MP29-18 PREFABRICATION OF NEUROMUSCULAR JUNCTION FOR ACCELERATED RECOVERY OF MUSCLE FUNCTION

Abstract

You have accessJournal of UrologyTrauma/Reconstruction: Ureter, Bladder, External Genitalia and Urotrauma II1 Apr 2015MP29-18 PREFABRICATION OF NEUROMUSCULAR JUNCTION FOR ACCELERATED RECOVERY OF MUSCLE FUNCTION In Kap Ko, Sang Jin Lee, John Jackson, Anthony Atala, and James Yoo In Kap KoIn Kap Ko More articles by this author , Sang Jin LeeSang Jin Lee More articles by this author , John JacksonJohn Jackson More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and James YooJames Yoo More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.621AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Surgical reconstruction involving muscle requires integration of nervous tissue for functional restoration. Failure of neural integration leads to atrophy and the tissue often becomes non-functional. Therefore, muscle innervation is a critical process in the recovery of function, this process is time sensitive and relies heavily on the host tissue microenvironment. Acceleration of nerve integration by facilitating the formation for neuromuscular junctions (NMJ) may be an option to restore muscle function after reconstruction. Toward this goal, we investigated whether muscle fibers cultured in vitro could form NMJ that would lead to rapid innervation to muscle tissue in vivo. METHODS Myotubes formed from muscle precursor cells (C2C12) were co-cultured with neuroblastoma cells (NG108), grown with conditioned medium derived from NG108, or treated with neurotrophic factors (agrin-containing medium). To confirm acetylcholine receptor (AChR) expression on the surface of myotubes, 10 μM of alpha-bungarotoxin (α-BTX) conjugated with fluorescent dye was added to the culture and visualized with a fluorescent microscope. AChR expressing myotubes were counted from randomly selected areas (n=9) in culture microplates. AChR expression on myotubes in a 3-D culture system was assessed. RESULTS Agrin treatment significantly increased the percentage of AChR expressing myotubes from 15% to 100%. Treatment with conditioned medium (CM) derived from NG108 cells in differentiation medium enhanced AChR expression by up to 50%; however, when CM was added to growth medium, AChR expression on the surface of myotubes was not affected. Treatment with agrin also increased AChR expression on myotubes grown in a 3-D culture system consisting of fibrin gel. CONCLUSIONS This study shows that expression of AchR can be controlled by the use of nerve, conditioned medium derived from neural cells or neurotrophic factors (agrin). These results suggest that formation of NMJ may allow for more effective neural integration and accelerate the recovery of muscle function. Furthermore, rapid integration of neural tissue would decrease the incidence of muscle atrophy and scarring. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e346 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information In Kap Ko More articles by this author Sang Jin Lee More articles by this author John Jackson More articles by this author Anthony Atala More articles by this author James Yoo More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...