显微CT在骨腱结合部损伤中的应用与低强度循环载荷对损伤的影响
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摘要
髌骨髌腱骨腱结合部损伤是常见的运动损伤,是当今体育科学界届的热点问题。在整个对髌骨髌腱骨腱结合部损伤机理及治疗手段的研究中,动物实验是其中重要的环节。本研究在Nakama循环载荷动物模型以及前阶段实验的基础上,研究阈下强度运动对骨腱结合部损伤的影响,并通过实验探讨Micro-CT技术计算纤维软骨带厚度的可行性,同时使用Micro-CT技术计算实验组和对照组的骨量和骨小梁变化。
16只成年雌性新西兰大白兔,随机筛选分组分为4周循环载荷组(n=8),8周循环载荷组(n=8),左后肢进行实验,右后肢为空白对照。实验过程中,前4周两组实验动物统一接受强度为7.17N/S的肌肉收缩刺激,一周3次,一次2小时,共24小时。实验4周后,4周循环载荷组停止实验,8周循环载荷组将强度改为4.2N/S,实验时间不变。8周后统一进行取材,将取材后组织首先进行Micro-CT进行扫描图像重建,随后进行固定、脱钙、脱水、石蜡包埋、切片和H&E染色制成组织切片,使用图像采集系统进行组织学分析。
结果：对比对照组,8周循环载荷组和4周循环载荷组均在髌腱出现了玻璃样变,纤维紊乱；潮线出现“涨潮”、模糊乃至消失,纤维软骨带增厚,显示损伤模型建立成功。H&E染色测量纤维软骨带厚度8周循环载荷组和4周循环载荷组没有显著差异,Micro-CT计算纤维软骨带厚度结果与H&E染色测量结果未见相关性,通过Micro-CT计算骨量和骨小梁结果显示实验组和对照组没有显著差异。结论：①对髌骨骨腱结合部损伤实施阈下强度载荷训练和休息的干预手段可以改变骨的微细结构,而不能改变骨腱结合部的组织变化,骨的重建和恢复能力要强于骨腱结合部；②Micro-CT测定法可以更好描述纤维软骨带厚度的假设不成立。技术由于技术特点的限制,不能准确的计算骨腱结合部纤维软骨带的面积。
Patellar tendon junction injuries is very common disease in athlete and normal people, it is hot issues of sport medicine. Animal experiment is an important part of reseach the patellar tendon junction injuries mechanism and treatment. In this study, Nakama cyclic loading and the former stage of experimental animal models, based on the study of subliminal intensity exercise on bone tendon junction injuries, and by experimental calculation of Micro-CT technology feasibility with thickness of fibrocartilage, while the Micro-CT technical computing the experimental group and control group trabecular bone volume and bone changes.
16matual female New Zealand rabbits were randomly divided into4week cyclic loading group (n=8),8week cyclic loading group (n=8), the experiment left hind, right hind limb of the control group. Two Experiment groups of animals receiving a unified strength7.17N/S stimulation of muscle contraction in first4weeks, trained three times a week,2hours per time, total4hours. After4weeks,4weeks experimental cyclic loading group stopped and the8week cyclic loading group adjust the cyclic load4.2N'/S, experimental time constant.8weeks after the reunification of the subjects, the organization will be drawn after the first Micro-CT scan image reconstruction, followed by fixed, decalcified, dehydrated, embedded in paraffin, sliced and made H&E stained tissue sections, and using the image acquisition system for histological analysis.
Result:Comparing the control group,8weeks and4weeks group cyclic loading cyclic loading in the patellar tendon group there were hyaline, fibrous disorder;tide line of a"rising tide"and disappeared, thickening with fibrous cartilage, showing damage model was successfully created. H&E stained measuring microscopic cartilage with a thickness of8weeks cyclic loading group and4-week cycle of load groups were not significantly different, Micro-CT calculations fibrocartilage zone thickness results with H&E staining measurements no correlation, by Micro-CT calculation of bone mass and trabecular bone showed the experimental group and control group were not significantly different. Conclusion:①The low intensity cyclic loading training and rest can change the bone micro construction, but can not change the bone tendon junction injury in histologyily. The bone have better reconstruction and recovery abitly than the bone tendon junction;②Micro-CT can not accurately calculate the bone tendon junction fibrocartilage zone of the area.
16只成年雌性新西兰大白兔,随机筛选分组分为4周循环载荷组(n=8),8周循环载荷组(n=8),左后肢进行实验,右后肢为空白对照。实验过程中,前4周两组实验动物统一接受强度为7.17N/S的肌肉收缩刺激,一周3次,一次2小时,共24小时。实验4周后,4周循环载荷组停止实验,8周循环载荷组将强度改为4.2N/S,实验时间不变。8周后统一进行取材,将取材后组织首先进行Micro-CT进行扫描图像重建,随后进行固定、脱钙、脱水、石蜡包埋、切片和H&E染色制成组织切片,使用图像采集系统进行组织学分析。
结果：对比对照组,8周循环载荷组和4周循环载荷组均在髌腱出现了玻璃样变,纤维紊乱；潮线出现“涨潮”、模糊乃至消失,纤维软骨带增厚,显示损伤模型建立成功。H&E染色测量纤维软骨带厚度8周循环载荷组和4周循环载荷组没有显著差异,Micro-CT计算纤维软骨带厚度结果与H&E染色测量结果未见相关性,通过Micro-CT计算骨量和骨小梁结果显示实验组和对照组没有显著差异。结论：①对髌骨骨腱结合部损伤实施阈下强度载荷训练和休息的干预手段可以改变骨的微细结构,而不能改变骨腱结合部的组织变化,骨的重建和恢复能力要强于骨腱结合部；②Micro-CT测定法可以更好描述纤维软骨带厚度的假设不成立。技术由于技术特点的限制,不能准确的计算骨腱结合部纤维软骨带的面积。
Patellar tendon junction injuries is very common disease in athlete and normal people, it is hot issues of sport medicine. Animal experiment is an important part of reseach the patellar tendon junction injuries mechanism and treatment. In this study, Nakama cyclic loading and the former stage of experimental animal models, based on the study of subliminal intensity exercise on bone tendon junction injuries, and by experimental calculation of Micro-CT technology feasibility with thickness of fibrocartilage, while the Micro-CT technical computing the experimental group and control group trabecular bone volume and bone changes.
16matual female New Zealand rabbits were randomly divided into4week cyclic loading group (n=8),8week cyclic loading group (n=8), the experiment left hind, right hind limb of the control group. Two Experiment groups of animals receiving a unified strength7.17N/S stimulation of muscle contraction in first4weeks, trained three times a week,2hours per time, total4hours. After4weeks,4weeks experimental cyclic loading group stopped and the8week cyclic loading group adjust the cyclic load4.2N'/S, experimental time constant.8weeks after the reunification of the subjects, the organization will be drawn after the first Micro-CT scan image reconstruction, followed by fixed, decalcified, dehydrated, embedded in paraffin, sliced and made H&E stained tissue sections, and using the image acquisition system for histological analysis.
Result:Comparing the control group,8weeks and4weeks group cyclic loading cyclic loading in the patellar tendon group there were hyaline, fibrous disorder;tide line of a"rising tide"and disappeared, thickening with fibrous cartilage, showing damage model was successfully created. H&E stained measuring microscopic cartilage with a thickness of8weeks cyclic loading group and4-week cycle of load groups were not significantly different, Micro-CT calculations fibrocartilage zone thickness results with H&E staining measurements no correlation, by Micro-CT calculation of bone mass and trabecular bone showed the experimental group and control group were not significantly different. Conclusion:①The low intensity cyclic loading training and rest can change the bone micro construction, but can not change the bone tendon junction injury in histologyily. The bone have better reconstruction and recovery abitly than the bone tendon junction;②Micro-CT can not accurately calculate the bone tendon junction fibrocartilage zone of the area.
引文
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[2]王人卫.优秀羽乒排网运动员运动创伤流行病学调查研究.中国运动医学杂志,2009,vol.28(4)：24-27
[3]陈东风,郑小飞.军事训练所致髌尖末端病的流行病学调查.中国现代医学杂志,2003,Vol.13(12)：54-55
[4]Julie Agel, David E. Olson, Randall Dick, et al. Descriptive Epidemiology of Collegiate Women's Basketball Injuries:National Collegiate Athletic Association Injury Surveillance System,1988-1989 Through 2003-2004.Journal of Athletic Training,2007,vol.42(2):202-210
[5]Benjamin, M,et al.Structure-function relationships in tendons:a review. J.Anat,2008,vol.20(4):23-35
[6]<运动解剖学>编辑委员会.运动解剖学,北京：人民体育出版社,1999：21-74
[7]于长隆.末端病.中国运动医学杂志,1984,vol.3(3)：180-186
[8]Ryuichi Gejo,Yuji Morita,Isao Matsushita,et al.Joint Gap Changes with Patellar Tendon Strain and Patellar Position During TKA.The Association of Bone and Joint Surgeons.2008.vol.15(3):143-149
[9]Benjamin, M.Theskeletal attachment of tendons—tendon'entheses'.J. Anat,2002,vol.11(9):57-74
[10]R..Cooper and S.Misol.Tendon and ligament insertion.A light and electron microscopic study J Bone Joint Surg Am,1970,vol.52(1):1-20
[11]Becker W.Die tendonpathien.Stuttgart:Thieme,1978:1-96
[12]J.L.Cook, J.A.Feller,S.F.Bonar,et al.Abnormal tenocyte morphpology is more prevalent than collagen disruption in asymptomatic athletes'patellar tendon.J Orhop Res,2004,vol.22(2):334-338
[13]N.Maffulli,V.Barras,et al.Light mircoscopic histology of Achilles tendon ruptures.A comparison with unruputered tendon.J Bone Joint Surg Am,2000,vol.28(6):857-863
[14]于长隆.髌尖末端结构对于生物意义.北京医学院报,1983,vol.15(4)：267
[15]P.Malliaras, J.L.Cook, P M Kent. Anthropometric risk factors for patellar tendon injury among volleyball players. Br J Sports Med,2007,vol.40(18):259-263
[16]Randall Dick, Jay Hertel, Julie Agel,et al. Descriptive Epidemiology of Collegiate Men's Basketball Injuries:National Collegiate Athletic Association Injury Surveillance System,1988-1989 Through 2003-2004. Journal of Athletic Training,2007,42(2):194-201
[17]陈世益.腱病的基础与临床若干研究.国外医学,骨科学分册,2005,vol.26(02)：83-86
[18]Havard Visnes, Roald Bahr. The evolution of eccentric training as treatment for patellar tendinopathy (jumper's knee):a critical review of exercise programmes. Br J Sports Med,2007, vol.41(12):217-223
[19]杜亚雯,王武韶,刘兆林.髌腱末端病二维超声与临床、X线综合研究.中国超声医学杂志,1999,vol.15(2)：133-134
[20]A Shalabi,L Svensson, M Kristoffersen-Wiberg et al. Tendon injury and repair after core biopsies in chronic Achilles tendinosis evaluated by serial magnetic resonance Imaging. Br J Sports Med,2004,vol.38(35):606-612
[21]M.Benjamin, H.Toumi, J.R.Ralphs et al. Where tendons and ligaments meet bone: attachment sites('entheses')in relation to exercise and/or mechanical load. J.Anat,2006 vol.208:471-490
[22]Esra Circi, Sercan Akpinar, Cenk Balcik et al. Biomechanical and histological comparison of the influence of oestrogen deficient state on tendon healing potential in rats. International Orthopaedics (SICOT),2009,vol.33:1461-1466
[23]扈盛,胡亚哲.末端病潮线涨潮的力学分析.中国运动医学杂志.2002,vol.21(4)：346--348.
[24]王恒俊,赵力.髌腱痛诊断与治疗的研究进展.医学综述,2008,vol.14(4)：567-569
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