兔股骨髁临界性骨缺损动物模型制备及临界骨缺损值

《中国组织工程研究》 Chinese Journal of Tissue Engineering Research·研究原著·徐石庄，男，1986年生，江苏省连云港市人，汉族，徐州医科大学在读硕士，主治医师，主要从事骨关节疾病研究。

通讯作者：赵凤朝，博士，副教授，徐州医科大学，江苏省徐州市 221000文献标识码:B投稿日期：2019-09-09 送审日期：2019-09-10 采用日期：2019-10-19 在线日期：2020-01-04Xu Shizhuang, Master candidate, Attending physician, Department of Orthopedics, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu Province, ChinaCorresponding author: Zhao Fengchao, MD, Associate professor, Department of Orthopedics, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu Province, China兔股骨髁临界性骨缺损动物模型制备及临界骨缺损值徐石庄1，王 进2，潘文振1，刘 磊1，杨冠杰1，赵凤朝1 (1徐州医科大学附属医院骨科，江苏省徐州市 221000；2苏州大学附属张家港医院，江苏省苏州市 215000)DOI:10.3969/j.issn.2095-4344.2614 ORCID: 0000-0002-1257-965X(徐石庄)文章快速阅读：文题释义：临界性骨缺损：首先定义为自然状况下骨缺损不进行任何处理无法自愈的最短的骨缺损尺寸。

随后考虑到观察实验动物完整的生命周期是非常困难的，将临界性骨缺损值定义为在实验期间物种不能自行愈合的最短骨缺损尺寸。

动物模型：是在医学研究中建立的模拟人类疾病表现的动物，骨组织工程中建立临床相关的测试动物模型来研究材料的生物相容性、降解、力学性能以及与宿主组织的相互作用，是体外实验和人体临床试验之间的关键一步。

摘要背景：兔股骨远端骨缺损模型被研究者们广泛用于骨缺损替代骨组织工程材料的测试，但对于兔股骨髁圆柱形骨缺损模型的大小文献报道不一，直径分布在5-9 mm ，深度8-12 mm ，目前尚无统一的标准。

目的：建立兔股骨髁不同尺寸骨缺损模型，确定兔股骨髁临界性骨缺损尺寸。

方法：6月龄雄性新西兰白兔18只，随机分为3组，每组各6只，分别建立骨缺损模型，骨缺损直径依次为5，6，7 mm ，深度均为10 mm ，双侧手术，共计12侧。

分别于术后第1天及术后第4，8，12周行CT 扫描及三维重建，CT-Hedberg 评分评价骨缺损愈合情况；于术后12周处死新西兰白兔，取出股骨髁缺损样本，通过大体观察和苏木精-伊红染色分析缺损区愈合情况。

实验方案经徐州医科大学实验动物道德伦理委员会批准。

结果与结论：①术后所有兔均存活，术后12周大体观察示：直径5 mm 组缺损由新生骨组织充填，股骨髁塑形良好，骨缺损基本完全修复；直径6 mm 组、直径7 mm 组骨缺损区可见明显凹陷，新生骨组织较少，骨缺损未修复；②CT 图像示：术后第4，8周，直径5 mm 组缺损区逐渐减小，断端桥接；直径6 mm 、直径7 mm 组缺损区仅周边有少量新生骨长入，缺损面积较前稍减小；术后第12周可见直径5 mm 组皮质骨结构完整、连续，骨缺损基本完全修复；直径6 mm 组骨缺损部分修复；直径7 mm 组缺损未修复，仍可见明显缺损空腔存在；③CT-Hedberg 评分显示，术后各时间点直径6 mm 组评分显著低于直径5 mm 组(P < 0.05)；与直径7 mm 组比较差异无显著性意义(P > 0.05)；④组织学结果示：术后12周直径5 mm 组缺损区出现排列不规则的骨小梁结构，并可见大量新生骨组织填充，其他2组在骨缺损周边可见部分新生骨小梁存在，但缺损区新生骨组织填充较少；⑤结果说明，在12周的实验观察期内，在缺损深度同为10 mm 的条件下，直径>6 mm 的股骨髁缺损未能自行愈合，而直径<6 mm 的股骨髁缺损基本完全修复。

此结果符合临界骨缺损的标准，故直径6 mm 可作为兔股骨髁临界骨缺损值。

关键词：兔；股骨髁；临界性骨缺损；缺损尺寸；动物模型 中图分类号：R446；R496；R318Preparing an animal model of critical femoral defect in rabbit femoral condyle and the critical bone defect sizeXu Shizhuang 1, Wang Jin 2, Pan Wenzhen 1, Liu Lei 1, Yang Guangjie 1, Zhao Fengchao 1 (1Department of Orthopedics, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu Province, China; 2Zhangjiagang Hospital of Soochow University, Suzhou 215000, Jiangsu Province, China)徐石庄，王进，潘文振，刘磊，杨冠杰，赵凤朝. 兔股骨髁临界性骨缺损动物模型制备及临界骨缺损值[J].中国组织工程研究，2020，24(20):3191-3195. DOI:10.3969/j.issn.2095-4344.2614AbstractBACKGROUND: Rabbit model of distal femoral bone defect has been widely used to test bone tissue engineering materials for bone defects. However, there is no uniform standard for the size of the cylindrical bone defect model of the rabbit femoral condyle, which ranges 5-9 mm in diameter and 8-12 mm in depth.OBJECTIVE: To establish the bone defect model of adult rabbit femoral condyle with different sizes and to determine the critical bone defect size of the femoral condyleMETHODS: Eighteen male New Zealand White rabbits aged 6 months were randomly divided into three groups according to the diameter of bone defect: 5 mm diameter group, 6 mm diameter group, and 7 mm diameter group. The defect depth was 10 mm. These rabbits underwent bilateral radial surgery, a total of 12 sides. Computed Tomography (CT) scan and three-dimensional reconstruction were performed at 1 day, 4, 8, 12 weeks after surgery. The CT-Hedberg score was used to evaluate the healing of bone defects. The rabbits were sacrificed at 12 weeks after surgery, and the femoral condyle specimens were taken out. Healing of the defect was analyzed by gross observation and hematoxylin-eosin staining. The study protocol was approved by the Animal Ethics Committee of Xuzhou Medical University.RESULTS AND CONCLUSION: All rabbits survived after surgery. The gross observation showed that the defect of 5 mm diameter group was filled with new bone tissue, the femoral condyle was well shaped, and the bone defect was completely repaired. In 6 mm and 7 mm diameter groups, depressed deformation was obviously observed in the defect area, with less new bone tissue, and the defect was was not repaired. The CT images showed that the defect area of 5 mm diameter group gradually decreased, and the broken ends of the defect were bridged. In the defect area of 6 mm and 7 mm diameter groups, only a small amount of new bone tissue was implanted, and the defect area was slightly reduced. At the 12th week after surgery, the cortical bone structure of 5 mm diameter group was intact and continuous, the femoral condyle was well shaped, and the bone defect was completely repaired. The defects of 6 mm and 7 mm diameter groups were partially or not repaired, and the defect cavity was still visible in the 7 mm diameter group. The CT-Hedberg scores of 6 mm diameter group were significantly lower than those of 5 mm diameter group at different time points (P < 0.05), and there was no significant difference in the CT-Hedberg scores between 6 mm and 7 mm diameter groups (P > 0.05). Histological results showed that there were irregular trabecular structures in the defect area of 5 mm diameter group, with a large amount of new bone tissue. In the other two groups, there were some new bone trabeculae around the bone defect, but the defect area was less filled with new bone tissue. During the 12-week observation period, the femoral condyle defect with a diameter of > 6 mm and a depth of 10 mm could not heal spontaneously, while the defect with a diameter of < 6 mm could be completely repaired, which met the criteria of critical bone defect. Therefore, the diameter of < 6 mm could be used as the critical bone defect size of rabbit femoral condyle.Key words: rabbit; femoral condyle; critical bone defect; defect size; animal model0 引言Introduction在全膝关节翻修或者复杂初次膝关节置换中经常会遇到股骨远端骨缺损的情况，如果股骨远端骨缺损得不到处理将会导致膝关节翻修和置换手术的失败[1]。