期刊
LARYNGOSCOPE
卷 128, 期 11, 页码 E386-E392出版社
WILEY
DOI: 10.1002/lary.27439
关键词
Microscopy; fluorescence; sciatic nerve; peripheral nerve injuries; nerve regeneration; mice; transgenic; guided tissue regeneration; nerve guidance conduit; confocal; disease models; animal; prosthesis implantation; materials testing; axons; neuronal outgrowth
资金
- Dartmouth SYNERGY 2016 Translational Pilot Grant
- NIDCD NIH [T32DC000020]
Objective Methods Use of cell culture and conventional in vivo mammalian models to assess nerve regeneration across guidance conduits is resource-intensive. Herein we describe a high-throughput platform utilizing transgenic mice for stain-free axon visualization paired with rapid cryosection techniques for low-cost screening of novel bioengineered nerve guidance conduit performance. Interposition repair of sciatic nerve transection in mice expressing yellow fluorescent protein in peripheral neurons (Thy1.2 YFP-16) was performed with various bioengineered neural conduit compositions using a rapid sutureless entubulation technique under isoflurane anesthesia. Axonal ingrowth was assessed at 3 and 6 weeks using epifluorescent microscopy following cryosectioning. Results Conclusion Mean procedure time (incision-to-closure) was less than 21/2 minutes. Direct operational costs of a 3-week experiment was calculated at $21.47 per animal. Tissue processing steps were minimized to aldehyde fixation, cryoprotection and sectioning, and rapid fluorescent dye staining for conduit visualization. Fluorescent microscopy readily resolved robust axonal sprouting at 3 weeks, with clear elucidation of ingrowth-permissive, semipermissive, or restrictive nerve guidance conduit environments. A rapid and cost-efficient in vivo platform for screening of nerve guidance conduit performance has been described. Level of Evidence NA. Laryngoscope, E392-E392, 2018
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