小鼠颅骨缺损后骨形成过程的初步研究-第三军医大学学报.DOCVIP

Wnt信号参与小鼠颅骨缺损后骨形成过程的初步研究 徐 伟1，谢杨丽1，翁土军1，金 旻1，罗凤涛1，黄俊兰1，谭乔燕1，杜晓兰1，陈 林1,2 (400042 重庆，第三军医大学大坪医院野战外科研究所：创伤实验室，骨代谢修复中心，创伤烧伤与复合伤国家重点实验室1，康复科2) ［摘要］目的 利用改良的小鼠颅骨缺损模型，活体动态观察颅骨缺损后骨形成过程，并对Wnt信号相关基因进行检测，为颅骨再生修复机制及促进修复措施研究提供新的数据。方法 用直径2mm的金刚石钻头于小鼠右侧颅顶骨建立颅骨缺损模型，术后2周和4周通过Micro-CT及组织病理切片来观察缺损部位新生骨生长情况，测量其面积大小，定量分析缺损部位的骨形成能力。同时提取缺损后不同时间点缺损周围骨组织RNA，定量PCR检测成骨分化相关基因Cbfa1、OP和OC,以及Wnt通路基因Wnt3a、β-catenin及下游转录因子cyclin D1、Tcf1的表达。结果 术后2周Micro-CT扫描示缺损愈合百分比为（20.1±6.0）％；术后4周缺损进一步愈合，缺损愈合百分比为（30.1±5.0）％。定量PCR结果显示Cbfa1和OP在缺损后2周表达较高，而OC则在缺损后4周达到高峰。Wnt3a、β-catenin、cyclin D1和Tcf1的mRNA表达水平在缺损后2周和4周均较缺损前明显升高。结论 成功改良制作了小鼠颅骨缺损模型，并通过活体Micro-CT和组织病理染色对颅骨缺损后的修复过程进行了动态观察，进一步发现Wnt信号通路的激活可能参与颅骨缺损后骨形成过程。 ［关键词］颅骨缺损；骨形成；Wnt信号 ［中图法分类号］R-332；R683.5 ［文献标志码］ A Wnt signaling pathway participate in the bone formation process after calvarial defect Xu Wei 1, Xie Yangli 1,Weng Tujun1 ,Jin Min1, Luo Fengtao1 , Huang Junlan1 , Tan Qiaoyan1 , Du Xiaolan1 , Chen Lin1,2 ( 1：State Key Laboratory of Trauma，Burns and Combined Injury，Laboratory of Trauma，Institute of Surgery Research，Daping Hospital，Third Military Medical University，Chongqing，400042，ChinaDepartment of Rehabilitation Medicine , Daping Hospital，Third Military Medical University，Chongqing400042，China) ［Abstract］Objective To provide experimental basis for study of regeneration and repair of calvaria through observing bone formation dynamically and detecting Wnt signal-related genes using mouse calvarial defects models. Methods Calvarial defect of 2mm diameter was made in mouse right parietal bone using a diamond-coated trephine bit. Two and four weeks after defect creation, Micro-CT combined with histopathologically staining were utilized to observe calvarial bone formation. New bone areas were measured to quantitatively evaluate calvarial bone regeneration ability. Total RNA of the defect site was extracted at different time points before and after injury and then real time PCR was employed to evaluate the mRNA temporal expression patt