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异种纤维蛋白生物聚合物与单根缝线相关联,可使周围神经损伤后神经肌肉接头恢复到成熟模式。

Heterologous fibrin biopolymer associated to a single suture stitch enables the return of neuromuscular junction to its mature pattern after peripheral nerve injury.

机构信息

Graduate Program in Surgery and Translational Medicine, Medical School, São Paulo State University (Unesp), Botucatu, São Paulo, Brazil; Department of Structural and Functional Biology (Anatomy Sector), Institute of Biosciences, São Paulo State University (Unesp), Botucatu, São Paulo, Brazil.

Department of Structural and Functional Biology (Anatomy Sector), Institute of Biosciences, São Paulo State University (Unesp), Botucatu, São Paulo, Brazil.

出版信息

Injury. 2021 Apr;52(4):731-737. doi: 10.1016/j.injury.2020.10.070. Epub 2020 Oct 17.

Abstract

Denervation leads to severe atrophy of neuromuscular junction (NMJ) structure including decrease of the expression of fundamental proteins. Up to now, conventional suture has been the gold standard method used to correct this injury. Fibrin sealant is one of the alternatives proposed to optimize this method. This study verified if the association of fibrin sealant - Heterologous Fibrin Biopolymer (HFB) and a single suture stitch promotes return of morphology and NMJ structure to mature pattern after peripheral nerve injury. Forty Wistar rats were distributed into 4 groups: Sham-Control (SC), Denervated-Control (DC), Suture-Lesion (SL) and Suture-Lesion + HFB (SFS). In SC group only the right sciatic nerve identification was done. In DC, SL and SFS groups fixation of nerve stumps on musculature immediately after neurotmesis was performed. After seven days, stump reconnection with 3 stitches in SL and a single stitch associated with HFB in SFS were done. After sixty days right soleus muscles were prepared for nicotinic acetylcholine receptors (nAChRs) and nerve terminal confocal analyses, and for nAChRs (α1, ε e γ), S100, Agrin, LRP-4, MMP-3, Rapsyn western blotting analyses. SC group presented normal morphology. In DC group it was observed flattening of NMJ, fragmentation of nAChRs and tangled nerve terminals. The majority of the parameters of SL and SFS groups presented values in between SC and DC groups. There was an increase of relative planar area in these groups (SL and SFS) highlighting that there was less nAChRs fragmentation and the values of protein expression showed return of nAChRs to mature pattern. Use of HFB associated with a single suture stitch decreased surgical time, minimized suture injuries, did not alter nerve regeneration and presented potential to reestablish the NMJ apparatus. These consolidated results encourage surgeons to develop future clinical trials to install definitively this new approach both for reconstructive surgery and neurosurgery.

摘要

去神经支配导致神经肌肉接头(NMJ)结构严重萎缩,包括基本蛋白表达减少。到目前为止,传统缝线一直是纠正这种损伤的金标准方法。纤维蛋白胶是优化这种方法的替代方案之一。本研究验证了纤维蛋白胶-异源纤维蛋白生物聚合物(HFB)与单缝线缝合是否促进周围神经损伤后形态和 NMJ 结构恢复成熟模式。将 40 只 Wistar 大鼠分为 4 组:Sham-Control(SC)、Denervated-Control(DC)、Suture-Lesion(SL)和 Suture-Lesion+HFB(SFS)。在 SC 组中,仅对右侧坐骨神经进行识别。在 DC、SL 和 SFS 组中,神经横断后立即将神经残端固定在肌肉上。七天后,在 SL 中进行 3 针缝合重新连接神经残端,在 SFS 中进行单针缝合并与 HFB 结合。六十天后,准备右侧比目鱼肌进行烟碱型乙酰胆碱受体(nAChRs)和神经末梢共聚焦分析,以及 nAChRs(α1、εeγ)、S100、Agrin、LRP-4、MMP-3、Rapsyn 免疫印迹分析。SC 组表现出正常形态。在 DC 组中,观察到 NMJ 扁平化、nAChRs 碎片化和神经末梢纠结。SL 和 SFS 组的大多数参数值介于 SC 和 DC 组之间。这些组的相对平面面积增加(SL 和 SFS),突出显示 nAChRs 碎片化减少,并且蛋白表达值表明 nAChRs 恢复成熟模式。使用 HFB 与单缝线缝合相结合,减少了手术时间,最小化了缝线损伤,不改变神经再生,并有可能重建 NMJ 装置。这些综合结果鼓励外科医生开展未来的临床试验,为重建手术和神经外科安装这种新方法。

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