Name: canalostomy as a surgical approach to local drug delivery into the inner ears of adult and neonatal mice
Uploaded: 2018-05-25T13:00:00
Description: Watch this Scientific Journal Video about Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice at JoVE.com

Dette arbejde blev støttet af den National Natural Science Foundation of China (grant numre 81570912, af 81771016, 81100717).

Alle procedurer og animalske operationer blev gennemført efter retningslinjerne i Animal Care og brug Udvalget af Capital medicinske universitet i Kina.
1. enhed præparater
<ol><li>For at gøre injektion kanyle (figur 1A), Tilslut polyimid slanger (indre diameter 114.3 µm, ydre diameter 139.7 µm, længde ~ 3 cm) for polyethylen slanger (indre diameter 280 µm, ydre diameter 640 µm, længde ~ 40 cm). Bruger superlim, forsegle forbindelsen med mindst tre prog...

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	<li>Stachler, R. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+practice+guideline:+sudden+hearing+loss.">Clinical practice guideline: sudden hearing loss.</a> Otolaryngol Head Neck Surg. 146 (3 Suppl), S1-S35 (2012).</li><li>Lentz, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rescue+of+hearing+and+vestibular+function+by+antisense+oligonucleotides+in+a+mouse+model+of+human+deafness.">Rescue of hearing and vestibular function by antisense oligonucleotides in a mouse model of human deafness.</a> Nat Med. 19 (3), 345-350 (2013).</li><li>Rivera, T., Sanz, L., Camarero, G., Varela-Nieto, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+delivery+to+the+inner+ear:+strategies+and+their+therapeutic+implications+for+sensorineural+hearing+loss.">Drug delivery to the inner ear: strategies and their therapeutic implications for sensorineural hearing loss.</a> Curr Drug Deliv. 9 (3), 231-242 (2012).</li><li>El Kechai, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+local+drug+delivery+to+the+inner+ear.">Recent advances in local drug delivery to the inner ear.</a> Int J Pharm. 494 (1), 83-101 (2015).</li><li>Akil, O., Rouse, S. L., Chan, D. K., Lustig, L. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+method+for+virally+mediated+gene+delivery+to+the+mouse+inner+ear+through+the+round+window+membrane.">Surgical method for virally mediated gene delivery to the mouse inner ear through the round window membrane.</a> J Vis Exp. (97), e52187 (2015).</li><li>Ahmed, H., Shubina-Oleinik, O., Holt, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+Gene+Therapies+for+Genetic+Hearing+Loss.">Emerging Gene Therapies for Genetic Hearing Loss.</a> J Assoc Res Otolaryngol. 18 (5), 649-670 (2017).</li><li>Murillo-Cuesta, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Comparative+Study+of+Drug+Delivery+Methods+Targeted+to+the+Mouse+Inner+Ear:+Bullostomy+Versus+Transtympanic+Injection.">A Comparative Study of Drug Delivery Methods Targeted to the Mouse Inner Ear: Bullostomy Versus Transtympanic Injection.</a> J Vis Exp. (121), e54951 (2017).</li><li>Stevens, S. M., Brown, L. N., Ezell, P. C., Lang, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Mouse+Round-window+Approach+for+Ototoxic+Agent+Delivery:+A+Rapid+and+Reliable+Technique+for+Inducing+Cochlear+Cell+Degeneration.">The Mouse Round-window Approach for Ototoxic Agent Delivery: A Rapid and Reliable Technique for Inducing Cochlear Cell Degeneration.</a> J Vis Exp. (105), e53131 (2015).</li><li>Salt, A. N., Plontke, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+local+drug+delivery+to+the+inner+ear.">Principles of local drug delivery to the inner ear.</a> Audiol Neurootol. 14 (6), 350-360 (2009).</li><li>Akil, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restoration+of+hearing+in+the+VGLUT3+knockout+mouse+using+virally+mediated+gene+therapy.">Restoration of hearing in the VGLUT3 knockout mouse using virally mediated gene therapy.</a> Neuron. 75 (2), 283-293 (2012).</li><li>Pan, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+therapy+restores+auditory+and+vestibular+function+in+a+mouse+model+of+Usher+syndrome+type+1c.">Gene therapy restores auditory and vestibular function in a mouse model of Usher syndrome type 1c.</a> Nat Biotechnol. 35 (3), 264-272 (2017).</li><li>Kilpatrick, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adeno-associated+virus-mediated+gene+delivery+into+the+scala+media+of+the+normal+and+deafened+adult+mouse+ear.">Adeno-associated virus-mediated gene delivery into the scala media of the normal and deafened adult mouse ear.</a> Gene Ther. 18 (6), 569-578 (2011).</li><li>Izumikawa, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+hair+cell+replacement+and+hearing+improvement+by+Atoh1+gene+therapy+in+deaf+mammals.">Auditory hair cell replacement and hearing improvement by Atoh1 gene therapy in deaf mammals.</a> Nat Med. 11 (3), 271-276 (2005).</li><li>Chang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virally+mediated+Kcnq1+gene+replacement+therapy+in+the+immature+scala+media+restores+hearing+in+a+mouse+model+of+human+Jervell+and+Lange-Nielsen+deafness+syndrome.">Virally mediated Kcnq1 gene replacement therapy in the immature scala media restores hearing in a mouse model of human Jervell and Lange-Nielsen deafness syndrome.</a> EMBO Mol Med. 7 (8), 1077-1086 (2015).</li><li>Chen, Z., Mikulec, A. A., McKenna, M. J., Sewell, W. F., Kujawa, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+intracochlear+drug+delivery+in+the+mouse.">A method for intracochlear drug delivery in the mouse.</a> J Neurosci Methods. 150 (1), 67-73 (2006).</li><li>Kawamoto, K., Oh, S. H., Kanzaki, S., Brown, N., Raphael, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+functional+and+structural+outcome+of+inner+ear+gene+transfer+via+the+vestibular+and+cochlear+fluids+in+mice.">The functional and structural outcome of inner ear gene transfer via the vestibular and cochlear fluids in mice.</a> Mol Ther. 4 (6), 575-585 (2001).</li><li>Bowers, W. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurotrophin-3+transduction+attenuates+cisplatin+spiral+ganglion+neuron+ototoxicity+in+the+cochlea.">Neurotrophin-3 transduction attenuates cisplatin spiral ganglion neuron ototoxicity in the cochlea.</a> Mol Ther. 6 (1), 12-18 (2002).</li><li>Wang, G. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adeno-associated+virus-mediated+gene+transfer+targeting+normal+and+traumatized+mouse+utricle.">Adeno-associated virus-mediated gene transfer targeting normal and traumatized mouse utricle.</a> Gene Ther. 21 (11), 958-966 (2014).</li><li>Isgrig, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapy+Restores+Balance+and+Auditory+Functions+in+a+Mouse+Model+of+Usher+Syndrome.">Therapy Restores Balance and Auditory Functions in a Mouse Model of Usher Syndrome.</a> Mol Ther. 25 (3), 780-791 (2017).</li><li>Gassner, D., Durham, D., Pfannenstiel, S. C., Brough, D. E., Staecker, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Canalostomy+as+a+surgical+approach+for+cochlear+gene+therapy+in+the+rat.">Canalostomy as a surgical approach for cochlear gene therapy in the rat.</a> Anat Rec (Hoboken). 295 (11), 1830-1836 (2012).</li><li>Yamasoba, T., Yagi, M., Roessler, B. J., Miller, J. M., Raphael, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inner+ear+transgene+expression+after+adenoviral+vector+inoculation+in+the+endolymphatic+sac.">Inner ear transgene expression after adenoviral vector inoculation in the endolymphatic sac.</a> Hum Gene Ther. 10 (5), 769-774 (1999).</li><li>Xia, L., Yin, S., Wang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inner+ear+gene+transfection+in+neonatal+mice+using+adeno-associated+viral+vector:+a+comparison+of+two+approaches.">Inner ear gene transfection in neonatal mice using adeno-associated viral vector: a comparison of two approaches.</a> PLoS One. 7 (8), e43218 (2012).</li><li>Chien, W. W., McDougald, D. S., Roy, S., Fitzgerald, T. S., Cunningham, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cochlear+gene+transfer+mediated+by+adeno-associated+virus:+Comparison+of+two+surgical+approaches.">Cochlear gene transfer mediated by adeno-associated virus: Comparison of two surgical approaches.</a> Laryngoscope. 125 (11), 2557-2564 (2015).</li><li>Zhu, B. Z., Saleh, J., Isgrig, K. T., Cunningham, L. L., Chien, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hearing+Loss+after+Round+Window+Surgery+in+Mice+Is+due+to+Middle+Ear+Effusion.">Hearing Loss after Round Window Surgery in Mice Is due to Middle Ear Effusion.</a> Audiol Neurootol. 21 (6), 356-364 (2017).</li><li>Wang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+postnatal+virus+inoculation+into+the+scala+media+achieved+extensive+expression+of+exogenous+green+fluorescent+protein+in+the+inner+ear+and+preserved+auditory+brainstem+response+thresholds.">Early postnatal virus inoculation into the scala media achieved extensive expression of exogenous green fluorescent protein in the inner ear and preserved auditory brainstem response thresholds.</a> J Gene Med. 15 (3-4), 123-133 (2013).</li><li>Lee, M. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+of+human+embryonic+stem+cells+implanted+in+the+guinea+pig+auditory+epithelium.">Survival of human embryonic stem cells implanted in the guinea pig auditory epithelium.</a> Sci Rep. 7, 46058 (2017).</li><li>Ishimoto, S., Kawamoto, K., Kanzaki, S., Raphael, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+transfer+into+supporting+cells+of+the+organ+of+Corti.">Gene transfer into supporting cells of the organ of Corti.</a> Hear Res. 173 (1-2), 187-197 (2002).</li><li>Okada, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+transfer+targeting+mouse+vestibule+using+adenovirus+and+adeno-associated+virus+vectors.">Gene transfer targeting mouse vestibule using adenovirus and adeno-associated virus vectors.</a> Otol Neurotol. 33 (4), 655-659 (2012).</li><li>Suzuki, J., Hashimoto, K., Xiao, R., Vandenberghe, L. H., Liberman, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cochlear+gene+therapy+with+ancestral+AAV+in+adult+mice:+complete+transduction+of+inner+hair+cells+without+cochlear+dysfunction.">Cochlear gene therapy with ancestral AAV in adult mice: complete transduction of inner hair cells without cochlear dysfunction.</a> Sci Rep. 7, 45524 (2017).</li><li>Guo, J. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cochleovestibular+gene+transfer+in+neonatal+mice+by+canalostomy.">Cochleovestibular gene transfer in neonatal mice by canalostomy.</a> Neuroreport. 28 (11), 682-688 (2017).</li><li>Beyea, J. A., Agrawal, S. K., Parnes, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmastoid+semicircular+canal+occlusion:+a+safe+and+highly+effective+treatment+for+benign+paroxysmal+positional+vertigo+and+superior+canal+dehiscence.">Transmastoid semicircular canal occlusion: a safe and highly effective treatment for benign paroxysmal positional vertigo and superior canal dehiscence.</a> Laryngoscope. 122 (8), 1862-1866 (2012).</li><li>Naples, J. G., Eisen, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+History+and+Evolution+of+Surgery+on+the+Vestibular+Labyrinth.">The History and Evolution of Surgery on the Vestibular Labyrinth.</a> Otolaryngol Head Neck Surg. 155 (5), 816-819 (2016).</li><li>Hamilton, L., Keh, S., Spielmann, P. M., Hussain, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+we+do+it:+locating+the+posterior+semicircular+canal+in+occlusion+surgery+for+refractory+benign+paroxysmal+positional+vertigo:+a+cadaveric+temporal+bone+study.">How we do it: locating the posterior semicircular canal in occlusion surgery for refractory benign paroxysmal positional vertigo: a cadaveric temporal bone study.</a> Clinical Otolaryngology. 41 (2), 190-193 (2016).</li><li>Jung, J. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=siRNA+targeting+Hes5+augments+hair+cell+regeneration+in+aminoglycoside-damaged+mouse+utricle.">siRNA targeting Hes5 augments hair cell regeneration in aminoglycoside-damaged mouse utricle.</a> Mol Ther. 21 (4), 834-841 (2013).</li><li>Kamiya, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cell+transplantation+accelerates+hearing+recovery+through+the+repair+of+injured+cochlear+fibrocytes.">Mesenchymal stem cell transplantation accelerates hearing recovery through the repair of injured cochlear fibrocytes.</a> Am J Pathol. 171 (1), 214-226 (2007).</li><li>Pfannenstiel, S. C., Praetorius, M., Plinkert, P. K., Brough, D. E., Staecker, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bcl-2+gene+therapy+prevents+aminoglycoside-induced+degeneration+of+auditory+and+vestibular+hair+cells.">Bcl-2 gene therapy prevents aminoglycoside-induced degeneration of auditory and vestibular hair cells.</a> Audiol Neurootol. 14 (4), 254-266 (2009).</li><li>Kawamoto, K., Izumikawa, M., Beyer, L. A., Atkin, G. M., Raphael, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+hair+cell+regeneration+in+the+mouse+utricle+following+gentamicin+ototoxicity.">Spontaneous hair cell regeneration in the mouse utricle following gentamicin ototoxicity.</a> Hear Res. 247 (1), 17-26 (2009).</li><li>Wang, G. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+signaling+and+Atoh1+expression+during+hair+cell+regeneration+in+the+mouse+utricle.">Notch signaling and Atoh1 expression during hair cell regeneration in the mouse utricle.</a> Hear Res. 267 (1-2), 61-70 (2010).</li><li>Pietola, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HOX-GFP+and+WOX-GFP+lentivirus+vectors+for+inner+ear+gene+transfer.">HOX-GFP and WOX-GFP lentivirus vectors for inner ear gene transfer.</a> Acta Otolaryngol. 128 (6), 613-620 (2008).</li><li>Han, J. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgene+expression+in+the+guinea+pig+cochlea+mediated+by+a+lentivirus-derived+gene+transfer+vector.">Transgene expression in the guinea pig cochlea mediated by a lentivirus-derived gene transfer vector.</a> Hum Gene Ther. 10 (11), 1867-1873 (1999).</li></ol>

I denne undersøgelse viste vi, at narkotika levering af canalostomy resulterede i omfattende udbredelse af reagens i hele cochlea og vestibulære slutningen-organer. Som en indre øre gen leveringsmetode resulterede canalostomy i normal god landbrugspraksis udtryk for indre ørerne af voksne og neonatal mus med minimal skade til høring og vestibulære funktion. Desuden kan flere injektioner let udføres i samme dyr.
En af de største styrker i canalostomy er, at det forårsager minimal skade...

Sensorineural høre tab og vestibulær dysfunktion påvirker et betydeligt antal patienter og er tæt forbundet med indre øre lidelser. Levering af terapeutiske lægemidler i det indre øre viser løfte til behandling af indre øre lidelser. En systemiske eller lokale tilgang kan bruges til at levere narkotika ind i det indre øre. Nogle indre øre sygdomme behandles med held med systemisk drug administration, såsom idiopatisk pludselige høretab, som er almindeligt behandles med systemisk steroid1. Derudover viste Lentz et al. , at systemisk administration af antisense oligonukleotid var i stand til at forbedre hørelse og balance funktioner i Ush1c mutant musen model2. Men en stor del af indre øre sygdomme er ikke effektivt behandles systemisk drug administration på grund af blod-labyrint barriere, som begrænser narkotika adgang til det indre øre3,4. Derimod kan lokale drug delivery strategier behandle indre øre lidelser mere effektivt. Ja, det indre øre er potentielt et ideelt mål for lokale medicinafgivelse; Det er fyldt med væske, som fremmer formidling af stoffet efter one-site diffusion eller injektion, og det er relativt isoleret fra nærliggende organer, som begrænser bivirkninger5,6.
Lokale drug delivery strategier omfatter intratympanic og intralabyrinthine metoder. Effektiviteten af den intratympanic rute bygger i vid udstrækning på narkotikamisbrug permeabilitet gennem det runde vindue membran (RWM) og opholdstid af lægemidlet på RWM3,4,7,8. Det er således ikke egnet til leveringen af narkotika eller reagenser, der ikke kan trænge igennem RWM. Intralabyrinthine metoder indebærer podning af medicin direkte ind i det indre øre, resulterer i en høj dosis og udbredt distribution. Men intralabyrinthine metoder kræver delikat operationer og er invasiv, fører til skader på indre øre funktion. I øjeblikket, bruges intralabyrinthine injektion af narkotika kun i dyreforsøg, som det ikke er påvist for at være tilstrækkeligt sikker til brug i mennesker9. Derfor, kirurgiske procedurer bør forenkles, og nedsættes risikoen for skade for at oversætte intralabyrinthine tilgange til klinikken.
Flere intralabyrinthine metoder er blevet vurderet i dyr ved injektion gennem RWM5,10,11 og ind i scala media12,13,14, scala tympani 15 , 16, scala forhallen17, halvrunde kanaler16,18,19,20og endolymphatic sac21. Hver af disse metoder har fordele og ulemper6. Levering gennem RWM er atraumatisk i neonatal mus5,22. Dog er milde høretab observeret hos voksne mus efter RWM injektion23, muligvis på grund af mellemøret effusion efter kirurgi24. Scala media injektion, der indebærer injektion af reagenset direkte i af endolymphatic rum sensoriske epitel, opnår en høj reagens koncentration i målet ende-organer12,14, 25 , 26. men denne tilgang kræver en kompliceret procedure og resulterer i betydelig udvidelse af høringen tærskel, hvis udført senere end postnatal dag 5 (P5)25,27, hvilket begrænser dets anvendelse.
I forhold til de ovennævnte intralabyrinthine tilgange, forårsager canalostomy minimal skade det indre øre, især i voksen mus16,18,28,29,30, som er vigtig for vurderingen af beskyttende effekter og translationel aspekter. Derudover i gnavere, er halvrunde kanaler placeret ud over bulla, som letter kirurgiske procedurer og forebygger forstyrrelser af mellemøret under kirurgi. I klinikken bruges halvrunde kanalen operationer for genstridig benign paroxysmal positional vertigo31,32,33, tyder på den kliniske feasibility af canalostomy. Da det først blev beskrevet af Kawamoto mfl. 16 i 2001, canalostomy har været brugt til at levere forskellige reagenser, såsom virale vektorer, siRNA, stamceller og aminoglycosid, i den murine indre øre18,19,28,29 ,34,35,36,37. Podning af adeno-associeret virus (AAV) vektorer af canalostomy aktiverer overekspression af udefrakommende gener i sensoriske epitel og primære neuroner i cochlea og vestibulære slutningen-organer18,28, 29,30. Whirlin genterapi af canalostomy genskaber balancen funktion og forbedrer hørelse i en musemodel af menneskelige Usher syndrom19, tyder på, at canalostomy er nyttigt for studier af genterapi for genetiske cochleovestibular sygdomme. Transplantation af mesenkymale stamceller af canalostomy resulterer i reorganiseringen af cochlear fibrocytes og hørelse opsving i en rotte model af akut sensorineural hearing tab35. Derudover canalostomy kan bruges til at indføre aminoglycosider i det indre øre at etablere vestibulære læsioner18,34,38, og flere injektioner kan udføres, hvis det kræves18 , 34.
I denne artikel beskriver vi i detaljer, canalostomy teknikker i voksen og neonatal mus. Vi inokuleres forskellige reagenser, herunder hurtig-grønne farvestof og AAV serotype 8 (AAV8), sammen med de grønne fluorescerende proteiner (NGL) genet (AAV8-NGL) og streptomycin, i mus indre øre til at vurdere de umiddelbare og langsigtede resultater efter canalostomy.

<table>
	<tbody>
		<tr>
			<td>Polymide Tubing</td>
			<td>A-M Systems</td>
			<td>823400</td>
			<td></td>
		</tr>
		<tr>
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			<td>Scientific Commodities Inc.</td>
			<td>BB31695-PE/1</td>
			<td></td>
		</tr>
		<tr>
			<td>10&#956;l Microsyringe</td>
			<td>Hamilton Company</td>
			<td>80001</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine HCL</td>
			<td>Sigma-Aldrich Co. Llc.</td>
			<td>X-1251</td>
			<td></td>
		</tr>
		<tr>
			<td>Operating Miroscope</td>
			<td>Carl Zeiss Optical LLC.</td>
			<td>Pico</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Forceps</td>
			<td>Dumont Dumostar</td>
			<td>10576</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast-green Dye</td>
			<td>Sigma-Aldrich Co. Llc.</td>
			<td>F7252</td>
			<td></td>
		</tr>
		<tr>
			<td>AAV8-GFP</td>
			<td>BioMiao Biological Technology Co. Ltd (Beijing, China)</td>
			<td>20161101</td>
			<td>Titer: 2&#215;10e12 vg/mL</td>
		</tr>
		<tr>
			<td>Streptomycin Sulfate</td>
			<td>Sigma-Aldrich Co. Llc.</td>
			<td>S9137</td>
			<td></td>
		</tr>
		<tr>
			<td>Microinjection Pump</td>
			<td>Stoelting Co.</td>
			<td>789100S</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric Pad</td>
			<td>Pet Fun</td>
			<td>11072931136</td>
			<td></td>
		</tr>
		<tr>
			<td>1 cc Syringe</td>
			<td>Mishawa Medical Industries Ltd. (Shanghai, China)</td>
			<td>2011-3151258</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine HCL</td>
			<td>Gutian Pharmaceutical Co., Ltd. (Fujian, China)</td>
			<td>H35020148</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric Animal Clipper</td>
			<td>Codos Electrical Appliances Co., Ltd. (Guangdong, China)</td>
			<td>CP-8000</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Pellet</td>
			<td>Yatai Healthcare Ltd. (Henan, China)</td>
			<td>Yu-2008-1640081</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Shanghai Pudong Jinhuan Medical Products Co., Ltd. (Shanghai, China)</td>
			<td>Hu-2013-2650207</td>
			<td></td>
		</tr>
		<tr>
			<td>Eye Ointment</td>
			<td>Beijing Shuangji Pharmaceutical Ltd. (Beijng China)</td>
			<td>H11021270</td>
			<td></td>
		</tr>
	</tbody>
</table>

canalostomy as a surgical approach to local drug delivery into the inner ears of adult and neonatal mice

Lokal levering af terapeutiske lægemidler i det indre øre er en lovende behandling for indre øre sygdomme. Injektion gennem halvrunde kanaler (canalostomy) har vist sig at være en nyttig tilgang til lokale medicinafgivelse ind i det indre øre. Målet med denne artikel er at beskrive i detaljer, de kirurgiske teknikker, der er involveret i canalostomy i både voksen og neonatal mus. Som anført af fast-grønne farvestof og adeno-associeret virus serotype 8 med grøn fluorescerende proteiner-gen, canalostomy lettes bred fordeling af injicerede reagenser i cochlea og vestibulære slutningen-organer med minimal skade til høring og vestibulære funktion. Operationen blev succesfuldt implementeret i både voksen og neonatal mus; faktisk, flere operationer kan udføres hvis det kræves. Afslutningsvis, canalostomy er en effektiv og sikker tilgang til medicinafgivelse indre ører af voksne og neonatal mus og kan bruges til at behandle menneskelige indre øre sygdomme i fremtiden.

Hurtig-grønne farvestof var tilført PSC af voksne og neonatal mus at evaluere dens umiddelbare distribution i det indre øre. Farvestoffet blev opdaget i hele cochlea, forhallen og halvcirkelformede kanaler umiddelbart efter operationen (figur 4).
For at vurdere sikkerheden og effektiviteten af canalostomy for indre øre gen levering, blev AAV8-NGL sprøjtet ind i det indre øre for voksne og neon...

Watch this Scientific Journal Video about Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice at JoVE.com

Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice

Canalostomy som en kirurgisk tilgang til lokale Drug Delivery indre ører af voksne og Neonatal mus

Her beskriver vi canalostomy procedure, som tillader lokale medicinafgivelse indre ører af voksne og neonatal mus gennem halvrunde kanalen med minimal skade til høring og vestibulære funktion. Denne metode kan bruges til podes virale vektorer, farmaceutiske produkter og små molekyler i mus indre øre.

Local delivery of therapeutic drugs into the inner ear is a promising therapy for inner ear diseases. Injection through semicircular canals (canalostomy) ...

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