Röhner Eric, Hoff Paula, Pfitzner Tilman, Seeger Joern Bengt, Buttgereit Frank, Perka Carsten, Matziolis Georg
Department of Orthopaedic, Center for Musculoskeletal Surgery, Charit´e University Hospital, Berlin, Germany.
J Invest Surg. 2012 Oct;25(5):311-6. doi: 10.3109/08941939.2011.648718.
In septic joint surgery, treatment with antiseptics is a standard procedure. Postinfectious degeneration of cartilage often results in early arthritis that necessitates joint replacement, even in young patients. It is still unclear whether antiseptics have only a toxic effect on chondrocytes or also induce late cell death through apoptosis. We hypothesized that commonly used antiseptics (polyhexanide, hydrogen peroxide, and chlorhexidine) induce different stages of apoptosis on human chondrocytes. Human chondrocytes were isolated and cultured. Polyhexanide, hydrogen peroxide, and chlorhexidine were added to the monolayer cultures. Early and late apoptotic cells were analyzed using flow cytometric detection of Annexin V, active caspases, and 7AAD, and fluorescence microscopy using Annexin V and propidium iodide staining. Flow cytometric analysis revealed an increase of Annexin V and active caspases expression of human chondrocytes after incubation with polyhexanide, hydrogen peroxide, and chlorhexidine. Fluorescence microscopy demonstrated a high number of apoptotic cells. Our data show that polyhexanide and chlorhexidine only promote the early stages of apoptosis without any differences in toxicity. Conversely, hydrogen peroxide induces early and later stages of apoptosis on primary human chondrocytes in vitro.
在脓毒性关节手术中，使用防腐剂进行治疗是一种标准程序。感染后软骨退变常导致早期关节炎，即使在年轻患者中也需要进行关节置换。目前尚不清楚防腐剂是否仅对软骨细胞具有毒性作用，还是也通过凋亡诱导晚期细胞死亡。我们推测常用的防腐剂（聚己缩胍、过氧化氢和氯己定）会在人软骨细胞上诱导不同阶段的凋亡。分离并培养人软骨细胞。将聚己缩胍、过氧化氢和氯己定添加到单层培养物中。使用膜联蛋白V、活性半胱天冬酶和7-氨基放线菌素D的流式细胞术检测以及使用膜联蛋白V和碘化丙啶染色的荧光显微镜分析早期和晚期凋亡细胞。流式细胞术分析显示，与聚己缩胍、过氧化氢和氯己定孵育后人软骨细胞的膜联蛋白V和活性半胱天冬酶表达增加。荧光显微镜显示大量凋亡细胞。我们的数据表明，聚己缩胍和氯己定仅促进凋亡的早期阶段，在毒性方面没有任何差异。相反，过氧化氢在体外原代人软骨细胞上诱导凋亡的早期和晚期阶段。
