【作者】 李思鸿；

【导师】 李康华； 雷光华；

【作者基本信息】 中南大学 ， 外科学， 2009， 博士

【摘要】 第一章前交叉韧带功能性分束的生物力学实验研究目的通过测量前交叉韧带各束在膝关节屈伸过程中的生物力学变化,探讨其相应的功能及分类,为后续实验及临床工作提供理论依据。方法6具新鲜成人尸体膝关节标本,依据ACL在膝关节中的位置将ACL分为:前内侧区纤维束(前内侧束)、前外侧区纤维束(前外侧束)、后内侧区纤维束(后内侧束)、后外侧区纤维束(后外侧束)。在膝关节标本上施加800N轴向载荷下,分别测试膝关节0°、30°、60°、90°状态下ACL各纤维束应变。比较各束应变的差异,采用样品聚类方法进行分类。结果1、膝关节0°位,ACL后外侧束、前外侧束应变均大于后内侧束、前内侧束应变,均有显著性差异,均P＜0.05;后外侧束与前外侧束应变无显著性差异,P＞0.05;后内侧束与前内侧束应变无显著性差异,P＞0.05。30°位和90°位,ACL后内侧束、前内侧束应变均大于后外侧束、前外侧束应变,均有显著性差异,均P＜0.05;后内侧束与前内侧束应变无显著性差异,P＞0.05;后外侧束与前外侧束应变无显著性差异,P＞0.05。60°位,应变大小依次为:后内侧束、前内侧束、后内侧束、前内侧束,束间应变比较均有显著性差异,均P＜0.05。2、前内侧束与后内侧束应变变化一致,依序0°、30°、60°、90°逐渐增大,均有显著性差异,均P＜0.05;前外侧束应变在膝0°、30°、60°、90°均无显著性差异,均P＞0.05。后外侧束应变在膝关节60°位大于0°、30°、90°,均有显著性差异,均P＜0.05;0°与90°应变无显著性差异,P＞0.05;0°、90°应变均大于30°,差异均有显著性,均P＜0.05。3、聚类分析:后内侧束与前内侧束归为一类,后内侧束与前内侧束归为一类。结论(1)前交叉韧带内存在不同功能的纤维束,后外侧区纤维和前外侧区纤维主要维持膝关节伸直稳定性;后内侧区纤维和前内侧区纤维主要维持膝关节屈曲稳定性。(3)结合聚类分析结果ACL可分为前内侧束、后外侧束两功能束。第二章前交叉韧带断裂对内侧胫骨平台影响的生物力学研究目的探讨ACL部分断裂和完全断裂对膝关节内侧胫骨平台的生物力学影响,为ACL损伤后继发骨关节炎的防治提供理论依据。方法新鲜成人尸体膝关节标本10具,依照测试先后顺序分为ACL完整组、AMB切断组、PLB切断组和ACL完全切断组。在膝关节标本上施加200～800N轴向载荷,分别测量各组膝关节0°、30°、60°、90°位,内侧胫骨平台前、中、后各部的应变,比较各组应变的差异。实验数据采用SPSS13.0统计软件进行统计学分析,检验水准为a=0.05。结果1.ACL完整组:①0°位各级载荷下,前部应变与中部应变,无显著性差异,p＞0.05;前部、中部应变均大于后部,有显著性差异,p＜0.05。②30°位,中部应变最大其次为前部,后部最小,三者差异有显著性,p＜0.05。③60°位,前部均为拉应变,中部、后部均为压应变。中部应变与后部应变差异无显著性,p＞0.05。④90°位,前部为拉应变,后部、中部为压应变。后部应变大于中部应变,差异有显著性,p＜0.05。2.AMB断裂组①0°位各级载荷下,前部、中部、后部应变与完整组应变无显著性差异,p＞0.05。②30°位各级载荷下,前部、中部应变与完整组应变无显著性差异,p＞0.05;200N后部应变与完整组应变无显著性差异,p＞0.05,400N～800N载荷下后部应变大于完整组应变,有显著性差异,p＜0.05。③60°位各级载荷下,前部为拉应变,中部、后部为压应变。200N载荷下前部、中部、后部应变与完整组应变无显著性差异,p＞0.05。400N～800N载荷下前部、中部、后部应变绝对值大于完整组,差异有显著性,p＜0.05。④90°位各级载荷下,前部为拉应变,后部、中部为压应变。200N载荷下前部、中部、后部应变与完整组应变无显著性差异,p＞0.05。400N～800N载荷下前部、中部、后部应变绝对值大于完整组,差异有显著性,p＜0.05。3.PLB断裂组:①0°位,200N载荷下前部、中部、后部应变与完整组应变无显著性差异,p＞0.05。400N～800N载荷下前部、中部、后部应变均大于完整组,有显著性差异,p＜0.05。②30°位,各级载荷下前部、后部应变均与完整组无显著性差异,p＞0.05。200N～600N中部应变与完整组无显著性差异,p＞0.05。800N载荷下中部应变小于完整组,差异有显著性,p＜0.05。③60°位,前部为拉应变,中部、后部为压应变。各级载荷下前部、中部、后部应变与完整组应变无显著性差异,p＞0.05。④90°位,前部为拉应变,后部、中部为压应变。各级载荷下,载荷下前部、中部应变均与完整组无显著性差异,p＞0.05。200N、400N后部压应变与完整组无显著性差异,p＞0.05。600N、800N后部应变大于完整组,有显著性差异,p＜0.05。4.全断组:各级载荷下0°、30°、60°、90°位,前部、中部、后部,各部位间应变均大于完整组,有显著性差异,p＜0.05。结论(1)ACL完全断裂对内侧胫骨平台(前、中,后)各部的应变均有影响。(2)AMB断裂在屈曲位(30°、60°、90°)对内侧胫骨平台各部的应变有影响。(3)PLB断裂在膝关节伸直位(0°)对内侧胫骨平台各部的应变均有影响。提示AMB在屈曲位起稳定作用,PLB在伸直位起稳定作用。第三章前交叉韧带断裂对内侧胫骨平台软骨组织学影响的动物试验研究目的研究兔前交叉韧带断裂后不同时期内侧胫骨平台软骨组织结构的变化及IL-1β、MMP-13的表达,探讨前交叉韧带断裂后内侧胫骨平台关节软骨退变的生物学机制。方法48只雄性新西兰大白兔随机分为四组,行后腿单侧ACL切断,对侧假手术自身对照。分别于造模1、3、6、8周各处死12只。观察内侧胫骨平台大体形态变化,取内侧胫骨平台软骨HE染色,病理光镜观察。免疫组化检测IL-1β、MMP-13的表达。结果1.大体观察:在四个时间观测点上,实验组内侧胫骨平台软骨逐渐出现色泽改变、局灶充血、表面磨损、软骨表面龟裂、溃疡形成。2.HE染色:自第三周开始出现基质染色不均、软骨细胞排列紊乱、软骨细胞增生的异常。3.IL-1β表达情况:实验组第1、3、6、8周IL-1β表达均高于对照组,均有显著性差异,均p＜0.05。对照组第1、3、6、8周IL-1β表达无显著性差异,均p＞0.05。实验组IL-1β表达阳性率第3周高于第1周和第8周,有显著性差异,p＜0.05;实验组第6周高于第1周和第8周,有显著性差异,p＜0.05;实验组第3周和第6周IL-1β表达阳性率无显著性差异,p＞0.05;实验组第8周IL-1β表达阳性率高于第1周,有显著性差异,p＜0.05。4.MMP-13表达情况:实验组第1、3、6、8周MMP13表达均高于对照组,均有显著性差异,均p＜0.05。对照组第1、3、6、8周MMP13表达均无显著性差异,均p＞0.05。实验组第6周MMP13阳性表达率高于第1周、第8周,均有显著性差异,均p＜0.05;实验组第3周MMP13阳性表达率高于第1周,有显著性差异,p＜0.05;第3周与第6周、第8周MMP13阳性表达率均无显著性差异,均p＞0.05;第1周与第8周阳性率无显著性差异,p＞0.05。结论(1)ACL断裂可引起内侧胫骨平台软骨退变。(2)ACL断裂后,软骨组织IL-1β、MMP13表达增高提示IL-1β、MMP13可能是内侧胫骨平台软骨退变,OA形成的病因之一。更多还原

【Abstract】 Chapter1 The biomechanical study of ACL bundles’ functional classificationObjective:To investigate the biomechanical function and classification of the ACL bundles and provide biomechanical evidence for the later experiments and clinical therapy.Methods:6 fresh cadaveric knees from adult human beings were utilized in this investigation.We classified ACL into four bundles on the basis of ACL location in knee:anteromedial area fiber bundle(anteromedial bundle)、anterolateral area fiber bundle(anterolateral bundle)、posteromedial area bundle (posteromedial bundle) and posterolateral area bundle(posterolateral bundle). The strain on the ACL bundles was measured when the knees were applied with 800N axial loading force in 0°、30°、60°、90°、positions.The bundles were functional classified by cluster analysis.Results:1.In 0°position,the strain on the posterolateral and anterolateral bundles was significantly larger than the posteromedial and anteromedial bundles(P＜0.05),There were no significant diference between the posterolateral and anterolateral bundles(P＞0.05) and so did the posteromedial and anteromedial bundles(P＞0.05);In 30°and 90°position,the strain on the posteromedial and anteromedial bundles was significantly larger than the posterolateral and anterolateral bundles(P＜0.05),There were no significant difference between the posteromedial and anteromedial bundles(P＞0.05) and so did the posterolateral and anterolateral bundles(P＞0.05);In 60°position,the strain on the posteromedial bundle was the largest,and then the anteromedial、posterolateral、anterolateral bundle,the differences among bundles were all significant(P＜0.05).2.The strain on the posteromedial and anteromedial bundles was increased by degrees when the knee flexed 0°、30°、60°、90°,and all the difference were significant(P＜0.01);There were no significant changes of the strain on the anterolateral bundle among different angles(P＞0.05).The strain on the posterolateral bundle in 60°position was larger than 0°、30°、90°positions,the differences were significant(P＜0.05);The strain in 30°position was significantly smaller than 0°and 90°positions(P＜0.05),but the difference of strain between 0°and 90°positions was not significant(P＞0.05). 3.Cluster analysis classfied the anteromedial bundle and the posteromedial bundle as one class and the anterolateral and posterolateral bundles as the other class.Conclusion:1.The posterolateral area and anterolateral area fiber bundles mainly maintain stability of the knee in extended positions.The posteromedial area and anteromedial area fiber bundles mainly maintain stability of the knee in flexed positions.2.Combined with the cluster analysis results,ACL may be classified into the anteromedial functional fiber bundle and the posterolateral functional fiber bundle. Chapter 2 The biomechanicaI innuence of ACL rupture on the medial tibial plateauObjective:To investigate the biomechanical influences of partial and total ACL rupture on the medial tibial plateau and provide theoretic evidence for prevention and cure ostarthritis.Methods:10 fresh cadaveric knees from adult human beings were classified into four group on the basis of test sequence:ACL intact group、AMB broken group、PLB broken group and ACL total broken group.The knees were applied with 200N～800N axial loading force when they flexed 0°、30°、60°、90°.The strain on the medial tibial plateau was measured and analyzed by SPSS for windows 13.0,the statistic significance was set atα=0.05.Results:1.In ACL intact group:①In 0°position,the strain of anterior part and the middle part of tibial plateau were not significantly different(P＞0.05) and were larger than posterior part(P＜0.05).②In 30°positions,the largest strain was on the middle part of the medial tibial plateau,then the posterior part and the smallest on the anterior part,and the differences among the parts were all significant(P＜0.05).③In 60°position,anterior part showed to be tensile strain while middle part and posterior part compressive strain,the strain of anterior part and the middle part of tibial plateau were not significantly different(P＞0.05).④In 90°position,anterior part showed to be tensile strain while middle part and posterior part compressive strain,the strain of posterior part and the middle part of tibial plateau were not significantly different(P＞0.05).2.In AMB broken group:①In 0°position,The strain on the anterior part、middle part and posterior part were not significantly different from that in ACL intact group(P＞0.05).②In 30°positions,the strain on the anterior part and the middle part was not significantly different from that in ACL intact group(P＞0.05);Under 400N～800N loading,the stain on the posterior part was larger than that in ACL intact group,the difference was significant(P＜0.05).③In 60°position,anterior part showed to be tensile strain while middle part and posterior part compressive strain.Under 200N loading, the strain on the posterior part was not significantly different from ACL intact grou(P＞0.05).Under 400N～800N loading,the stain on anterior part、middle part and posterior part were larger than that on ACL intact group,the difference was significant(P＜0.05).④In 90°position,anterior part showed to be tensile strain while middle part and posterior part compressive strain.Under 200N loading,the strain on the anterior part、middle part and posterior part were not significantly different from ACL intact group(P＞0.05).Under 400N～800N loading,the stain on anterior part、middle part and posterior part were larger than that in ACL intact group,the difference was significant(P＜0.05).3.In PLB broken group:①In 0°position,Under 200N loading,the strain on the anterior part、middle part and posterior part were not significantly different from ACL intact group(P＞0.05).Under 400N～800N loading,the stain on anterior part、middle part and posterior part were larger than that on ACL intact group,the difference was significant(P＜0.05).②In 30°positions,expect the strain on the middle part was smaller than that in ACL intact group under 800N loading(the difference was significant(P＜0.05) ) the strain on the anterior、middle and posterior parts was not significantly different from that in ACL intact group(P＞0.05).③In 60°positions,The strain on the anterior、middle and posterior parts was not significantly different from that in ACL intact group (P＞0.05).④In 90°positions,expect the strain on the middle part was larger than that in ACL intact group under 600N,800N loading(the difference was significant(P＜0.05) ) the strain on the anterior、middle and posterior parts was not significantly different from that in ACL intact group(P＞0.05).4.In ACL total broken group:In all condition the strain on the anterior、middle and posterior parts was larger than that in ACL intact group,the difference was significant(P＜0.05)Conclusion:1.ACL total rupture may cause abnormal load on the medial tibial plateau in all the positions.2.AMB rupture may cause abnormal load on the all part of the tibial plateau in flexed positions(30°、60°,90°).3.PLB rupture may cause abnormal load on all parts of the medial tibial plateau in extended positions(0°). Chapter 3 The histological influence of ACL rupture on the medial tibial plateauObjective:Observe tissue construction changes and IL-1、MMP-13 express in the medial tibial plateau cartilage,to further explore the degeneration mechanism of the medial tibial plateau articula cartilage after ACL rupture.Methods:48 male New Zealand rabbits were randomly divided into four groups,and all were under one side posterior leg ACL cut and the opposite side as the control.HE staining and immunohistochemical methods were used. Tissue construction and IL-1β、MMP-13 expression changes of the medial tibial plateau cartilage were observed 1、3、6、8 weeks later.Results:1.Gross observation:As the time lasted,the medial tibial plateau cartilage had the color changed、the gloss decreased、the surface abrased and even had ulcer on the cartilage.2.Routine HE staining:There are abnormal cartilage surface and cell disposition after 3 weeks.3.IL-1βexpression:All the experimental groups had higher IL-1βexpression than that in control groups (P＜0.01).In experimental groups,IL-1βexpressed significantly lower in 1-week group than that in 3、6、8-week groups(p＜0.05);In 6-week group,IL-1βexpressed higher than that in 3-week group and 8-week group(P＜0.01).IL-1βexpressed higher in 8-week group than that in 3-week group(P＜0.01).There were no signifigant differences among the control groups(P＞0.05).4.MMP-13 expression:All the experimental groups had higher IL-1βexpression than that in control groups(P＜0.01).In experimental groups,MMP-13 expressed significantly lower in 1-week group than that in 3、6、8-week groups(p＜0.01);In 6-week group,MMP-13 expressed higher than that in 3-week group and 8-week group(P＜0.01).There was no significant difference between 3-week group and 8-week group(P＞0.05).There were no signifigant differences among the control groups(P＞0.05).Conclusion:1.ACL rupture may cause cartilage degeneration on the medial tibial plateau.2.The increased IL-1βand MMP-13 expression suggest that IL-1β、MMP-13 may participate in cartilage degeneration on the medial tibial plateau after ACL rupture.更多还原