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Thy1-GFP大鼠面神经再生的宏观体内成像

Macroscopic in vivo imaging of facial nerve regeneration in Thy1-GFP rats.

作者信息

Placheta Eva, Wood Matthew D, Lafontaine Christine, Frey Manfred, Gordon Tessa, Borschel Gregory H

机构信息

Division of Plastic and Reconstructive Surgery, Medical University of Vienna, Vienna, Austria.

Division of Plastic and Reconstructive Surgery, The Hospital for Sick Children, Toronto, Ontario, Canada.

出版信息

JAMA Facial Plast Surg. 2015 Jan-Feb;17(1):8-15. doi: 10.1001/jamafacial.2014.617.

Abstract

IMPORTANCE

Facial nerve injury leads to severe functional and aesthetic deficits. The transgenic Thy1-GFP rat is a new model for facial nerve injury and reconstruction research that will help improve clinical outcomes through translational facial nerve injury research.

OBJECTIVE

To determine whether serial in vivo imaging of nerve regeneration in the transgenic rat model is possible, facial nerve regeneration was imaged under the main paradigms of facial nerve injury and reconstruction.

DESIGN, SETTING, AND PARTICIPANTS: Fifteen male Thy1-GFP rats, which express green fluorescent protein (GFP) in their neural structures, were divided into 3 groups in the laboratory: crush-injury, direct repair, and cross-face nerve grafting (30-mm graft length). The distal nerve stump or nerve graft was predegenerated for 2 weeks. The facial nerve of the transgenic rats was serially imaged at the time of operation and after 2, 4, and 8 weeks of regeneration. The imaging was performed under a GFP-MDS-96/BN excitation stand (BLS Ltd).

INTERVENTION OR EXPOSURE

Facial nerve injury.

MAIN OUTCOME AND MEASURE

Optical fluorescence of regenerating facial nerve axons.

RESULTS

Serial in vivo imaging of the regeneration of GFP-positive axons in the Thy1-GFP rat model is possible. All animals survived the short imaging procedures well, and nerve regeneration was followed over clinically relevant distances. The predegeneration of the distal nerve stump or the cross-face nerve graft was, however, necessary to image the regeneration front at early time points. Crush injury was not suitable to sufficiently predegenerate the nerve (and to allow for degradation of the GFP through Wallerian degeneration). After direct repair, axons regenerated over the coaptation site in between 2 and 4 weeks. The GFP-positive nerve fibers reached the distal end of the 30-mm-long cross-face nervegrafts after 4 to 8 weeks of regeneration.

CONCLUSIONS AND RELEVANCE

The time course of facial nerve regeneration was studied by serial in vivo imaging in the transgenic rat model. Nerve regeneration was followed over clinically relevant distances in a small number of experimental animals, as they were subsequently imaged at multiple time points. The Thy1-GFP rat model will help improve clinical outcomes of facial reanimation surgery through improving the knowledge of facial nerve regeneration after surgical procedures.

LEVEL OF EVIDENCE

NA.

摘要

重要性

面神经损伤会导致严重的功能和美学缺陷。转基因Thy1-GFP大鼠是面神经损伤与重建研究的一种新模型,通过转化型面神经损伤研究有助于改善临床治疗效果。

目的

为了确定在转基因大鼠模型中对神经再生进行连续活体成像是否可行,在面神经损伤与重建的主要模式下对面神经再生进行成像。

设计、场所和参与者:15只在神经结构中表达绿色荧光蛋白(GFP)的雄性Thy1-GFP大鼠在实验室中被分为3组:挤压伤组、直接修复组和跨面神经移植组(移植长度30毫米)。远端神经残端或神经移植体预先变性2周。在手术时以及再生2、4和8周后对转基因大鼠的面神经进行连续成像。成像在GFP-MDS-96/BN激发架(BLS有限公司)下进行。

干预或暴露

面神经损伤。

主要结局和测量指标

再生面神经轴突的光学荧光。

结果

在Thy1-GFP大鼠模型中对GFP阳性轴突的再生进行连续活体成像是可行的。所有动物都很好地耐受了简短的成像程序,并且在临床相关距离上对神经再生进行了跟踪。然而,为了在早期时间点对面神经再生前沿进行成像,远端神经残端或跨面神经移植体的预先变性是必要的。挤压伤不适合使神经充分预先变性(以及通过沃勒变性使GFP降解)。直接修复后,轴突在2至4周内在吻合部位再生。再生4至8周后,GFP阳性神经纤维到达30毫米长的跨面神经移植体的远端。

结论与意义

通过在转基因大鼠模型中进行连续活体成像研究了面神经再生的时间进程。在少数实验动物中,在临床相关距离上对神经再生进行了跟踪,因为随后在多个时间点对它们进行了成像。Thy1-GFP大鼠模型将通过增进对手术后面神经再生的了解,有助于改善面部恢复手术的临床治疗效果。

证据水平

无。

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