这项工作得到了中国国家自然科学基金资助82100982 F.L.的支持。

本研究中的动物程序由四川大学华西口腔医学院伦理委员会审查和批准（WCHSIRB-D-2017-041）。本研究使用成年C57BL / 6小鼠（参见 材料表）。该方案增加了右上颌第一磨牙 （M1） 的机械负荷，用于近中运动，其中通过骨吸收和骨形成的耦合诱导高度协调的骨重塑过程（图 1）。
1.术前准备
<ol><li>手术项目<ol><li>为手术准备以下手术物品:手术平台（图2A），紧固件（图2B），手术器械（图2C 和 补充图S1），正畸用品（图2C）和牙齿修复用品（图2D）。 注意: 定制的螺旋弹簧是定制的，当拉伸到 10 毫米时提供 10cN 的力。</li>
	</ol></li>
	<li>灭菌<ol><li>通过高压灭菌对手术器械进行消毒，并用紫外线照射对所有手术物品进行消毒至少30分钟。</li>
	</ol></li>
	<li>麻醉<ol><li>通过腹腔注射给予氯胺酮（100mg / kg）和地西泮（5mg / kg）麻醉小鼠。</li>
		<li>用棉棒将兽医软膏涂抹在小鼠的眼睛上，以避免眼睛干涩。</li>
		<li>只有当用镊子捏住老鼠的脚趾时鼠标没有反应时，才进行手术。</li>
	</ol></li>
</ol>2.手术过程
<ol><li>使用胶带将麻醉小鼠的四肢以仰卧位展开并用胶带粘贴到手术平台上。</li>
	<li>在头部上方的两侧钉一根 27 G 针，在腋窝下方的两侧钉另一根 27 G 针。</li>
	<li>用橡皮筋缠绕在上面的两根针和上门牙上，用另一根橡皮筋缠绕在另外两根针和下门牙上。改变针头位置以控制张开程度和嘴的方向（图3A）。 注意: 对于正畸牙齿移动操作，在 buccinator 完全收紧之前，请保持嘴巴最大张度。应将舌头拉向非手术侧，以暴露手术区域并防止缺血。</li>
	<li>弯曲 1.5 cm 304 不锈钢丝的 304 mm 端，并用弯曲的眼镊子将弯曲端从颊侧推过 M1 和上颌第二磨牙 （M2） 之间的相邻空间（图 3B）。当从腭侧看到结扎线的腭端时，将其拉出至其长度的一半左右，并将其穿过定制螺旋弹簧的一端。</li>
	<li>用结扎线的两端在上颌骨 M1 的近中方向打一个方形结，直到弹簧牢固地固定在牙齿上（图 3C）。减去多余的电线。</li>
	<li>同样，将第二根 3 cm 304 不锈钢丝穿过螺旋弹簧的另一端。</li>
	<li>用棉球清洁并擦干门牙表面。用棉棒在所有这些表面上涂上粘合剂并光固化。</li>
	<li>将第二根不锈钢丝推过上颌门牙之间的相邻空间，并在唇部打一个滑结（图3D）。减去多余的金属丝，使金属丝的其余部分靠近齿面。</li>
	<li>注入光固化树脂覆盖结和门牙;光固化树脂（图3E）。</li>
</ol>3.术后管理
<ol><li>手术后，腹膜内注射0.05mg/kg丁丙诺啡用于术后镇痛。</li>
	<li>将麻醉的小鼠放在37°C恒温电热毯上。当小鼠通过行走恢复意识时，将其放回单独的笼子中。</li>
	<li>由于手术后门牙的功能有限，请仅用软食代替常规硬饲料。</li>
	<li>每天检查正畸矫治器。如果在检查过程中观察到任何影响正畸力传导的情况，例如弹簧变形、弹簧松动和装置脱落，则应将小鼠排除在实验之外。</li>
	<li>为了保持实验的可比性，每天评估手术后小鼠的体重。任何体重减轻超过其术前体重30%的小鼠都必须被排除在实验之外。</li>
</ol>

小鼠上颌骨正畸模型骨重塑分析指南

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作者声明没有利益冲突。

本文试图逐步描述小鼠上颌骨模型上最简单的正畸牙齿运动方案，以研究机械负荷诱导的骨重塑的潜在机制。除了骨重塑的研究外，这种方法还有一些其他的主流应用:1）正畸牙齿运动加速的方法学研究;2）正畸根部吸收研究;3）正畸牙齿运动和疼痛的生物学机制;4）转基因模型研究。
与其他机械负荷相关治疗（如下颌牵引成骨术 37）相比，正畸牙齿运动是最简单、最温和的方法，不会造成伤口和出血。此外，鼠模型具有易于操作、时间更短、成本更低的优点 38.上颌骨模型可以在操作过程中提供宽广的视野和稳定的固定，并且在操作后舌头对器具的干扰最小14.
在此建立的模型的基础上，我们进一步描述了三个具有代表性的时间点。从术后第三天开始，可以从宏观上测量牙齿移动，并且移动距离随时间增加。术后第3天，通过牙周韧带纤维向骨骼增加机械负荷，骨骼无明显变化。术后第8天，骨重塑已经开始，骨吸收占主导地位，术后第14天骨形成占主导地位。该模型可以显示正畸牙治疗过程中不同阶段的骨重塑特征。
需要考虑一些关键的操作步骤。在协议步骤2.7之前，鼠标的头部应朝向操作员，以获得更好的手术视野。在协议步骤2.4之后，操作区域靠近门牙，鼠标尾部必须朝向操作员。当必须将不锈钢丝从颊侧推过M1和M2之间的相邻空间时，需要预弯以安全地定位目标区域并减少器械在口腔中占用的空间。弯曲角度应为>45°，以确保不锈钢丝在通过邻间空间时不会刺穿牙龈。以平行方式穿孔是阻力最小的方法。小角度的咬合线穿孔也可以通过光滑坚韧的牙齿表面引导到腭侧。弯曲的眼科镊子的尖端应夹紧弯曲处，以减少口腔内占用的空间并便于用力（补充图S2）。
由于不锈钢丝可能无法穿过上颌门牙之间的邻间空间，因此齿状眼镊有助于门牙分离。此外，不需要方形结，因为树脂粘接是这里的主要保留方法。滑结几乎可以在靠近齿面的地方打结，方形结会增加涂层树脂的体积。
但是，这种模式也有其缺点。由于口腔中存在异物的感觉，正畸矫治器可能会被小鼠破坏。臼齿侧的部分停留在咬合平面下方，难以破坏。然而，下切牙正好咬在门牙侧的固定部分，包括螺旋弹簧的末端。因此，我们建议两个上切牙的所有表面都应用树脂包裹，以增加保持力。弹簧的门牙端 - 最薄弱的部分 - 可能被树脂覆盖（补充图S3）。总之，该协议逐步展示了在小鼠上颌骨模型上操作的正畸牙齿运动的细节。通过对每个步骤的明确解释和视觉演示，研究人员可以掌握该模型，并通过一些修改将其应用于他们的实验需求。

骨骼是一种高度活跃的重建组织，在个体的一生中会调整其大小、形状和特性 1,2。除了荷尔蒙、衰老、营养和其他生物或生化因素3外，机械负荷是最具决定性因素的观点也得到了普遍接受4,5。在一些机械负荷异常的情况下，骨吸收和骨形成之间的不平衡可能导致异常的骨重塑和骨骼疾病。长期卧床休息或在太空飞行中存在微重力条件下的废用性骨质疏松症和骨质流失等骨骼疾病与异常机械负荷密切相关6,7,8。
机械负荷也被用于治疗骨骼相关疾病，如牵引治疗和正畸治疗。牵引治疗已被用于发育性疾病，如颅缝早闭和下颌骨发育不全 9,10，而正畸治疗已被广泛用于矫正异常的牙齿位置和任何咬合不正11。正畸治疗的核心也是机械负荷的管理。当骨组织承受机械负荷时，通过骨吸收和骨形成的耦合诱导高度协调的骨重塑过程，这可以移动牙齿以达到正畸目的12,13。
虽然正畸治疗在临床实践中得到了广泛的应用，但由于我们对机械负荷的生物学效应的认识有限，正畸治疗的结果是不可控的。为了克服这些局限性，已经建立了几种动物模型，如小鼠、大鼠、兔、猫、狗、猴和猪，以研究机械负荷诱导的骨重塑的潜在机制（表1）14,15,16,17,18,19,20,21,22,23,24，25、26、27、28、29、30、31、32。狗、猴子、猪等大型动物在正畸手术中比小动物有一些优势——它们有更多类似人类的牙齿和牙列，因此外科手术很容易在人类身上复制。此外，广阔的视野可以降低手术难度，并可以应用各种正畸方案33,34。然而，大型动物很难获得，导致与样本量相关的挑战，并且它们受到伦理限制35.此外，常规的提取程序和复杂的仪器使实验难以进行，因此很少使用大型动物。
在这种情况下，啮齿动物主要用于建立正畸模型。在这些模型中，与小鼠相比，大鼠和兔子的操作难度较低，牙齿运动方案更多。然而，小鼠模型具有独特的优势，即有大量转基因小鼠可用，这对于研究潜在机制尤为重要36。然而，小鼠模型是最难操作的模型，因为它的体积很小。回顾目前的方法，将第一磨牙沿近中方向移动是正畸模型的唯一实用方法。两个装置主要用于移动齿形螺旋弹簧和松紧带。使用松紧带比较容易，但正畸力变化很大，很难获得稳定的结果。
Xu等[15 ]建立了下颌骨上有螺旋弹簧的小鼠模型。然而，由于下颌骨的活动性和舌头的阻塞性，上颌骨手术始终是术中和术后考虑的首选。Taddei 等人 16 在 10 年前描述了鼠上颌骨的更详细方案，应该添加更多视觉和透明细节。综上所述，该方案系统地描述了小鼠上颌骨模型中详细的正畸牙齿运动方案，以帮助研究人员以标准化的方式掌握建模方法，并实现不同研究之间的比较评估。

<table><tbody><tr><td>&lt;strong&gt;Experimental Models: Mouse Lines&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>C57/B6J&amp;nbsp;</td><td>Gempharmatech Experimental Animals Company&amp;nbsp;</td><td>C57/B6J</td><td></td></tr><tr><td>&lt;strong&gt;Critical Commercial Assays&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Hematoxylin and Eosin Stain Kit</td><td>Biosharp</td><td>BL700B</td><td></td></tr><tr><td>Masson&amp;rsquo;s Trichrome Stain Kit</td><td>Solarbio</td><td>G1340</td><td></td></tr><tr><td>&lt;strong&gt;Instruments&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>27 G needle</td><td>Chengdu Xinjin Shifeng Medical Apparatus &amp;amp; Instruments Co. LTD.</td><td>SB1-074(IV)</td><td></td></tr><tr><td>Adhesives</td><td>Minnesota Mining and Manufacturing Co., Ltd.</td><td>41282</td><td></td></tr><tr><td>Corkboard</td><td>DELI Group Co., Ltd.</td><td>8705</td><td></td></tr><tr><td>Cotton balls</td><td>Haishi Hainuo Group Co.,&amp;nbsp; Ltd.</td><td>20120047</td><td></td></tr><tr><td>Cotton sticks</td><td>Lakong Medical Devices Co., Ltd.</td><td>M6500R</td><td></td></tr><tr><td>Customized coil spring</td><td>Chengdu Mingxing Spring Co., Ltd.</td><td>1109-02</td><td></td></tr><tr><td>Forceps</td><td>Chengdu Shifeng Co., Ltd.</td><td>none</td><td></td></tr><tr><td>Light-cured fluid resin</td><td>Shofu Dental Trading (SHANGHAI) Co., Ltd.</td><td>518785</td><td></td></tr><tr><td>Light curer</td><td>Liang Ya Dental Equipment Co., Ltd.</td><td>LY-A180</td><td></td></tr><tr><td>Medical adhesive tapes&amp;nbsp;</td><td>Haishi Hainuo Group Co.,&amp;nbsp; Ltd.</td><td>0008-2014</td><td></td></tr><tr><td>Medical non-woven fabric</td><td>Henan Yadu Industrial Co., Ltd.</td><td>01011500018</td><td></td></tr><tr><td>Needle holders</td><td>Chengdu Shifeng Co., Ltd.</td><td>none</td><td></td></tr><tr><td>Rubber bands</td><td>Haishi Hainuo Group Co.,&amp;nbsp; Ltd.</td><td>32X1</td><td></td></tr><tr><td>Surgical scissors</td><td>Chengdu Shifeng Co., Ltd.</td><td>none</td><td></td></tr><tr><td>Tweezers</td><td>Chengdu Shifeng Co., Ltd.</td><td>none</td><td></td></tr></tbody></table>

the establishment of a murine maxillary orthodontic model

由于缺乏建立小鼠上颌骨正畸模型的可重复方案，我们提出了一个可靠且可重复的方案，为研究人员提供分析机械负荷相关骨重塑的可行工具。除了不同类型的原理图、操作照片和视频外，本研究还提供了详细的流程图。我们对 11 只成年宽型 C57/B6J 小鼠进行了该方案，并在术后第 3、8 和 14 天收获了样品。显微CT和组织病理学数据已经证明了使用该协议进行牙齿移动与骨重塑相结合的成功。此外，根据第3天、第8天和第14天的显微CT结果，我们将骨建模分为三个阶段:准备阶段、骨吸收阶段和骨形成阶段。这些阶段有望帮助关注不同阶段的研究人员合理地设置样本采集时间。该协议可以为研究人员提供一种工具，以进行骨重塑的再生分析。

我们已经对 11 只成年雄性小鼠（C57/BL6,3 个月大）进行了 OTM 手术。他们在手术后第 3、8 和 14 天被安乐死。在这些实验中，右上颌侧是手术侧，而左上颌侧是控制侧。显微CT显示，术后第3、8和14天，M1和M2之间的距离在时间上连续增加:分别为30μm、70μm和110μm（图4）。由于机械负荷，低密度牙周韧带在远端变宽，在根部近中侧变窄（图5）。此外，牙周韧带是连续的，任何根部均未发生吸收。这些结果证明，使用该协议物理移动 M1 是可行且安全的。
此外，我们还分析了 M1 根部内的骨骼区域，参数如 图 6 所示。与对照侧相比，第 8 天手术侧的骨体积和骨矿物质密度百分比显着下降（图 6A、B）。相比之下，与第8天的手术侧相比，第3天和第14天手术侧的骨体积百分比显着增加（图6A）。这些结果表明，在术后第 3 天之前，骨重塑是不活跃的。术后第 3 天后，骨吸收开始主导骨重塑过程。术后第8天后，骨形成在骨重塑中占据优势，牙槽骨几乎恢复到生理水平，这也意味着牙齿运动几乎停止。到本方案的第14天，封闭在M1根部的骨重塑经历了三个阶段，大致可分为准备阶段，骨吸收阶段和骨形成阶段。因此，研究人员可以使用该模型研究骨重塑的不同阶段。
图 7 显示了苏木精-伊红染色和 Masson-trichrome 染色的结果。我们选择了 M1 的近中颊根 （MB） 和远端颊根 （DB） 之间的牙槽骨作为感兴趣的区域。MB 远端和 DB 近中端的牙周韧带是感兴趣骨区域力传递的前沿。每组的对照侧表现出相似的表现:这些牙周韧带与排列的波浪状纤维和纺锤形细胞共享相似的宽度，并且牙槽骨的表面是完整的线性的。这表明，在生理条件下，封闭在M1根部的牙周组织没有受到不平衡和过度的机械负荷。
术后第3天，牙周韧带纤维在张力侧被紧绷，而牙周韧带纤维则被压缩，形态模糊。在压力最大的区域注意到透明化。牙槽骨的表面仍然保持其两侧的完整性。与显微CT结果一致，在术后最初3天，M1通过压迫压力侧的牙周韧带在牙槽窝内移动，而尚未观察到骨吸收或形成。
在手术后的第8天，两侧的牙周韧带显示出与第3天相同的特征，尽管牙槽骨的表面开始看起来很粗糙。此外，骨髓腔扩大，骨小梁数量似乎减少，如CT数据所示。因此，在术后第8天，骨重塑的组织病理学表型显示骨吸收增加。牙槽骨也表明 M1 高速移动。
术后第14天，两侧牙周韧带的宽度似乎几乎相等。与术后第8天相比，牙槽骨的表面变得更加粗糙。然而，在对照侧，骨骼恢复到生理水平，CT数据也表明了这一点。这一阶段表明，骨骼形成在骨骼建模过程中占主导地位。由于机械载荷在操作时仅施加一次，因此载荷随着移动距离的增加而减小。随着牙槽骨恢复正常，M1的运动也停止了。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/66033/66033fig01.jpg"> 图 1:牙齿运动的示意图。 当对臼齿施加机械负荷时，可以定义牙槽骨重塑的拉伸和压缩侧。粗箭头表示机械载荷的方向。细箭头表示骨重塑前沿的拉伸侧和压缩侧。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/66033/66033fig02.jpg"> 图 2:手术项目。 （一）手术平台:用医用无纺布包裹的发泡板或软木板。（B） 紧固件:（2） 两根橡皮筋，（3） 胶带，（4） 四根 27 G 针。（C） 手术器械和正畸用品:（5） 手术剪刀，（6） 眼科镊子，（7） 针架，（8） 304 不锈钢丝，以及 （9） 定制螺旋弹簧。白色矩形是指定制的螺旋弹簧。弹簧的放大版本显示在 附图S1中。（D）牙齿修复用品:（10）气泵瓶，（11）光固化剂，（12）棉球，（13）棉棒，（14）光固化液树脂，（15）粘合剂。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig02large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/66033/66033fig03.jpg"> 图3:手术过程。 （A） 将鼠标固定在手术平台上。（B） 将 304 不锈钢丝从颊侧推过 M1 和 M2 之间的相邻空间。（地下1层）添加了原理图以帮助理解。（C） 螺旋弹簧固定在 M1 上，并且在 M1 处不会发生咬合干扰。（C1级）添加了原理图以帮助理解。（D） 螺旋弹簧的另一端固定在同侧上切牙上。（第1天）添加了原理图以帮助理解。（E） 涂上流体树脂将门牙和不锈钢包裹在一起。（F） 所有正畸矫治器的最终视图。缩写:M1 = 上颌第一磨牙;M2 = 上颌第二磨牙。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig03large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/66033/66033fig04.jpg"> 图4:代表性显微CT三维图像及M1运动不同阶段的统计分析。 （A） 在生理情况下，M1 和 M2 之间没有空间。（B-D）M1开始移动，移动距离随时间推移，根据M1和M2之间的相互位置关系而增加。红色框表示 M1 和 M2 之间的距离。黑色箭头表示机械载荷的方向。（E）M1移动距离的统计分析。缩写:M1 = 上颌第一磨牙;M2 = 上颌第二磨牙;OTM = 正畸牙齿运动。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig04large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/66033/66033fig05.jpg"> 图 5:M1 运动不同阶段的水平和矢状面的代表性显微 CT 二维图像。 （A，B） 在生理情况下，低密度牙周韧带是等肌的，连续占据一定的空间而不是被压缩，牙槽骨表面完整线性。（C，D）牙周韧带在远端变宽，在根部近中侧变窄，可以在手术后第 3 天观察到。（E-H）在手术后第 8 天和第 14 天，由于骨的吸收和沉积，牙周韧带开始恢复，牙槽骨表面变得粗糙。黄色箭头是指受压的牙周韧带。红色箭头是指牙槽骨的粗糙表面，用于骨的吸收和沉积。* P < 0.05;P < 0.005。单因素方差分析。数据±标准差、n ≥ 3 的平均值。比例尺 = 100 μm。 缩写: M1 = 上颌第一磨牙;M2 = 上颌第二磨牙;OTM = 正畸牙齿运动。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig05largev2.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/66033/66033fig06.jpg"> 图 6:显微 CT 对 M1 运动不同阶段 M1 根部封闭的牙槽骨进行统计分析。 （A） 第 8 天骨容量百分比显着下降表明第 3 天和第 8 天之间的活跃骨吸收。第 14 天骨体积百分比的显着增加表明在第 8 天和第 14 天之间有活跃的骨形成。（B） 与对照组相比，第 8 天骨矿物质密度的显着差异。也支持上述结论。（C-E）使用三个补充指标进行评价。几乎没有发现显著差异，但趋势仍然支持上述结论。*P < 0.05。单因素方差分析。数据是平均值±标准差、n ≥ 3。缩写:M1 = 上颌第一磨牙;OTM = 正畸牙齿运动;BV/TV = 骨容量百分比;BMD = 骨矿物质密度;Tb. N = 小梁数;Tb. Th = 小梁厚度;Tb. Sp = 小梁分离。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig06large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/66033/66033fig07.jpg"> 图7:M1运动不同阶段的苏木精-伊红染色和Masson-trichrome染色的代表性结果。 （A，B） 在生理条件下，牙周韧带纤维受到一定的力，呈明显的波浪状形状，如"~"，牙槽骨表面完整线性。当M1受到机械载荷时，（C，E，G，I，K，M）纤维在张力侧被紧紧拉伸，而（D，F，H，J，L，N）牙周韧带纤维被压缩，形态模糊。（C-N）随着骨建模的进行，牙槽骨的表面变得越来越不平整。比例尺 = 20 μm。 缩写: M1 = 上颌第一磨牙;OTM = 正畸牙齿运动。<a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/66033fig07large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td>物种</td>
			<td>动牙</td>
			<td>锚地</td>
			<td>装置</td>
			<td>移动方向</td>
			<td>参考</td>
		</tr><tr><td>鼠科的</td>
			<td>第一磨牙</td>
			<td>门牙</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>14,15</td>
		</tr><tr><td></td>
			<td>第一磨牙</td>
			<td>第二磨牙</td>
			<td>松紧带</td>
			<td>内侧</td>
			<td>16</td>
		</tr><tr><td>鼠</td>
			<td>第一磨牙</td>
			<td>迷你植入物</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>17</td>
		</tr><tr><td></td>
			<td>第一磨牙</td>
			<td>门牙</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>18</td>
		</tr><tr><td></td>
			<td>第二和第三磨牙</td>
			<td>对侧同义齿</td>
			<td>弹簧膨胀装置</td>
			<td>颊的</td>
			<td>19</td>
		</tr><tr><td></td>
			<td>第一磨牙</td>
			<td>第二磨牙</td>
			<td>正畸线</td>
			<td>内侧</td>
			<td>20</td>
		</tr><tr><td>兔</td>
			<td>第一前磨牙</td>
			<td>门牙</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>21</td>
		</tr><tr><td></td>
			<td>第一前磨牙</td>
			<td>迷你植入物</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>22</td>
		</tr><tr><td></td>
			<td>板牙</td>
			<td>对侧同义齿</td>
			<td>螺旋弹簧</td>
			<td>末梢部的</td>
			<td>23</td>
		</tr><tr><td></td>
			<td>板牙</td>
			<td>对侧同义齿</td>
			<td>欧米茄环</td>
			<td>末梢部的</td>
			<td>24</td>
		</tr><tr><td>狗</td>
			<td>第二前磨牙和第一磨牙</td>
			<td>迷你植入物</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>25</td>
		</tr><tr><td></td>
			<td>第二前磨牙</td>
			<td>犬</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>26</td>
		</tr><tr><td></td>
			<td>第一前磨牙</td>
			<td>迷你植入物</td>
			<td>松紧带</td>
			<td>末梢部的</td>
			<td>27</td>
		</tr><tr><td></td>
			<td>侧切牙</td>
			<td>犬</td>
			<td>松紧带</td>
			<td>末梢部的</td>
			<td>28</td>
		</tr><tr><td>猪</td>
			<td>第一磨牙</td>
			<td>乳第三磨牙和微型植入物</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>29</td>
		</tr><tr><td></td>
			<td>第一磨牙</td>
			<td>第二磨牙</td>
			<td>正畸线</td>
			<td>颊的</td>
			<td>30</td>
		</tr><tr><td>猴子</td>
			<td>中切牙</td>
			<td>第一磨牙、前磨牙、犬牙和侧切牙</td>
			<td>螺旋弹簧和正畸钢丝</td>
			<td>唇</td>
			<td>31</td>
		</tr><tr><td>猫</td>
			<td>犬</td>
			<td>迷你植入物</td>
			<td>螺旋弹簧</td>
			<td>内侧</td>
			<td>32</td>
		</tr></tbody></table>表1:现有动物正畸模型的总结。 该表列出了专注于简单正畸牙齿运动的常规实验动物的常用模型。它们始终由三个元件组成:目标动齿、锚固和用于增加机械载荷的连接装置。通过改变这三个要素，已经衍生出各种正畸程序。已排除具有多颗牙齿的复杂正畸牙齿运动。
补充图S1:弹簧的放大版本。 （A） 无机械载荷和 （B） 有机械载荷。比例尺 = 5 毫米。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/S-FIGURE.1.zip" target="_blank">请点击这里下载此文件。</a>
补充图S2:用镊子夹紧结扎线的方法。 在协议步骤 2.4 中，此处显示了在穿刺前夹住结扎线弯曲的最安全、最方便的方法。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/S-FIGURE.2.zip" target="_blank">请点击此处下载此文件。</a>
补充图S3:树脂涂层的范围。 在协议步骤2.9中，弹簧的门牙端（A）没有树脂覆盖物，（B）如图所示。不得将树脂添加到弹性部分。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/66033/S-FIGURE.3.zip" target="_blank">请点击此处下载此文件。</a>

Watch this Scientific Journal Video about Author Spotlight: Insights into an Efficient Murine Maxillary Orthodontic Model Protocol at JoVE.com

作者聚焦:深入了解高效的小鼠上颌正畸模型方案

在这里，我们逐步演示了在小鼠上颌骨模型上操作的可管理的正畸牙齿运动方案。通过对每个步骤的明确解释和视觉演示，研究人员可以掌握该模型，并通过一些修改将其应用于他们的实验需求。

小鼠上颌骨正畸模型的建立

Due to the lack of reproducible protocols for establishing a murine maxillary orthodontic model, we present a reliable and reproducible protocol to ...

正如我们所回顾的，小鼠上颌正畸模型被广泛使用，但只是简单地描述。基于当前方法，很难在不同研究之间重复和比较实验数据。因此，该协议将帮助研究人员以标准化的方式掌握建模方法，并实现不同研究之间的比较评估。该协议实际上是迄今为止在小鼠中最简单的建模方法，我们的视频展示了许多实用的建模细节。这种方法以鼠为主，狭窄的操作空间肯定会给初学者带来很大的挑战。一定程度的练习可能会有所帮助。该协议以最简单的方式提供正畸负荷，因此该协议的实验将节省时间且易于重复。对于新手来说也有很多优势，例如手术视野开阔，视觉外科手术。零基学习者可以通过我们的协议更容易掌握这个模型。

the-establishment-of-a-murine-maxillary-orthodontic-model

Nanyang Technological University

Research

JoVE Journal

Medicine

The Establishment of a Murine Maxillary Orthodontic Model

580 次观看.  Sichuan University. 正如我们所回顾的，小鼠上颌正畸模型被广泛使用，但只是简单地描述。基于当前方法，很难在不同研究之间重复和比较实验数据。因此，该协议将帮助研究人员以标准化的方式掌握建模方法，并实现不同研究之间的比较评估。该协议实际上是迄今为止在小鼠中最简单的建模方法，我们的视频展示了许多实用的建模细节。这种方法以鼠为主，狭窄的操作空间肯定会给初?...

视频: 作者聚焦:深入了解高效的小鼠上颌正畸模型方案

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

科研

医学

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable cell lines are inoculated either by intravesical instillation (model of nonmuscle invasive growth) or intramural injection into the bladder wall (model of invasive growth). Both procedures are complex and highly time-consuming. Additionally, the superficial model has its shortcomings due to the lack of cell lines that are tumorigenic following instillation. Intramural injection, on the other hand, is marred by the invasiveness of the procedure and the associated morbidity for the host mouse.
With these shortcomings in mind, we modified previous methods to develop a minimally invasive approach for creating orthotopic bladder cancer xenografts. Using ultrasound guidance we have successfully performed percutaneous inoculation of the bladder cancer cell lines UM-UC1, UM-UC3 and UM-UC13 into 50 athymic nude. We have been able to demonstrate that this approach is time efficient, precise and safe. With this technique, initially a space is created under the bladder mucosa with PBS, and tumor cells are then injected into this space in a second step. Tumor growth is monitored at regular intervals with bioluminescence imaging and ultrasound. The average tumor volumes increased steadily in in all but one of our 50 mice over the study period.
In our institution, this novel approach, which allows bladder cancer xenograft inoculation in a minimally-invasive, rapid and highly precise way, has replaced the traditional model.

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

微创建立小鼠原位膀胱移植瘤的

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

肝转移的临床前小鼠模型

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

3D Imaging of PDL Collagen Fibers during Orthodontic Tooth Movement in Mandibular Murine Model

Orthodontic tooth movement is a complex biological process of altered soft and hard tissue remodeling as a result of external forces. In order to understand these complex remodeling processes, it is critical to study the tooth and periodontal tissues within their 3D context and therefore minimize any sectioning and tissue artefacts. Mouse models are often utilized in developmental and structural biology, as well as in biomechanics due to their small size, high metabolic rate, genetics and ease of handling. In principle this also makes them excellent models for dental related studies. However, a major impediment is their small tooth size, the molars in particular. This paper is aimed at providing a step by step protocol for generating orthodontic tooth movement and two methods for 3D imaging of the periodontal ligament fibrous component of a mouse mandibular molar. The first method presented is based on a micro-CT setup enabling phase enhancement&#160;imaging of fresh collagen tissues. The second method is a bone clearing method using ethyl cinnamate that enables imaging through the bone without sectioning and preserves endogenous fluorescence. Combining this clearing method with reporter mice like Flk1-Cre;TdTomato provided a first of its kind opportunity to image the 3D vasculature in the PDL and alveolar bone.

We present a protocol for generating orthodontic tooth movement in mice and methods for 3D visualization of the collagen fibers and blood vessels of periodontal ligament without sectioning.

曼迪布勒穆林模型中矫形牙齿运动期间 PDL 胶原蛋白纤维的 3D 成像

Orthodontic tooth movement is a complex biological process of altered soft and hard tissue remodeling as a result of external forces. In order to ...

开发用于研究口鼻瘘的有效小鼠模型

Establishment of an Oronasal Fistula Mice Model

This study presents a method utilizing heated ophthalmologic cautery to develop a viable model for investigating oronasal fistulas. C57BL/6 mice were used to establish the oronasal fistula (ONF) model. To create the ONF, the mice were anesthetized, immobilized, and their hard palates were exposed. During the surgical procedure, a 2.0 x 1.5 mm full-thickness mucosal injury was induced in the midline of the hard palate using ophthalmologic cautery. It was crucial to control the size of the ONF and minimize bleeding in order to ensure the success of the experiment. Verification of the ONF model's effectiveness was conducted on the 7th-day post-operation, encompassing both anatomical and functional assessments. The presence of the nasal septum within the oral cavity and the outflow of sterile water from the nostrils upon injection into the oral cavity confirmed the successful establishment of the ONF model. The model demonstrated a practical and successful oronasal fistula, characterized by a low mortality rate, significant weight changes, and minimal variation in ONF size. Future studies may consider adopting this methodology to elucidate the mechanisms of palate wound healing and explore novel treatments for oronasal fistulas.

作者聚焦：研究小鼠口鼻瘘模型的伤口愈合 

This article outlines a step-by-step procedure for establishing a mice model with an oronasal fistula. The oronasal fistula was created by employing heated ophthalmologic cautery to damage the midline portion of the hard palate, resulting in the formation of an opening between the oral and nasal cavities.

口鼻瘘小鼠模型的建立

This study presents a method utilizing heated ophthalmologic cautery to develop a viable model for investigating oronasal fistulas. C57BL/6 mice were used ...

使用新型 Mouth-Gag 创建牙髓炎症小鼠模型 

Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research

Pulpitis, a common cause of natural tooth loss, leads to necrosis and loss of bioactivity in the inflamed dental pulp. Unraveling the mechanisms underlying pulpitis and its efficient treatment is an ongoing focus of endodontic research. Therefore, understanding the inflammatory process within the dental pulp is vital for improving pulp preservation. Compared to other in vitro experiments, a murine pulpitis model offers a more authentic and genetically diverse context to observe the pathological progression of pulpitis. However, using mice, despite their cost-effectiveness and accessibility, poses difficulties due to their small size, poor coordination, and low tolerance, complicating intraoral and dental procedures. This protocol introduces a novel design and application of a mouth-gag to expose mouse pulp, facilitating more efficient intraoral procedures. The mouth-gag, comprised of a dental arch readily available to most dentists and can significantly expedite surgical preparation, even for first-time procedures. Micro-CT, hematoxylin-eosin (HE) staining, and immunofluorescence staining were used to identify changes in morphology and cell expression. The aim of this article is to help researchers establish a more reproducible and less demanding procedure for creating a pulp inflammation model using this novel mouth-gag.

作者聚焦：通过改进的小鼠模型增强牙髓研究

This article presents a streamlined protocol for establishing a pulpitis model in mice using an innovative mouth-gag, followed by subsequent histological analysis.

小鼠上颌骨正畸模型的建立

副本

摘要

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用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

曼迪布勒穆林模型中矫形牙齿运动期间 PDL 胶原蛋白纤维的 3D 成像

口鼻瘘小鼠模型的建立

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口鼻瘘小鼠模型的建立

基于新型口堵的鼠牙髓暴露模型的建立及其在牙髓炎研究中的应用

基于新型口堵的鼠牙髓暴露模型的建立及其在牙髓炎研究中的应用