution to 2.9% after 60 minutes of methylprednisolone exposure (8 mg/mL) (n = 4, P < .05). The combination of 8 mg/mL of methylprednisolone and 1% lidocaine further reduced viability to 1.0% after 60 minutes (n = 4, P < .05).
These results show a dose- and time-dependent decrease in chondrocyte viability after exposure to clinically relevant doses of methylprednisolone. The combination of methylprednisolone and lidocaine was toxic, with virtually no cells surviving after treatment. In addition, methylprednisolone did not mitigate the inflammatory effects of IL-1beta; rather, it further potentiated the chondrotoxicity.
Intra-articular injections of corticosteroids and local anesthetics are widely used in clinical practice. This in vitro study provides information on the potential effects of these drugs on articular cartilage.
本研究检测了牛关节软骨细胞在暴露于甲基强的松龙、甲基强的松龙与利多卡因联合使用以及在模拟炎症环境中的甲基强的松龙后的活力。
将牛关节软骨细胞悬浮于藻酸盐珠中,并在实验前于杜尔贝科改良伊格尔培养基/F-12中培养1周。将悬浮的软骨细胞暴露于0.9%盐溶液(阴性对照)、甲基强的松龙(4、8和16mg/mL)、含1%利多卡因的甲基强的松龙(8mg/mL)或在模拟炎症环境(白细胞介素[IL]1β暴露,10ng/mL)中的甲基强的松龙(8mg/mL)和盐溶液中15、30和60分钟。在暴露后1天、4天和7天使用膜联蛋白V和碘化丙啶进行流式细胞术检测以评估软骨细胞活力。
暴露于4、8和16mg/mL甲基强的松龙60分钟后1天,软骨细胞活力分别从盐溶液中的84%降至62%、38%和2.4%(n = 7,P <.05)。软骨毒性随着暴露于甲基强的松龙的时间增加以及暴露后时间的增加而增加。在IL-1β激活的软骨细胞中,暴露于甲基强的松龙(8mg/mL)60分钟后,活力从盐溶液中的76%降至2.9%(n = 4,P <.05)。60分钟后含8mg/mL甲基强的松龙和1%利多卡因的联合用药使活力进一步降至1.0%(n = 4,P <.05)。
这些结果表明,暴露于临床相关剂量的甲基强的松龙后,软骨细胞活力呈剂量和时间依赖性下降。甲基强的松龙和利多卡因联合用药具有毒性,治疗后几乎没有细胞存活。此外,甲基强的松龙并未减轻IL-1β的炎症作用;相反,它进一步增强了软骨毒性。
关节内注射皮质类固醇和局部麻醉剂在临床实践中广泛使用。这项体外研究提供了这些药物对关节软骨潜在影响的信息。