【作者】 黄跃平；

【导师】 李康华； 胡建中；

【作者基本信息】 中南大学 ， 外科学， 2009， 博士

【摘要】 前交叉韧带(anterior curciate ligament,ACL)是维持膝关节稳定的重要结构,其作用主要表现为控制胫骨的前移和对膝内外翻和旋转运动的调节。股骨内髁是膝关节的重要组成部分。临床上ACL损伤十分常见,常导致膝关节继发性损伤。然而,ACL损伤是否影响股骨内髁软骨,目前研究甚少。本研究运用生物力学测定方法并结合聚类统计学分析方法对ACL各纤维束进行功能性分束;并在此基础上,运用生物力学、组织学方法观测ACL损伤后股骨内髁不同部位生物力学特性及病理学改变;同时检测IL-β、MMP-13表达。为ACL损伤是否对股骨内髁产生影响提供理论依据。第一章前交叉韧带功能性分束的生物力学研究目的探讨ACL功能性分束,为后续研究提供基础。方法新鲜成人正常膝关节标本6具,取ACL胫骨止点前方两侧、股骨止点后方两侧作为应变测量点,静态应变测试仪平衡标定后,在800N轴向载荷下,测试膝关节0°、30°、60°、90°状态下测量ACL前内侧束、前外侧束、后内侧束、后外侧束应变值。运用聚类方法进行功能性分束。结果1、膝关节0°位: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;ACL后外侧束、前外侧束应变大于内侧束、前内侧束应变,均有显著性差异,均P＜0.05。30°位和90°位: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;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。60°位:ACL应变后内侧束最大后依次为、前内侧束、后外侧束、前外侧束,各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:后外侧束与前外侧束归为一类,后内侧束与前内侧束归为一类。结论膝伸直位ACL后外侧区纤维束和前外侧区纤维束应变大于后内侧区纤维束和前内侧区纤维束。膝屈曲位后内侧区纤维束和前内侧纤维束应变大于后外侧区纤维束和前外侧区纤维应变,说明后外侧区纤维束和前外侧区纤维束主要维持膝关节伸直稳定性,后内侧区纤维束和前内侧区纤维束主要维持膝关节屈曲稳定性。结合聚类分析结果,证明ACL可分为前内、后外两功能性纤维束。第二章前交叉韧带断裂对股骨内髁生物力学影响的实验研究目的探讨ACL断裂对股骨内髁生物力学的影响,为防治股骨内髁继发性损伤提供依据和参考。方法新鲜成人尸体膝关节标本10具,分为ACL完整组、前内侧束(AMB)切断组、后外侧束(PLB)切断组和ACL完全切断组。先将ACL完整组(10具)分别在膝关节0°、30°、60°、90°位轴向加载200～800N,测量股骨内髁关节面的应变;再随机将标本造模成AMB切断组和PLB切断组(各5具),在上述条件下测试应变;最后将ACL完全切断组(10具)在上述条件下测试应变;并分别对数据进行统计分析。结果1.膝关节0°位,200N和400N载荷下,股骨内髁前部、中部、后部的应变在PLB切断组与ACL全断组、ACL完整组与AMB切断组之间无显著性差异,P＞0.05;600N、800N载荷下,股骨内髁前部、后部的应变在各实验组间均有显著性差异,P＜0.05,中部应变在各实验组间无显著性差异,P＞0.05:相同载荷下,前部、中部应变值表现为全断组＞PLB切断组＞AMB切断组＞完整组,后部表现为完整组＞AMB切断组＞PLB切断组＞全断组。2.膝关节30°位,200N和400N载荷下,股骨内髁前部、中部、后部的应变在ACL完整组与PLB切断组、AMB切断组与ACL全断组之间无显著性差异,均P＞0.05;600N、800N载荷下各实验组间均有显著性差异,均P＜0.05,相同载荷下应变值表现为全断组＞AMB切断组＞PLB切断组＞完整组。3.膝关节60°,200N和400N、600N、800N载荷下,股骨内髁前部、中部、后部的应变各实验组间均有显著性差异,均P＜0.05;相同载荷下应变值表现为全断组＞AMB切断组＞PLB切断组＞完整组。4.膝关节90°,在200N、400N、600N、800N载荷下,股骨内髁前部、中部、后部的应变各实验组间均有显著差异,均P＜0.05;前部应变表现为完整组＞PLB切断组＞AMB切断组＞全断组,中部、后部表现为全断组＞AMB切断组＞PLB切断组＞完整组。结论1.PLB切断在膝伸直位对股骨内髁前、中、后各部位的应变均有影响。2.AMB切断在膝屈曲位时对股骨内髁前、中、后各部位的应变均有影响。3.ACL完全切断,对股骨内髁前、中、后各部位的应变均有影响。第三章前交叉韧带断裂对股骨内髁软骨的组织学影响目的观察ACL断裂后股骨内髁软骨组织HE染色改变与IL-1β、MMP-13的表达变化,探讨ACL断裂对股骨内髁软骨组织学影响。方法48只雄性家兔随机分为四组,兔后腿膝关节配对为实验侧和对照侧,自身对照,实验侧行ACL切断。造模后于第1、3、6、8周各随机处死12只。观察各时间点股骨内髁软骨组织结构HE染色与IL-1β、MMP-13表达的变化。结果1.大体观察:随着ACL断裂时间周期的延长,实验组股骨内髁软骨逐渐出现色泽改变、表面光泽减退、表面出现不平整甚至溃疡。2.组织HE染色观察:第1周实验组与对照组比较无显著性差异,P＞0.05,其他各实验组均大于对照组,差异均具有显著性,均P＜0.01;自第三周开始出现软骨表面及软骨细胞排列的异常。Mankin评分实验组随时间推移而增高,各实验组间比较均有显著性差异,均P＜0.01;3.IL-1β表达:第1、3、6、8周实验组均高于正常对照组,均有显著性差异,均P＜0.05。实验组第1周低于3、6、8周,均有显著性差异,均P＜0.05;第6周表达高于第3、8周,均有显著性差异,均P＜0.01;第8周表达高于第3周,有显著性差异,P＜0.01。正常对照组各周间比较均无显著性差异,均P＞0.05。4.MMP-13表达:第1、3、6、8周实验组均高于正常对照组,均有显著性差异,均P＜0.01。实验组第1周低于第3、6、8周,均有显著性差异,均P＜0.01;第6周表达高于第3、8周,均有显著性差异,均P＜0.01;第8周表达与第3周比较无显著性差异,P＞0.05。正常对照组各周间比较均无显著性差异,均P＞0.05。结论1.证实ACL断裂继发引起股骨内髁软骨组织退变。2.ACL断裂后股骨内髁软骨IL-1β、MMP-13表达递提示IL-1β、MMP-13可能是ACL断裂后股骨内髁软骨退变因素之一。更多还原

【Abstract】 Anterior cruciate ligament of knee joint stability is important to maintain the structure,its role is mainly expressed in the anterior tibial control and turned inside and outside of the knee and rotation adjustment.Within the femoral condyle is an important component of the knee.ACL injury clinically are common,often leads to secondary injury in the knee.However,ACL injury would not affect the femoral condyle with the cartilage,little current research.In this study,the use of bio-mechanical method combined with statistical analysis of clustering methods on the fiber bundle ACL functional splitter;and on this basis,the use of biomechanical and histological methods of observation after the ACL injury in different parts of femoral condyles in biomechanics characteristics and pathological changes;At the same time,detection of IL-β,MMP-13 expression.For ACL injury in femoral condyle of the impact and provide a theoretical basis. Chapter1 The biomechanical study of ACL bundles’ functional classificationObjective:To investigate the functional bundle division of the ACL.Methods:6 fresh cadaveric knees from adult human beings were used.Selected the bilateral anterior part of the ACL tibial insertion and the bilateral posterior part of the ACL femoral insertion 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:Extension after ACL knee lateral fiber bundle and the former lateral strain than the fiber bundle fiber bundle after the medial area of the medial preoptic area and the fiber bundle.Knee flexion after the medial area of fiber bundles and fiber bundles anteromedial lateral strain after more than fiber bundles and fiber strain anterolateral area shows that after the lateral fiber bundle and the former major lateral fibers to maintain the knee joint extended stability of fibers after the medial area of the medial preoptic area and the main fiber bundle to maintain the stability of knee flexion.Combining the results of cluster analysis that can be divided into pre-ACL,the posterolateral fiber bundles of the two functions. Chapter 2 The biomechanical influence of ACL rupture on the Within the femoral condyleObjective:To investigate the biomechanical influences of partial and total ACL rupture on the Within the femoral condyle and provide theoretic evidence for ACL repair and rehabilitation.Methods:10 fresh cadaveric knees from adult human beings were divided into 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 Within the femoral condyle was measured and analysed.Results:In 0°position,the straining of the anterior part,middle part and posterior part on the Within the femoral condyler shows no significant difference between PLB rupture group and ACL complete rupture group,complete ACL group and AMB rupture group under the load of 200N and 400N,P＞0.05;significant difference is showed among the straining of the anterior part and posterior part on the Within the femoral condyler under the loads of 600N and 800N,P＜0.05,the straining of middle part on the Within the femoral condyler shows no significant difference among all the groups under the loads of 600N and 800N,P＞0.05;under the same load and angle of flexion,the correlateion of the absolute value of straining in every group of the anterior part and posterior part on the Within the femoral condyler increases in this way of ACL complete rupture group＞PLB rupture group＞AMB rupture group＞complete ACL group,But,middle part on the Within the femoral condyler increases in this way of ACL complete rupture group＞AMB rupture group＞PLB rupture group＞complete ACL group.In 30°position,the straining of the anterior part,middle part and posterior part on the Within the femoral condyler shows no significant difference between complete ACL group and PLB rupture group,AMB rupture group and ACL complete rupture group under the load of 200N and 400N,P＞0.05;significant difference is showed among all the groups under the loads of 600N and 800N,P＜0.05;under the same load and angle of flexion,the correlateion of the absolute value of straining in every group increases in this way of A CL complete rupture group＞AMB rupture group＞PLB rupture group＞complete ACL group.In 60°position,the straining of the anterior part and middle part and posterior part on the Within the femoral condyler shows significant difference among all experiemental groups under the load of 200N and 400N and 600N and 800N,P＜0.05;under the same load and angle of flexion, the correlateion of the absolute value of straining in every group increases in this way of A CL complete rupture group＞AMB rupture group＞PLB rupture group＞complete ACL group.In 90°position,the straining of the anterior part and middle part and posterior part on the Within the femoral condyler shows significant difference among all experiemental groups under the load of 200N and 400N and 600N and 800N,P＜0.05. under the same load and angle of flexion,the correlateion of the absolute value of straining in every group of the anterior part on the Within the femoral condyler increases in this way of A CL complete ACL group＞PLB rupture group＞AMB rupture group＞complete rupture group, but,every group of middle part and posterior part on the Within the femoral condyler increases in this way of A CL complete rupture group＞AMB rupture group＞PLB rupture group＞complete ACL group.Conclusion:1.PLB cut off in the knee in extension position of the femoral condyle before,during and after each part of the strain affects both.2.AMB cut off in the knee flexion with the femoral condyle on before, during and after each part of the strain affects both.3.ACL completely cut off,with the condyle of the femur before,during and after each part of the strain affects both. Chapter 3 The histological influence of ACL rupture on the Within the femoral condyler cartilageObjective:To investigate the tissue construction changes and IL-1、MMP-13 express in the Within the femoral condyler cartilage,to further explore the influence of ACL rupture on the Within the femoral condyler cartilage.Methods:48 male 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 lateral tibial plateau cartilage were observed 1、3、6、8 weeks later.Results:1.Gross observation:As the time lasted,the Within the femoral condyler 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.The Mankin scores in experimental groups increased as the time lasted,the differences among the groups were all significant(P＜0.01); there was no significant difference between 1-week group and control group(P＞0.05),the Mankin scores in the other experimental groups were significantly higher than that in control group(P＜0.01).3.IL-1β expression: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.Confirmed ACL rupture with secondary femoral condyle caused by degeneration of cartilage tissue.Femur fracture within2.ACL condylar cartilage after IL-1β,MMP-13 expression prompt delivery IL-1β,MMP-13 may be broken after ACL femoral condyle cartilage degeneration in one of the factors.更多还原