title: Angiogenesis in tissue-engineered nerves evaluated objectively using MICROFIL perfusion and micro-CT scanning.
abstract: Angiogenesis is a key process in regenerative medicine generally, as well as in the specific field of nerve regeneration. However, no convenient and objective method for evaluating the angiogenesis of tissue-engineered nerves has been reported. In this study, tissue-engineered nerves were constructed in vitro using Schwann cells differentiated from rat skin-derived precursors as supporting cells and chitosan nerve conduits combined with silk fibroin fibers as scaffolds to bridge 10-mm sciatic nerve defects in rats. Four weeks after surgery, three-dimensional blood vessel reconstructions were made through MICROFIL perfusion and micro-CT scanning, and parameter analysis of the tissue-engineered nerves was performed. New blood vessels grew into the tissue-engineered nerves from three main directions: the proximal end, the distal end, and the middle. The parameter analysis of the three-dimensional blood vessel images yielded several parameters, including the number, diameter, connection, and spatial distribution of blood vessels. The new blood vessels were mainly capillaries and microvessels, with diameters ranging from 9 to 301 μm. The blood vessels with diameters from 27 to 155 μm accounted for 82.84% of the new vessels. The microvessels in the tissue-engineered nerves implanted in vivo were relatively well-identified using the MICROFIL perfusion and micro-CT scanning method, which allows the evaluation and comparison of differences and changes of angiogenesis in tissue-engineered nerves implanted in vivo.
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu Province, China; Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Hong-Kui
authors.initials: HK
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Ya-Xian
authors.initials: YX
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Cheng-Bin
authors.initials: CB
authors.lastname: Xue
authors.affiliation: Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Zhen-Mei-Yu
authors.initials: ZM
authors.lastname: Li
authors.affiliation: Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Huang
authors.affiliation: Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Ya-Hong
authors.initials: YH
authors.lastname: Zhao
authors.affiliation: Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Yu-Min
authors.initials: YM
authors.lastname: Yang
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu Province, China; Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China.
authors.forename: Xiao-Song
authors.identifier: 0000-0002-2562-6275
authors.initials: XS
authors.lastname: Gu
keywords: MICROFIL perfusion; Schwann cell; angiogenesis; chitosan nerve conduit; micro-CT; nerve regeneration; neural regeneration; skin-derived precursor; three-dimensional reconstruction; tissue-engineered nerve