A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice

Mitch R. Paro; Daylin Gamiotea Turro; Michael Mcgonnigle; Ketan R. Bulsara; Rajkumar Verma

doi:10.3791/63951

JoVE Journal
Neuroscience

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Neuroscience

A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice

小鼠脑中动脉闭塞诱导脑卒中后脑肌粘连血管病治疗模型

Full Text

3,206 Views

06:54 min

June 22, 2022

DOI: 10.3791/63951-v

Mitch R. Paro*¹, Daylin Gamiotea Turro*¹, Michael Mcgonnigle¹, Ketan R. Bulsara², Rajkumar Verma¹

¹Department of Neuroscience,UConn School of Medicine, ²Division of Neurosurgery,UConn School of Medicine

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a protocol for encephalomyosynangiosis (EMS) using a vascular temporalis muscle flap on ischemic brain tissue to treat acute ischemic stroke. The efficacy in promoting angiogenesis was evaluated through a transient middle cerebral artery occlusion model in mice.

Key Study Components

Area of Science

Neuroscience
Stroke Recovery
Angiogenesis

Background

Stroke injuries damage the vascular network in the brain, making early restoration critical for recovery.
EMS surgery enhances the vascular network by increasing angiogenesis.
This method reduces reliance on pharmacological interventions for stroke recovery.
EMS may also provide insights into neuroangiogenesis and neurogenesis following stroke.

Purpose of Study

To assess the effectiveness of EMS in increasing blood flow to ischemic brain tissue.
To evaluate the cellular and molecular changes following EMS surgery.
To explore EMS's potential as a therapeutic intervention for malignant stroke.

Methods Used

The main platform used was a transient middle cerebral artery occlusion model in mice.
Key biological model involved the temporalis muscle grafted onto the pial surface of ischemic brain tissue.
Following 60 minutes of occlusion, mice underwent EMS surgery involving a craniotomy and myotomy.
Recovery monitoring occurred post-surgery, with evaluations at 7 and 21 days for muscle cell health and angiogenesis.

Main Results

EMS significantly increased blood vessel area and density in the perilesional cortex post-stroke.
Angiogenesis was indicated by elevated FGF acidic protein and reduced osteopontin levels a few weeks post-surgery.
Although a 10%-11% mortality rate was noted, EMS did not increase this mortality, suggesting a favorable tolerance.

Conclusions

This study demonstrates EMS as a viable surgical method to enhance stroke recovery and angiogenesis.
These findings improve understanding of therapeutic strategies for stroke and other acute brain injuries.
EMS has prospects for broader applications in treating various acute neurological conditions.

Frequently Asked Questions

What are the advantages of using the EMS model?

The EMS model allows for rapid restoration of vascular networks in the ischemic brain, promoting effective recovery from stroke and reducing the need for drugs.

How is the EMS surgery implemented?

EMS surgery involves grafting a temporalis muscle flap onto the ischemic brain's pial surface, facilitated by a series of surgical steps including craniotomy and myotomy.

What kind of outcomes can be obtained from this study?

Outcomes include assessments of angiogenesis through molecular readouts and evaluation of muscle cell survival and recovery post-surgery.

Can the EMS method be adapted for other types of injuries?

Yes, EMS has potential applications beyond stroke, including treatment for other acute brain injuries such as hematomas.

What are some limitations of the EMS approach?

While EMS is promising, it requires surgical skill and practice, as well as careful management of post-operative recovery in animal models.

该方案旨在提供脑肌粘连血管病的方法 - 在缺血性脑组织的皮瓣上移植血管颞肌瓣 - 用于治疗非烟雾病急性缺血性中风。该方法在增加血管生成方面的功效是在小鼠中使用瞬时大脑中动脉闭塞模型进行评估的。

中风损伤会破坏大脑中的血管网络，血管供应的早期恢复对于中风恢复是必要的。EMS手术通过增加大脑中的血管生成来迅速恢复血管网络。EMS手术提供了一种实现脑血管生成的安全方法，最终将改善流向缺血组织的血流。

这种方法可以减少对药物干预的需求。EMS作为缺血性中风的治疗方法具有治疗潜力。特别是，它可以应用于恶性卒中的情况，其中半颅骨切除术是常规的。

EMS 可促进卒中迅速恢复。EMS手术可以提供对神经血管生成机制的见解，并且可以将其应用扩展到神经发生研究，神经发生通常在血管生成之后。虽然任何外科手术都需要大量的实践，但是我相信任何有啮齿动物手术经验的研究人员都可以轻松学习和进行EMS手术。

手术前通过高压灭菌对所有器械进行消毒。用 70% 乙醇消毒操作表面，并用电加热垫将其加热至 37 摄氏度。异氟诱导麻醉后，将鼠标放在操作表面上的左侧。

通过用钝钳捏脚趾来验证麻醉深度。然后用眼药膏润滑双眼。接下来，用电动剃须刀在手术区域剃毛，并用70%乙醇清洁手术区域，然后使用提供溶液。

皮下注射单剂量0.25%布比卡因，作为手术部位术前镇痛。然后以所需的放大倍率设置手术显微镜。在大脑中动脉闭塞60分钟后，将小鼠随机分为仅大脑中动脉闭塞或大脑中动脉闭塞以及脑肌粘连血管闭塞组。

对于接受脑肌粘连血管病的群体，用剪刀做一个10至15毫米的皮肤切口，从1到2毫米的嘴部延伸到右耳，从1到2毫米的尾部延伸到右眼。然后使用夹子缩回皮瓣，目视识别颞肌和颅骨。用剪刀和展开技术将颞肌从颅骨上直接解剖。

接下来，沿着肌肉的尾缘进行两到三毫米的肌切开术，以促进腹侧反射。之后，使用微型钻头在反射的颞肌下方的颅骨上进行直径约5毫米的开颅手术。接下来，用镊子取出硬脑膜，格外小心地露出大脑的痔疮表面，以免意外伤害大脑。

如文中所述缝合切口后，将鼠标放回笼中并监测直至从麻醉中恢复。然后将鼠标放回其外壳设施。颞肌移植在移植和对照肌肉手术后7天显示肌肉细胞短暂损伤，肌细胞存活率约为71%和97%，但移植肌和对照肌之间的这种差异消失，肌肉在手术后21天完全恢复。

在仅脑肌粘连血管和大脑中动脉闭塞模型中，颞肌移植物粘附在皮质表面，表明手术、移植物植入和粘合成功。脑肌粘连血管病显著增加中风后病灶周围皮层的血管表面积和综合密度。蛋白阵列结果显示，脑卒中后21 d脑肌粘连管闭塞组的FGF酸性蛋白水平显著升高，骨桥蛋白水平降低，提示血管生成和神经保护得到改善。

大脑中动脉闭塞手术后21天，小鼠死亡率为10%至11%。然而，闭塞手术后的脑肌粘连血管病并没有增加死亡率，表明其耐受性。首先，当您解剖颞肌时，请采取预防措施以避免损伤。

第二，用微型钻头打开头骨。第三，在用镊子去除硬脑膜期间。EMS可用于其他急性损伤，如血性脑损伤。

如果成功的EMS不仅可以改善中风的结果，还可以改善其他急性脑损伤的益处。EMS将为恶性中风的新治疗方法打开大门。它将提供改善血管生成的新方法，并可能揭示成人神经发生，这通常取决于灌注良好的组织。

View the full transcript and gain access to thousands of scientific videos

Explore More Videos

神经科学第184期