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人工蜘蛛丝在体外支持和引导神经突生长。

Artificial spider silk supports and guides neurite extension in vitro.

机构信息

Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.

Experimental Traumatology Unit, Department of Neuroscience, Biomedicum B8 Karolinska Institutet, Stockholm, Sweden.

出版信息

FASEB J. 2021 Nov;35(11):e21896. doi: 10.1096/fj.202100916R.

Abstract

Surgical intervention with the use of autografts is considered the gold standard to treat peripheral nerve injuries. However, a biomaterial that supports and guides nerve growth would be an attractive alternative to overcome problems with limited availability, morbidity at the site of harvest, and nerve mismatches related to autografts. Native spider silk is a promising material for construction of nerve guidance conduit (NGC), as it enables regeneration of cm-long nerve injuries in sheep, but regulatory requirements for medical devices demand synthetic materials. Here, we use a recombinant spider silk protein (NT2RepCT) and a functionalized variant carrying a peptide derived from vitronectin (VN-NT2RepCT) as substrates for nerve growth support and neurite extension, using a dorsal root ganglion cell line, ND7/23. Two-dimensional coatings were benchmarked against poly-d-lysine and recombinant laminins. Both spider silk coatings performed as the control substrates with regards to proliferation, survival, and neurite growth. Furthermore, NT2RepCT and VN-NT2RepCT spun into continuous fibers in a biomimetic spinning set-up support cell survival, neurite growth, and guidance to an even larger extent than native spider silk. Thus, artificial spider silk is a promising biomaterial for development of NGCs.

摘要

外科干预使用自体移植物被认为是治疗周围神经损伤的金标准。然而,一种能够支持和引导神经生长的生物材料将是一种有吸引力的替代方法,可以克服自体移植物的可用性有限、采集部位发病率和神经不匹配等问题。天然蜘蛛丝是构建神经引导导管(NGC)的一种很有前途的材料,因为它能够使绵羊长达 cm 的神经损伤再生,但医疗器械的监管要求需要使用合成材料。在这里,我们使用重组蜘蛛丝蛋白(NT2RepCT)和一种携带来自 vitronectin 的肽的功能化变体(VN-NT2RepCT)作为神经生长支持和神经突延伸的底物,使用背根神经节细胞系 ND7/23。二维涂层与聚-d-赖氨酸和重组层粘连蛋白进行了基准测试。两种蜘蛛丝涂层在增殖、存活和神经突生长方面的表现都与对照底物一样。此外,NT2RepCT 和 VN-NT2RepCT 在仿生纺丝装置中纺成连续纤维,在更大程度上支持细胞存活、神经突生长和引导,甚至超过天然蜘蛛丝。因此,人工蜘蛛丝是开发 NGC 的一种很有前途的生物材料。

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