小鼠股骨骨折合并坐骨神经损伤的标准化动物模型构建与评价  

Construction and evaluation of a standardized animal model of femoral fracture combined with sciatic nerve injury in mice

作　　者：丁云[1] 隋钧皓 侯宜锦 章浩[1] DING Yun;SUI Jun-hao;HOU Yi-jin;ZHANG Hao(Department of Orthopaedics Trauma,The First Affiliated Hospital of Naval Medical University(Second Military Medical University),Shanghai 200433,China;Institute of Biomedical Engineering,School of Health Science and Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China)

机构地区：[1]海军军医大学(第二军医大学)第一附属医院创伤骨科,上海200433 [2]上海理工大学健康科学与工程学院生物医学工程研究所,上海200093

出　　处：《海军军医大学学报》2023年第2期214-221,共8页Academic Journal of Naval Medical University

基　　金：国家自然科学基金(81702666)。

摘　　要：目的建立并评价小鼠股骨骨折合并坐骨神经损伤的标准化动物模型。方法将96只雄性C57BL/6J小鼠随机分为两组:股骨骨折合并坐骨神经损伤组(48只)和股骨骨折组(48只)。股骨骨折合并坐骨神经损伤组小鼠通过止血钳钳夹致左侧坐骨神经损伤,并于同侧进行股骨开放性骨折联合髓内针固定,制作小鼠股骨骨折合并坐骨神经损伤模型;股骨骨折组小鼠仅游离左侧坐骨神经,不做钳夹处理,同时进行左侧股骨开放性骨折联合髓内针固定。术后仔细观察两组小鼠的行为学变化。于术后第3、5、7、10、14、18天取两组各36只小鼠的左侧坐骨神经,每个时间点每组6只,通过免疫组织化学染色观察坐骨神经的组织学变化。于术后第7、14、21、28天通过股骨X线片、Micro-CT、H-E染色及番红固绿染色分析骨折愈合情况。结果成功建立小鼠股骨骨折合并坐骨神经损伤模型,行为学观察与评估发现术后第14天股骨骨折合并坐骨神经损伤组小鼠步态基本恢复正常;坐骨神经免疫组织化学染色显示,术后第14天股骨骨折合并坐骨神经损伤组小鼠的坐骨神经纤维连续性基本恢复;Micro-CT检查显示在各时间点,股骨骨折合并坐骨神经损伤组的骨体积均小于股骨骨折组(P均<0.01),骨体积百分比也均低于股骨骨折组(P均<0.01)。骨切片组织形态学结果显示,股骨骨折合并坐骨神经损伤组小鼠术后第7天骨折断端几乎不存在骨痂;术后第14天出现少量软骨痂,主要由未分化的间充质干细胞构成;术后第21天软骨痂开始钙化,骨痂中存在部分肥大的软骨细胞;术后第28天骨折断端有新生骨形成,含有大量肥大的软骨细胞、成骨细胞与骨细胞。结论小鼠股骨骨折合并坐骨神经损伤模型构建成功,该模型愈合过程良好,并较单纯股骨骨折的愈合有所延迟,可较好地模拟临床骨折合并周围神经损伤后的骨修复及愈合过程。Objective To establish and evaluate a standardized animal model of femoral fracture combined with sciatic nerve injury in mice.Methods A total of 96 male C57BL/6J mice were randomly divided into 2 groups:femoral fracture combined with sciatic nerve injury group(n=48)and femoral fracture group(n=48).The left sciatic nerves were crushed by hemostatic forceps,and the ipsilateral femurs were cut andfixed with intramedullary nails in the bone marrow.And then,the mouse model of femoral fracture combined with sciatic nerve injury was established in femoral fracture combined with sciatic nerve injury group.Mice in the femoral fracture group underwent fracture of the left femurs by intramedullary nailsfixation,and the left sciatic nerves were dissociated,but not clamped.Behavioral observation of postoperative mice was performed in both groups.The left sciatic nerves of mice were collected in both groups(36 mice each)on the 3^(rd),5^(th),7^(th),10^(th),14^(th)and 18^(th)d after operation,with 6 mice in each group at each time point,and the histological changes of the sciatic nerves were observed by immunohistochemical staining.On the 7^(th),14^(th),21stand 28^(th)d after operation,the fracture healing was analyzed by femoral X-ray radiographs,micro-CT,hematoxylin-eosin staining and safranine-fast green staining.Results A mouse model of femoral fracture combined with sciatic nerve injury was successfully established.Behavioral observation and assessment revealed that the gait of mice in the femoral fracture combined with sciatic nerve injury group basically returned to normal on the 14thd after operation.The immunohistochemical results of sciatic nerves showed that the structural continuity of sciatic nervefibers was basically recovered in the mice of femoral fracture combined with sciatic nerve injury group on the 14thd after operation.Micro-CT showed that the bone volumes of femoral fracture combined with sciatic nerve injury group were significantly lower than those of femoral fracture group at all time points(all P<0.01

关 键 词：股骨骨折 坐骨神经损伤 动物模型 骨折愈合 

分 类 号：R683.421[医药卫生—骨科学]