中文摘要：

目的探讨骨关节炎（OA）膝关节正常及退变区域原位软骨细胞自发性[Ca^（2＋）]i（细胞内游离钙离子浓度）信号特征与差异。方法以西南医院关节外科提供的OA患者膝关节置换软骨组织作为研究对象,根据软骨区域分正常组与OA组。使用Fluo-8 AM钙离子探针以及荧光显微镜观测法对组织中原位软骨细胞在不同Ca^（2＋）浓度环境下自发性[Ca^（2＋）]i信号进行观测;使用图像处理软件与统计学单因素方差分析法对检测结果进行分析。结果正常与OA软骨细胞在0 mM与4 mM Ca^（2＋）环境中均产生自发性[Ca^（2＋）]i信号,且具向相邻细胞传递特性。正常组处于4 mM Ca^（2＋）浓度中,表层及中层软骨细胞自发性[Ca^（2＋）]i信号各项指标与深层软骨细胞比较存在统计学差异。（1）表层vs.深层,峰值量级：（1.73±0.13）vs.（2.90±0.25）;响应率：（15.08%±8.29%）vs（69.65%±5.21%）;峰值数目：（0.17±0.09）vs（0.95±0.08）,P〈0.05。（2）中层vs深层,峰值量级：（2.03±0.76）vs（2.90±0.25）;响应率：（36.75%±6.73%）vs（69.65%±5.21%）;峰值数目：（0.61±0.10）vs（0.95±0.08）,P〈0.05。但该差异在0 mM Ca^（2＋）环境中不存在。OA组在0 mM或4 mM Ca^（2＋）环境中各层软骨细胞[Ca^（2＋）]i信号均不存在差异;但其中层与深层软骨细胞[Ca^（2＋）]i信号易受胞外Ca^（2＋）浓度影响。（1）中层,4 mM vs 0 mM：峰值量级：（2.3±0.11）vs.（1.86±0.11）、响应率：（53.88%±8.21%）vs.（26.50%±8.89%）、峰值数目：（0.84±0.94）vs.（0.28±0.07）,P〈0.05;（2）深层,4 mM vs 0 mM：峰值数目：（0.59±0.11）vs.（0.21±0.06）,P〈0.05。在4 mM Ca^（2＋）环境中,OA组表层与中层细胞[Ca^（2＋）]i信号明显强于正常组：（1）表层,OA vs正常：峰值量级：（2.62±0.51）vs（1.73±0.13）,P〈0.05;（2）中层,OA vs.正常：峰值量级：（2.3±0.11）vs.（2.03±0.76?

英文摘要：

Objective To analyze the pattern difference of spontaneous[Ca~（2＋）]i（ Intracellular free calcium concentration） signal in normal and osteoarthritis（ OA） in-situ chondrocytes of human knee osteoarthritis articular cartilage. Methods Cartilage samples form OA patients who had knee replacement surgery in department of joint surgery,the 1st affiliated hospital of third Military Medical University were separated into normal and OA groups. Spontaneous [Ca~（2＋）]isignal was recorded with Fluo-8 AM calcium indicator and fluorescence microscope. Data were analyzed with image processing software and one-way one-factor analysis of variance（ ANOVA） analysis. Results Both normal and OA chondrocytes were able to display spontaneous [Ca~（2＋）]isignal,which usually propagated to neighboring chondrocytes. In the normal chondrocytes,when 4 mM Ca~（2＋）was presented,differences in spontaneous [Ca~（2＋）]isignal pattern were observed among different zones（ deep,middle and superficial）.（ 1） superficial vs deep,fold increment：（ 1. 73 ± 0. 13） vs（ 2. 90 ± 0. 25）,percentage of responding cells：（ 15. 08% ± 8. 29%） vs（ 69. 65% ± 5. 21%）,peak number（ 0. 17 ± 0. 09） vs（ 0. 95 ± 0. 08）,P〈0. 05.（ 2） middle vs deep,fold increment：（ 2. 03 ± 0. 76） vs（ 2. 90 ± 0. 25）,percentage of responding cells：（ 36. 75% ± 6. 73%） vs（ 69. 65% ± 5. 21%）,peak number：（ 0. 61 ± 0. 10） vs（ 0. 95 ± 0. 08）,P〈0. 05. The difference was not observed in calcium free environment. In the OA chondrocytes,whether in 0 mM or in 4 mM Ca~（2＋）environment,the zonal difference in spontaneous [Ca~（2＋）]isignal pattern was not observed,but the spontaneous [Ca~（2＋）]isignal intensity of the middle and deep zones was more easily affected by the extracellular Ca~（2＋）concentration.（ 1） Middle,4 mM vs 0 mM,fold increment：（ 2. 3 ± 0. 11） vs（ 1. 86 ±0. 11）,percentage of responding cells：（ 53. 88% ± 8. 21%） vs（ 26. 50% ± 8