三足负重犬股骨头坏死模型的生物力学相关性研究

目的研究股骨头坏死病程各阶段的生物力学改变,探讨其生物力学发生作用的机制。方法取杂种犬24只,固定一侧前肢建立三足负重犬模型。随机取三足负重犬一侧后肢为实验侧,在股骨头内注射无水酒精,致股骨头坏死;对侧为对照侧,在股骨头内注入等量生理盐水。造模后1、3、6及12周处死动物,每组6只,分别对两组股骨头行大体观察以及放射学、组织学、生物力学检测。结果术后3周股骨头坏死侧点压硬度及中部松质骨弹性模量相对对照侧分别下降29%和32.9%,此时仅MRI可见股骨头出现低密度坏死区,组织学主要表现为骨坏死。术后6、12周股骨头坏死侧点压硬度相对对照侧分别下降了45.5%和48.7%,中部松质骨弹性模量相对对照侧分别下降了34.1%和32.4%。6周时坏死侧X线可见股骨头内密度不均,组织学表现为坏死和修复反应并存,坏死区向软骨下骨区进展。12周时坏死侧X线可见股骨头负重区下出现局部骨密度减低区,组织学出现关节软骨面塌陷和关节间隙狭窄。结论生物力学是股骨头病程进展的重要影响因素;病程进展中,特别是修复期力学性能的显著下降可能是后期塌陷的最直接原因。治疗股骨头坏死不仅应促进骨修复,更应提供病变区一个有利、稳定的生物力学环境。     

骨内注射无水乙醇建立兔股骨头坏死模型

背景：目前还难以建立合适的早期股骨头坏死的动物模型。 目的：建立一个简单、标准和可靠的股骨头坏死动物模型用于实验研究。 方法：X射线透视下将新西兰兔右侧股骨头中心钻孔并注入无水乙醇,左侧不做处理做对照。经过2,4和6周麻醉下处死动物获取股骨头。 结果与结论：大体及X射线观察显示,造模侧股骨头第2周关节软骨颜色变暗、骨质密度不均匀;第6周,关节面有轻微凹陷,骨质低密度影较前进一步增大。MRI显示,造模侧股骨头第2周,T1加权股骨头负重区显示线样或不规则低信号,T2加权呈高信号;第6周,股骨头变性,软骨下骨折,关节面塌陷,新月体形成。对照侧第2,4,6周股骨头结果均常。组织病理学观察显示,造模侧股骨头第2周后,骨细胞核固缩、变性坏死。结果证实,2周后兔股骨头均发生了部分坏死,股骨头坏死的完整的外观形态,大体循环和关节软骨与人类早期股骨头坏死是相似的,提示实验建立了良好的股骨头坏死动物模型。     

液氮冷冻兔股骨头坏死模型制备新方法及可靠性评价

目的 建立一种可靠的并可用于治疗和研究新的液氮冷冻法制成的兔股骨头坏死动物模型。方法 采用成年新西兰大白兔21只,无菌条件下手术分离臀肌,切断股骨头圆韧带,显露股骨头。用医用棉签蘸取液氮,对股骨头进行3次冷冻3次复温,制备兔双侧股骨头坏死模型。于术后3、7d及2、4、6、8周进行X线摄片,股骨头大体形态和组织病理学观察。结果 X线摄片结果显示,制模2周时股骨头密度增高;4周时出现密度不均影像;6周时外形出现不规则变化,边缘有透亮区;8周时开始塌陷,关节间隙增大,骺板模糊。股骨头大体形态随时间推移,其受损程度呈加重演变特点。制模3 d的股骨头组织切片可见软骨细胞和骨细胞坏死;2周见骨小梁断裂、排列紊乱;4周见髓腔脂肪细胞坏死,内有新生血管及增生纤维组织;6周时出现匍行附着;而8周可见新骨沉积性生长,骺板处细胞挤压、变形。结论 本实验中建立的液氮冷冻股骨头坏死模型新方法具有创伤性小、贴近人股骨头坏死病理演变规律的优点,为干细胞移植等治疗和研究提供了一种新的模型制备方法。     

Experimental rat model for alcohol‐induced osteonecrosis of the femoral head

Alcohol‐induced osteonecrosis of the femoral head (ONFH) is observed in alcohol abusers and patients with alcoholic fatty liver disease. It has been reported that Toll‐like receptor 4 (TLR4) signalling plays a crucial role in the pathogenesis of alcoholic fatty liver disease. We previously reported a corticosteroid‐induced ONFH rat model, and suggested that TLR4 signalling contributes to the pathogenesis of ONFH. Thus, it is thought that the pathogenesis of alcohol‐induced ONFH is probably similar to that of corticosteroid‐induced ONFH. The aim of this study was to develop a new animal model for alcohol‐induced ONFH and to evaluate the relationship between the pro‐inflammatory response via TLRs and the development of ONFH in rats. Male Wistar rats were fed a Lieber–DeCarli liquid diet containing 5% ethanol (experimental group) or dextran (control group) for 1–24 weeks. Histopathological and biochemical analyses were performed. Feeding the ethanol‐containing liquid diet resulted in the development of ONFH with hepatic steatosis, hepatic dysfunction and hyperlipidaemia, whereas feeding the dextran‐containing diet did not cause ONFH. However, we could not recognize any relationship between the pro‐inflammatory response via TLR4 and the development of alcohol‐induced ONFH. Thus in this study we have developed a new rat model for alcohol‐induced ONFH based on the feeding of an ethanol liquid diet. ONFH was observed within seven days from the start of feeding with 5% ethanol‐containing liquid diet. Although this was linked to hepatic steatosis, a TLR4 association was not a feature of this model.     

个体股骨头坏死三维有限元模型的建立与应用

同时基于个体股骨头坏死患者的X-ray、CT和MRI图像,采用图像配准和融合技术对包含坏死股骨头的髋关节进行三维重建,获取具有高度几何相似性的三维有限元网格模型。选择1例中年女性股骨头坏死患者,分别获取X-ray、CT和MRI三套图像,采用Mimics 13.1和Pro/E 5.1软件分别基于这三套数据建立相关三维实体模型,经图像投影转换后,确定图像之间的匹配点,进行二维图像配准,配准后对成功融合的图像进行三维有限元网格模型显示。建立了具有良好几何相似性的髋关节三维有限元网格模型,包括正常皮质骨、松质骨、关节软骨和股骨头坏死区、断裂骨小梁等六部份,较真实地反映了包含坏死股骨头的髋关节的形态特征及毗邻关系,为进一步的生物力学分析和手术模拟提供了较理想的研究平台。     

Weight bearing does not contribute to the development of osteonecrosis of the femoral head

The hip joint is one of the major structures in the human body and the resultant force acting through the hip joint is 300% of body weight. Therefore, weight bearing, as a cause of ischaemia, may contribute to the development of non‐traumatic osteonecrosis of the femoral head (ONFH). However, it remains unclear whether weight bearing is related to the development of non‐traumatic ONFH. Therefore the aim of this study was to clarify the role of weight bearing in the development of non‐traumatic ONFH. Non‐weight‐bearing (NWB) rats were tail suspended to prevent any weight coming to bear on the hindlimbs from day 1 to the time of sacrifice. The weight‐bearing (WB) group rats were also housed individually, although without tail suspension. All rats were injected with lipopolysaccharide and methylprednisolone to promote the development of non‐traumatic ONFH. All animals were sacrificed three weeks after the final methylprednisolone injection. Histopathological analysis was performed. Osteonecrosis of the femoral head was observed not only in the NWB but also in the WB rats; however, no osteonecrosis of the humeral head was observed in either group. We confirmed that non‐traumatic ONFH developed in NWB rats, suggesting that weight bearing does not contribute to the development of non‐traumatic ONFH in rats.     

股骨头坏死动物模型评价方法的优缺点及展望

背景:股骨头坏死发病机制仍需要进一步研究,因此需要建立一个能够高度模拟人类股骨头坏死的动物模型,而如何正确评价股骨头坏死动物模型,是建立股骨头坏死动物模型的前提条件。目的:归纳总结近几年国内外对股骨头坏死动物模型的评价方法,综述各评价方法的优缺点,并探索新的评价方法,为动物模型的建立提供参考。方法:以“股骨头坏死,动物模型,评价方法”为检索词检索CNKI中国知网、万方、维普数据库,以“osteonecrosis of femoral head,femoral head necrosis,animal model,evaluation methods”为检索词检索PubMed、Web of Science及Medline数据库,检索2010年1月至2020年9月发表关于股骨头坏死动物模型评价方法的文献。结果与结论:①根据纳入标准通过阅读文献进行初步筛选,共获得51篇文章进行综述,其中中文文献33篇,英文文献18篇;②近年来,随着影像技术的发展和新技术的引入,股骨头坏死动物模型评估方法的选择增多,而选择合理的评价方式能减少建立动物模型过程中的工作量,并为后续研究提供参考,因此需要继续完善和改进。     

计算机软件测定股骨头坏死的体积及其意义

目的评估计算机软件测定股骨头坏死体积比的准确性。方法 20个比格犬(40髋)通过液氮冷冻法制成双侧股骨头坏死模型,对股骨头坏死区域体积比通过二种不同的方法来计算:①通过液体测量法测定股骨头体积和坏死股骨头体积,二者相除即为体积比;②将坏死模型的MRI图像以Dicom导入Minics,重建坏死区与股骨头的三维图像,通过逆向工程软件Geomagic处理后,导入Ansys软件划分单元,计算所有坏死单元的体积与整个股骨头单元体积比。结果计算机软件计算的体积比结果为(26.4±10.2)%,实体测定的体积比为(25.8±10.6)%,通过SPPS11.0分析,无统计学差异。结论计算机软件能准确评估股骨头坏死区域体积比,是一个可以提倡的新方法。     

Effect of bone mesenchymal stem cells transplantation on the micro-environment of early osteonecrosis of the femoral head

Autologous implantation of bone mesenchymal stem cells (BMSCs) has achieved promising clinical efficacy for the treatment of early-stage osteonecrosis of the femoral head (ONFH). However, the underlying mechanisms are not completely elucidated. Here, we investigated the effect of BMSCs on the early ONFH in vitro and in vivo. In co-cultured system, primary BMSCs enhanced the activity and inhibited the apoptosis of primary OB. The concentrations of VEGF and BMP-2 in the co-cultured medium were significantly higher than those without co-culture. Importantly, BMSCs implantation increased OB, capillaries and VEGF and BMP-2 expressions of the necrotic areas of femoral head in the ONFH rabbits. In conclusion, our results indicated that BMSCs treated the early ONFH possibly through increasing OB and capillaries, as well as VEGF and BMP-2 expression in the femoral head. These results provided possible mechanisms for the treatment of early-stage ONFH with BMSCs transplantation.     

构建双足类大型鸟类鸸鹋股骨头坏死模型

BACKGROUND: There are many studies on the establishment of animal models of femoral head necrosis, but it is difficult to simulate an ideal animal model of femoral head necrosis, especially collapse models. OBJECTIVE: To establish femoral head necrosis model of double-foot large bird emu so as to simulate femoral head necrosis in human.   METHODS: A total of 30 adult emus were frozen in liquid nitrogen and received radiofrequency heating alternately to simulate femoral head necrosis models. After 3 cycles of freezing, local frozen in liquid nitrogen and method of radiofrequency heating was alternated for closed loop liquid nitrogen freezing injury and radiofrequency heating injury. At 6, 12 and 16 weeks after model establishment, gross observation, X-ray, histology examination and MRI were conducted. Effects of model induction of avascular necrosis of the femoral head were evaluated.  RESULTS AND CONCLUSION: After liquid nitrogen cold-hot alternating method, emu at 12-18 weeks gradually suffered from femoral head necrosis signs and imaging findings. X-ray films, histological examination and MRI examination showed the avascular necrosis of femoral head was consistent with the pathological process of human femoral head necrosis, and the model had good repeatability. These results indicate that the use of liquid nitrogen cold-hot alternating method has successfully established the model of femoral head necrosis, with a good collapse rate, and can be used for mechanism research and treatment evaluation of osteonecrosis.      

Treatment of osteonecrosis of the femoral head with VEGF165 transgenic bone marrow mesenchymal stem cells in mongrel dogs

We evaluated the efficacy of vascular endothelial growth factor 165 (VEGF(165)) transgenic bone marrow mesenchymal stem cells (BMSCs) for the repair of early-stage osteonecrosis of the femoral head (ONFH) in mature mongrel dogs. This animal model was surgically established by femoral neck osteotomy and subsequent repinning. Twenty-seven dogs (54 hips) were divided into 3 equal-sized groups: a pCI-neo-VEGF(165) BMSC group, a pCI-neo BMSC group and a core decompression-alone group. The lipofectamine was used to introduce the VEGF(165) gene into the BMSCs. After core decompression, transgenic and non-transgenic autologous BMSCs were implanted. Therapeutic efficacy, including new bone formation and neovascularization in the femoral head, was examined by computed radiography, single-photon emission computed tomography, histological and histomorphometric analysis and immunofluorescent staining for von Willebrand factor in pathological sections. The femoral osteotomy site healed completely by the 4th week after the osteotomy surgery and regions of histologically evident osteonecrosis were found 12 weeks later. A regular arrangement of trabeculae and obvious bone regeneration were observed in the animals receiving implanted VEGF-transgenic BMSCs. The quantity of newly generated capillaries was significantly increased in the pCI-neo-VEGF(165) BMSC group, but there was no significant difference between the pCI-neo BMSC group and the core decompression-alone group. These results demonstrated that VEGF(165) transgenic autologous BMSCs enhanced bone reconstruction and blood vessel regeneration in the ONFH model. Compared with non-transgenic BMSCs, this approach could provide advanced benefits in the treatment of ONFH.     

Micro-CT-based bone ceramic scaffolding and its performance after seeding with mesenchymal stem cells for repair of load-bearing bone defect in canine femoral head

Osteonecrosis of the femoral head is a debilitating and painful orthopedic condition characterized by joint collapse. Salvage of the femoral head is highly desirable to preserve the contour and mechanical properties and prevent joint collapse. This study aimed to develop a new tissue-engineering approach for treatment of large bone defect in femoral head, that is, after osteonecrosis. The biphasic calcium phosphate (BCP) ceramic scaffolds were fabricated by a 3D gel-lamination technique based on micro-computed tomography (micro-CT) images of the cancellous bone microarchitecture of femoral heads. After seeding with autologous bone marrow-derived mesenchymal stem cells (BMSCs) in vitro, the cell-scaffold composite was implanted into a bone defect surgically induced in canine femoral head via trapdoor procedure, which was a common procedure for treatment of osteonecrosis. A total of 24 adult dogs were randomly divided into three groups (n = 8 each) for implantation of the BCP scaffold with or without with BMSCs, and also the autologous bone chips for comparisons. All animals were sacrificed at 30 weeks postoperatively and processed for radiological and histological evaluations. The contour of the femoral head was well preserved with implantation of BCP scaffolds with or without BMSCs, whereas joint collapse was found after treatment with autologous bone chips. The osteointegration and new bone formation was significantly greater with BCP scaffold implantation with than without BMSC seeding and showed greater strength and compressive modulus in the repair site. Micro-CT-based bone ceramic scaffolds seeding with BMSC might be a promising way to repair bone defects in the femoral head.     

Computer-assisted image analysis of the rat postosteonecrotic remodeled femoral head.

Osteonecrosis of rat femoral heads was induced by stripping the periosteum of the neck and cutting the ligamentum teres. The epiphyseal marrow and bone were necrotic on the 5th postoperative day. Specimens obtained 18 and 36 days postoperatively showed fibrous and hematopoietic-fatty tissue in the intertrabecular spaces, osteoclastic bone resorption, osteogenesis, and degeneration of the joint cartilage. Morphometrically, the means of the height-to-length ratios of the control, 6-day, 18-day, and 36-day femoral heads were 0.26, 0.28, 0.48, and 0.29, respectively. The shape factor of the femoral heads of the control rats was higher than 0.81 in 80% of the cases, while those of rats killed on the 6th, 18th, and 36th postoperative day were higher than 0.81 in 65, 60, and 50% of cases, respectively. Statistically, the means of the height-to-length ratios and the values of the shape factors of the femoral heads of the rats killed 18 days postoperatively differed significantly from those of the other three groups of rats. The quantitatively gauged data of the remodeled epiphyses negate the authors' subjective impression concerning early flattening of the femoral heads after surgically produced osteonecrosis.     

计算机手术导航系统对股骨头表面假体植入精确度的研究

目的研究非影像手术导航系统对股骨头表面假体植入位置的影响。方法25例股骨头坏死患者（共32髋）。其中男性16例，女性9例，年龄（36±7）岁。随机分为2组，计算机导航手术组（导航组）11例（16髋），在导航系统下进行股骨头表面置换术，设定髋最佳植入柄颈角为0°，柄干角为140°；对照组14例（16髋）采用传统手术。术后测定假体植入的角度，进行手术评估。结果术后导航组植入股骨假体柄干角为（140．5±1．7）°，对照组为（135．9±6．5）°，导航组的股骨假体柄颈角为（O．30±1．40）°，对照组柄颈角为（-4．93±4．30）°，两组差异具有统计学意义（P〈0．05）。Harris评分，导航组优14髋，良2髋；对照组优9髋，良4髋，中3髋。导航组无一例发生肢体缩短、旋转、内外翻畸形，对照组2例发生短缩畸形，1例内翻15°畸形。随访（14±4）个月，均无假体松动、脱位。结论在非影像手术导航系统下手术操作可以精确地植入假体，减少假体松动的发生,具有重要的临床价值。     

Development of a Clinically Relevant Animal Model for the Talar Osteonecrosis in Sheep

There are a lot of reports and reviews about osteonecrosis of the talus (ONT), yet reports about the animal model of ONT to evaluate proper therapeutic approaches are rarely heard. In our study, a novel animal model was established. Pure ethanol was injected into the cancellous bone of sheep''s talus. Macroscopic observation, X-ray, CT and histology were performed at two, four, 12 and 24 weeks postoperatively. It was revealed that the trabeculae of talar head began to change their structure after two weeks postoperatively compared to the normal talus. The ONT was obvious at the end of the fourth week, and their outstanding feature was the damage of trabeculae bone and formation of cavities. CT scans and pathological changes of the subjects all showed characteristics of the early stage of osteonecrosis, also the sections of the specimens confirmed necrosis of tali. By 12 weeks, the phenomenon of necrosis still existed but fibrous tissue proliferated prominently and bone reconstruction appeared in certain area. Most specimens (3/4) got late stage necrosis which presented as synarthrosis in X-ray and mass proliferation of fibrous tissue in histology at the end of 24 weeks. The novel animal model of ONT was successful, and it is inclined to deteriorate without any intervention. The study provides us a new way to evaluate various treatments on ONT in laboratory, which may eventually pave way to clinical applications.     

液氮冷冻致兔股骨头坏死病理学特征研究

目的 探讨液氮冷冻法制备兔股骨头坏死动物模型的病理学特征.方法 采用成年新西兰大白兔,无菌条件下手术显露股骨头;采用棉签蘸取液氮,对股骨头软骨面负重区冷冻、复温以制备股骨头坏死模型.于制模后第3、7天及第2、4、6、8周随机选择模型兔,手术取下股骨头,进行股骨头纵切面肉眼观察,常规石蜡包埋、切片,经HE染色后光镜观察.结果 股骨头纵切面4周前各观察时间点,股骨头局灶性坏死区域渐增.制模术后第3天兔股骨头组织切片镜下可见软骨细胞及骨细胞坏死;术后第7天见坏死区空骨陷窝较多;术后第2周见关节软骨缺损,软骨下区域小血管充血,骨小梁开始断裂、排列紊乱;术后第4周见关节软骨剥脱,髓腔内脂肪细胞坏死,并现增生纤维组织;术后第6周见股骨头塌陷,并出现纤维化骨的匍伏附着;术后第8周见新骨沉积性生长,骺板细胞挤压、变形.结论 本研究建立的液氮冷冻致兔股骨头坏死模型具有典型的病理演变特征,贴近已知人类股骨头坏死病理演变规律,是干细胞移植等治疗研究的理想模型.     

体外冲击波疗法结合同种异体骨钉治疗成人股骨头缺血性坏死

目的探讨体外冲击波疗法（ESWT）结合同种异体骨钉治疗成人股骨头缺血性坏死的疗效。方法2003年7月～2007年1月,采用体外冲击波或结合同种异体皮质骨钉植入术治疗54例（84髋）股骨头缺血性坏死患者并随访观察。男41例,女13例;年龄20～61岁,平均30岁。按Ficat分期,Ⅱ期55髋,Ⅲ期29髋。其中40髋（Ⅱ期27髋,Ⅲ期13髋）,采用ESWT结合同种异体骨钉治疗,治疗后6、12、24个月复查双髋X线、MRI及髋关节Harris评分（HHS）。44髋（Ⅱ期28髋,Ⅲ期16髋）患者采用单纯ESWT治疗。对两组治疗结果进行统计学分析。结果随访24～29个月,平均25．7个月。二者治疗后Harris评分分别为83．71±18．76和57．20±12．82,二者比较差异有统计学意义妒〈0．01）：影像学X光片评估：ESWT结合同种异体骨钉术后随访股骨头塌陷12髋,塌陷率为30％,单纯ESWT治疗随访股骨头塌陷25髋,塌陷率为56．8％。结论体外冲击波疗法结合同种异体骨钉是治疗中期ONFH的方法之一,它不但具有冲击波疗法促进骨愈合作用,而且复合骨钉移植可起到骨传导及骨支撑作用,近期可以减少股骨头塌陷发生。     

Osteonecrosis in systemic lupus erythematosus: Systematic insight from the epidemiology,pathogenesis, diagnosis and management

Osteonecrosis (ON) is one of the serious and well recognized complicationscausing severe pain and disability in patients with systemic lupus erythematosus (SLE), and its manifestation and pathogenesis are only partially understood. This review provide an update of the recent progress in etiology, pathogenesis, diagnosis, and treatment of systemic lupus erythematosus related osteonecrosis (SLE-ON). Despite the concomitant use of corticosteroids, alcohol and obesity, the dysregulated immune micro-environment and the complex pathogenesis of SLE synergistically play important roles in the development of ON. Osteonecrosis of femoral head (ONFH) is the most often involved joint in SLE. The current classification and staging system of ONFH is based on imaging techniques, particularly relating to MRI and CT, for the identification and quantification of necrotic lesions. For SLE-ONFH patients, both SLE specific clinical symptoms and ONFH imaging findings should be comprehensively evaluated. Even though advances concerning bone grafting and arthroplasty procedures have resulted in improved clinical outcomes, early pharmacological treatment at the pre-collapse stage may prevent joint collapse and reduce the joint arthroplasty rate, and this needs to be accounted. Although some progress has been made, considerably more research is needed before we fully understand SLE-ONFH. Future treatments of SLE-ONFH may involve genetic or cell-based therapies that target potential biomarkers, and this will lead to effective measures for saving thefunction of hip joint and preventing osteonecrosis.     

Finite Element Analysis on the Diagnosis and Treatment of Osteonecrosis of Femoral Head

Developing the radiographic images from two to three-dimensional, finite element analysis(FEA) technology can set up the model, predicting diagnosis, treatment design, as well as surgical plan. FEA provides an accurate three-dimensional finite element biomechanical study in osteonecrosis of femoral head(ONFH). The papers in the latest 5 years related to femoral head osteonecrosis and finite element analysis application are concentrated on. We summarize the latest research progress and problems, including the applied research carried out in the femoral head osteonecrosis clinical cases,innovational skills, so as to point out the direction of future research in FEA.     

骨内压在激素诱导型股骨头坏死病程发展中的价值

目的动态观察激素诱导型股骨头坏死演变过程中骨内压的改变和组织病理变化的关系,评价骨内压在早期诊断股骨头坏死中的价值。方法健康成年大耳白兔,随机分为2组:实验组每天1次肌注2．5mg/ kg地塞米松(Dexamethasone DEX)。对照组肌注等量生理盐水。在实验的各个时相点分别对实验组和对照组的股骨头行骨内压测定;观察病理组织学变化、股骨头血管形态的变化、股骨头组织的超微结构。结果实验组各时相点股骨头软骨下区平均骨陷窝空虚率随着时间延长而增高;用DEX后,股骨头骺部血管面积进行性下降;电镜下可见骨细胞经历变性、坏死、溶解过程;实验组股骨头骨内压在各个时相点随时间进程先升后降。结论股骨头坏死早期骨内压增高明显,可作为早期诊断股骨头坏死的指标。