VEGF抗体抑制磨损颗粒诱导骨溶解的实验研究  

INHIBITORY EFFECTS OF VASCULAR ENDOTHELIAL GROWTH FACTOR ANTIBODY ON WEAR PARTICLEINDUCED OSTEOLYSIS

作　　者：戴闽[1] 钟艳春[1] 宗凌[2] 杨小刚[1] 程明[1] 杨康骅[1] 

机构地区：[1]南昌大学第一附属医院骨科,南昌330006 [2]中山大学附属第一医院耳鼻喉科医院

出　　处：《中国修复重建外科杂志》2012年第6期647-651,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(30760255)~~

摘　　要：目的通过观察局部注射VEGF及VEGF抗体对小鼠植骨气囊模型中磨损颗粒诱导骨溶解的影响,探讨VEGF在人工关节无菌性松动中的作用。方法取人工髋关节翻修术取出的金属关节假体柄,参照真空球磨法体外制备磨损颗粒,PBS配制成浓度为10 mg/mL颗粒悬液。取8～10周龄雌性昆明小鼠50只,体重约25 g。10只作为气囊植骨模型颅骨供体。剩余40只小鼠随机分为4组(n=10),分别为空白对照组(A组)、颗粒组(B组)、VEGF刺激组(C组)和VEGF抑制组(D组);小鼠背部皮下注射无菌空气制备气囊,第8天切开气囊植入颅骨骨片,制备植骨气囊模型。于植骨后第1天B、C、D组气囊内注入0.5 mL颗粒悬液,A组注入0.5 mL PBS。气囊制备期间第6、7天及植骨后隔天,C、D组气囊内分别注射0.2 mL重组人VEGF和Bevacizumab溶液,A、B组注射0.2 mL生理盐水。植骨2周后取囊壁连同骨片行HE染色、实时荧光定量PCR及ELISA检测。结果各组小鼠均存活至实验完成。大体观察见A组气囊红肿程度轻,新生血管少;B、C、D组气囊明显红肿,可见较多渗出及新生血管,其中C组最重,B组次之,D组介于A、B组间。组织学及分子生物学检测发现,B组囊壁明显炎性反应及骨溶解,囊壁厚度、细胞密度及TNF-α、IL-1β、VEGF表达均较A组明显增加(P<0.05);C组囊壁炎性反应及骨溶解最显著,以上观察指标均高于B组(P<0.05);D组囊壁亦可见炎性反应及骨溶解,但以上指标均低于B组(P<0.05),高于A组(P<0.05)。结论 VEGF在人工关节无菌性松动中具有促进炎性反应及骨溶解作用,局部给予VEGF抗体可抑制磨损颗粒诱导的骨溶解。Objective To observe the effect of local injection of vascular endothelial growth factor（VEGF） and VEGF antibody on the wear particle-induced osteolysis in the mouse air pouch model and to investigate the role of VEGF in the process of aseptic loosening of prosthesis.Methods The stem of metal hip prosthesis was obtained from the revision surgery.Metallic wear particles were made by vacuum ball milling.Wear particles suspension was prepared into the concentration of 10 mg/mL with PBS.Fifty female Kunming mice（aged 8-10 weeks,weighing about 25 g） were selected.Of 50 mice,10 were used as the donors of bone graft,the other 40 were equally divided into control group（group A）,particle group（group B）,VEGF group（group C）,and VEGF inhibited group（group D）.Air pouches were made on the back of 40 mice by injecting sterile air subcutaneously.At 8th day,a graft of calvaria from the donor mice was implanted in air pouch.In groups B,C,and D,0.5 mL wear particles suspension was injected into the air pouches,and in group A,0.5 mL PBS was injected.Once a day at 6th and 7th days during the air pouch preparation and one time every two days after bone implantation,0.2 mL recombinant human VEGF（rhVEGF） and VEGF antibody（Bevacizumab） were injected into the air pouches in groups C and D,respectively.In group A and group B,0.2 mL saline was injected.Pouch tissues and bone were harvested at 2 weeks after bone implantation for HE staining,real-time uorescent quantitative PCR and ELISA analyses.Results All mice survived to the end of experiment.The gross observation showed that there were mild redness,swelling,and less neovascularization in air pouches in group A.There were obvious redness,swelling,and more exudative and neovascularization in groups B,C,and D,most obvious in group C,the next in group B,then in group D.The histological and molecular biological analysis showed that in ammatory responses and osteolysis were obvious in group B and the pouch membrane thickness,the cell density,transforming growth

关 键 词：人工关节无菌性松动 VEGF VEGF抗体 骨溶解 小鼠 

分 类 号：R687.4[医药卫生—骨科学]