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探索猪神经移植物的创新去细胞化方案:一种用于周围神经修复的转化方法。

Exploring an innovative decellularization protocol for porcine nerve grafts: a translational approach to peripheral nerve repair.

作者信息

Muratori Luisa, Crosio Alessandro, Ronchi Giulia, Molinaro Debora, Tos Pierluigi, Lovati Arianna B, Raimondo Stefania

机构信息

Department of Clinical and Biological Sciences, Neuroscience Institute Cavalieri Ottolenghi (NICO), University of Turin, Turin, Italy.

UOC Traumatology-Reconstructive Microsurgery, Department of Orthopedics and Traumatology, CTO Hospital, Turin, Italy.

出版信息

Front Neuroanat. 2024 Mar 19;18:1380520. doi: 10.3389/fnana.2024.1380520. eCollection 2024.

Abstract

INTRODUCTION

Peripheral nerves are frequently affected by lesions caused by traumatic or iatrogenic damages, resulting in loss of motor and sensory function, crucial in orthopedic outcomes and with a significant impact on patients' quality of life. Many strategies have been proposed over years to repair nerve injuries with substance loss, to achieve musculoskeletal reinnervation and functional recovery. Allograft have been tested as an alternative to the gold standard, the autograft technique, but nerves from donors frequently cause immunogenic response. For this reason, several studies are focusing to find the best way to decellularize nerves preserving either the extracellular matrix, either the basal lamina, as the key elements used by Schwann cells and axons during the regenerative process.

METHODS

This study focuses on a novel decellularization protocol for porcine nerves, aimed at reducing immunogenicity while preserving essential elements like the extracellular matrix and basal lamina, vital for nerve regeneration. To investigate the efficacy of the decellularization protocol to remove immunogenic cellular components of the nerve tissue and to preserve the basal lamina and extracellular matrix, morphological analysis was performed through Masson's Trichrome staining, immunofluorescence, high resolution light microscopy and transmission electron microscopy. Decellularized porcine nerve graft were then employed in vivo to repair a rat median nerve lesion. Morphological analysis was also used to study the ability of the porcine decellularized graft to support the nerve regeneration.

RESULTS AND DISCUSSION

The decellularization method was effective in preparing porcine superficial peroneal nerves for grafting as evidenced by the removal of immunogenic components and preservation of the ECM. Morphological analysis demonstrated that four weeks after injury, regenerating fibers colonized the graft suggesting a promising use to repair severe nerve lesions. The idea of using a porcine nerve graft arises from a translational perspective. This approach offers a promising direction in the orthopedic field for nerve repair, especially in severe cases where conventional methods are limited.

摘要

引言

外周神经经常受到创伤性或医源性损伤所导致的病变影响,从而造成运动和感觉功能丧失,这对骨科治疗结果至关重要,并对患者的生活质量产生重大影响。多年来,人们提出了许多策略来修复伴有实质缺损的神经损伤,以实现肌肉骨骼的再支配和功能恢复。异体神经移植已被作为金标准自体移植技术的替代方法进行了测试,但供体神经常常会引发免疫反应。因此,多项研究致力于寻找最佳方法来使神经脱细胞,同时保留细胞外基质和基膜,因为它们是雪旺细胞和轴突在再生过程中所利用的关键要素。

方法

本研究聚焦于一种用于猪神经的新型脱细胞方案,旨在降低免疫原性,同时保留对神经再生至关重要的细胞外基质和基膜等关键要素。为了研究该脱细胞方案去除神经组织免疫原性细胞成分以及保留基膜和细胞外基质的效果,通过马松三色染色、免疫荧光、高分辨率光学显微镜和透射电子显微镜进行了形态学分析。然后将脱细胞猪神经移植物用于修复大鼠正中神经损伤的体内实验。形态学分析还用于研究猪脱细胞移植物支持神经再生的能力。

结果与讨论

脱细胞方法有效地制备了用于移植的猪腓浅神经,去除免疫原性成分并保留细胞外基质证明了这一点。形态学分析表明,损伤四周后,再生纤维在移植物中定植,这表明其在修复严重神经损伤方面具有广阔的应用前景。使用猪神经移植物的想法源于转化医学的视角。这种方法在骨科领域为神经修复提供了一个有前景的方向,特别是在传统方法受限的严重病例中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cfb8/10985228/801332a2fad9/fnana-18-1380520-g001.jpg

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