血管内皮生长因子在磨损颗粒诱导骨溶解中的作用研究

Role of Vascular Endothelial Growth Factor in Wear Debris-induced Osteolysis

目的运用小鼠植骨气囊模型,观察局部注射血管内皮生长因子(VEGF)及VEGF抑制剂bevacizumab对磨损颗粒诱导骨溶解的影响,进一步探讨VEGF在人工关节无菌性松动中的作用。方法将40只小鼠按随机数字表法分成空白对照组、颗粒组、VEGF刺激组、VEGF抑制组,每组10只。在小鼠背部皮下注射无菌空气形成气囊,继而将近交系小鼠(10只)颅骨片植入气囊。在颗粒组、VEGF刺激组、VEGF抑制组小鼠气囊内注入磨损颗粒,空白对照组则注入PBS。气囊植骨后隔天分别在VEGF刺激组和VEGF抑制组小鼠气囊内注入重组人血管内皮生长因子(rh-VEGF)和bevacizumab进行干扰,颗粒组和空白对照组则注入生理盐水。植骨2周后取囊壁和植入颅骨组织进行HE染色检测炎症反应情况及骨片吸收情况;采用抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞的分化成熟情况;取组织匀浆液应用ELISA方法检测炎性细胞因子IL-1β、TNF-α浓度;qRT-PCR进一步检测炎性细胞因子IL-1β、TNF-αmRNA表达水平。结果磨损颗粒诱导囊壁产生明显炎症反应及骨溶解。颗粒组囊壁厚度、细胞密度及TNF-α、IL-1β等炎性因子表达均较空白对照组明显增加(均P<0.05),局部注射VEGF进一步促进磨损颗粒诱导囊壁产生炎症反应及骨溶解,而bevacizumab则使囊壁炎症反应及骨溶解受到了明显抑制。结论磨损颗粒在体内可以诱导VEGF的表达,而VEGF抑制剂可抑制其诱导的炎症反应及骨溶解。

Objective To observe the effects of local injection of vascular endothelial growth factor（VEGF） and VEGF inhibitor bevacizumab on the wear debris-induced osteolysis by using mouse air pouch model,and to investigate the role of VEGF in aseptic loosening of artificial joints.Methods Forty mice were randomly divided into control group,particle group,VEGF group and VEGF inhibitor group,10 in each group.Air pouches were developed by hypodermic injection of sterile air into the back of mice,followed by implantation of bone grafts of calvaria from syngeneic mouse（n=10）donors.Mice in particle group,VEGF group and VEGF inhibitor group were given the injection of wear particles into air pouches.Mice in control group were given PBS.After bone implantation,recombinant human VEGF（rh-VEGF） and VEGF inhibitor bevacizumab were injected into air pouches in VEGF group and VEGF inhibitor group,respectively.Normal saline was injected into air pouches in particle group and control group.Pouch wall and implanted calvaria tissues were harvested 2 weeks after bone implantation for molecular and histological analysis.Inflammatory response and bone osteolysis were detected by HE staining,osteoclast differentiation and maturation by tartrate-resistant acid phosphatase staining,IL-1β and TNF-α concentrations by ELISA,and IL-1β and TNF-α expression by qRT-PCR.Results Compared with control group,wear particles significantly induced inflammatory response and bone osteolysis,and obviously increased the thickness of pouch wall,cell density and the expression of TNF-α and IL-1β（P〈0.05）.The injection of VEGF into the air pouches promoted the inflammatory response and bone osteolysis induced by wear particles,but the bevacizumab inhibited the role of wear particles.Conclusion Wear particles can induce VEGF expression in vivo,and VEGF inhibitor can effectively inhibit wear particle-induced inflammatory response and osteolysis.