VEGF对小鼠髁突关节软骨影响的体外实验研究  

Effect of VEGF on condylar articular cartilage of mouse in vitro

作　　者：曲鹏宇[1] 张善勇[2] 胡温庭[1] 范宝婷 李慧萍[2] 孙守福 

机构地区：[1]潍坊医学院口腔医学院,山东潍坊261053 [2]上海交通大学医学院附属第九人民医院.口腔医学院口腔外科上海市口腔医学重点实验室,上海200011

出　　处：《中国口腔颌面外科杂志》2017年第2期109-114,共6页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81371168;81671010);上海交通大学医学院转化医学创新基金(15ZH2007)

摘　　要：目的:通过研究不同浓度VEGF作用下离体培养小鼠髁突关节软骨的改变,探讨VEGF对离体培养小鼠髁突骨关节炎的直接影响。方法:取4周龄C57雄性小鼠髁突126个,随机分为21小组,每组6个。加入不同浓度(100 ng/m L、500 ng/m L、1μg/m L、2μg/m L)的VEGF进行离体培养。在不同的时间点(1、2、4、7 d)获取样本后,采用H-E染色、番红O快绿复合染色观察样本髁突的形态结构,并通过Mankin评分评价样本软骨改变;采用免疫组织化学方法观察各组样本中VEGFR2、MMP9、MMP13、TRANCE的表达。采用SPSS 23.0软件包对结果进行统计学分析。结果:H-E染色显示,与不作任何处理的空白对照组相比,加入VEGF离体培养的实验组小鼠髁突软骨结构破坏,肥大细胞增生。番红O快绿复合染色显示,与未加入VEGF的对照组相比,实验组小鼠髁突蛋白多糖含量降低,软骨发生退行性改变。对番红O快绿复合染色采用Mankin评分系统评分,结果表明,不同VEGF刺激时间组内,随着浓度的增高,Mankin评分增高,差异显著(P<0.05)。不同VEGF浓度组内,随着刺激时间的增加,Mankin评分增高,差异显著(P<0.05)。免疫组织化学显示,离体培养时加入VEGF的实验组,VEGFR2、MMP9、MMP13、TRANCE表达量较对照组显著增多。结论:在离体培养条件下,VEGF可直接引起小鼠颞下颌关节发生骨关节炎。PURPOSE： To explore the direct influence of VEGF on temporomandibular joint osteoarthritis, and the transformations of the condylar cartilage of mouse after cultivated in vitro under different concentrations of VEGF. METHODS： One hundred and twenty-six condyles from 4-week-old C57 male mouse were divided into 21 groups randomly. VEGF with different concentrations （100 ng/mL, 500 ng/mL, 1 μg/mL, 2 μg/mL） was added to the culture of the condyles. Samples were obtained at different time （1, 2, 4, 7 days） H-E staining and safranin O fast green staining were performed to observe the morphology of the condyle cartilages. Mankin＇s scoring system was used to evaluate the changes of the condyle cartilages and immunohistochemistry was carried out to observe the expression of VEGFR2 MMPg, MMP13 and TRANCE. The data were analyzed with SPSS 23.0 software package. RESULTS： H-E staining indicated that, compared with the blank control group, the experimental group added with VEGF showed destruction of the condylar cartilages and proliferation of the hypertrophic cells; safranin O-fast green staining showed that the experimental group had decreased contents of proteoglycan and degeneration changes in the condylar cartilage; Mankin＇s scoring system showed that Mankin＇s score was increased with increasing concentration and stimulation time of VEGF in each group, the differences was statistically significant （P〈0.05）; immunohistochemistry showed that the positive expression of VEGFR2,MMP9, MMP13, TRANCE in the experimental group was significantly increased compared with the control group. CONCLUSION： VEGF can cause temporomandibular joint osteoarthritis of mouse in vitro culture.

关 键 词：小鼠髁突 颞下颌关节骨关节炎 离体培养 

分 类 号：R782.6[医药卫生—口腔医学]