Experimental animal models of osteonecrosis of the femoral head: classification and experimental applications

doi:10.3969/j.issn.2095-4344.2014.36.024

Abstract

Abstract:

BACKGROUND: So far there is not an ideal animal model for clinical research of osteonecrosis of the femoral head, which hinders its development.
OBJECTIVE: To compare the merits and characteristics of different methods and various animal models, and to provide a reference for selection of models of osteonecrosis of the femoral head.
METHODS: Literature search was performed in databases of PubMed, EMBASE, Wanfang, and China National Knowledge Infrastructure, published from 2006 to 2014 by computers. The key words were “Osteonecrosis of the Femoral Head, Animal model, Vivo Experiment, Legg Calve Perthes disease, Osteoarthritis, Collapse of the Femoral Head, Trauma, Corticosteroid, Alcohol, Liquid Nitrogen” in English and Chinese, respectively. Finally, 48 articles were included.
RESULTS AND CONCLUSION: Traumatic osteonecrosis of the femoral head models simulates the most common clinical type, which is more effective in Legg Calve Perthes disease. Glucocorticoid-induced osteonecrosis of the femoral head models is more suitable for etiology, pathology and comprehensive research. Physical osteonecrosis of the femoral head models could be used for evaluating treatment methods, such as surgery. The ideal animal models should be similar with human in anatomy, histology and biomechanics; pathology, pathophysiology, and radiographic progression of osteonecrosis of the femoral head can simulate the whole process of human osteonecrosis of the femoral head, and it is easy to be obtained, cost-effective and good repeatable.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: models, animal, femur head necrosis, femur head

CLC Number:

R318

Wang Yan-mao, Zhang Chang-qing. Experimental animal models of osteonecrosis of the femoral head: classification and experimental applications[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(36): 5879-5884.

Figures/Tables 1

2.1 创伤性股骨头坏死模型临床上，股骨颈骨折所造成的股骨头缺血是股骨头坏死发病的重要原因之一，对实验动物进行手术，例如结扎股骨颈、切除股骨头圆韧带等，从而阻断股骨头血供模拟临床上的发病过程是创伤性股骨头坏死模型的主要方法。小猪模型是一个经常被用作骨坏死研究的动物模型。Zhang等[4]发现在未成熟小猪的解剖结构中，股骨干和近端骨骺间没有血管交通支，为结扎股骨颈造成股骨头缺血坏死提供了理论基础。Aruwajoye等[5]通过在股骨颈部紧密结扎并切断圆韧带的方法在8只五六周大的Yorkshire小猪单侧未成熟的股骨头上建模，而另一侧作为对照组。术后2周，对其中3只的坏死股骨头进行苏木精-伊红染色和TUNEL标记法，另外5只进行结合定量散射电子显像、纳米压痕技术、微裂纹评估进行检测；组织学结果显示在骨小梁及骨髓区分布有大量的坏死细胞，纳米压痕技术软骨下区微裂纹的密度显著增加。此模型是研究LCPD的经典模型，可用于发病机制及防治措施的探索。Shapiro等[6]用同样的方法成功地构造了小猪股骨头坏死模型，并且证实了坏死的股骨头会继发性地引起相邻髋臼的畸形，同时为髋臼发育畸形这类至今了解甚少的疾病提供了研究模型。而Kim等[7]在上述方法造模的同时，对小猪的一侧后肢进行截肢以避免负重，结果发现局部的避免负重可以减少坏死股骨头的畸形，并促进血管的再生及骨骺中血栓的吸收，对临床的治疗有一定的参考意义。有学者用3个月大的新西兰兔造模[8]，手术切除髋关节囊，完全分离环状韧带及软组织，切断股骨颈基底部，用不可吸收线将分离的股骨头固定在髋臼上以模拟股骨头坏死早期临床阶段，通过CT、MRI及组织学方法发现骺板两侧有坏死斑的分布，而软骨保存，这与人类早期股骨头坏死相似。Hofstaetter等[9]使用成年的新西兰兔建造股骨头坏死，实验者同时破坏了股骨头的髓内供血及髓外供血：髓内向股骨颈钻孔到达髓腔，髓外环形损伤股骨颈的骨膜的同时结扎圆韧带。术后使用影像学及组织学的方法进行评价，6个月未发现股骨头塌陷，12个月只有13.3%的动物出现了股骨头塌陷，并且只有软骨下骨的骨折，股骨头大体形态并没有显著改变，12个月的造模周期也比较难以接受。范猛等[10]用手术方式将SD大鼠的髋关节脱位，切断圆韧带，并破坏股骨颈近端骨膜上的血供，通过影像学及多个组织学方法证实了股骨头出现塌陷，骨小梁结构改变，破骨细胞明显活跃，成功建造了大鼠创伤性股骨头坏死模型。Gao等[11]通过低速钻头对狗的股骨颈造成Garden IV型骨折，从术后2个月的X射片上观察到未经治疗的股骨颈骨折造成了典型的股骨头坏死。Hang等[12]使用手术方式对狗造成股骨颈骨折及髋关节脱位，并通过3枚克氏针进行内固定的方式建立了股骨头坏死的动物模型。术后4周X射线显示骨折愈合，骨密度均匀，未出现硬化带及新月征或者塌陷；但苏木精-伊红染色显示有坏死骨小梁并伴微结构的改变。这种方法模拟了临床上常见的股骨颈骨折手术后出现股骨头坏死的过程，曾有学者做过类似研究，但结果不一，重复性较差。创伤性股骨头坏死动物模型在LCPD的造模上具有重要作用，但对于成人的股骨头坏死，其造模操作复杂、周期长、重复性差、成功率低等缺点较为明显，并不是理想的股骨头坏死模型。 2.2 激素性股骨头坏死的模型当今，许多疾病，例如系统性红斑狼疮、肾病综合征、支气管哮喘等都需要长期大量的服用激素进行治疗，但其带来的并发症如股骨头坏死等同样是显而易见的。Tong等[13]通过基因芯片技术发现激素性股骨头坏死小鼠有多种基因的表达发生了显著变化，这些改变的基因包括：血管形成基因、凋亡基因、信号转导基因、氧化应激相关基因等类型，因此可以得出结论是有多种基因参与到了激素性股骨头坏死形成的过程中。 2.2.1 单纯激素性股骨头坏死模型大剂量及长时间使用糖皮质激素是引起股骨头坏死的一个高危因素，因此国内外有学者给予兔注射甲基强的松龙建模。Yamaguchi等[14]对30只日本白兔单次肌肉注射20 mg/kg的甲强龙，注射后3周发现股骨及肱骨干骺端有73.3%的兔发生了骨坏死，动物死亡率为0%；Iwakiri等[15]运用相同的方法通过影像学及组织学方法发现有83%的股骨头坏死率。而Kuribayashi等[16]则表示4周股骨头坏死率为70%，但动物死亡率为20%。在兔的激素性骨坏死模型中，骨坏死的病灶还会出现在肱骨近端等其余长骨处，不仅仅局限于股骨头，而其股骨头坏死多为早期表现，这些兔的模型均不是理想的股骨头坏死实验动物模型，但可以为股骨头坏死的发病机制研究作为参考。 Nozaki等[17]对自发性高血压脑卒中大鼠运用甲基强的松龙皮下注射(1 000 mg/60 kg类似于人类激素冲击疗法的剂量)，组织学检测股骨头坏死的发生率要显著大于对照组(P < 0.001)。Tian等[18]对SD大鼠连续8周注射甲基强的松龙(20 mg/kg, 1次/周)，也成功构建了大鼠的股骨头坏死模型，并证实了激素引导的股骨头坏死模型中，Toll样受体4信号通路(TLR4 signaling pathway)干扰了正常的免疫反应，从而进一步说明了免疫反应的破坏在股骨头坏死中扮演了重要的角色。大鼠有90%的基因与人类一致，喂养时占地面积小，较为经济，而且从解剖学上看来，大鼠的股骨头也同样具有颈干角和前倾角，在生物力学方面与人类相似。但大鼠的股骨头坏死激素模型也有着其局限性：造模周期较长，股骨头形态较难形成塌陷等等。 Erken等[19]通过对25只Leghorn鸡每周肌肉注射 3 mg/kg甲基强的松龙建立了的股骨头坏死模型，造模的14周过程中有48%的动物死亡，组织学发现了软骨下骨小梁的破坏，但未出现股骨头的塌陷(图1)。鸡与人类同为双足行走动物，股骨头的生物力学相近，这是相较于其他四足行走动物的优势，其股骨头坏死的病理病生过程与人类也相似。但动物死亡率极高，坏死大体表现不典型，基因与人类相差甚远，这些因素造成了鸡作为股骨头坏死模型运用较少的原因。"

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