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I-Gene vectors yonyango lwe-cystic fibrosis lung disease kufuneka ijolise kwiindlela zomoya eziqhubayo ngenxa yokuba i-peripheral lung transduction ayiboneleli ngonyango. ixesha labathwali bemizila yemfuza kwiindlela zomoya libalulekile kodwa kunzima ukulifumana.IGene carrier-conjugated magnetic particles ezinokuthi zibhekiswe kumphezulu wendlela zomoya zingaphucula ukujoliswa kwengingqi.Ngenxa yemiceli mngeni ye vivo visualization, ukuziphatha kwaloo masuntswana mancinane kazibuthe kumphezulu wendlela yomoya phambi kobuso bemagnethi esetyenzisiweyo ayiqondwa kakuhle.Injongo yolu phononongo yolu phononongo yayikukusebenzisa i-synchromotion ye-imaging series kwi-vinchromotion ye-imaging series kukusebenzisa i-synchromotion ye-imaging series. Amasuntswana kuqhoqhoqho lweempuku ezinesthetized ukuhlola amandla kunye neepatheni zokuziphatha komntu kunye nobuninzi besuntswana kwi-vivo.Siye saphinda savavanya ukuba ngaba ukuhanjiswa kwamasuntswana emagnethi yelentiviral kubukho bemagnethi kuya konyusa ukusebenza kakuhle kogqithiso kwi-trachea yempuku.I-Synchrotron X-ray imaging ityhila indlela yokuziphatha kwamasuntswana omagnetic kwi-vivory fields lula. irhuqe kumphezulu womoya ophilayo kunye nemagnethi, kodwa ngexesha lothutho, iidiphozithi zigxininiswe kwintsimi yembono apho intsimi yemagneti inamandla kakhulu.Ubuchule boTshintsho lwaye lwandiswa ngokuphindwe kathandathu xa amasuntswana emagnethi yelentiviral ahanjiswa kubukho bemagnethi.Ngokudibeneyo, ezi ziphumo zibonisa ukuba amasuntswana emagnethi kunye nemagnethi anokuba ziindlela ezixabisekileyo ekujoliseni ekuphuculeni i-victor yomoya.
I-Cystic fibrosis (CF) ibangelwa ukuguquguquka kwemfuza enye ebizwa ngokuba yi-CF transmembrane conductance regulator (CFTR) .Iprotheyini ye-CFTR yijelo le-ion elikhoyo kwiiseli ezininzi ze-epithelial kuwo wonke umzimba, kubandakanywa neendlela zomoya eziqhubayo, indawo enkulu ye-CF pathogenesis.Iziphene zeCFTR zikhokelela ekuthuthweni kwamanzi okungaqhelekanga, ukunciphisa ulwelo lwamanzi kunye nokunciphisa umoya we-airway we-SL. umaleko.Oku kwakhona kuphazamisa isakhono senkqubo yothutho mucociliary (MCT) ukucima amasuntswana ephefumle kunye pathogens ukusuka airways.Injongo yethu kukuphuhlisa lentiviral (LV) gene unyango ukuhambisa ikopi echanekileyo yofuzo CFTR kunye nokuphucula ASL, MCT, kunye nempilo emiphungeni, kwaye ukuqhubeka nokuphuhlisa ubugcisa obutsha ezikwaziyo ukumeta-mitha paravo1.
I-LV vectors ingomnye wabaviwa abakhokelayo kwi-CF yonyango ye-gene ye-airway gene, ngokukodwa ngenxa yokuba banokudibanisa ngokusisigxina i-gene yonyango kwiiseli ze-basal ye-airway (iiseli ze-airway stem) .Oku kubalulekile kuba banokubuyisela i-hydration eqhelekileyo kunye ne-mucus clearance ngokuhlula kwi-gene esebenzayo-echanekileyo ye-CF-ehambelana ne-airway ye-airway ehambelana neeseli ze-airway, okubangela ukuba i-vector iqhube i-airway, ekhokelela ekuqhubeni iiseli ze-airway, okubangela ukuba i-vector iqhube ubomi bonke. Isifo se-CF emiphunga siqala.Ukuhanjiswa kwe-vector enzulu kwimiphunga kunokubangela ukuguqulwa kwe-alveolar, kodwa le nto ayinayo inzuzo yokwelapha kwi-CF.Nangona kunjalo, ulwelo olunjengama-gene carriers lufudukela kwi-alveoli ngokuphefumlelwa emva kokunikezelwa kwe-3,4 kunye neengqungquthela zonyango zihlanjululwa ngokukhawuleza kwi-transduction yomlomo we-LV ukuya kwiiseli ezijoliswe ngqo kwi-MCT ehambelana ngqo ne-MCT. vumela ukuthathwa kweselula - "ixesha lokuhlala" i-5 - eyancipha ngokulula ukuhamba komoya oqhelekileyo wengingqi kunye nokulungelelaniswa kwe-particle mucus capture kunye ne-MCT.Kuba i-CF, ukukwazi ukwandisa ixesha lokuhlala kwe-LV ngaphakathi kwe-airway kubalulekile ukufezekisa amanqanaba aphezulu okutshintshwa kulo mmandla, kodwa kude kube ngumngeni.
Ukuze sinqobe lo mqobo, sicebisa ukuba i-LV magnetic particles (MPs) inokunceda ngeendlela ezimbini ezihambelanayo.Okokuqala, ziyakwazi ukukhokelwa ngamagnetic kwindawo yomoya ukuze kuphuculwe ukujoliswa kunye nokunceda iinqununu ze-gene carrier zihlala kwindawo efunwayo yomoya; kunye ne-ASL) ukuya kwi-cell layer 6.MPs zisetyenziswe ngokubanzi njengezithuthi ezijoliswe kuzo zokuhambisa iziyobisi xa zibophelela kwii-antibodies, iziyobisi ze-chemotherapeutic, okanye ezinye iimolekyuli ezincinci ezincamathela kwi-cell membranes okanye zibophe kwi-cell cell receptors ezifanelekileyo kwaye ziqokelele kwiindawo ze-tumor phambi kombane omileyo. IMagnetic Fields for Cancer Treatment 7. Ezinye iindlela “zehyperthermal” zijolise ekufudumezeni iiMPs xa zisesichengeni samagnetic oscillating, ngaloo ndlela zitshabalalisa iiseli zethumba.Umgaqo wokudluliselwa kwemagnethi, apho imagnethi isetyenziswa njengearhente yodluliselo ukuphucula ukudluliselwa kweDNA kwiiseli, isetyenziswa ngokuqhelekileyo kwi-vitro usebenzisa uluhlu lwe-non-viral kunye ne-viral cell cell cell. Imagnetotransfection isekiwe, kunye nokuhanjiswa kwe-vitro ye-LV-MPs kumgca weseli ye-bronchial epithelial phambi kommandla wemagnethi omileyo, ukwandisa ukusebenza kakuhle kodluliselo nge-186-fold xa kuthelekiswa neVector ye-LV kuphela. I-LV-MP iye yasetyenziswa kwimodeli ye-in vitro CF, apho ugqithiso lwemagnethi lwandise ukudluliselwa kwe-LV kwindawo yobuso bomoya-olwelwe-20 kujongano lwe-air-liquid-CF I-sputum10.Nangona kunjalo, kwi-vivo magnetotransfection yamalungu ifumene ingqwalasela encinci kwaye ihlolwe kuphela kwizifundo zezilwanyana ezimbalwa11,12,13,14,15, ngakumbi kwimiphunga16,17.Nangona kunjalo, amathuba okudluliselwa kwamagnetic kunyango lwe-CF lung acacile.Tan et al. Ukuhanjiswa kwe-nanoparticle pulmonary kuya kuvula indlela yezicwangciso zexesha elizayo ze-CFTR zokuphefumla ukuphucula iziphumo zekliniki kwizigulana ze-CF”6.
Ukuziphatha kwamasuntswana amancinci amagnetic kwiindawo zomoya phambi kobuso bomhlaba osetyenzisiweyo kunzima ukukubona kunye nokufunda, kwaye ngaloo ndlela kuqondwa kakuhle.Kwezinye izifundo, siphuhlise i-synchrotron-propagation-based based phase-contrast X-ray imaging (PB-PCXI) indlela yokungabonakali kunye nokulinganisa umzuzu kwi-vivo utshintsho kwi-ASL ye-ASL kunye ne-MAC20 yokulinganisa ngokuthe ngqo kwi-ASL kunye ne-M20 ye-ASL kunye ne-Mhydration ye-M20, kunye ne-MAC20. isetyenziswe njengesalathisi sokuqala sokusebenza konyango.Ukongezelela, indlela yethu yokuvavanya i-MCT isebenzisa i-10-35 µm i-diameter particles eyenziwe nge-alumina okanye i-high refractive index glass njengoko iimpawu ze-MCT ezibonakalayo zisebenzisa i-PB-PCXI21.Zombini iindlela zobugcisa zifanelekile ukubonwa kweentlobo zeentlobo zamasuntswana, kuquka i-MP.
Ngenxa yesisombululo sayo esiphezulu sesithuba kunye nesexeshana, i-PB-PCXI-based ASL kunye ne-MCT yohlalutyo lweendlela zifaneleke kakuhle ukuhlola amandla kunye neepateni zokuziphatha okukodwa kunye nobuninzi besuntswana kwi-vivo ukusinceda siqonde kwaye sandise ubuchule bokuhanjiswa kofuzo lwe-MP. Indlela esiyisebenzisayo apha isuka kwizifundo zethu zisebenzisa i-SPring-8 BL20B yedosi elandelwayo kwidosi ebonwayo kwi-shammom yedosi ebonwayo kwi-shammom yedosi ebonwayo kwi-sham yonikezelo lwedosi. Iimpumlo zempumlo kunye nemiphunga yomoya yeempuku ukunceda ukucacisa iipatheni zethu zokuchazwa kwemfuza ezingeyoyunifomu ezibonwa kwi-gene yethu yedosi yezilwanyana ze-3,4.
Injongo yolu phononongo yayikukusebenzisa i-synchrotron PB-PCXI ukubona ngeso lengqondo iintshukumo ze-vivo zothotho lwamaLungu ePalamente kuqhoqhoqho lweempuku eziphilayo. Ezi zifundo zokucinga ze-PB-PCXI zenzelwe ukuvavanya uluhlu lwamaLungu ePalamente, amandla ombane, kunye neendawo zokumisela impembelelo yazo kwintshukumo yeMP. Kuluhlu lwesibini lwezifundo, siye safuna ukusebenzisa olu lungelelwaniso lufanelekileyo ukubonisa ipateni yoguqulelo oluphuma kunikezelo lwe-vivo lwe-LV-MPs kwindlela yomoya yempuku, ngokusekwe kwingcinga yokuba ukuhanjiswa kwe-LV-MPs kuphuculo lomoya ekujoliswe kuko kuya kuphucula umgangatho weLV-MPs kwindlela yomoya.
Zonke izifundo zezilwanyana zenziwa ngokwemigaqo evunyiweyo yiYunivesithi yaseAdelaide (M-2019-060 kunye ne-M-2020-022) kunye neKomiti ye-SPring-8 Synchrotron Animal Ethics Committee.Uvavanyo lwenziwa ngokwemigaqo ye-ARRIVE.
Yonke imifanekiso ye-X-ray yenziwe kwi-BL20XU ye-beamline kwi-SPring-8 synchrotron eJapan, isebenzisa ukuseta okufana noko kuchazwe ngaphambili21,22.Ngokufutshane, ibhokisi yokuhlola ibekwe kwi-245 m ukusuka kwindandatho yokugcina i-synchrotron.Umgama we-sample-to-detector we-0.6 m isetyenziselwa izifundo ze-particle ye-imaging ye-particle ye-vi. I-monochromatic beam energy ye-25 keV isetyenzisiwe.Imifanekiso ibanjwe kusetyenziswa i-X-ray converter ephezulu (SPring-8 BM3) idityaniswe ne-sCMOS detector.Umguquleli uguqula i-X-rays ekukhanyeni okubonakalayo usebenzisa i-10 µm engqindilili ye-scintillator (Gd3Al2Ga3C eqondiswe kwi-sensor ye-mic × 12), ethi emva koko iqondise i-microscope × (NA 0.3) I-mm.Ubude be-exposure ye-100 ms bukhethwe ukwandisa umlinganiselo we-signal-to-noise of particles magnetic ngaphakathi nangaphandle kwendlela yomoya ngelixa unciphisa izinto ezihambayo ezibangelwa ukuphefumla. Kwizifundo ze-vivo, i-shutter ye-X-ray ekhawulezayo ifakwe kwindlela ye-X-ray ukunciphisa umthamo we-radiation ngokuthintela umqadi we-X-ray phakathi kwe-exposure beam.
I-LV carrier ayizange isetyenziswe kuyo nayiphi na i-SPring-8 PB-PCXI yokucinga ngezifundo ngenxa yokuba igumbi lokucinga le-BL20XU alikho i-Biosafety Level 2 eqinisekisiweyo. Endaweni yoko, sikhethe uluhlu lwamaLungu e-MP abonakaliswe kakuhle kubathengisi ababini bezorhwebo-egubungela uluhlu lobukhulu, izixhobo, ukugxilwa kwentsimbi, kunye nezicelo ngaphakathi kwe-glass motion , i-capilla ephilayo ichaphazela njani i-MP. iindlela zomoya. phezu komhlaba.MPs ubungakanani ukusuka 0.25 ukuya 18 μm kwaye zenziwe ngezinto ezahlukeneyo (bona Itheyibhile 1), kodwa ukubunjwa isampuli nganye, kuquka ubungakanani amasuntswana magnetic ngaphakathi MP, akwaziwa.Ngokusekelwe kwizifundo zethu MCT olubanzi 19, 20, 21, 23, 24, silindele ukuba MPs kwi-5 μ umphezulu womoya njengoko kubonwa umphezulu we-trache emoyeni encinci. ukukhupha izakhelo ezilandelelanayo ukubona ukubonakala okuphuculweyo kwentshukumo yeMP.IMP eyodwa eyi-0.25 μm-ubungakanani incinci kunesisombululo sesixhobo sokucinga, kodwa iPB-PCXI kulindeleke ukuba ibone umahluko wevolumu kunye nokushukuma kolwelo olungaphezulu apho zifakwe khona emva kokumiswa.
Iisampulu zeMP nganye kwiTheyibhile 1 zalungiswa kwi-20 μl yeglasi ye-capillaries (iDrummond Microcaps, PA, USA) kunye nobubanzi obungaphakathi be-0.63 mm. Amasuntswana eCorpuscular afumaneka emanzini, ngelixa amasuntswana e-CombiMag afumaneka kwi-fluid yobunikazi bomenzi. ityhubhu nganye igcwele isiqingatha kunye nolwelo (malunga nesampulu ye-1) 1) .I iicapillaries zeglasi zabekwa ngokuthe tye kwinqanaba lesampulu kwibhokisi yokucinga, ngokulandelelana, kwaye yabeka imiphetho ye-fluid.A 19 mm ububanzi (28 mm ubude) iqokobhe lenikeli elinqabileyo lomhlaba neodymium yentsimbi yeboron (NdFeB) imagnethi (N35, ikati no. idityaniswe kwinqanaba lokuguqulela elahlukileyo ukufezekisa Guqula indawo yayo ukude ngexesha lokucinga.Ukufumana umfanekiso weX-ray iqala xa umazibuthe ubekwe malunga ne-30 mm ngaphezu kwesampulu, kwaye imifanekiso ifunyenwe ngesantya seefreyimu ezi-4 ngomzuzwana.Ngethuba lomfanekiso, umazibuthe wasondezwa kufutshane netyhubhu yeglasi ye-capillary (malunga ne-1 mm kude) kwaye emva koko iguqulelwe ecaleni kwetyhubhu yamandla kwaye iguqulelwe kwimibhobho yokuqina.
Ukusekwa kwe-in vitro imaging equlethe iisampuli ze-MP kwii-capillaries zeglasi kwisampulu ye-xy translation stage.
Emva kokuba ukubonakala kwe-in vitro yama-MPs kusungulwe, i-subset yabo yavavanywa kwi-vivo kwi-wistar-type female albino Wistar rats (~ iiveki ze-12 ubudala, ~ 200 g) .0.24 mg / kg medetomidine (Domitor®, Zenoaq, Japan), 3.2 mg / kg midazolam (Dormicum®, I-Astella, i-Astella, i-Astella yaseJapan) kunye ne-butor (I-Vetorphale®, i-Meiji Seika) Iigundane zenziwa i-anesthetized ngomxube we-Pharma), eJapan) nge-injection ye-intraperitoneal. I-22 .I-plate ye-imaging yabe ifakwe kwinqanaba lokuguqulela isampula kwibhokisi ye-imaging kwi-angle encinci ukulungelelanisa i-trachea ngokuthe tye kumfanekiso we-X-ray, njengoko kuboniswe kuMfanekiso 2a.
(a) Ukuseta umfanekiso we-vivo kwibhokisi yokucinga ye-SPring-8, indlela ye-X-ray iphawulwe ngomgca obomvu odayiweyo.
Inkqubo yepompo yesirinji elawulwa kude (UMP2, i-World Precision Instruments, iSarasota, FL) isebenzisa i-100 μl yeglasi yesirinji idibaniswe ne-PE10 tubing (OD 0.61 mm, i-ID 0.28 mm) ngenaliti ye-30 ye-Ga. Phawula ityhubhu ukuqinisekisa ukuba i-tip ikwi-tube echanekileyo xa ufaka umbhobho we-microET pump echanekileyo. I-syringe plunger yarhoxiswa ngelixa i-tip ye-tube igxininiswe kwisampuli ye-MP ukuba ihanjiswe.I-tube yokuhambisa elayishiweyo emva koko yafakwa kwi-tube endotracheal, ibeka incam ngaphakathi kweyona nxalenye inamandla okulindelekileyo kwintsimi yethu yamagnetic.Ukufumana umfanekiso walawulwa ngokusebenzisa umtshina wokuphefumula oxhunywe kwi-Arduino yethu esekelwe kwibhokisi yexesha, kunye nebhokisi yokuvula iqondo lokushisa, kunye neempawu zokuvula i-respiration ukufunyanwa) zirekhodwe kusetyenziswa iPowerlab kunye neLabChart (i-AD Instruments, eSydney, e-Australia) 22. Xa i-imaging Xa i-enclosure ingafikeleleki, iikhamera ezimbini ze-IP (i-Panasonic BB-SC382) zibekwe malunga ne-90 ° komnye nomnye kwaye zisetyenziselwa ukubeka esweni indawo yemagnethi ngokubhekiselele kwi-trachea, i-miniging . izinto zakudala, umfanekiso omnye wafunyanwa ngomphefumlo ngamnye ngexesha lokuhamba kolwandle.
Imagnethi idityaniswe kwinqanaba lesibini elinokuthi libekwe kude ukusuka ngaphandle kwendawo yokucinga.Izikhundla ezahlukeneyo zemagnethi kunye nolungelelwaniso zavavanywa, kubandakanya: Inyuswe kwi-engile emalunga ne-30 ° ngaphezulu koqhoqhoqho (uqwalaselo oluboniswe kwiMizobo 2a kunye ne-3a); omnye umazibuthe ngaphezu kwesilwanyana kunye nomnye ongezantsi, kunye neepali ezibekwe ukutsala (Umfanekiso 3b); omnye umazibuthe phezu kwesilwanyana kunye nomnye ongezantsi, kunye neepali ezimiselwe ukugxotha (Umfanekiso 3c); kunye nemagnethi enye ngaphezulu kunye ne-perpendicular kuqhoqhoqho (Umfanekiso 3d) Emva kokuba isilwanyana kunye nomazibuthe ziqwalaselwe kwaye iMP eza kuvavanywa ilayishwe kwimpompo yesirinji, hambisa i-50 μl idosi kwireyithi ye-4 μl / isekhondi ngelixa ufumana imifanekiso.Imagnethi ihanjiswa emva naphambili kunye okanye ngokuqhubekayo ukuya kwi-trachea.
Ulungelelwaniso lwemagnethi kumfanekiso we-vivo (a) umazibuthe omnye ngentla koqhoqhoqho kwi-engile emalunga nama-30°, (b) iimagnethi ezimbini ezisetelwe ukutsala, (c) iimagnethi ezimbini ezisetelwe ukugxotha, (d) umazibuthe omnye ngentla kunye ne-perpendicular kuqhoqhoqho. icala.Imagnethi ishukunyiswa kubude bendlela yomoya okanye ekhohlo kwaye ekunene ngaphezu koqhoqhoqho kwicala lomqadi weX-reyi.
Siphinde safuna ukufumanisa ukubonakala kunye nokuziphatha kweengqungquthela kwiindlela zomoya ngokungabikho kokuphefumla okudidayo kunye nokunyakaza kwentliziyo.Ngoko ke, ekupheleni kwexesha lokucinga, izilwanyana zabulawa ngabantu ngenxa ye-pentobarbital overdose (i-Somnopentil, i-Pitman-Moore, i-Washington Crossing, i-USA; ~ 65 mg / kg i-ip, intliziyo yayeka i-ip kunye ne-ibeat kanye, izilwanyana zaye zayeka i-ip). inkqubo yokucinga yaphindwa, yongeza idosi eyongezelelweyo yeMP ukuba akukho MP ibonakala kumoya womoya.
Imifanekiso efunyenweyo yayiyi-flat-field kunye ne-dark-field yalungiswa kwaye emva koko ihlanganiswe kwi-movie (iifreyimu ezingama-20 ngomzuzwana; 15-25 × isantya esiqhelekileyo kuxhomekeke kwisantya sokuphefumula) usebenzisa iskripthi esibhalwe kwi-MATLAB (R2020a, iMathworks).
Zonke izifundo zokuhanjiswa kwe-LV gene vector zaqhutywa kwiLebhu yeZilwanyana zoPhando lweZilwanyana kwiYunivesithi yaseAdelaide kwaye zijolise ekusebenziseni iziphumo zovavanyo lwe-SPring-8 ukuvavanya ukuba ukuhanjiswa kwe-LV-MP kubukho bemagnethi kunokuphucula ukuhanjiswa kofuzo kwi-vivo. umazibuthe.
I-LV gene vectors yenziwe ngokusebenzisa iindlela ezichazwe ngaphambili 25, 26. I-LacZ vector ivakalisa i-nuclear-localized beta-galactosidase gene eqhutywe ngumgqugquzeli we-MPSV (LV-LacZ), ovelisa umkhiqizo ohlaza okwesibhakabhaka kwiiseli ezitshintshiweyo, ezibonakala kwimida yezicubu zemiphunga kunye namacandelo ee-tissue. i-hemocytometer ukubala i-titer kwi-TU / ml. Abathwali be-cryopreserved kwi--80 ° C, bancibilika ngaphambi kokusetyenziswa, kwaye baboshwe kwi-CombiMag ngokuxuba kwi-1: umlinganiselo we-1 kunye nokufukamela kwiqhwa ubuncinane imizuzu engama-30 ngaphambi kokunikezelwa.
Iigundane eziqhelekileyo ze-Sprague Dawley (n = 3 / iqela, ~ 2-3 zaye zafakwa i-anesthetized intraperitoneally kunye nomxube we-0.4 mg / kg medetomidine (Domitor, Ilium, Australia) kunye ne-60 mg / kg i-ketamine (I-Ilium, e-Australia) inyanga ubudala) ip) inaliti kunye ne-non-ukuhlinzwa yokuhlinzwa i-cannulation yomlomo i-Galaway i-air cannulation yomlomo16. ifumana i-LV transduction, yalungiswa kusetyenziswa iprothokholi yethu echazwe ngaphambili yokuphazamiseka komatshini, apho umphezulu womoya woqhoqhoqho wakhuhlwa nge-axially ngebhasikithi yocingo (N-Circle, Nitinol Tipless Stone Extractor NTSE-022115) -UDH, Cook Medical, USA) 30 s28.Ikhabhinethi yetracheal yenziwa malunga nemizuzu yokhuseleko ye-LV-MPological emva kwemizuzu yokhuseleko lwe-LV-0MP.
Intsimi yamagnetic esetyenzisiweyo kolu vavanyo yacwangciswa ngendlela efanayo kwi-vivo ye-X-ray ye-imaging study, kunye nemagnethi efanayo ebanjwe ngaphezu kwe-trachea usebenzisa iziqeshana ze-distillation stent (Figure 4) .I-50 μl umthamo (2 × 25 μl aliquots) ye-LV-MP yanikezelwa kuqhoqhoqho (n = i-tiptte yezilwanyana ezichazwe ngaphambili) kusetyenziswa iqela lokulawula i-tippt njengezilwanyana ezichazwe ngaphambili. = Izilwanyana ezi-3) zifumene i-LV-MPs efanayo ngaphandle kokusetyenziswa kwemagnethi.Emva kokuba ukunyuswa kugqityiwe, i-cannula isuswe kwi-tube ye-ET kwaye isilwanyana sikhutshwe.Imagnethi ihlala kwindawo ye-10 imizuzu, emva koko isuswe.Iigundane zifumana i-dose engaphantsi kwe-meloxicam (1 ml / kg) (Ilium, i-Australia ye-injection ye-amativersp ye-anesthesia ye-amatizo i-hydrochloride (i-Antisedan, i-Zoetis, i-Australia) .Iigundane zigcinwe zifudumele kwaye zibekwe esweni de zifumaneke ngokupheleleyo kwi-anesthesia.
Isixhobo sokuhanjiswa kwe-LV-MP kwikhabhinethi yokhuseleko lwebhayoloji.I-hub ye-grey ye-Luer ekhanyayo ye-tube ye-ET ingabonwa iphuma emlonyeni kunye ne-gel incam ye-pipette eboniswe kumfanekiso ifakwe kwi-ET tube ukuya kubunzulu obufunayo kwi-trachea.
Iveki enye emva kwenkqubo ye-LV-MP ye-dosing, izilwanyana zabulawa ngabantu nge-100% ye-CO2 inhalation kunye ne-LacZ ibonakaliso yavavanywa kusetyenziswa unyango lwethu oluqhelekileyo lwe-X-gal.Izindandatho ezintathu ze-caudal ezininzi ze-cartilaginous zisuswe ukuze kuqinisekiswe ukuba nawuphi na umonakalo owenziwe ngomatshini okanye ukugcinwa kwamanzi kwi-endotracheal tube placement ayizange ifakwe kuhlalutyo. isitya esinerabha ye-silicone (i-Sylgard, i-Dow Inc) isebenzisa inaliti ye-Minutien (iZixhobo zeSayensi eziFine) ukujonga umphezulu we-luminal. Ukuhanjiswa kunye nepateni yeeseli ezitshintshiweyo ziqinisekiswe ngokufota okungaphambili kusetyenziswa i-Nikon microscope (SMZ1500) ngekhamera ye-DigiLite kunye nesofthiwe ye-TCapture (Tucsen Photonics, e-China). ububanzi boqhoqhoqho), kunye nobude bonke bomqhoqho owenziwe inyathelo ngenyathelo, ukuqinisekisa ukuhlangana okwaneleyo phakathi komfanekiso ngamnye ukuvumela umfanekiso "ukuthunga".Imifanekiso esuka kuqhoqhoqho ngamnye yadityaniswa ibe ngumfanekiso omnye odibeneyo kusetyenziswa uMhleli weMifanekiso eDityanisiweyo v2.0.3 (uPhando lweMicrosoft) kusetyenziswa i-algorithm yentshukumo ecwangcisiweyo. automated MATLAB script (R2020a, MathWorks) njengoko kuchaziwe ngaphambili, usebenzisa izicwangciso ze-0.35
I-trachea nganye yayifakwe kwiparafini kwaye amacandelo e-5 μm ayesikiwe.Amacandelo athintelwa ngokungathathi hlangothi obomvu okukhawulezayo kwi-5 min kunye nemifanekiso yafunyanwa kusetyenziswa i-microscope ye-Nikon Eclipse E400, ikhamera ye-DS-Fi3 kunye ne-software ye-NIS yokubamba i-software (inguqulo 5.20.00).
Lonke uhlalutyo lwamanani lwenziwa kwi-GraphPad Prism v9 (i-Software ye-GraphPad, Inc.) .Ukubaluleka kwezibalo kubekwe kwi-p ≤ 0.05.I-Normality yaqinisekiswa kusetyenziswa uvavanyo lwe-Shapiro-Wilk, kwaye ukungafani kwi-LacZ staining kwavavanywa ngokusebenzisa i-t-test engabonakaliyo.
AmaLungu ePalamente amathandathu achazwe kwiThebhile 1 ahlolwe kusetyenziswa i-PCXI, kwaye ukubonakala kuchazwe kwiThebhile 2. Ii-MP1 ze-polystyrene ezimbini (MP1 kunye ne-MP2; i-18 μm kunye ne-0.25 μm, ngokulandelanayo) zazingabonakali phantsi kwe-PCXI, kodwa ezinye iisampuli zazichonga (imizekelo iboniswe kwi-5% (i-5) ye-MP1 kunye ne-MP2 ye-5. I-0.25 μm kunye ne-0.9 μm, ngokulandelelanayo) zibonakala kancinci.Nangona ziqulathe amasuntswana amancinci avavanyiweyo, iMP5 (98% Fe3O4; 0.25 μm) yayiyeyona ivakaliswe kakhulu.Imveliso yeCombiMag MP6 inzima ukuyibona.Kuzo zonke iimeko, ukukwazi kwethu ukubona i-MP5 kwi-translath ye-translath iphuculwe kakhulu I-capillary.Xa imagnethi isuka kwi-capillary, amasuntswana anwetshiwe kwiintambo ezide, kodwa njengoko iimagnethi zasondela kwaye amandla ombane asanda, i-particle strings yanciphisa njengoko amasuntswana afudukela kwindawo ephezulu ye-capillary (jonga i-Supplementary Video S1: i-MP4), inyusa I-particle density ye-conpillary isuswe kwi-surface, i-capilla magnetti isuswe kwi-surface. iyancipha kwaye amalungu ePalamente aphinde alungelelanise kwiintambo ezide ezisuka kumphezulu we-capillary (jonga i-Supplementary Video S2: MP4) .Emva kokuba umazibuthe eyeka ukuhamba, amasuntswana aqhubeka ehamba ixesha elifutshane emva kokufikelela kwindawo elinganayo.Njengoko i-MP ibheka kwaye isuka kumphezulu we-capillary, i-particle ye-drag ye-drag idonsa i-particle.
Ukubonakala kweMP phantsi kwe-PCXI kuyahluka kakhulu phakathi kweisampulu.(a) MP3, (b) MP4, (c) MP5 kunye (d) MP6.Yonke imifanekiso eboniswe apha ithathwe ngemagnethi ebekwe malunga ne-10 mm ngaphezulu kwe-capillary.Izangqa ezinkulu ezibonakalayo ngamaqamza omoya avaleleke kwi-capillaries, ebonisa ngokucacileyo i-black and white imaging edge iqulethe i-edge yesigaba esibomvu se-imaging edge. Ukwandiswa.Qaphela ukuba iidayamitha zeschematics zemagnethi kuwo onke amanani azinakulinganiswa kwaye zimalunga namaxesha angama-100 amakhulu kunoko kubonisiwe.
Njengoko umazibuthe uguqulelwa ngasekhohlo nasekunene ecaleni komphezulu we-capillary, i-engile yomtya weMP iyatshintsha ukuze ilungelelanise umazibuthe (jonga umfanekiso 6), ngaloo ndlela ichaza imigca yomhlaba wemagnethi.Kwi-MP3-5, emva kokuba i-chord ifikelela kwi-engile yomgubasi, amasuntswana atsalwa kumphezulu we-capillary. Ividiyo eyoNgezelelweyo ye-S3: MP5) .Oku kubonakala ngakumbi xa kusenziwa umfanekiso kufutshane nesiphelo se-capillary, nto leyo ebangela ukuba amalungu ePalamente adibanise kwaye agxininise kujongano lolwelo-moya.Iinxalenye zeMP6, ekwakunzima ukuziqonda kuneMP3-5, azizange zitsalwe njengoko umazibuthe wayehamba ecaleni kwecapillary, kodwa iMP, ishiya icandelo levidiyo eyongezwayo kwinxalenye yeVidiyo. S4: MP6) . Kwezinye iimeko, xa i-magnetic field esetyenzisiweyo yancitshiswa ngokuhambisa umazibuthe umgama omkhulu ukusuka kwindawo yomfanekiso, nawaphi na amaLungu ePalamente aseleyo ehla ngokuthe chu ukuya kumphezulu wetyhubhu ngokuxhuzula ngelixa ehlala kumtya (jonga iVidiyo eyoNgezelelweyo S5: MP3).
I-engile yomtya weMP iyatshintsha njengoko umazibuthe eguqulelwa ekunene phezu kwe-capillary.(a) MP3, (b) MP4, (c) MP5 kunye (d) ne-MP6. Ibhokisi ebomvu iqulathe ukwandiswa kokwahlula-hlula.Qaphela ukuba iividiyo ezongezelelweyo zinolwazi njengoko zityhila ubume bamasuntswana abalulekileyo kunye nolwazi oluguqukayo olungenakubonwa kule mifanekiso imileyo.
Iimvavanyo zethu zibonise ukuba ukuhambisa umazibuthe ngokuthe chu emva naphambili ecaleni koqhoqho kuququzelela ukubonwa kweMP kumxholo wentshukumo entsonkothileyo kwi-vivo. Uvavanyo lwe-vivo aluzange lwenziwe njengoko amaso e-polystyrene (MP1 kunye neMP2) ayengabonakali kwi-capillary.Nganye kwi-MPs eseleyo emine yavavanywa kwi-vivo kunye nemagnethi ende ye-axis eqwalaselweyo malunga ne-angle ye-30 ye-axis eqwalaselwe ngaphezulu kwe-angle ye-30 Amanani 2b kunye ne-3a), njengoko oku kubangele amatyathanga amade eMP kwaye kwasebenza ngakumbi kunokuba uqwalaselo lwemagnethi lunqanyulwe.MP3, MP4 kunye neMP6 azizange zibonwe kuqhoqhoqho lwaso nasiphi na isilwanyana esiphilayo.Xa iimpuku zomoya zifanekiselwa emva kokuba izilwanyana zibulewe ngobuntu, amasuntswana ahlala engabonakali naxa umthamo owongezelelweyo wongezwa kusetyenziswa i-iron oxide kunye ne-MP6 ebonakalayo yongezwa kuphela nge-iron oxide kunye ne-MP. ngoko ke yasetyenziswa ukuvavanya nokubonisa indlela yokuziphatha yeMP.
Ukubeka umazibuthe phezu koqhoqhoqho ngexesha lonikezelo lweMP kubangele abaninzi, kodwa ingengabo bonke, amaLungu ePalamente agxininiswe kwintsimi yokujonga. Ukukhangela kude kuqhoqhoqho emva konikezelo lweMP, amanye amaLungu ePalamente afunyenwe kufutshane necarina, ecebisa ukuba amandla omhlaba kazibuthe ayenganelanga ukuqokelela nokugcina onke amaLungu ePalamente, njengoko ayehanjiswa kummandla wamandla ombane wemagnethi ngexesha lenkqubo yolwelo.
Imifanekiso esuka ku (a) phambi kunye (b) emva kokunikezelwa kweMP5 kuqhoqhoqho lwempuku esandul’ ukubulawa kunye nomazibuthe ebekwe ngokuthe ngqo ngentla kwendawo yokucinga. Indawo enomfanekiso iphakathi kwamakhonkco amabini e-cartilage. Phambi kokuhanjiswa kweMP, kukho ulwelo oluthile kwindlela yomoya. Ibhokisi ebomvu iqulethe ukwandiswa kokwahluka-hlukileyo.
Ukuguqulela umazibuthe ecaleni koqhoqhoqho kwi-vivo kubangele ukuba ikhonkco leMP litshintshe i-angle ngaphakathi kwendawo yomoya ngendlela efana naleyo ibonwa kwi-capillaries (jonga umfanekiso we-8 kunye neVidiyo eyoNgezelelweyo ye-S7: MP5) .Nangona kunjalo, kuphononongo lwethu, amaLungu ePalamente awakwazanga ukutsalwa kumphezulu wendlela yomoya ephilayo njengoko ebenako kunye neecapillaries. Kwakhona kwafumanisa ukuba umtya wamasuntswana ubonakala utshintsha ubunzulu bomgangatho wolwelo lwamanzi xa umazibuthe ushukunyiswa ngobude ecaleni koqhoqhoqho, kwaye uyanda xa umazibuthe ushukunyiswa ngqo phezulu kwaye umtya wamasuntswana ujikeleziswa kwindawo ethe nkqo (jonga iVidiyo eyoNgezelelweyo S7). : MP5 ngo-0:09, ekunene ezantsi).Ipatheni yeempawu zentshukumo yatshintsha xa umazibuthe waguqulelwa kumphezulu woqhoqhoqho ecaleni (oko kukuthi, ukuya ngasekhohlo okanye ngasekunene kwesilwanyana kunokuba kubude boqhoqhoqho).Amasuntswana ayesabonakala ngokucacileyo njengoko eshukuma, kodwa xa umazibuthe wasuswa kuqhoqhoqho, iincam zesuntswana leVidiyo seeSupplement:5MP 0:08).Oku kuhambelana nokuziphatha kweMP esiyibonileyo phantsi kwemagnethi esetyenziswayo kwicapillary yeglasi.
Umzekelo wemifanekiso ebonisa iMP5 kuqhoqhoqho lwempuku ephilayo ene-anesthetized.(a) Umazibuthe usetyenziselwa ukufumana imifanekiso engasentla nasekhohlo kuqhoqhoqho, emva koko (b) emva kokuba umazibuthe usiwe ekunene.Ibhokisi ebomvu iqulathe ukwandiswa kokwahluka kokwandiswa.Le mifanekiso isuka kwividiyo eboniswe kwiVidiyo eyoNgezelelweyo S7:MP5.
Xa izibonda ezimbini zicwangciswe kwicala elingasentla-emzantsi ngasentla nangaphantsi kwe-trachea (oko kukuthi ukutsala; Umzobo 3b), i-MP chords yabonakala ixesha elide kwaye ibekwe ecaleni kwendlela yoqhoqhoqho kunokuba i-dorsal tracheal surface (jonga i-Supplementary Video S9: MP5) .Nangona kunjalo, ukugxininiswa okuphezulu kwamasuntswana, kungabonakali indawo ye-trachea ye-trachea. unikezelo xa isixhobo ezimbini-umazibuthe kusetyenziswa, nto leyo eyenzeka ngokuqhelekileyo xa isixhobo umazibuthe enye isetyenzisiwe.Kwathi xa umazibuthe enye iqwalaselwe ukugxotha izibonda umva (Fig. 3c), inani lamasuntswana abonakalayo kwinkalo yembono ayizange ibonakale ukwanda emva konikezelo.Ukuseta zombini ulungelelwaniso lwemagnethi-mbini lucelomngeni ngenxa yokutsalwa kwemagnethi ephezulu okanye ukuseta amandla emagnethi. Imagnethi enye ihambelana nomoya kodwa idlula kwindlela yomoya kwii-degrees ze-90 ukwenzela ukuba imigca yentsimi iwele udonga lwe-tracheal orthogonally (Fig. 3d), i-orientation eyenzelwe ukugqiba ukuba i-particle aggregation eludongeni lwecala inokubonwa. uqwalaselo lokuqhelaniswa (Umfanekiso 3a) wakhethwa kwizifundo ze-vivo gene carrier.
Xa isilwanyana sasifanekiselwa ngokuphindaphindiweyo emva kokubulawa komntu, ukungabikho kokunyakaza kwezicubu eziphazamisayo kuthetha ukuba imigca emifutshane kunye nefutshane yayinokuqondwa kwibala elicacileyo le-interchondral, "i-wobbly" ngokuhambelana nentshukumo yokuguqulela umazibuthe. Nangona kunjalo, awukabuboni ngokucacileyo ubukho kunye nokushukuma kwamasuntswana eMP6.
I-LV-LacZ i-titer yayiyi-1.8 × 108 TU / ml, kwaye emva kwe-1: 1 ixutywe ne-CombiMag MP (MP6), izilwanyana zafumana i-50 μl idosi ye-tracheal ye-9 × 107 TU / ml LV isithuthi (okt 4.5 × 106 TU / rat). ) .Kwezi zifundo, endaweni yokuguqulela umazibuthe ngexesha lomsebenzi, silungise umazibuthe kwindawo enye ukugqiba ukuba i-LV transduction (a) inokuphuculwa xa kuthelekiswa nokuhanjiswa kwe-vector ngokungabikho kwendawo yamagnetic, kwaye (b) inokujoliswa kwiiseli ze-Airway zigqithiselwa kwimimandla ekujoliswe kuyo yamagnetic yomoya ophezulu.
Ubukho bamagnethi kunye nokusetyenziswa kwe-CombiMag edibeneyo kunye ne-LV vectors ayizange ibonakale ibe nemiphumo emibi kwimpilo yezilwanyana, njengoko yenza i-protocol yethu yokuhanjiswa kwe-vector ye-LV.Imifanekiso yangaphambili yommandla we-tracheal ephantsi kokuphazamiseka komatshini (i-Supplementary Fig. 1) ibonise ukuba kukho amanqanaba aphezulu kakhulu okudluliselwa kwiqela lezilwanyana xa iphathwa nge-magneto9 encinci ye-LV-MP. isixa se-LacZ esiluhlaza sasikhona kwiqela lolawulo (umzobo 9b) .Ubungakanani beendawo eziqhelekileyo ze-X-Gal ezingcoliswe zibonise ukuba ulawulo lwe-LV-MP phambi kwendawo yamagnetic luvelise ukuphuculwa kwe-6-fold (Fig. 9c).
Umzekelo wemifanekiso ehlanganisiweyo ebonisa ukuhanjiswa koqhoqhoqho nge-LV-MP (a) kubukho bommandla wemagnethi kunye (b) nokungabikho kwemagnethi.(c) Uphuculo olubalulekileyo ngokwezibalo kwindawo eqhelekileyo yeLacZ yotshintsho ngaphakathi kuqhoqhoqho xa usebenzisa umazibuthe (*p = 0.029, t-test, n = 3 ngeqela ngalinye, ithetha ± SEM).
Amacandelo abomvu abomvu angathathi hlangothi (umzekelo oboniswe kwi-Supplementary Fig. 2) ubonise iiseli ze-LacZ-stained ekhoyo kwipatheni efanayo kunye nendawo njengoko kuchazwe ngaphambili.
Umceli mngeni ophambili wonyango lwejene yomoya uhlala uchanekileyo kwindawo echanekileyo yamasuntswana othwala ukuya kwimimandla enomdla kunye nokufikelela kumanqanaba aphezulu okuhanjiswa kakuhle kwimiphunga eshukumayo phambi kokuhamba komoya kunye ne-mucus esebenzayo. ukuphucula ukuhanjiswa kunenzuzo xa kuthelekiswa nezinye iindlela zokuhanjiswa kwemfuza ezifana ne-electroporation, njengoko inokudibanisa ukulula, ukusebenza kakuhle kweendleko, ukuhanjiswa kwendawo, ukonyuka kobuchule, kunye namaxesha amafutshane okufukamela, kwaye mhlawumbi i-dose encinci ye-carrier10. Nangona kunjalo, i-vivo deposition kunye nokuziphatha kwamasuntswana magnetic kwi-airways phantsi kwempembelelo ye-magnetic forces yangaphandle ayizange ichazwe ngendlela ye-vivo ye-feasis eneneni ayizange ichazwe. amanqanaba kwimizila yomoya ephilayo.
Uvavanyo lwethu lwe-in vitro synchrotron PCXI lubonise ukuba onke amasuntswana esiwavavanyeyo, ngaphandle kwe-polystyrene MP, abonakala kwi-imaging setup esiyisebenzisayo.Phambi kobume bemagnethi, iiMPs zenza imitya ubude bayo bunxulumene nohlobo lwamasuntswana kunye namandla emagnethi (oko kukuthi, ukusondela kunye nokushukuma kwemagnethi) ukuphembelela indawo yayo yasekhaya yamagnetic.Ezi ndawo zahlukeneyo zibangela ukuba amanye amasuntswana afanayo adityaniswe kwaye aqhagamshele, kunye neentshukumo ezinjengomtya weqela ngenxa yemikhosi yasekhaya evela kumandla asekuhlaleni anomtsalane kunye nenyanyekayo yamanye amasuntswana.
I-Schematic ebonisa (a,b) izitimela ze-particle ezenziwe ngaphakathi kwee-capillaries ezizaliswe ngamanzi kunye (c, d) nomoya ogcwele umoya we-trachea.Qaphela ukuba i-capillaries kunye ne-trachea ayitsalwanga kwisikali.I-Panel (a) iqulethe inkcazo ye-MP, equkethe i-Fe3O4 i-particle ehlelwe kwiintambo.
Xa umazibuthe washukunyiswa ngaphezu kwe-capillary, i-engile yomtya wamasuntswana wafikelela kwinqanaba elibalulekileyo le-MP3-5 equlathe i-Fe3O4, emva koko umtya wamasuntswana akaphindanga wahlala kwindawo yokuqala, kodwa washukuma kumphezulu ukuya kwindawo entsha.umabuthe.Le mpembelelo inokwenzeka ngenxa yokuba umphezulu we-capillary weglasi ugudile ngokwaneleyo ukuvumela le ntshukumo ukuba yenzeke. Amasuntswana ayemancinci, aneengubo ezahlukeneyo zokugquma okanye iintlawuliso zomphezulu, okanye ulwelo lomthwali womnini luchaphazele amandla abo okuhamba.Umahluko womfanekiso wamasuntswana e-CombiMag nawo ubuthathaka, nto leyo ebonisa ukuba ulwelo kunye namasuntswana anokuba noxinaniso olufanayo kwaye ngenxa yoko azihambisani ngokulula. Ayimangalisi into yokuba amandla emagnethi kunye nomgama phakathi kwemagnethi kunye nendawo ekujoliswe kuyo Kubaluleke kakhulu. Zithatyathwe kunye, ezi ziphumo zikwacebisa ukuba, ngelixa oomazibuthe banokubamba amaLungu ePalamente amaninzi ahamba kwindawo ekujoliswe kuyo, akunakwenzeka ukuba imagnethi inokuthenjwa ukuhambisa amasuntswana e-CombiMag kumphezulu wetrachea. umthi.
Xa amasuntswana ahanjiswa emzimbeni, kunzima ukuchonga kumxholo wezicubu zomzimba ezihambayo ezintsonkothileyo, kodwa ukukwazi ukuzibhaqa kwaphuculwa ngokuguqulela umazibuthe ngokuthe tye ngaphezu koqhoqhoqho ukuze "ujike" imitya yeMP.Nangona umfanekiso ophilayo unokwenzeka, kulula ukuqonda ukunyakaza kwamasuntswana xa isilwanyana sithe saye sabulawa ngobuntu kwindawo ye-imaging, nangona i-magnig ye-MP ibulewe ngokubanzi, le ndawo yoxinaniso yeMP yayibulewe ngokubanzi. ezinye iinqununu zazivame ukufunyanwa ngokuqhubekayo kunye ne-trachea.Ngokwahlukileyo kwizifundo ze-in vitro, iinqununu azikwazi ukutsalwa ecaleni koqhoqhoqho ngokuguqulela imagnethi.Oku kufunyaniswayo kuhambelana nendlela i-mucus egubungela ngayo umphezulu we-trachea ngokuqhelekileyo iqhubekisela phambili amaqhekeza afakwe ngaphakathi, ukuwafaka kwi-mucus kwaye emva koko ihlanjululwe yi-mucociliary clearance mechanism.
Sacinga ukuba ukusetyenziswa kweemagnethi ukukhanga ngaphezulu nangaphantsi kwetrachea (Fig. 3b) kunokubangela ukuba kubekho indawo elinganayo yamagnetic, kunokuba imagnethi igxininiswe kakhulu kwindawo enye, enokuthi ikhokelele ekuhanjisweni okufanayo kwamasuntswana.Nangona kunjalo, uphononongo lwethu lwangaphambili aluzange lufumane ubungqina obucacileyo bokuxhasa le hypothesis. amasuntswana ngaphezulu ukubekwa kwindawo umfanekiso.Ezi zinto zifunyanisiweyo ezimbini zibonisa ukuba ukuseta ezimbini-umazibuthe ayiphuculi kakhulu ulawulo lwendawo MP ekujoliswe, kwaye isiphumo amandla anamandla kazibuthe kunzima ukuqwalasela, ukwenza le ndlela ingaphantsi practical.Similarly, ukuqhelanisa umazibuthe ngasentla kunye ngokusebenzisa trachea (Fig. 3d) kwakhona akazange ukwandisa inani amasuntswana eenguqu ezigciniweyo ezinokuthi zigcinwe kwindawo enempumelelo ngenxa yokuba isiphumo somfanekiso singagcinwa. kumandla ombane asezantsi ngaphakathi kwendawo yokubeka.Ngoko ke, uqwalaselo lwemagnethi ye-engile ye-30-degree eyodwa (Umfanekiso 3a) ithathwa njengeyona ndlela ilula nesebenzayo yovavanyo lwe-vivo.
Uphononongo lwe-LV-MP lubonise ukuba xa iivektha ze-LV zidityanisiwe kunye ne-CombiMag kwaye zihanjiswa emva kokuphazamiseka ngokomzimba kubukho bomhlaba wamagnetic, amanqanaba okutshintshwa anyuswe kakhulu kuqhoqhoqho xa kuthelekiswa nolawulo.Ngokusekelwe kwizifundo ze-synchrotron imaging kunye neziphumo ze-LacZ, i-magnetic field yayibonakala ikwazi ukugcina i-LV ngaphakathi kwe-trachea kunye nokunciphisa inani leepuleti eziphuculweyo ngokukhawuleza. kunokukhokelela ekusebenzeni okuphezulu ngelixa kuncitshiswa i-titers ezinikezelweyo, i-off-target transduction, i-inflammation and immune side effects, kunye neendleko ze-gene carrier carrier.Kubaluleke kakhulu, ngokutsho komenzi, i-CombiMag ingasetyenziselwa kunye nezinye iindlela zokudlulisa i-gene, kubandakanywa nezinye i-viral vectors (ezifana ne-AAV) kunye ne-nucleic acids.
Ixesha lokuposa: Jul-16-2022
