获得性异位骨化动物模型的系统综述

doi:10.3969/j.issn.1006-9771.2022.04.009

摘要/Abstract

摘要：

目的系统综述获得性异位骨化(AHO)相关动物模型,为研究疾病防治方案提供可靠造模方法。方法检索PubMed、Web of Science、中国知网、万方数据库自建库至2021年11月AHO动物模型相关文献,筛选并提取文献关键内容,采用文献归纳法对各型AHO动物模型进行分析与评价。结果最终共纳入20篇动物实验研究相关文献,可归纳为创伤后和神经源性两大类异位骨化动物模型,用于模拟AHO的发生发展过程。创伤后异位骨化动物模型主要有肌肉损伤、跟腱切断、肌肉损伤联合关节制动、髋关节损伤、异位植入、爆炸伤和烧伤7种。神经源性异位骨化动物模型主要包括脊髓损伤和脑外伤。目前实验室中常用跟腱切断法和成骨因子植入法造模,此类方法可靠、易行、成功率高,但无法准确解释临床中复杂条件下异位骨化的发病机制。基于爆炸伤、烧伤、神经损伤等条件下造模方法正在完善,为临床研究某些特殊病因所致异位骨化的分子生物学机制及防治方案提供了支持。结论目前存在的几种造模方法各有利弊,但均无法完整复制人类异位骨化的全部特征,因此,临床上在判定模型的选择方面尚无统一标准。针对疾病的不同病因,选用合适的动物模型,对不同类型AHO在早期制定有效干预措施至关重要。

关键词: 异位骨化, 获得性异位骨化, 动物模型, 系统综述

Abstract:

Objective To systematically summarize the animal models of acquired heterotopic ossification (AHO), and provide reliable modeling methods for the study of disease prevention and treatment programs.Methods Literatures about the animal models of AHO were researched from PubMed, Web of Science, CNKI and Wanfang Database till November, 2021. The important contents of the literatures were extracted, and the animal models of various types of AHO were evaluated and analyzed by literature induction.Results A total of 20 literatures related to animal experiments were included, which could be divided into two types: post-traumatic and neurogenic heterotopic ossification animal models, which were used to simulate the occurrence and development of AHO. Currently, seven different animal models were commonly used to study post-traumatic heterotopic ossification, such as muscle injury, achilles tenotomy, muscle injury combined with joint immobilization, hip injury, heterotopic implantation, blast injury and burn. The studies of neurogenic heterotopic ossification animal models mainly included spinal cord injury and traumatic brain injury. At present, the methods of achilles tenotomy and osteogenic factor implantation were commonly used in the laboratory, and with the advantages of reliability, feasibility and high success rate; however, they could not accurately explain the pathogenesis of heterotopic ossification under complicated clinical conditions. Therefore, the improvement of modeling methods based on explosion injury, burn, nerve injury and other conditions became the basis for clinical research of molecular biological mechanism, prevention and treatment of heterotopic ossification.Conclusion Current modeling methods have their own advantages and disadvantages, but none of them can completely replicate all the characteristics of human heterotopic ossification. Therefore, there is no unified standard in the selection of animal model in clinic. According to different etiology of the disease, the selection of appropriate animal models is crucial to study effective intervention for different types of AHO in the early stage.

Key words: heterotopic ossification, acquired heterotopic ossification, animal model, systematic review

蔡童欣,雷敏聪,周逸钧,孟殿怀. 获得性异位骨化动物模型的系统综述[J]. 《中国康复理论与实践》, 2022, 28(4): 429-438.

CAI Tongxin,LEI Mincong,ZHOU Yijun,MENG Dianhuai. Animal models of acquired heterotopic ossification: a systematic review[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(4): 429-438.

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纳入文献的基本特征"

纳入文献	国家	动物种属	n	模型类别	模型设计	造模周期	成功率	主要评估指标
Walton等^[12] (1983)	美国	绵羊	42	肌肉损伤模型	反复钝力作用于羊大腿深层肌肉致钝挫伤诱导异位骨化。	3~12周	16.6%	组织学分析
Moore-Lotridge等^[13](2019)	美国	ABCC6缺陷小鼠	101	肌肉损伤模型	心脏毒素注射到双侧小腿后室肌诱导肌肉损伤,并抑制巨噬细胞介导的纳米羟基磷灰石消退。	4周	80%	X线显微CT 组织学分析能量色散 X 射线
Yu等^[15](2021)	中国	雄性SD大鼠	80	跟腱切断模型	第1组：右后肢跟腱切断。第2组：仅同类皮肤切口。	10周	第1组：100%。第2组：0%。	X线组织学分析实时定量PCR
Tsailas等^[16](2009)	希腊	雄性新西兰白兔	18	肌肉损伤联合关节制动模型	石膏固定右膝和踝关节于过伸位+强制暴力活动按摩膝关节,每次5 min,每周6 d,持续5周。	10周	100%	X 线
Anthonissen等^[19](2015)	德国	雄性Wistar大鼠	20	髋关节损伤模型	右侧髋关节大转子尖端内侧钻通髓腔,股骨管扩孔至2 mm,钳夹臀大肌、臀中肌3 min。	12周	90%	显微CT 改良Brooker评分
Anthonissen等^[20](2016)	德国	雄性Wistar大鼠	50	髋关节损伤模型	第1组：右髋关节手术+额外的关节周围肌肉损伤。第2组：右髋关节手术+保护关节周围肌肉。	12周	第1组：96%,且程度更重。第2组：86.9%。	显微CT 改良Brooker评分
Li等^[11](2019)	美国	雄性C57BL/6 J小鼠	4	异位植入模型	双小腿肌注含BMP-2的水凝胶,右侧小腿肌注射心脏毒素。	2周	左侧：100%。右侧：100%,且更明显。	显微CT 组织学分析实时定量PCR 蛋白质印迹
Zotz等^[23](2012)	巴西	雄性Wistar大鼠	12	异位植入模型	双侧髂嵴收集自体骨髓0.35 mL分别注入其双侧股四头肌中。	5周	100%	血清碱性磷酸酶组织学分析火焰光度法评估股四头肌中钙含量
Polfer等^[26](2015)	美国	雄性SD大鼠	41	爆炸伤模型	第1组：单纯暴露于爆炸超压 (120±7) kPa中。第2组：股骨骨折和周围肌肉挤压伤后,损伤区股骨截肢。第3组：1+2组干预措施。	2~4周	第1组：0%。第2组：66.6%。第3组：100%。	X线显微CT 组织学分析
Epperson等^[25](2021)	美国	雌性兰布莱绵羊	8		利用AID模拟军事场景爆炸,同时模拟战场中创伤因素(如骨损伤、止血带、细菌、负压创伤治疗)。	24周	63%	X线显微CT 组织学分析电子背散射分析
Pavey等^[28](2015)	美国	雄性SD大鼠	48		爆炸伤模型+截肢部位接种1×10⁶个细菌菌落。第1组：接种耐甲氧西林金黄色葡萄球菌。第2组：接种鲍曼不动杆菌。第3组：接种不含细菌的载体。	12周	第1组：100%,且HO程度重。第2组：100%。第3组：100%。	显微CT 组织学分析
Tannous等^[29](2011)	美国	雄性SD大鼠	12		局部暴露于水下烈性炸药所致高能爆炸中,行指定部位(7只前肢,5只后肢)爆炸截肢。	4~24周	100%	X线
Peterson等^[33](2015)	美国	C57BL/6 小鼠	50	烧伤模型	60 ℃铝块置于小鼠背部17秒,制造30%体表面积皮肤烫伤,后行左下肢跟腱切断术。	9~15周	100%	显微CT
Debaud等^[35](2017)	法国	雌性C57B16小鼠	21	脊髓损伤模型	第1组：脊髓损伤手术+右后肢外周神经切断+术中双侧腘绳肌心脏毒素注射。第2组：相同的手术程序+术后第10天双侧心脏毒素注射。第3组：假脊髓损伤手术+右后肢外周神经切断+术中双侧腘绳肌心脏毒素注射。	4周	第1组：100%,且失神经支配侧异位骨化体积显著增加; 第2组：0%; 第3组：50 %。	显微CT 组织学分析实时定量PCR
Genêt等^[37](2015)	美国	雌性C57BL/6小鼠	113		T₇~T₈间脊髓横断,且右侧腘绳肌内注射心脏毒素。	2周	99.12%,仅发生在注射心脏毒素侧肢体。	X线显微CT 组织学分析实时定量PCR
Kang等^[38](2014)	美国	雄性C57/BL6小鼠	24		实验组：T₁₀节段脊髓完全性损伤,后肢四头肌注射不同剂量(0 g、0.25 g、0.5 g) BMP-2。对照组：假脊髓损伤+相应剂量BMP-2肌注。	2周	0.5 µg BMP-2可显著增加四头肌异位骨化含量。 0.25 µg BMP-2仅使实验组产生异位骨化。 0 µg BMP-2不能诱发异位骨化。	显微CT 组织学分析实时定量PCR
Shi等^[42](2017)	中国	雄性 SD大鼠	66	脑损伤模型	重物撞击顶叶区制造轻度创伤性脑损伤结合右后肢跟腱切断术。	10周	100%	X线组织学分析
Ju等^[43](2019)	中国	雄性SD大鼠	44		联合脑外伤+烧伤+跟腱切断术。	10周	100%	X线组织学分析实时定量PCR Western Blotting检测
Brady等^[44](2020)	澳大利亚	雄性SD大鼠	36		第1组：三重损伤(脑外伤+股骨骨折+肌肉损伤)。第2组：单纯外周损伤(假脑外伤+股骨骨折+肌肉损伤)。第3组：假损伤对照组(假脑外伤+假股骨骨折+假肌肉损伤)。	6周	第1组：70%。第2组：20%。第3组;0%。	显微CT 组织学分析实时定量PCR Western blotting检测
Anthonissen等^[46](2020)	德国	雄性Wistar大鼠	40		第1组：单纯行髋关节手术。第2组：髋关节手术+中度脑外伤。第3组：髋关节手术+重度脑外伤。第4组：仅重度脑外伤。	12周	第1组：90%。第2、3组：100%,且3组比2组骨化体积增加。第4组：0%。	显微CT 组织学分析

表1

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