腺病毒介导NELL1促进骨溶解后的高质量骨再生  

Adenovirus-mediated NELL1 promotes bone regeneration in calvarial osteolysis model

作　　者：郭旭[1,3] 彭江[1] 王玉[1] 赵斌[1] 袁玫[1] 孟昊业[1] 张莉[1] 张新立 许文静[1] 汪爱媛[1] 卢世璧[1] 

机构地区：[1]解放军总医院骨科研究所,北京100853 [2]加利佛尼亚大学口腔颅面外科研究所,加利佛尼亚90095 [3]河北沧州市中心医院,061001

出　　处：《中国矫形外科杂志》2012年第9期834-837,共4页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:30872621)国家自然科学基金项目(编号:30872633);国家863主题项目(编号:2012AA020502)

摘　　要：[目的]研究具有特异性成骨作用的NELL1因子对磨屑所致骨溶解的影响,为骨溶解病的治疗提供一种新方案。[方法]建立聚乙烯(Polyethylene,PE)颗粒诱导的颅骨骨溶解模型,68只Balb/c小鼠随机分成4组,即假手术组:术后1 d颅顶皮下注射0.1 ml生理盐水;颗粒对照组:植入10 mg PE颗粒,术后1 d注射0.1 ml生理盐水;实验组(Ad-GFP-NELL1组):植入10 mg PE颗粒,术后1 d注射0.1 ml(109 pfu)NELL1荧光重组腺病毒载体(Ad-GFP-NELL1);安慰剂组(Ad-GFP组):植入10 mg PE颗粒,术后1 d注射0.1 ml(109pfu)荧光重组腺病毒载体(Ad-GFP)。术后10 d采用活体荧光成像检测NELL1及GFP表达情况,并于处死后进行Micro-CT、组织学及生物力学的检测。[结果]活体荧光证实实验组与安慰剂组小鼠颅顶有NELL1及GFP的高度表达,Micro-CT 3D图像观察实验组小鼠的颅骨骨溶解所致骨量丢失明显减少,所选兴趣区(ROI)的骨体积(BV)、骨密度(BMD)、骨小梁厚度(Tb.Th.3D)、骨表面积体积比(BS/BV)均明显优于对照组及安慰剂组。组织学切片观察到实验组的骨再生明显增多。另外,实验组小鼠的颅骨弹性模量[(2.50±0.12)MPa]也明显优于对照组[(0.60±0.20)MPa]及安慰剂组[(0.82±0.09)MPa](OP<0.01)。[结论]腺病毒介导表达的成骨因子NELL1能通过促进高质量的骨再生平衡磨屑所致骨溶解的骨量丢失,是治疗骨溶解的一种潜在性解决方案。[ Objective] To investigate the therapeutic effects of a craniosystosis -associated molecule, NEL -like molecule -1 （NELL1 ; NEL, a protein strongly expressed in neural tissue encoding epidermal growth factor like domain） on osteolysis induced by polyethylene （PE） -particle debris. [ Methods ] A murine calvarial osteolysis model was established with in vivo adenovirus （Ad） - mediated gene transfer. In total, 68 female Balb/c mice were randomly assigned to 4 groups for treatment one day post - operation： SHAM surgery with 0. 1 ml saline alone, PE particles with O. 1 ml saline, and PE - particles andadenovirus - green fluorescent protein with NELL1 （Ad -GFP - NELL1, O. 1 ml） or Ad - GFP （0. lml） at 1×10^9 plaque -forming units/ml diluted in saline. GFP and NELL1 delivery in vivo after injection were certified by optical imaging at 10 days post -operation, and then all mice were sacrificed for analysis by 3 -D micro -computed tomography （micro -CT）, histology and biomechanical testing. [ Results] Exogenous NELL1 and GFP were expressed in the head for at least l0 days after Ad - GFP - NELL1 injection. Serial 3 - D micro - CT images and testing of bone volume, bone mineral density, trabecular thickness, bone surface density revealed the new bone promoted with Ad - GFP - NELL1 injection could almost compensate the PE -induced osteolysis and regenerate significantly better than with PE and/or Ad - GFP treatment. The elastic modulus was significantly greater with Ad - GFP - NELL1 than with PE and/or Ad - GFP （P 〈 0. 01 ） . [Conclusion] Ad- GFP- NELL1 gene transfer effectively reversed the calvarial osteolysis model by promoting bone regeneration, and it may be a novel treatment for osteolysis.

关 键 词：NELL1 骨溶解 磨屑 骨再生 Micro—CT 

分 类 号：R687.4[医药卫生—骨科学]