{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T20:01:32Z","timestamp":1782417692042,"version":"3.54.5"},"reference-count":45,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T00:00:00Z","timestamp":1725408000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016221","name":"Rowan University","doi-asserted-by":"publisher","award":["16000-85003-13"],"award-info":[{"award-number":["16000-85003-13"]}],"id":[{"id":"10.13039\/100016221","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016221","name":"Rowan University","doi-asserted-by":"publisher","award":["16000-85003-13"],"award-info":[{"award-number":["16000-85003-13"]}],"id":[{"id":"10.13039\/100016221","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Application of polyethylene glycol (PEG) to a peripheral nerve injury at the time of primary neurorrhaphy is thought to prevent Wallerian degeneration via direct axolemma fusion. The molecular mechanisms of nerve fusion and recovery are unclear. Our study tested the hypothesis that PEG alters gene expression in neural and muscular environments as part of its restorative properties. Lewis rats underwent unilateral sciatic nerve transection with immediate primary repair. Subjects were randomly assigned to receive either PEG treatment or standard repair at the time of neurorrhaphy. Samples of sciatic nerve distal to the injury and tibialis muscle at the site of innervation were harvested at 24 hours and 4 weeks postoperatively. Total RNA sequencing and subsequent bioinformatics analyses were used to identify significant differences in differentially expressed genes (DEGs) and their related biological pathways (p&lt;0.05) in PEG-treated subjects compared to non-PEG controls. No significant DEGs were identified in PEG-treated sciatic nerve compared to controls after 24 hours, but 1,480 DEGs were identified in PEG-treated tibialis compared to controls. At 4 weeks, 918 DEGs were identified in PEG-treated sciatic nerve, whereas only 3 DEGs remained in PEG-treated tibialis compared to controls. DEGs in sciatic were mostly upregulated (79%) and enriched in pathways present during nervous system development and growth, whereas DEGs in muscle were mostly downregulated (77%) and related to inflammation and tissue repair. Our findings indicate that PEG application during primary neurorrhaphy leads to significant differential gene regulation in the neural and muscular environment that is associated with improved functional recovery in animals treated with PEG compared to sham non-PEG controls. A detailed understanding of key molecules underlying PEG function in recovery after peripheral nerve repair may facilitate amplification of PEG effects through systemic or focal treatments at the time of neurotmesis.<\/jats:p>","DOI":"10.1371\/journal.pone.0304773","type":"journal-article","created":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T17:38:14Z","timestamp":1725471494000},"page":"e0304773","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["An analysis of differential gene expression in peripheral nerve and muscle utilizing RNA sequencing after polyethylene glycol nerve fusion in a rat sciatic nerve injury model"],"prefix":"10.1371","volume":"19","author":[{"given":"Samantha N.","family":"Weiss","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph M.","family":"Legato","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yichuan","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Courtney N.","family":"Vaccaro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renata Pellegrino","family":"Da Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Miskiel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grace V.","family":"Gilbert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hakon","family":"Hakonarson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A.","family":"Fuller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3799-4355","authenticated-orcid":true,"given":"Russell J.","family":"Buono","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,9,4]]},"reference":[{"issue":"3","key":"pone.0304773.ref001","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.hcl.2013.04.002","article-title":"Nerve physiology: mechanisms of injury and recovery","volume":"29","author":"RM Menorca","year":"2013","journal-title":"Hand Clin"},{"key":"pone.0304773.ref002","doi-asserted-by":"crossref","first-page":"551","DOI":"10.4103\/neuroindia.NI_987_16","article-title":"Peripheral nerve injuries: A retrospective survey of 1124 cases","volume":"65","author":"J.A. Kouyoumdjian","year":"2017","journal-title":"Neurol. India"},{"key":"pone.0304773.ref003","doi-asserted-by":"crossref","first-page":"905","DOI":"10.3171\/jns.2001.94.6.0905","article-title":"Evaluation of iatrogenic lesions in 722 surgically treated cases of peripheral nerve trauma","volume":"94","author":"T. Kretschmer","year":"2001","journal-title":"J. Neurosurg"},{"issue":"5","key":"pone.0304773.ref004","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1097\/PHM.0b013e31815e6370","article-title":"The incidence of peripheral nerve injury in extremity trauma","volume":"87","author":"CA Taylor","year":"2008","journal-title":"Am. J. Phys. Med. Rehabil"},{"issue":"3","key":"pone.0304773.ref005","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1016\/j.jhsa.2009.12.009","article-title":"Treatment of acute peripheral nerve injuries: current concepts","volume":"35","author":"J. Isaacs","year":"2010","journal-title":"J Hand Surg Am"},{"key":"pone.0304773.ref006","volume-title":"Green\u2019s Operative Hand Surgery","author":"D.P. Green","year":"2011","edition":"6"},{"key":"pone.0304773.ref007","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.bjps.2014.09.043","article-title":"Prognostic factors for outcome after median, ulnar, and combined median-ulnar nerve injuries: A prospective study","volume":"68","author":"C.A. Hundepool","year":"2015","journal-title":"J. Plast. Reconstr. Aesthet. Surg"},{"key":"pone.0304773.ref008","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1159\/000484174","article-title":"Predictors of Surgical Outcomes of Traumatic Peripheral Nerve Injuries in Children: An Institutional Experience","volume":"53","author":"B.I. Devi","year":"2018","journal-title":"Pediatr. Neurosurg"},{"key":"pone.0304773.ref009","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/B978-0-12-420045-6.00007-9","article-title":"The role of timing in nerve reconstruction","volume":"109","author":"L.B. Dahlin","year":"2013","journal-title":"Int. Rev. Neurobiol"},{"key":"pone.0304773.ref010","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.expneurol.2017.01.009","article-title":"Surgical repair in humans after traumatic nerve injury provides limited functional neural regeneration in adults","volume":"290","author":"W.A. Palispis","year":"2017","journal-title":"Exp. Neurol"},{"key":"pone.0304773.ref011","first-page":"704589","article-title":"Sensory recovery outcome after digital nerve repair in relation to different reconstructive techniques: Meta-analysis and systematic review","volume":"2013","author":"F.J. Paprottka","year":"2013","journal-title":"Plast. Surg. Int"},{"key":"pone.0304773.ref012","doi-asserted-by":"crossref","first-page":"243","DOI":"10.5435\/00124635-200007000-00005","article-title":"Peripheral nerve injury and repair","volume":"8","author":"S.K. Lee","year":"2000","journal-title":"J. Am. Acad. Orthop. Surg"},{"issue":"6","key":"pone.0304773.ref013","doi-asserted-by":"crossref","first-page":"1419","DOI":"10.1097\/00006534-200105000-00016","article-title":"Clinical outcome fol- lowing nerve allograft transplantation","volume":"107","author":"S.E. MacKinnon","year":"2001","journal-title":"Plast. Reconstr. Surg"},{"issue":"2","key":"pone.0304773.ref014","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1097\/01.prs.0000172896.86594.07","article-title":"Median and ulnar nerve injuries: a meta-analysis of predictors of motor and sensory recovery after modern microsurgical nerve repair","volume":"116","author":"ACJ Ruijs","year":"2005","journal-title":"Plast Reconstr Surg"},{"key":"pone.0304773.ref015","first-page":"369","article-title":"Experiments on the Section of the Glosso-Pharyngeal and Hypoglossal Nerves of the Frog, and Observations of the Alterations Produced Thereby in the Structure of Their Primitive Fibres","volume":"76","author":"A. Waller","year":"1851","journal-title":"Edinb. Med. Surg. J"},{"key":"pone.0304773.ref016","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1242\/jeb.19.1.14","article-title":"The Rate of Regeneration of Nerve","volume":"19","author":"E. Gutmann","year":"1942","journal-title":"J. Exp. Biol"},{"key":"pone.0304773.ref017","doi-asserted-by":"crossref","DOI":"10.1093\/med\/9780195169904.001.0001","volume-title":"Nerve Repair","author":"T.M. Brushart","year":"2011"},{"key":"pone.0304773.ref018","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1007\/BF01148493","article-title":"The role of non-resident cells in Wallerian degeneration","volume":"13","author":"W. Beuche","year":"1984","journal-title":"J. Neurocytol"},{"key":"pone.0304773.ref019","doi-asserted-by":"crossref","first-page":"3521","DOI":"10.1113\/JP270874","article-title":"The repair Schwann cell and its function in regenerating nerves","volume":"594","author":"K.R. Jessen","year":"2016","journal-title":"J. Physiol"},{"key":"pone.0304773.ref020","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1111\/j.1460-9568.2009.06847.x","article-title":"Chemical communication between regenerating motor axons and Schwann cells in the growth pathway","volume":"30","author":"G. Vrbova","year":"2009","journal-title":"Eur. J. Neurosci"},{"key":"pone.0304773.ref021","volume-title":"Principles of Neural Science","author":"E.R. Kandel","year":"2013","edition":"5"},{"key":"pone.0304773.ref022","doi-asserted-by":"crossref","first-page":"e56484","DOI":"10.1371\/journal.pone.0056484","article-title":"Effect of delayed peripheral nerve repair on nerve regeneration, Schwann cell function and target muscle recovery","volume":"8","author":"S. Jonsson","year":"2013","journal-title":"PLoS ONE"},{"key":"pone.0304773.ref023","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00421-006-0256-z","article-title":"The denervated muscle: Facts and hypotheses. A historical review","volume":"98","author":"M. Midrio","year":"2006","journal-title":"Eur. J. Appl. Physiol"},{"key":"pone.0304773.ref024","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.jss.2018.07.013","article-title":"Fusogens: Chemical agents that can rapidly restore function after nerve injury","volume":"233","author":"S.A. Abdou","year":"2019","journal-title":"J. Surg. Res"},{"key":"pone.0304773.ref025","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/0006-8993(86)91617-3","article-title":"Reconnection of severed nerve axons with polyethylene glycol","volume":"367","author":"G.D. Bittner","year":"1986","journal-title":"Brain Res"},{"issue":"47","key":"pone.0304773.ref026","doi-asserted-by":"crossref","first-page":"15790","DOI":"10.1523\/JNEUROSCI.4155-10.2010","article-title":"A model for sealing plasmalemmal damage in neurons and other eukaryotic cells","volume":"30","author":"C.S. Spaeth","year":"2010","journal-title":"J. Neurosci"},{"key":"pone.0304773.ref027","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1002\/jnr.23022","article-title":"Cellular mechanisms of plas- malemmal sealing and axonal repair by polyethylene glycol and methylene blue","volume":"90","author":"C.S. Spaeth","year":"2012","journal-title":"J. Neursosci. Res"},{"key":"pone.0304773.ref028","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1016\/j.jss.2012.03.049","article-title":"Hydrophilic polymers enhance early functional outcomes after nerve autografting","volume":"177","author":"K.W. Sexton","year":"2012","journal-title":"J. Surg. Res"},{"key":"pone.0304773.ref029","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1002\/jnr.23685","article-title":"The curious ability of polyethylene glycol fusion technologies to restore lost behaviors after nerve severance","volume":"94","author":"G.D. Bittner","year":"2015","journal-title":"J. Neurosci. Res"},{"key":"pone.0304773.ref030","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.4103\/1673-5374.187019","article-title":"Repair of traumatic plasmalemmal damage to neurons and other eukaryotic cells","volume":"11","author":"G.D. Bittner","year":"2016","journal-title":"Neural Regen. Res"},{"key":"pone.0304773.ref031","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jneumeth.2018.12.015","article-title":"Polyethylene glycol (PEG) and other bioactive solutions with neurorrhaphy for rapid and dramatic repair of peripheral nerve lesions by PEG-fusion","volume":"314","author":"CL Ghergherehchi","year":"2019","journal-title":"J Neurosci Methods"},{"key":"pone.0304773.ref032","doi-asserted-by":"crossref","first-page":"1223","DOI":"10.1002\/jnr.24225","article-title":"Polyethylene glycol solutions rapidly restore and maintain axonal continuity, neuromuscular structures and behaviors lost after sciatic nerve transections in female rats","volume":"96","author":"M. Mikesh","year":"2018","journal-title":"J. Neurosci. Res"},{"key":"pone.0304773.ref033","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1002\/jnr.23704","article-title":"Effects of extracellular calcium and surgical techniques on restoration of axonal continuity by polyethylene glycol fusion following complete cut or crush severance of rat sciatic nerves","volume":"94","author":"C.L. Ghergherehchi","year":"2016","journal-title":"J. Neurosci. Res"},{"issue":"1","key":"pone.0304773.ref034","doi-asserted-by":"crossref","first-page":"53","DOI":"10.4103\/1673-5374.224363","article-title":"Conundrums and confusions regarding how PEG-fusion produces excellent behavioral recovery after peripheral nerve injuries","volume":"13","author":"G.D. Bittner","year":"2017","journal-title":"Neural Regen. Res"},{"issue":"6","key":"pone.0304773.ref035","doi-asserted-by":"crossref","first-page":"1478","DOI":"10.3390\/ijms20061478","article-title":"Polyethylene Glycol: The Future of Posttraumatic Nerve Repair? Systemic Review","volume":"20","author":"AM Paskal","year":"2019","journal-title":"Int J Mol Sci"},{"issue":"5","key":"pone.0304773.ref036","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1002\/jnr.23023","article-title":"Rapid, effective, and long- lasting behavioral recovery produced by microsutures, methylene blue, and polyethylene glycol after completely cutting rat sciatic nerves","volume":"90","author":"GD Bittner","year":"2012","journal-title":"J Neurosci Res"},{"key":"pone.0304773.ref037","first-page":"22","article-title":"Microarray and qPCR Analyses of Wallerian Degeneration in Rat Sciatic Nerves","volume":"11","author":"S Yi","year":"2017","journal-title":"Front. Cell. Neurosci"},{"key":"pone.0304773.ref038","doi-asserted-by":"crossref","first-page":"69","DOI":"10.3389\/fnmol.2018.00069","article-title":"The Whole Transcriptome Involved in Denervated Muscle Atrophy Following Peripheral Nerve Injury","volume":"11","author":"J Weng","year":"2018","journal-title":"Front. Mol. Neurosci"},{"key":"pone.0304773.ref039","doi-asserted-by":"crossref","first-page":"1243","DOI":"10.1002\/jnr.24227","article-title":"Polyethylene glycol treated allo- grafts not tissue matched nor immunosuppressed rapidly repair sciatic nerve gaps, maintain neuromuscular functions, and restore voluntary behaviors in female rats","volume":"96","author":"M. Mikesh","year":"2018","journal-title":"J. Neurosci. Res"},{"issue":"12","key":"pone.0304773.ref040","doi-asserted-by":"crossref","first-page":"2468","DOI":"10.1002\/jnr.24720","article-title":"Polyethylene glycol-fusion repair of sciatic allografts in female rats achieves immunotolerance via attenuated innate and adaptive responses","volume":"98","author":"Dec","year":"2020","journal-title":"J Neurosci Res"},{"issue":"1","key":"pone.0304773.ref041","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1186\/s12974-020-01953-8","article-title":"Coding transcriptome analyses reveal altered functions underlying immunotolerance of PEG-fused rat sciatic nerve allografts","volume":"17","author":"TA Smith","year":"2020","journal-title":"J Neuroinflammation"},{"issue":"1","key":"pone.0304773.ref042","doi-asserted-by":"crossref","first-page":"e5535","DOI":"10.1097\/GOX.0000000000005535","article-title":"Fibrin Glue Acutely Blocks Distal Muscle Contraction after Confirmed Polyethylene Glycol Nerve Fusion: An Animal Study","volume":"12","author":"AH Fisher","journal-title":"Plast Reconstr Surg Glob Open"},{"issue":"12","key":"pone.0304773.ref043","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"MI Love","year":"2014","journal-title":"Genome Biol"},{"issue":"1","key":"pone.0304773.ref044","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/nar\/gkn923","article-title":"Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists","volume":"37","author":"DW Huang","year":"2009","journal-title":"Nucleic Acids Res"},{"key":"pone.0304773.ref045","doi-asserted-by":"crossref","first-page":"100043","DOI":"10.1016\/j.bioflm.2021.100043","article-title":"Do results obtained with RNA-sequencing require independent verification?","volume":"3","author":"T. Coenye","year":"2021","journal-title":"Biofilm"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0304773","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T17:38:41Z","timestamp":1725471521000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0304773"}},"subtitle":[],"editor":[{"given":"Jianxun","family":"Ding","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,9,4]]},"references-count":45,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2024,9,4]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0304773","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,4]]}}}