Name: the establishment of a murine mandibular molar extraction socket healing model
Uploaded: 2023-01-13T13:40:00
Description: Watch this Scientific Journal Video about The Establishment of a Murine Mandibular Molar Extraction Socket Healing Model at JoVE.com

这项工作得到了国家自然科学基金81825005（L.Y.）、82201045（F.Y.）和82100982（F.L.）以及四川省科技计划2021JDRC0144（F.L.），2022JDRC0130（F.Y.）的支持。

本研究中的所有动物手术均由四川大学华西口腔医学院伦理委员会审核批准（WCHSIRB-D-2017-041）。从商业来源获得的成年C57BL / 6小鼠（见 材料表）用于本研究。
1.术前准备
<ol><li>仪器准备<ol><li>准备各种一次性注射器针头（26G，25G，23G;见 材料表）用作电梯。将针头弯曲约20°-40°，如图 1所示。</li>
		<li>准备带齿的眼...

<ol>
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鼠窝愈合模型是揭示骨愈合和再生潜在机制的重要方法，最终解决临床挑战。现有的研究已经证明了门牙拔除模型和上颌磨牙拔除模型的可能性，而研究尚未使用下颌第一磨牙模型13，17，18。然而，门牙对啮齿动物的生活至关重要，它们的损伤可能是致命的。此外，由于上颌骨中的骨骼比下颌骨更松质，因此在愈合过程中，潜...

拔牙后的牙槽愈合是一种常见的临床情况，在不期望的愈合下，这可能导致不适当的并发症，例如牙槽出血、干槽症，甚至颌骨骨髓炎1，2，3。这些合并症可能会损害患者的生活质量，更糟糕的是，由于大量骨质流失，对假体康复构成重大挑战4。虽然牙槽愈合阶段已经阐明，但在遇到各种预后挑战时，它们不足以指导拔牙手术后的临床护理4。
已经进行了多项基于动物模型的研究，以更好地了解插座愈合过程中的潜在机制并避免上述情况。Sp7是成骨细胞分化的主要调节剂，在骨骼发育、骨止血和骨再生中起着至关重要的作用5，6。合理的牙槽愈合模型可以显示Sp7创伤后在骨再生中的冗余。此外，与长骨骨折愈合不同，只有一个单一的成骨过程，即膜内骨化，涉及拔牙窝7的愈合过程。这使得动物拔牙模型成为研究基于种植体的疗法的最佳选择，因为种植体骨整合遵循相同的成骨规则8。
几十年来，拔牙模型已经在大鼠、兔子和狗身上进行，因为这些物种的大牙齿便于在9、10、11 上操作。然而，鉴于对转基因的蓬勃发展的需求以及作为人类更具适应性的遗传背景，小鼠越来越多地被用来建立拔牙模型。然后，研究人员可以使用基因组修饰的小鼠来揭示特定细胞群在插座愈合过程中的作用，而不是仅观察表型12。在鼠牙拔牙窝模型中，已有研究证明了鼠上颌牙和门牙拔牙窝的建立和愈合过程13，14，15，16。然而，预后的愈合模式以及检测和观察时间点可能因方案而异。这为学者建立鼠窝愈合模型提供了一个普遍标准。
本研究旨在为上述问题构建一个切实可行的鼠窝愈合模型。小鼠的下颌磨牙与上颌磨牙和门牙相比具有独特的形态特征，具有独特的优缺点。由于专注于小鼠下颌骨的模型目前是基于真空的，因此该协议试图提供一种完成的方法来提取小鼠的下颌第一磨牙。我们希望该协议能够启发基础研究人员新的想法，以揭示插座愈合的潜在机制并指示临床护理。

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			<td>23/25/26 G needle</td>
			<td>Chengdu Xinjin Shifeng Medical Apparatus &#38; Instruments Co. LTD.</td>
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			<td>C57/B6J&#160;</td>
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			<td>Rabbit polyclonal&#160;</td>
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			<td>Tweezers</td>
			<td>Chengdu Xinjin Shifeng Medical Apparatus &#38; Instruments Co. LTD.</td>
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the establishment of a murine mandibular molar extraction socket healing model

本研究介绍了鼠下颌骨磨牙拔除模型的发展，为研究牙槽骨再生和膜内骨化提供实用的模型。使用C57 / J6小鼠提取下颌第一磨牙以建立该模型。他们分别在手术后1周和4周被安乐死，并收获双侧下颌骨。随后进行连续立体采集、组织学评估和免疫荧光染色，以证明手术成功。手术后，立体图像立即显示一个空的提取窝。手术后1周的苏木精和伊红（H&E）和术后4周的Masson染色显示，原始根部的区域分别部分和完全充满骨小梁。免疫荧光染色显示，与稳态侧相比，术后1周Sp7表达增加，提示牙槽窝成骨剧烈。所有这些结果都证明了一种可行的鼠牙拔牙槽愈合模型。即将进行的揭示颚骨缺陷愈合或牙槽愈合机制的研究可以采用这种方法。

为了阐明该方法的实际应用，提取两只健康C57BL / 6小鼠（3个月大，均为雌性）的右下颌第一磨牙，并分别随访1周和4周。未受损的左下颌骨被用作健康对照。 图1A 显示了手术器具的具体特征，包括26-23 G针和带齿眼科镊子。26 G 针被精确移除并弯曲。25 G 针在针点处弯曲约 25°。23 G 针是手术的关键，在针点处分别弯曲约 25° 和 35°（图 1B1，B2</strong...

Watch this Scientific Journal Video about The Establishment of a Murine Mandibular Molar Extraction Socket Healing Model at JoVE.com

The Establishment of a Murine Mandibular Molar Extraction Socket Healing Model

该协议演示了如何提取小鼠下颌第一磨牙的分步细节。它为专注于颚骨愈合和再生的研究人员提供了一种替代方法。

小鼠下颌磨牙拔牙窝愈合模型的建立

This study introduces the development of a molar extraction model in the murine mandible to provide a practicable model for studying alveolar bone ...

该协议提供了一种鼠下颌磨牙拔除模型，该模型在可用的骨再生研究中很少引入。开发鼠下颌磨牙拔除模型是必要的，因为下颌骨在不同干预组中更具可比性和一致性。这种技术的含义不会扩展到任何疾病的治疗。然而，它可能有助于临床愈合和颌骨再生，通过帮助回顾这一过程背后的机制。开发该技术是为了帮助研究，揭示可用愈合和再生的机制。遗憾的是，这种方法不能应用于其他系统。通过用镊子内侧握住臼齿开始消除麻醉小鼠的远端阻力。然后用力将 23 号针插入远端根部的颊牙槽骨并呈现间隔。接下来，更改为 25 号针以继续间隔扩展。小心翼翼地向根尖周区域前进，同时缓慢地向前旋转针头，将根部从牙槽窝中压出。为了消除内侧阻力，将23号针插入根叉并抬起咬合的磨牙。紧紧握住臼齿后，再取一根 23 号针，用力将其插入舌内侧牙周膜以形成间隔。然后使用25号针，缓慢地向前和向颊旋转。如果一些潜在的障碍阻碍了臼齿的脱位，请使用 26 号针穿透根尖并重复操作。在最后拔牙过程中，拔牙，确保牙冠高于咬合面，并且可以清楚地看到两个完整的牙根。成功拔出下颌第一磨牙后，涂干棉止血，重新定位舌头，给予镇痛，并将鼠标放在恒温加热垫上，直到它从麻醉中恢复过来。拔牙后牙槽立即充满凝块。观察插座的愈合过程一周。形成了一些海绵状的小梁骨，但凝块仍然存在。在愈合过程中，两周后插座充满海绵骨，表明再生完成。成骨细胞分化Sp7的主要调节因子在骨形成前部边缘的骨髓细胞中广泛表达。在体内平衡状态下，小梁骨一致，与散落如岛的骨髓细胞块汇合。在手术后一周，许多表达Sp7的细胞充满了拔牙槽，周围撒有新形成的小梁骨。在手术后四周，病情逆转并转变为大部分合并为小梁骨。表达Sp7的细胞的活性下降到接近体内平衡状态的水平。也可以进行演示新生儿再生过程。这种技术可以通过提供一种新的小鼠模型来帮助研究人员揭示可用愈合和再生过程的潜在机制。

Research

JoVE Journal

Biology

Murine Renal Transplantation Procedure

小鼠肾移植程序

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

科研

生物学

LAD-Ligation: A Murine Model of Myocardial Infarction

法援署结扎：有心肌梗死的小鼠模型

Research models of infarction and myocardial ischemia are essential to investigate the acute and chronic pathobiological and pathophysiological processes ...

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

生产转基因非洲爪蟾限制性内切酶介导的整合和核移植

Stable integration of cloned gene products into the Xenopus genome is necessary to control the time and place of expression, to express genes at later ...

Isolation of Murine Coronary Vascular Smooth Muscle Cells

鼠冠状动脉血管平滑肌细胞的分离

While the isolation and culture of vascular smooth muscle cells (VSMCs) from large vessels is well established, we sought to isolate and culture VSMCs ...

Filtration Isolation of Nucleic Acids:  A Simple and Rapid DNA Extraction Method

核酸的分离过滤：一种简单快速的DNA提取方法

FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad ...

A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle

小鼠下颌骨髁突的形态学及细胞分析方法

The temporomandibular joint (TMJ) has the capacity to adapt to external stimuli, and loading changes can affect the position of condyles, as well as the ...

The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury

The Murine Choline-Deficient, Ethionine-Supplemented CDE Diet Model of Chronic Liver Injury

慢性肝损伤小鼠胆碱缺乏、氨基补充的饮食模型

Chronic liver diseases, such as viral hepatitis, alcoholic liver disease, or non-alcoholic fatty liver disease, are characterized by continual ...

A Murine Model of Vertical Sleeve Gastrectomy

小鼠垂直套胃切除术的动物模型

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. ...

Establishment of an Extracellular Acidic pH Culture System

胞外酸性 pH 培养体系的建立

Conditions of the tumor microenvironment, such as hypoxia or nutrient starvation, play critical roles in cancer progression and malignancy. However, the ...

Establishment of a Clonal Culture of Unicellular Conjugating Algae

单细胞共轭藻类无性系培养的建立

It is essential to establish clonal cultures of microalgae for use in studies of various topics, such as physiology, genetics, taxonomy, and microbiology. ...