硫化铜@二氧化硅经光热作用建立大鼠膝骨关节炎模型  

A novel knee osteoarthritis model in rats induced by the photothermal effect of Cu_9S_5@SiO_2 nanoparticles

作　　者：张菲[1] 周峰[1] 邓国英[1] 王秋根[1] 王谦[1] 

机构地区：[1]上海交通大学附属第一人民医院创伤骨科,201600

出　　处：《国际骨科学杂志》2017年第4期264-270,共7页International Journal of Orthopaedics

基　　金：国家自然科学基金面上项目(81272002);上海市科委引导项目(15411968800);上海市第一人民医院优秀医学青年人才培养前期计划(061417)

摘　　要：目的探索采用硫化铜@二氧化硅(Cu_9S_5@SiO_2)经光热作用建立大鼠膝关节骨关节炎模型的方法。方法使用CCK-8试剂检测Cu_9S_5@SiO_2在应用浓度下对原代软骨细胞增殖能力的影响。将软骨细胞分为无材料组(对照组)、无材料+光照2 min组、材料+光照0min组、材料+光照1 min组以及材料+光照2 min组。材料组加入浓度为100μg/mL的Cu_9S_5@SiO_2。使用980 nm、0.72 W/cm2激光进行光照,使用流式细胞仪分析其凋亡情况,并使用逆转录聚合酶链式反应(RT-PCR)法检测样本中半胱氨酸蛋白酶-3(caspase-3)和白细胞介素-1β(IL-1β)的表达情况。将24只SD大鼠随机均分为对照组(a组)、Cu_9S_5@SiO_2+光照5 min组(b组)、Cu_9S_5@SiO_2+光照10 min组(c组)以及前交叉韧带切除组(d组)。980 nm、0.72 W/cm2激光照射相应时间,利用红外光谱仪监测膝关节温度变化并绘制升温曲线。4周后取材,通过RT-PCR和HE染色评估关节损伤情况。结果 Cu_9S_5@SiO_2浓度≤100μg/mL对于软骨细胞的生长无明显毒性。材料+光照1 min组以及材料+光照2 min组软骨细胞凋亡率明显增加,其IL-1β和caspase-3表达较对照组显著增高。c组大鼠膝关节照光前后升温明显,a组大鼠膝关节照光前后温度变化无明显差异。RT-PCR检测显示b、c组大鼠膝关节软骨、滑膜、半月板组织IL-1β表达较a组明显增高。HE染色结果显示b、c组大鼠膝关节结构紊乱、形态不规则,与a组差异明显。结论 Cu_9S_5@SiO_2可应用于SD大鼠膝关节骨关节炎模型的建立。Objective To establish a novel knee osteoarthritis（OA）model in rats induced by the photothermal effect of copper sulfide@SiO2（Cu9S5@SiO2）nanoparticles.Methods The effect of Cu9S5@SiO2 nanoparticles on the cell proliferation of the primary chondrocytes was investigated with the CCK-8 kit.Chondrocytes received no treatment（control group）,laser irradiation for 2 minutes,nanoparticle treatment,nanoparticle treatment with laser irradiation for 1 minute,or nanoparticle treatment with laser irradiation for 2 minutes.Nanoparticle treatment means that Cu9S5@SiO2nanoparticles were added into the culture medium at a concentration of 100μg/mL.Laser irradiation with the energy of 980 nm,0.72 W/cm^2 was adopted.Afterwards,the apoptosis ratio was detected by flow cytometry.The expression of caspase-3 and interleukin-1β（IL-1β）was detected by reverse transcription polymerase chain reaction（RT-PCR）.24 SD rats were divided into 4 groups：control group（group A）,Cu9S5@SiO2＋photothermy 5 min group（group B）,Cu9S5@SiO2＋photothermy 10 min group（group C）and anterior cruciate ligament removal group（group D）.The temperature change was monitored by infrared spectroscopy and the temperature curve was plotted under laser irradiation with the same energy（980 nm,0.72 W/cm^2）.After 4 weeks,the samples of right knee joint of SD rats were examined by means of RT-PCR and HE staining to evaluate the extent of damage.Results Cu9S5@SiO2 nanoparticles showed no obvious cytotoxity in chondrocytes within the concentration of 100μg/mL.The apoptosis ratio of cells treated by nanoparticle with laser irradiation for1 minute or 2 minutes increased significantly,and the expression of caspase-3 and IL-1β was increased significantly,compared with the control group.The temperature of knee in group C rose significantly while the change of temperature in group A showed no difference after laser irradiation.The RT-PCR showed that the expression of IL-1β in cartilage,synovium and meniscus of group B and C was increase

关 键 词：纳米材料 硫化铜 安全性 光热效应 骨关节炎 

分 类 号：R-332[医药卫生] R684.3