本研究得到了国家自然科学基金（第82074523,82104996号）的支持。

所有动物实验均经北京中医药大学动物伦理委员会评审批准。BUCM-4-2022010101-1097）。在该协议中，24 只 6 周龄的雄性新西兰兔被饲养在特定条件下，即 20-25 °C、50-60% 湿度和 12 小时光照/12 小时黑暗昼夜节律周期，可以自由获得常规食物饮食。通过结合深度麻醉和空气栓塞术对兔子进行麻醉和处死。疼痛是KOA的典型病理特征之一，也是评估KOA动物模型和干预方法的关键指标之一，因此在模型制备过程中不使用镇痛药。
1. KOA兔模型
<ol><li>用 3% 戊巴比妥钠 （30 mg/kg） 在耳缘静脉注射麻醉兔子。为了确认适当的麻醉水平，寻找角膜反射明显减弱或消失，并且用止血钳夹住皮肤时没有疼痛。麻醉期间，每15分钟在兔子的眼睛上滴2-3滴润滑剂，以防止兔子的眼睛变干。</li>
	<li>麻醉后，将每只兔子固定在仰卧位，将左后肢拉到完全伸展的位置。</li>
	<li>将每只兔子的左后肢固定在伸展位置。<ol><li>作为第一层，用医用胶带覆盖兔子的皮肤，从腹股沟到踝关节。</li>
		<li>作为第二层，将 36 毫米宽的双面泡沫胶带缠绕在医用胶带上，然后用聚合物绷带从腹股沟缠绕到踝关节。确保膝关节伸直 180°，踝关节背屈 60°。</li>
		<li>作为第三层，在膝关节和踝关节的前后用小夹板固定关节，并在最外层缠绕钢网以防止咬伤。露出兔子的脚趾，观察血液循环是否正常。</li>
	</ol></li>
	<li>将动物固定6周以建立KOA模型（图1）。 注意： 1） 在模型准备过程中，每隔一天检查一次模具。如果任何霉菌松动或脱落，请麻醉兔子，并在伸展位置重新固定左后肢。2）在笼子底部铺设保护垫，防止兔子的四肢被卡住而受伤。</li>
</ol>2. 针灸干预
注意：在针灸术干预开始之前，通过耳缘静脉注射用3%戊巴比妥钠（30mg / kg）麻醉兔子。
<ol><li>确定治疗点。<ol><li>剃掉兔子左后肢膝关节的皮毛。</li>
		<li>触诊兔膝关节内侧股肌肌腱插入、股直肌肌腱插入、股二头肌肌腱插入和鹅足滑囊。用无菌皮肤标记标记局部肌肉的病理性硬结。用医用碘伏和 75% 医用酒精交替对膝关节进行三次消毒。</li>
	</ol></li>
	<li>针灸手术<ol><li>保持针灸刀片平行于平行于肌腱和肢体纵轴的行进方向。</li>
		<li>用左手的拇指向下按压进入标记点的皮肤并横向移动，使拇指腹侧的血管和神经分开。</li>
		<li>右手握住针灸手柄，用小力快速按压，使针灸刀片瞬间穿过皮肤。缓慢地将针灸刀片推进到局部肌肉硬结处，并进行纵向切割和横向摆动。</li>
		<li>针灸手术完成后，再次对膝关节进行消毒并贴上创可贴。</li>
	</ol></li>
	<li>每周执行一次此操作，持续 4 周（图 2）。 注意：1）如果股内侧肌、股外侧肌、股直肌、股二头肌或鹅肌滑囊的肌腱插入处没有接触硬结或索状组织，则应使用针灸针直接释放其肌腱插入处。2）针灸干预时，不要将针灸组和模型组兔的左后肢固定在伸展位置。</li>
</ol>3. 股四头肌弹性模量
注：1）本实验采用实时横波弹性成像（SWE）超声诊断仪测量每组兔体内股四头肌的弹性模量。2）测试人员应为具有超声检测经验的超声医师。测量时，应将超声探头轻轻放在股四头肌的皮肤表面，以避免局部肌肉紧张。当动物处于安静状态，没有挣扎或活动时，需要进行测量。如果动物处于活动状态，请等到它平静后再进行测试。
<ol><li>剃掉皮毛，露出左后肢股四头肌区域的皮肤。</li>
	<li>使用传统的二维超声来定位股四头肌肌肉腹部并确定感兴趣区域 （ROI），设置为 1-2 厘米的深度。</li>
	<li>启动 SWE 模式 进行检查。<ol><li>将感兴趣区域均匀设置为直径为 2 mm 的圆形区域，并将感兴趣区域设置为距皮肤表面 ~0.5-1 cm 深。</li>
		<li>使用超声诊断仪产生声辐射力脉冲，刺激肌肉组织，获得组织弹性成像。</li>
		<li>等待 2-3 秒让图像稳定，然后冻结图像。激活仪器的 Q-BOX 功能以测量股四头肌的杨氏模量。</li>
		<li>等待系统自动计算 ROI 杨氏模量的最大值、最小值和平均值（单位：KPa）。为三个测量值选择三个相同深度的 ROI，并取平均值进行统计分析。 注意：测试人员应是经验丰富的超声检测超声医师。测量时，应将超声探头轻轻放在股四头肌的皮肤表面，以避免局部肌肉紧张。当动物处于安静状态，没有挣扎或活动时，需要进行测量。如果动物处于活动状态，请等到它平静后再进行测试。</li>
	</ol></li>
</ol>4. 测量股四头肌的收缩力
注：测量股四头肌收缩力后，在麻醉下通过空气栓塞对兔子实施安乐死。
<ol><li>用3%戊巴比妥钠（30mg / kg）在耳缘静脉注射麻醉兔子。为了确认已达到适当的麻醉水平，请寻找角膜反射明显减弱或消失，并且用止血钳夹紧皮肤时没有疼痛。麻醉期间，每15分钟在兔子的眼睛上滴2-3滴润滑剂，以防止兔子的眼睛变干。</li>
	<li>暴露股四头肌并连接张力换能器。<ol><li>切开髌骨下方的皮肤，沿肢体纵轴向上切至大腿根部，继续向上切开皮肤3-4厘米。小心地剥离皮肤和筋膜，露出肌肉。切开髌韧带，小心地将股四头肌与髂关节分开，保持股四头肌与髂骨相连。</li>
		<li>在髌骨和股四头肌之间的肌腱连接处结扎手术缝合线。在自然状态下将肌肉拉伸至全长，然后将其连接到张力传感器上。将肌肉上的结扎线与力传感器上的结扎线保持在一条直线上。</li>
		<li>将张力传感器固定到支架上。将张力传感器上的信号采集线连接到生物信号采集系统处理器。</li>
	</ol></li>
	<li>测量股四头肌的收缩性能。<ol><li>将电极平行于股四头肌腹部插入，并避免电极之间有任何接触。</li>
		<li>按下示波器按钮。调整力传感器在支架上的位置，使基线保持在零。选择波宽为5ms，延迟为10ms的刺激器的刺激参数。</li>
		<li>首先使用单个刺激，然后从零开始逐渐调整刺激强度，每次增量为 0.1 V。观察肌肉收缩曲线和收缩幅度的变化，直到确定股四头肌的最大单次收缩幅度（Pt）。记录下来以备后续统计。</li>
		<li>使用集群刺激，以诱发最大单次收缩幅度的刺激幅值为基线，持续刺激肌肉，逐渐增加刺激频率。观察肌肉收缩曲线的变化，直到确定股四头肌的最大收缩幅度 （Pt）。记录下来以备后续统计。 注意： 1） 每次肌肉收缩后，应给予肌肉 30 秒放松，肌肉缓冲溶液不断滴在肌肉上。2）手术过程中，通过监测兔子的眼睑反射、呼吸节律、肌肉松弛和皮肤捏合反应来判断麻醉状态。</li>
	</ol></li>
</ol>5.股四头肌肌腱的力学性能
<ol><li>预处理：在测试当天，用游标卡尺测量股四头肌肌腱的长度、宽度和厚度，并在疲劳试验机中安装专用防滑夹具。重复加载和卸载 15 倍进行预处理。</li>
	<li>应力松弛测试：使用0N至100N的传感器，以5mm/min的速度拉伸，直到达到所需长度，然后开始收集数据。从t（0）开始设置计算机数据采集时间，每0.1秒采集一次数据，持续1800秒。达到设定时间后，记录数据和曲线。</li>
	<li>拉伸试验：使用0N至100N的传感器，以5mm/min的速度将其拉伸至最大载荷，直到试样被拉开。试验后，计算试样的最大位移、极限载荷和刚度。</li>
</ol>6.关节接触面压力和软骨单位面积压力
<ol><li>将两侧的股骨和胫骨标本固定在固定装置上的笔直位置并进行预紧力测试。测量膝关节的大致范围，将压敏纸切成相同的形状，并用保鲜膜密封。</li>
	<li>将密封的压敏纸放在胫骨和股骨关节之间，以5mm/min的压力和50N的最大压力对膝关节进行压力测试。 保持压力 2 分钟，直到压力敏纸稳定着色时达到 50 N。</li>
	<li>2分钟后，取出压敏纸，将彩色表面固定在A4尺寸的纸上，并在放置比例的情况下获取图像。</li>
	<li>将图像上传到计算机。使用参考软件进行面积测量和不规则图形的多段测量。测量股骨和胫骨关节内外的压力并记录结果。</li>
</ol>7. Safranin O/Fast Green 膝关节软骨染色
<ol><li>针灸干预结束后，取软骨 - 软骨下骨复合物组织并将其嵌入石蜡中。将准备好的组织蜡块切片并准备载玻片。用环境脱蜡溶液（I）和环境脱蜡溶液（II）将制备的组织载玻片脱蜡，每次15分钟;然后，依次浸入二甲苯和无水乙醇（1：1），无水乙醇（I），95%乙醇，85%乙醇和75%乙醇中，每步2-5分钟;最后，将它们浸泡在蒸馏水中15分钟。</li>
	<li>进行染色。<ol><li>用Fast Green溶液染色载玻片1分钟。在此过程中，将载玻片从溶液中取出并在显微镜下观察它们，直到组织染成深绿色。</li>
		<li>分色：用超纯水冲洗多余的Fast Green溶液。将载玻片快速浸泡在1%醋酸溶液中5-10秒。 再次，用超纯水冲洗载玻片。</li>
		<li>将载玻片浸入Safranine O溶液中10-15分钟，直到软骨染成红色。</li>
	</ol></li>
	<li>脱水并澄清组织，密封载玻片，并在显微镜下观察它们。<ol><li>将载玻片依次浸泡在75%乙醇，85%乙醇，95%乙醇和100%乙醇中3-5秒。</li>
		<li>将载玻片连续浸入环境脱蜡溶液（I）和环境脱蜡溶液（II）中10分钟。取出载玻片，将中性树脂介质放在载玻片的前面，避开组织。将盖玻片的边缘放在载玻片上，然后慢慢放下以覆盖中性香脂。去除空气，避免气泡。擦去多余的二甲苯和中性香脂，让它在室温下静置过夜。</li>
		<li>在显微镜下观察载玻片并获取图像。对于每组，选择六个兔膝软骨样本，并为每个样本随机选择四个不同的观察区域进行评估。根据Mankin方法对每组的软骨组织学进行评分（表1）。</li>
	</ol></li>
</ol>8. 统计分析
<ol><li>将数据表示为平均值±标准差 （<img alt="Equation 1" src="/files/ftp_upload/65584/65584eq01.jpg" style="margin: auto;"> ± s）。</li>
	<li>执行单因素方差分析 （ANOVA） 和 LSD 检验，以确定多组比较的统计显着性。</li>
	<li>当 P < 为 0.05 时，考虑具有统计学意义的差异。</li>
</ol>

使用改进的 Videman 方法在兔子中建立 KOA 模型

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Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+acupotomy+therapy+on+tenderness+point+around+knee+joint+in+patients+with+knee+osteoarthritis:+a+randomized+controlled+clinical+trial.">Effects of acupotomy therapy on tenderness point around knee joint in patients with knee osteoarthritis: a randomized controlled clinical trial.</a> Journal of Chengdu University of Traditional Chinese Medicine. 33 (3), 3-5 (2010).</li><li>Zhao, Y., Dong, F. H., Zhang, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+soft+tissues+mechanical+changes+and+treatment+of+meridian+tendon+pain.">Analysis of soft tissues mechanical changes and treatment of meridian tendon pain.</a> Beijing Journal of Traditional Chinese Medicine. (9), 705-707 (2008).</li><li>Zhang, L. P., Cheng, F., Liu, D. Y., Zhu, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+progress+of+biomechanics+in+knee+osteoarthritis.">Application progress of biomechanics in knee osteoarthritis.</a> Chinese Archives of Traditional Chinese Medicine. 34 (7), 1644-1647 (2016).</li><li>Thomas, A. C., Sowers, M., Karvonen-Gutierrez, C., Palmieri-Smith, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lack+of+quadriceps+dysfunction+in+women+with+early+knee+osteoarthritis.">Lack of quadriceps dysfunction in women with early knee osteoarthritis.</a> J Orthop Res. 28 (5), 595-599 (2010).</li><li>Li, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+unloaded+muscle+atrophy+on+contractile+characteristics+of+hind-limb+skeletal+muscles+in+mice.">Effects of unloaded muscle atrophy on contractile characteristics of hind-limb skeletal muscles in mice.</a> Space Med Med Eng. 25 (5), 322-325 (2012).</li></ol>

作者没有需要披露的利益冲突。

合适的动物模型是实现实验目标和阐明特定科学问题的关键因素之一。本研究基于针灸学中的“宗进控制骨骼和润滑关节”和“机械失衡”理论，旨在解释针灸治疗中“调节肌肉和肌腱治疗骨骼疾病”治疗KOA背后的科学内涵。换句话说，针灸术通过调节膝关节周围软组织的力学特性来延缓退化并保护软骨，从而改善软骨的异常机械环境。KOA动物模型一般分为两类：自发模型和诱导模型。由于建模?...

膝骨关节炎 （KOA） 是最常见的骨关节炎形式，通常被认为是一种以关节软骨变性为特征的全关节疾病，临床表现为受累关节疼痛、肿胀和活动受限1。根据最近的流行病学统计数据，据报道，到 2020 年，全球 40 岁或以上的 KOA 影响了 6.541 亿人。KOA 的患病率和发病率随着年龄的增长而上升，在中老年人中最高，并且影响的女性多于男性2。由于全球人口老龄化和肥胖流行，KOA的患病率可能会增加，对全球公共卫生构成越来越大的威胁。年龄、性别、肥胖、创伤和其他与 KOA 相关的复杂危险因素都直接影响膝关节不稳定，使膝关节的生物力学失衡成为 KOA3 的主要原因之一。
在正常生理条件下，膝关节处于机械平衡状态，保证关节内的机械负荷均匀分布在软骨上。膝关节的任何机械失衡都会导致软骨异常应力，导致软骨退化和 KOA4 的发作。肌腱系统是维持膝关节机械平衡的主要动力系统。伸屈肌腱系统的协调运动可以均匀地分布在软骨表面的运动产生的负荷，避免局部软骨应力超出其生理负荷的代谢失衡，导致软骨流失5。肌肉力量下降是肌肉内运动障碍和软骨损伤的主要原因，这可能发生在有症状的 KOA 之前。 
KOA 还可诱发关节肌抑制 （AMI），表现为肌肉无力和膝关节周围肌肉力量下降6.在这些肌肉中，股四头肌群是唯一的膝伸肌，是维持膝关节稳定性的重要结构。研究表明，股四头肌横截面积和肌肉力量的减少与 KOA 进展显着正相关7。股四头肌力量的下降会影响步态模式、膝关节稳定性、运动模式和许多其他功能。此外，肌肉力量的下降会损害肌腱功能，表现为肌腱刚度、弹性模量和其他生物力学特性的降低8.在长期应变修复中，膝关节的肌肉和肌腱可能发生粘连、挛缩等变化，破坏其力学性能，引起关节不稳定，最终形成KOA病理变化的恶性循环。因此，对于KOA治疗来说，改善肌腱系统的机械性能并恢复关节机械平衡至关重要。
在 KOA 的原因中，生物力学失衡是膝关节疼痛、功能障碍、炎症性病变和软骨退行性变的主要诱发因素9。因此，治疗KOA的关键是恢复膝关节的生物力学平衡。针灸学认为KOA的病因和发病机制是“机械失衡”。当膝关节周围软组织的力学特性发生异常变化时，膝关节失去机械平衡，关节异常的机械应力环境加速退化，引起炎症刺激，进一步加重软组织粘连、挛缩，关节稳定性进一步下降。这种恶性循环最终发展成KOA。针灸术通过松弛软组织粘连和挛缩，以及降低肌肉和肌腱的压力集中，结合“调节肌肉和肌腱治疗骨骼疾病”的理论，改善软组织力学，“调节肌肉和肌腱”，平衡关节的机械应力，有效缓解软骨退化和“治疗骨骼疾病”10.在动物模型选择方面，基于本研究的目的，我们采用改良的Videman左后肢伸展固定方法制备了KOA模型。
本文详细介绍了使用改良的Videman左后肢伸展固定方法建立KOA模型以及针灸切开术的操作方法和注意事项。我们通过测试股四头肌和肌腱的机械性能以及检测关节软骨应力和形态的变化来证明针灸切开术的有效性。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Acupotomy</td>
			<td>Beijing Zhuoyue Huayou Medical Devices Co., Ltd.</td>
			<td></td>
			<td>0.4 x 40 mm</td>
		</tr>
		<tr>
			<td>Connect Cast Orthopedic Casting Tape</td>
			<td>Suzhou Connect Medical Technology Co.,Ltd.</td>
			<td>KCP06</td>
			<td>15.0 cm x 360 cm</td>
		</tr>
		<tr>
			<td>Double-sided Foam Tape</td>
			<td>Deli Group Co.,Ltd.</td>
			<td>NO.30416</td>
			<td>36 mm x 5 yard x 2.5 mm</td>
		</tr>
		<tr>
			<td>Environmental Dewaxing Solution</td>
			<td>Wuhan Servicebio Technology Co.,Ltd.</td>
			<td>G1128</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol absolute</td>
			<td>Beijing Hengkangda Medicine Co., Ltd.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fast Green solution</td>
			<td>Wuhan Servicebio Technology Co.,Ltd.</td>
			<td>G1031</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast grenn FCF</td>
			<td>Sigma,America</td>
			<td>2353-45-9</td>
			<td></td>
		</tr>
		<tr>
			<td>Fatigue testing machine</td>
			<td>BOSE, America</td>
			<td></td>
			<td>Bose Electro Force 3300</td>
		</tr>
		<tr>
			<td>Four-channel physiological recorder</td>
			<td>Chengdu Instrumeny Frctory</td>
			<td></td>
			<td>RM-6420</td>
		</tr>
		<tr>
			<td>FPD-305E</td>
			<td>Fuji, Japan</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>FPD-306E</td>
			<td>Fuji, Japan</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin solution</td>
			<td>Wuhan Servicebio Technology Co.,Ltd.</td>
			<td>G1005</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical iodophor disinfectant</td>
			<td>Shan Dong Lircon Medical Technology Co., Ltd.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Medical Tape</td>
			<td>Shandong Rongjian Sanitary Products Co., Ltd.</td>
			<td>200402</td>
			<td>1.5 x 500 cm</td>
		</tr>
		<tr>
			<td>Muscle tension transducer&#160;</td>
			<td>Chengdu Instrumeny Frctory</td>
			<td></td>
			<td>JH-2204005, 50 g</td>
		</tr>
		<tr>
			<td>Prescale</td>
			<td>Fuji, Japan</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Real-time SWE ultrasound diagnostic instrument</td>
			<td>SuperSonic Imagine SA,France</td>
			<td></td>
			<td>SuperSonic Imagine AixPlorer</td>
		</tr>
		<tr>
			<td>Rhamsan gum</td>
			<td>Wuhan Servicebio Technology Co.,Ltd.</td>
			<td>WG10004160</td>
			<td></td>
		</tr>
		<tr>
			<td>Safranine O</td>
			<td>Sigma,America</td>
			<td>477-73-6</td>
			<td></td>
		</tr>
		<tr>
			<td>Safranine O solution</td>
			<td>Wuhan Servicebio Technology Co.,Ltd.</td>
			<td>G1015</td>
			<td></td>
		</tr>
		<tr>
			<td>Statistical Package for the Social Sciences (SPSS)</td>
			<td>IBM, America</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

application of acupotomy in a knee osteoarthritis model in rabbit

膝骨关节炎（KOA）是骨科最常遇到的疾病之一，严重降低了KOA患者的生活质量。在几个致病因素中，膝关节的生物力学失衡是KOA的主要原因之一。针灸学认为，恢复膝关节的机械平衡是治疗KOA的关键。临床研究表明，针灸术可以通过减少膝关节周围肌肉和肌腱的粘连、软组织挛缩和应力集中点来有效减轻疼痛并改善膝关节活动度。 
在该协议中，我们使用改进的 Videman 方法通过将左后肢固定在笔直位置来建立 KOA 模型。我们详细概述了针灸术的操作方法和注意事项，并通过检测股四头肌和肌腱的力学性能以及软骨力学和形态学，结合“调节肌肉和肌腱治疗骨骼疾病”理论评估了针灸切开术的疗效。结果表明，针灸术通过调节膝关节周围软组织的力学性能，改善软骨应力环境，延缓软骨退化，对软骨具有保护作用。

Knee osteoarthritis (KOA) is the most frequent form of osteoarthritis, often recognized as a whole-joint disease characterized by articular cartilage degeneration, which manifests clinically as pain, swelling, and limited movement of the affected joints1. According to recent epidemiological statistics, KOA is reported to have affected 654.1 million individuals globally who were 40 years of age or older by 2020. The prevalence and incidence of KOA rise with age, are the highest in middle-aged and older adults, and affect more women than men2. The prevalence of KOA is likely to increase due to the aging population and obesity epidemic worldwide, posing a growing threat to global public health. Age, sex, obesity, trauma, and other complicated risk factors associated with KOA all directly impact knee instability, making a biomechanical imbalance in knee joints one of the primary causes of KOA3.
Under normal physiological conditions, the knee joint is in a state of mechanical balance, ensuring that the mechanical loads in the joint are evenly distributed on the cartilage. Any mechanical imbalance in the knee joint can lead to abnormal stress in cartilage, resulting in cartilage degeneration and the onset of KOA4. The muscle-tendon system is the main dynamic system that maintains the mechanical balance of the knee joint. The coordinated movement of the extensor and flexor muscle-tendon system can evenly distribute the load generated by the movement on the cartilage surface, avoiding the metabolic imbalance of local cartilage stresses beyond its physiological load that results in cartilage loss5. Decreased muscle strength is the main cause of intramuscular movement disorder and cartilage damage, which may occur before symptomatic KOA. 
KOA can also induce arthrogenous muscle inhibition (AMI), manifesting as muscle weakness and decreased muscle strength around the knee6. Among these muscles, the quadriceps femoris group functions as the only knee extensor, an important structure in maintaining knee joint stability. Studies have shown that a decrease in quadriceps cross-sectional area and muscle strength is significantly and positively correlated with KOA progression7. The decline in quadriceps strength affects the gait pattern, knee stability, movement patterns, and many other functions. Moreover, the decline in muscle strength impairs tendon function, manifested as a decrease in tendon stiffness, elastic modulus, and other biomechanical properties8. In long-term strain repair, changes such as adhesion and contracture may occur in the muscles and tendons of the knee joint, damaging their mechanical properties, causing joint instability, and ultimately forming a vicious cycle of pathological changes of KOA. It is, therefore, crucial for KOA treatment to improve the mechanical properties of the muscle-tendon system and restore the joint mechanical balance.
Among the causes of KOA, biomechanical imbalance is the main inducing factor for knee pain, dysfunction, inflammatory lesions, and cartilage degeneration9. Therefore, the key to treating KOA is to restore the biomechanical balance of the knee joint. Acupotomology believes that the etiology and pathogenesis of KOA are "mechanical imbalance." When the mechanical characteristics of the soft tissues around the knee change abnormally, the knee joint loses its mechanical balance, and the abnormal mechanical stress environment of the joint accelerates degeneration, causing inflammatory stimulation to further aggravate the soft tissue adhesions, contractures, and further decline in joint stability. This vicious cycle eventually develops into KOA. By loosening soft tissue adhesions and contractures, as well as reducing stress concentration in the muscles and tendons, acupotomy in conjunction with the theory of "Modulating Muscles and Tendons to Treat Bone Disorders" improves the soft tissue mechanics and "modulates muscles and tendons," which balances the mechanical stress of the joint, effectively alleviating cartilage degeneration and "treating bone disorders"10. In terms of animal model selection, based on the purpose of this study, we prepared the KOA model by the modified Videman method of left hindlimb extension immobilization.
This paper details the establishment of the KOA model using the modified Videman method of left hind limb extension immobilization and the method of operation and precautions of acupotomy. We demonstrate the effectiveness of acupotomy by testing the mechanical properties of quadriceps femoris and tendon and detecting changes in articular cartilage stress and morphology.

作者聚焦：研究针尖切开术治疗膝骨关节炎的作用机制

针灸术在兔膝骨关节炎模型中的应用

Our research scope is the study of the mechanism of action of acupotomy in the treatment of new osteoarthritis. The question we want to answer, the how acupotomy regulates the mechanical properties of the soft tissues around the knee to protect the cartilage. At present, micro CT, nuclear magnetic resonance, phenetole elemental analysis, and bio mechanical detection techniques can be used to promote research in this field.We found that acupotomy improved the mechanical properties of soft tissue by removing the excisions and pathological contractors around the knee, thereby playing a role in protecting cartilage. To a certain extent, this protocol explains the mechanism and reveals the scientific connotation of acupotomy in the treatment of KOA and fills the gap in the basic research of acupotomy tendon and bone treatment method. In the future, our laboratory will focus on the specific mechanism or action of the acupotomy tendon and bone control method on cartilage protection.

股四头肌和肌腱力学性能实验结果 为了评估针灸学对KOA兔股四头肌力学性能的影响，我们分别使用了实时横波弹性超声成像和肌肉张力换能器。与对照组相比，KOA组股四头肌Young模量降低（P < 0.05）。与KOA组相比，针灸组的Young模量增加（P < 0.05，图3A）。股四头肌收缩能力方面，与对照组相比，KOA组股四头肌单次收缩幅度和股四头肌强直收缩幅度...

Watch this Scientific Journal Video about Author Spotlight: Investigating the Mechanism of Action of Acupotomy in Treating Knee Osteoarthritis at JoVE.com

在该方案中，使用改良的Videman方法制备了膝骨关节炎模型，并详细介绍了针灸切开术的操作规程和注意事项。针灸切开术的有效性已通过测试股四头肌和肌腱的机械性能以及软骨的机械和形态特性得到证明。

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of ...

我们的研究范围是研究针灸术治疗新发骨关节炎的作用机制。我们想要回答的问题是，针灸术如何调节膝盖周围软组织的机械性能以保护软骨。目前，显微CT、核磁共振、苯乙醚元素分析、生物力学检测技术等技术可用于推动该领域的研究。我们发现针灸术通过去除膝盖周围的切除和病理收缩来改善软组织的机械性能，从而在保护软骨方面发挥作用。该方案在一定程度上阐明了针灸治疗KOA的机理，揭示了针灸治疗KOA的科学内涵，填补了针灸肌腱和骨治疗方法基础研究的空白。未来，我们的实验室将重点研究针灸肌腱和骨控制方法对软骨保护的具体机制或作用。

application-of-acupotomy-in-a-knee-osteoarthritis-model-in-rabbit

Research

JoVE Journal

Medicine

Application of Acupotomy in a Knee Osteoarthritis Model in Rabbit

680 次观看.  Beijing University of Chinese Medicine. 我们的研究范围是研究针灸术治疗新发骨关节炎的作用机制。我们想要回答的问题是，针灸术如何调节膝盖周围软组织的机械性能以保护软骨。目前，显微CT、核磁共振、苯乙醚元素分析、生物力学检测技术等技术可用于推动该领域的研究。我们发现针灸术通过去除膝盖周围的切除和病理收缩来改善软组织的机械性能，从而在保护软骨方面发挥作用。该方案在一定程度?...

视频: 作者聚焦：研究针尖切开术治疗膝骨关节炎的作用机制

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of people with KOA. Among several pathogenic factors, the biomechanical imbalance of the knee joint is one of the main causes of KOA. Acupotomology believes that restoring the mechanical balance of the knee joint is the key to treating KOA. Clinical studies have shown that acupotomy can effectively reduce pain and improve knee mobility by reducing adhesion, contracture of soft tissues, and stress concentration points in muscles and tendons around the knee joint. 
In this protocol, we used the modified Videman method to establish a KOA model by immobilizing the left hindlimb in a straight position. We have outlined the method of operation and the precautions related to acupotomy in detail and evaluated the efficacy of acupotomy in conjunction with the theory of "Modulating Muscles and Tendons to Treat Bone Disorders" through the detection of the mechanical properties of quadriceps femoris and tendon, as well as cartilage mechanics and morphology. The results show that acupotomy has a protective effect on cartilage by adjusting the mechanical properties of the soft tissues around the knee joint, improving the cartilage stress environment, and delaying cartilage degeneration.

In this protocol, a knee osteoarthritis model was prepared using the modified Videman method, and the operation procedures and precautions of acupotomy are detailed. The effectiveness of acupotomy has been demonstrated by testing the mechanical properties of quadriceps femoris and tendon and the mechanical and morphological properties of cartilage.

科研

医学

Establishing the Knee Osteoarthritis Rabbit Model and Applying Acupotomy for Modulating Muscles and Tendons

To begin, fix the anesthetized rabbit in a supine position. Then, pull and fix the left hind limb in the fully extended position. Using medical tape as the first layer, cover the skin from the groin to the ankle joint.For the second layer, wrap 36 millimeter-wide double-sided foam tape over the medical tape. Then, wrap a polymer bandage from the groin to the ankle joint. Ensure the knee joint is 180 degrees straight, and the ankle joint is dorsiflexed by 60 degrees.For the third layer, immobilize the joints with small splints at the front and back of the knee and ankle joints. Then, wrap steel mesh around the outermost layer to protect against bites. Ensure to keep toes exposed, to observe blood circulation.To determine points of acupotomy treatment, palpate and mark pathological indurations of knee joints with a sterile skin marker. Then, disinfect the knee joint three times with alternating rounds of iodophor and 75%ethanol. Now align an acuoptomy blade parallel to the travel direction of the tendon and the longitudinal axis of the limb.Using the left thumb, press down on the skin at the marker point, then shift laterally to facilitate the separation of blood vessels and nerves on the thumb's ventral side. With the acupotomy handle in the right hand, apply a quick gentle pressure, causing the acupotomy blade to penetrate the skin. Slowly advance the blade to muscle induration, making longitudinal cuts and swinging laterally.After acupotomy, disinfect the knee joint using iodophor and apply a bandaid.

建立膝骨关节炎兔模型并应用针灸术调节肌肉和肌腱

Measuring Mechanical Properties of Quadriceps Femoris Contraction Force in Rabbits

To begin, cut the skin of an anesthetized rabbit below the patella, following the length of the limb upwards to the thigh base, then continue upwards for three to four centimeters. Carefully peel off the skin and fascia, exposing the quadriceps muscle. Cut the patellar ligament and carefully separate the quadriceps from the iliac junction, while maintaining the quadriceps in connection with the ilicium.Now ligate the surgical sutures at the tendon junction between the patella and quadriceps muscle. Stretch the muscle to its full length in its natural state and attach it to the tension transducer, aligning the ligation line on the muscle with that on the transducer. Secure the tension transducer to the bracket.Connect the signal acquisition line on the transducer to the bio signal acquisition system processor. Insert the electrodes parallel to the quadriceps abdomen, avoiding contact between the electrodes. Next, press the oscilloscope button.Adjust the position of the force transducer on the bracket to maintain the baseline at zero. At this stage, set the stimulator stimulation parameters with a single stimulus, a wave width of five milliseconds and a delay of 10 milliseconds. Set the initial value of the stimulus amplitude to zero volt and set the automatic amplitude to increase by 0.1 volt each time, to gradually increase the stimulus intensity.Then start the stimulation while observing the changes in the muscle contractions curve and contraction amplitude, until the maximum signal twitch amplitude of the quadriceps is determined. Next, use a cluster stimulus and set the stimulus amplitude, causing a maximum single twitch as the baseline to continuously stimulate the muscle. Set the initial value of the stimulation frequency to 10 hertz, and set the automatic frequency to increase by one hertz each time to gradually increase the stimulus frequency.Finally, start the stimulation while observing the changes in the muscle contraction curve until the maximum tetanic contraction amplitude of the quadriceps is determined. Young's modulus, single contraction amplitude, and tetanic contraction amplitude of the quadriceps femoris were significantly reduced in knee osteoarthritis group compared to the control and Acupotomy group.

测量兔子股四头肌收缩力的机械性能

Safranin O/Fast Green Staining of Knee Joint Cartilage in Rabbits

After acupotomy intervention, embed the cartilage subchondral bone complex tissues in paraffin. Then, slice the tissue wax blocks and place them on the slides. Deparaffinize the slides with successive environmental dewaxing solutions for 15 minutes each.Then, dip the slides successively in xylene and anhydrous ethanol for two to five minutes each, followed by dehydration in decreasing ethanol concentrations. Stain the slide with fast green solution for one minute. After rinsing the excess fast green solution with ultrapure water, rinse slide quickly with 1%acetic acid solution several times for color separation.Again, rinse the slide with water as demonstrated before staining with safranin O solution for 10 to 15 minutes. Soak the slide in increasing ethanol concentrations for three to five seconds each, then, place the slide in successive xylene solution for 10 minutes each. Add a neutral resonance medium on the front of the slide, avoiding the tissue.Place the edge of the cover glass on the slide and slowly lower it to cover the neutral balsam. Wipe off the extra xylene and neutral balsam. After overnight drying, observe the slide under the microscope for cartilage histology scoring.In the control group, the cartilage surface was smooth and exhibited organized chondrocyte arrangement in all layers. In contrast, the cartilage surface was rough in the KOA group and the chondrocyte arrangement was disordered. Acupotomy improved the smoothness of the cartilage surface and preserved normal chondrocyte structure.The cartilage integrity score was significantly increased in the KOA group compared to the control and acupotomy group.