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Sau npe koj cov ntaub ntawv tshwj xeeb thiab cov tshuaj tshwj xeeb uas koj xav tau, thiab peb yuav phim cov ntaub ntawv koj muab nrog cov ntawv hauv peb lub ntaub ntawv loj thiab xa daim PDF rau koj ntawm email raws sijhawm.
Tswj kev txav mus los ntawm cov hlau nplaum oxide nanoparticles rau kev xa khoom ntawm cytostatics
Sau Toropova Y, Korolev D, Istomina M, Shulmeyster G, Petukhov A, Mishanin V, Gorshkov A, Podyacheva E, Gareev K, Bagrov A, Demidov O
Yana Toropova,1 Dmitry Korolev,1 Maria Istomina,1,2 Galina Shulmeyster,1 Alexey Petukhov,1,3 Vladimir Mishanin,1 Andrey Gorshkov,4 Ekaterina Podyacheva,1 Kamil Gareev,2 Alexei Bagrov,5 Oleg Demidov6,71Almazov National Medical Research Center ntawm Ministry of Health ntawm Russian Federation, St. Petersburg, 197341, Russian Federation; 2 St. Petersburg Electrotechnical University "LETI", St. Petersburg, 197376, Russian Federation; 3 Chaw rau Kev Kho Mob Tus Kheej, Almazov State Medical Research Center, Ministry of Health ntawm Russian Federation, St. Petersburg, 197341, Russian Federation; 4FSBI "Influenza Research Institute named after AA Smorodintsev" Ministry of Health ntawm Russian Federation, St. Petersburg, Russian Federation; 5 Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, Russian Federation; 6 RAS Lub Tsev Kawm Ntawv ntawm Cytology, St. Petersburg, 194064, Lavxias teb sab Federation; 7INSERM U1231, Kws Qhia Ntawv ntawm Tshuaj thiab Tshuaj, Bourgogne-Franche Comté University of Dijon, Fabkis Kev Sib Txuas Lus: Yana ToropovaAlmazov National Medical Research Centre, Ministry of Health ntawm Lavxias teb sab Federation, Saint-Petersburg, 197341, Lavxias teb sab Federation Tel +7 981 95264800 4997069 Email [email protected] Keeb Kwm: Ib txoj hauv kev zoo rau qhov teeb meem ntawm cytostatic toxicity yog kev siv cov magnetic nanoparticles (MNP) rau kev xa tshuaj. Lub Hom Phiaj: Siv cov kev xam los txiav txim siab qhov zoo tshaj plaws ntawm lub zog magnetic uas tswj MNPs hauv vivo, thiab los ntsuas qhov ua tau zoo ntawm magnetron xa MNPs rau cov qog nqaij hlav nas hauv vitro thiab hauv vivo. (MNPs-ICG) siv. Kev tshawb fawb txog qhov muaj zog ntawm lub zog luminescence hauv vivo tau ua tiav hauv cov nas qog nqaij hlav, nrog thiab tsis muaj lub zog magnetic ntawm qhov chaw uas xav paub. Cov kev tshawb fawb no tau ua tiav rau ntawm lub tshuab hydrodynamic uas tau tsim los ntawm Lub Tsev Kawm Ntawv ntawm Kev Tshawb Fawb Txog Kev Kho Mob ntawm Almazov State Medical Research Center ntawm Lavxias Ministry of Health. Qhov Tshwm Sim: Kev siv cov hlau nplaum neodymium tau txhawb nqa kev xaiv cov khoom sib sau ua ke ntawm MNP. Ib feeb tom qab kev siv MNPs-ICG rau cov nas uas muaj mob qog noj ntshav, MNPs-ICG feem ntau sib sau ua ke hauv lub siab. Thaum tsis muaj thiab muaj lub zog sib nqus, qhov no qhia txog nws txoj kev zom zaub mov. Txawm hais tias muaj kev nce ntxiv ntawm qhov fluorescence hauv qhov mob qog noj ntshav tau pom nyob rau hauv qhov muaj lub zog sib nqus, qhov muaj zog fluorescence hauv lub siab ntawm tus tsiaj tsis hloov pauv raws sijhawm. Xaus Lus: Hom MNP no, ua ke nrog lub zog sib nqus uas tau xam, tuaj yeem yog lub hauv paus rau kev tsim cov tshuaj cytostatic tswj hwm los ntawm magnetically rau cov nqaij mob qog noj ntshav. Cov Lus Tseem Ceeb: kev tshuaj xyuas fluorescence, indocyanine, hlau oxide nanoparticles, magnetron xa cov cytostatics, kev tsom mus rau qhov mob qog noj ntshav.
Cov kab mob qog nqaij hlav yog ib qho ntawm cov ua rau tuag thoob ntiaj teb. Tib lub sijhawm, qhov kev hloov pauv ntawm kev mob nkeeg thiab kev tuag ntawm cov kab mob qog nqaij hlav tseem muaj. 1 Cov tshuaj chemotherapy siv niaj hnub no tseem yog ib qho ntawm cov kev kho mob tseem ceeb rau ntau hom qog nqaij hlav. Tib lub sijhawm, kev tsim cov txheej txheem los txo qhov kev lom ntawm cov tshuaj cytostatics tseem ceeb. Ib txoj hauv kev cog lus los daws nws qhov teeb meem lom yog siv cov nano-scale carriers los tsom cov txheej txheem xa tshuaj, uas tuaj yeem muab cov tshuaj hauv zos hauv cov ntaub so ntswg qog nqaij hlav yam tsis nce lawv cov khoom sib sau ua ke hauv cov kabmob thiab cov ntaub so ntswg noj qab haus huv. concentration. 2 Txoj kev no ua rau nws muaj peev xwm txhim kho qhov ua tau zoo thiab kev tsom mus rau cov tshuaj chemotherapy ntawm cov ntaub so ntswg qog nqaij hlav, thaum txo lawv cov tshuaj lom hauv lub cev.
Ntawm ntau yam nanoparticles uas tau txiav txim siab rau kev xa cov neeg sawv cev cytostatic, cov nanoparticles sib nqus (MNPs) yog qhov txaus siab tshwj xeeb vim lawv cov tshuaj lom neeg, biological, thiab magnetic tshwj xeeb, uas ua kom lawv muaj peev xwm ua tau ntau yam. Yog li ntawd, cov nanoparticles sib nqus tuaj yeem siv ua lub tshuab cua sov los kho cov qog nrog hyperthermia (magnetic hyperthermia). Lawv kuj tuaj yeem siv ua cov neeg sawv cev kuaj mob (magnetic resonance diagnosis). 3-5 Siv cov yam ntxwv no, ua ke nrog qhov ua tau ntawm MNP sib sau ua ke hauv ib cheeb tsam tshwj xeeb, los ntawm kev siv lub zog sib nqus sab nraud, kev xa cov tshuaj npaj tshwj xeeb qhib kev tsim lub tshuab magnetron ntau yam los tsom cytostatics rau qhov chaw mob. Lub kaw lus zoo li no yuav suav nrog MNP thiab cov teb sib nqus los tswj lawv txoj kev txav mus los hauv lub cev. Hauv qhov no, ob qho tib si sab nraud magnetic teb thiab cov hlau nplaum cog rau hauv thaj chaw lub cev uas muaj cov qog tuaj yeem siv ua qhov chaw ntawm lub zog sib nqus. 6 Txoj kev thawj zaug muaj qhov tsis zoo loj heev, suav nrog qhov xav tau siv cov khoom siv tshwj xeeb rau kev tsom cov tshuaj sib nqus thiab qhov xav tau kev cob qhia cov neeg ua haujlwm kom ua phais. Ntxiv mus, txoj kev no raug txwv los ntawm tus nqi siab thiab tsuas yog tsim rau cov qog "surficial" ze rau ntawm qhov chaw ntawm lub cev. Lwm txoj hauv kev ntawm kev siv cov hlau nplaum cog ua rau qhov kev siv cov thev naus laus zis no nthuav dav, ua rau nws siv tau rau cov qog nyob rau ntau qhov chaw ntawm lub cev. Ob qho tib si cov hlau nplaum ib leeg thiab cov hlau nplaum sib xyaw ua ke rau hauv lub stent intraluminal tuaj yeem siv ua cov khoom cog rau kev puas tsuaj ntawm cov qog hauv cov kabmob khoob kom ntseeg tau tias lawv muaj zog. Txawm li cas los xij, raws li peb tus kheej txoj kev tshawb fawb tsis tau luam tawm, cov no tsis muaj hlau nplaum txaus kom ntseeg tau tias khaws cia MNP ntawm cov ntshav.
Qhov ua tau zoo ntawm kev xa tshuaj magnetron nyob ntawm ntau yam: cov yam ntxwv ntawm tus nqa hlau nplaum nws tus kheej, thiab cov yam ntxwv ntawm qhov chaw magnetic field (suav nrog cov qauv geometric ntawm cov hlau nplaum tas mus li thiab lub zog ntawm lub magnetic field uas lawv tsim). Kev txhim kho cov thev naus laus zis xa tshuaj magnetically guided cell inhibitor yuav tsum suav nrog kev txhim kho cov khoom nqa tshuaj magnetic nanoscale tsim nyog, kev ntsuas lawv txoj kev nyab xeeb, thiab kev tsim cov txheej txheem pom kev uas tso cai rau kev taug qab lawv cov kev txav mus los hauv lub cev.
Hauv kev tshawb fawb no, peb tau suav lej txog cov yam ntxwv ntawm lub zog sib nqus zoo tshaj plaws los tswj cov tshuaj uas nqa tau hauv lub cev los ntawm lub zog sib nqus nano. Qhov ua tau ntawm kev khaws MNP los ntawm phab ntsa ntawm cov hlab ntsha hauv qab kev cuam tshuam ntawm lub zog sib nqus uas siv nrog cov yam ntxwv suav no kuj tau kawm hauv cov hlab ntsha nas uas cais tawm. Tsis tas li ntawd, peb tau tsim cov conjugates ntawm MNPs thiab cov neeg sawv cev fluorescent thiab tsim ib txoj cai rau lawv pom hauv vivo. Hauv qab cov xwm txheej hauv vivo, hauv cov nas qauv qog, kev ua haujlwm ntawm MNPs hauv cov nqaij qog thaum muab tshuaj rau hauv lub cev hauv qab kev cuam tshuam ntawm lub zog sib nqus tau kawm.
Hauv kev tshawb fawb hauv vitro, peb siv cov MNP siv ua piv txwv, thiab hauv kev tshawb fawb hauv vivo, peb siv MNP uas tau coated nrog lactic acid polyester (polylactic acid, PLA) uas muaj cov neeg sawv cev fluorescent (indolecyanine; ICG). MNP-ICG suav nrog hauv Hauv rooj plaub, siv (MNP-PLA-EDA-ICG).
Cov khoom siv thiab cov khoom siv lub cev thiab tshuaj ntawm MNP tau piav qhia meej hauv lwm qhov chaw.
Yuav kom ua tau cov MNPs-ICG, PLA-ICG conjugates tau tsim ua ntej. Ib qho hmoov sib xyaw ua ke ntawm PLA-D thiab PLA-L nrog qhov hnyav molecular ntawm 60 kDa tau siv.
Vim tias PLA thiab ICG yog ob qho tib si acids, txhawm rau kom ua tau PLA-ICG conjugates, ua ntej yuav tsum ua kom muaj amino-terminated spacer ntawm PLA, uas pab ICG chemisorb rau lub spacer. Lub spacer tau ua kom siv ethylene diamine (EDA), carbodiimide txoj kev thiab carbodiimide uas yaj tau hauv dej, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC). Lub PLA-EDA spacer tau ua raws li nram no. Ntxiv 20-npaug molar ntau dhau ntawm EDA thiab 20-npaug molar ntau dhau ntawm EDAC rau 2 mL ntawm 0.1 g/mL PLA chloroform kua. Kev ua kom tau ua tiav hauv 15 mL polypropylene raj kuaj ntawm lub shaker ntawm qhov ceev ntawm 300 min-1 rau 2 teev. Cov txheej txheem ua kom tau pom hauv Daim Duab 1. Rov ua qhov kev ua kom tau nrog 200-npaug ntau dhau ntawm cov tshuaj reagents kom zoo dua cov txheej txheem ua kom tau zoo.
Thaum kawg ntawm kev tsim cov tshuaj, cov tshuaj tau centrifuged ntawm qhov ceev ntawm 3000 min-1 rau 5 feeb kom tshem tawm cov polyethylene derivatives precipitated ntau dhau. Tom qab ntawd, 2 mL ntawm 0.5 mg / mL ICG kua hauv dimethyl sulfoxide (DMSO) tau ntxiv rau 2 mL kua. Lub agitator raug kho ntawm qhov ceev stirring ntawm 300 min-1 rau 2 teev. Daim duab schematic ntawm cov conjugate tau txais tau qhia hauv Daim Duab 2.
Hauv 200 mg MNP, peb ntxiv 4 mL PLA-EDA-ICG conjugate. Siv lub LS-220 shaker (LOIP, Russia) los do cov tshuaj rau 30 feeb ntawm qhov zaus ntawm 300 min-1. Tom qab ntawd, nws tau ntxuav nrog isopropanol peb zaug thiab raug sib cais los ntawm hlau nplaum. Siv UZD-2 Ultrasonic Disperser (FSUE NII TVCH, Russia) los ntxiv IPA rau cov tshuaj rau 5-10 feeb nyob rau hauv kev ua ultrasonic tas mus li. Tom qab ntxuav IPA thib peb, cov dej nag tau ntxuav nrog dej distilled thiab rov ua dua hauv physiological saline ntawm qhov concentration ntawm 2 mg / mL.
Cov khoom siv ZetaSizer Ultra (Malvern Instruments, UK) tau siv los kawm txog qhov loj ntawm MNP uas tau txais hauv cov kua dej. Lub tshuab kuaj hluav taws xob xa hluav taws xob (TEM) nrog JEM-1400 STEM field emission cathode (JEOL, Nyiv Pooj) tau siv los kawm txog cov duab thiab qhov loj ntawm MNP.
Hauv kev tshawb fawb no, peb siv cov hlau nplaum tas mus li cylindrical (qib N35; nrog nickel tiv thaiv txheej) thiab cov qauv loj hauv qab no (qhov ntev ntawm txoj kab ntev × txoj kab uas hla lub tog raj kheej): 0.5 × 2 hli, 2 × 2 hli, 3 × 2 hli thiab 5 × 2 hli.
Kev tshawb fawb hauv vitro ntawm MNP kev thauj mus los hauv lub cev qauv tau ua tiav ntawm lub tshuab hydrodynamic uas tsim los ntawm Lub Tsev Kawm Ntawv ntawm Kev Tshawb Fawb Txog Kev Kho Mob ntawm Almazov State Medical Research Center ntawm Lavxias Ministry of Health. Qhov ntim ntawm cov kua dej uas ncig (dej distilled lossis Krebs-Henseleit kua) yog 225 mL. Cov hlau nplaum cylindrical magnetized axially siv ua cov hlau nplaum tas mus li. Muab lub hlau nplaum tso rau ntawm lub tuav 1.5 hli deb ntawm phab ntsa sab hauv ntawm lub raj iav nruab nrab, nrog nws qhov kawg tig mus rau qhov kev taw qhia ntawm lub raj (ntsug). Tus nqi ntws ntawm cov kua dej hauv lub voj kaw yog 60 L / h (sib raug rau qhov ceev ntawm linear ntawm 0.225 m / s). Krebs-Henseleit kua siv ua cov kua dej ncig vim nws yog ib qho piv ntawm plasma. Tus coefficient viscosity dynamic ntawm plasma yog 1.1–1.3 mPa∙s. 9 Tus nqi ntawm MNP adsorbed hauv lub teb magnetic yog txiav txim siab los ntawm spectrophotometry los ntawm qhov concentration ntawm hlau hauv cov kua dej ncig tom qab kev sim.
Ntxiv rau, kev tshawb fawb tau ua tiav ntawm lub rooj kho cov kua dej kom paub txog qhov permeability ntawm cov hlab ntsha. Cov khoom tseem ceeb ntawm kev txhawb nqa hydrodynamic tau qhia hauv Daim Duab 3. Cov khoom tseem ceeb ntawm hydrodynamic stent yog lub voj voog kaw uas ua rau pom qhov seem ntawm cov qauv vascular system thiab lub tank cia khoom. Kev txav mus los ntawm cov qauv kua dej raws li qhov nkhaus ntawm cov hlab ntsha module yog muab los ntawm lub twj tso kua mis peristaltic. Thaum lub sijhawm sim, tswj qhov vaporization thiab qhov kub thiab txias xav tau, thiab saib xyuas cov txheej txheem system (kub, siab, tus nqi ntws kua, thiab pH tus nqi).
Daim Duab 3 Daim duab thaiv ntawm qhov teeb tsa siv los kawm txog qhov permeability ntawm phab ntsa carotid artery. 1-lub tank cia khoom, 2-lub twj tso kua mis peristaltic, 3-lub tshuab rau kev qhia txog kev ncua uas muaj MNP rau hauv lub voj voog, 4-lub ntsuas dej ntws, 5-lub ntsuas siab hauv lub voj voog, 6-lub tshuab hloov pauv cua sov, 7-chav nrog lub thawv, 8-qhov chaw ntawm lub teb sib nqus, 9-lub npas nrog cov hydrocarbons.
Lub chamber uas muaj lub thawv muaj peb lub thawv: ib lub thawv loj sab nraud thiab ob lub thawv me me, uas cov caj npab ntawm lub voj voog nruab nrab hla dhau. Lub cannula raug ntxig rau hauv lub thawv me, lub thawv raug khi rau ntawm lub thawv me, thiab lub hau ntawm cannula raug khi nruj nrog ib txoj hlua nyias. Qhov chaw ntawm lub thawv loj thiab lub thawv me yog puv nrog dej distilled, thiab qhov kub tseem nyob ruaj khov vim yog kev sib txuas rau lub tshuab hloov pauv cua sov. Qhov chaw hauv lub thawv me yog puv nrog Krebs-Henseleit kua kom tswj tau qhov muaj sia nyob ntawm cov hlwb ntawm cov hlab ntsha. Lub tank kuj tseem puv nrog Krebs-Henseleit kua. Lub kaw lus muab roj (carbon) yog siv los ua pa cov kua hauv lub thawv me hauv lub tank cia thiab lub chamber uas muaj lub thawv (Daim Duab 4).
Daim Duab 4 Lub chamber uas lub thawv tau muab tso rau. 1-Cannula rau kev txo cov hlab ntsha, 2-Cham sab nraud, 3-Cham me me. Tus xub qhia qhov kev taw qhia ntawm cov kua qauv.
Txhawm rau txiav txim siab qhov ntsuas permeability ntawm phab ntsa ntawm cov hlab ntsha, cov hlab ntsha carotid nas tau siv.
Qhov kev qhia txog MNP suspension (0.5mL) rau hauv lub system muaj cov yam ntxwv hauv qab no: tag nrho cov ntim sab hauv ntawm lub tank thiab cov yeeb nkab txuas hauv lub voj voog yog 20mL, thiab qhov ntim sab hauv ntawm txhua lub chamber yog 120mL. Lub zog sab nraud magnetic teb yog lub hlau nplaum tas mus li nrog qhov loj me ntawm 2 × 3 hli. Nws tau teeb tsa saum toj ntawm ib qho ntawm cov chamber me me, 1 cm deb ntawm lub thawv, nrog ib kawg ntsib phab ntsa thawv. Qhov kub thiab txias khaws cia ntawm 37 ° C. Lub zog ntawm lub twj tso kua mis roller yog teeb tsa rau 50%, uas sib raug rau qhov ceev ntawm 17 cm / s. Ua ib qho kev tswj hwm, cov qauv tau coj mus rau hauv lub cell tsis muaj hlau nplaum tas mus li.
Ib teev tom qab siv cov tshuaj MNP uas muaj ntau npaum li cas, lawv tau coj ib qho kua los ntawm lub chamber. Cov khoom me me ntawd tau ntsuas los ntawm lub spectrophotometer siv Unico 2802S UV-Vis spectrophotometer (United Products & Instruments, USA). Thaum xav txog qhov absorption spectrum ntawm MNP suspension, kev ntsuas tau ua tiav ntawm 450 nm.
Raws li cov lus qhia ntawm Rus-LASA-FELASA, txhua tus tsiaj raug tsa thiab loj hlob hauv cov chaw tshwj xeeb uas tsis muaj kab mob. Kev tshawb fawb no ua raws li txhua txoj cai kev coj ncaj ncees rau kev sim tsiaj thiab kev tshawb fawb, thiab tau txais kev pom zoo los ntawm Almazov National Medical Research Center (IACUC). Cov tsiaj haus dej ad libitum thiab pub zaub mov tsis tu ncua.
Txoj kev tshawb fawb no tau ua rau 10 tus nas txiv neej uas muaj 12 lub lis piam uas tsis muaj zog tiv thaiv kab mob (NOD.Cg-Prkdcscid Il2rgtm1Wjl/Szj, Jackson Laboratory, USA) 10, uas tau muab tshuaj loog thiab hnyav 22 g ± 10%. Vim tias kev tiv thaiv kab mob ntawm cov nas uas tsis muaj zog tiv thaiv kab mob raug txo qis, cov nas uas tsis muaj zog ntawm cov kab no tso cai rau kev hloov pauv ntawm cov hlwb thiab cov nqaij ntawm tib neeg yam tsis muaj kev tsis lees txais kev hloov pauv. Cov nas uas yug los ntawm ntau lub tawb sib txawv tau raug xaiv los ua pawg sim, thiab lawv tau yug ua ke lossis raug tso rau hauv lwm pawg kom ntseeg tau tias muaj kev sib npaug ntawm cov kab mob me me.
Cov kab mob qog noj ntshav HeLa tib neeg siv los tsim kom muaj tus qauv xenograft. Cov hlwb tau cog qoob loo hauv DMEM uas muaj glutamine (PanEco, Russia), ntxiv nrog 10% fetal bovine serum (Hyclone, USA), 100 CFU/mL penicillin, thiab 100 μg/mL streptomycin. Cov kab mob hlwb tau muab los ntawm Gene Expression Regulation Laboratory ntawm Lub Tsev Kawm Ntawv Tshawb Fawb Txog Cell ntawm Lavxias Academy of Sciences. Ua ntej txhaj tshuaj, cov hlwb HeLa tau raug tshem tawm ntawm cov yas cog qoob loo nrog 1:1 trypsin:Versene kua (Biolot, Russia). Tom qab ntxuav, cov hlwb tau raug dai rau hauv nruab nrab kom txog thaum muaj 5 × 106 hlwb ib 200 μL, thiab diluted nrog basement membrane matrix (LDEV-FREE, MATRIGEL® CORNING®) (1:1, ntawm dej khov). Cov hlwb uas tau npaj tseg tau raug txhaj rau hauv daim tawv nqaij ntawm tus nas lub duav. Siv cov calipers hluav taws xob los saib xyuas kev loj hlob ntawm qog txhua 3 hnub.
Thaum lub qog loj txog 500 mm3, tau muab ib lub hlau nplaum ruaj khov tso rau hauv cov leeg nqaij ntawm cov tsiaj sim ze ntawm lub qog. Hauv pawg sim (MNPs-ICG + tumor-M), 0.1 mL ntawm MNP kua tau txhaj thiab raug rau lub zog sib nqus. Cov tsiaj tag nrho uas tsis tau kho tau siv ua cov kev tswj (keeb kwm yav dhau). Tsis tas li ntawd, cov tsiaj uas tau txhaj nrog 0.1 mL ntawm MNP tab sis tsis tau muab hlau nplaum tso rau hauv (MNPs-ICG + tumor-BM) tau siv.
Kev pom kev fluorescence ntawm cov qauv hauv vivo thiab hauv vitro tau ua tiav ntawm IVIS Lumina LT series III bioimager (PerkinElmer Inc., USA). Rau kev pom kev hauv vitro, ib qho ntim ntawm 1 mL ntawm cov khoom siv hluavtaws PLA-EDA-ICG thiab MNP-PLA-EDA-ICG conjugate tau ntxiv rau hauv cov qhov dej phaj. Xav txog cov yam ntxwv fluorescence ntawm ICG zas xim, lub lim dej zoo tshaj plaws siv los txiav txim siab qhov muaj zog ntawm cov qauv tau xaiv: qhov siab tshaj plaws excitation wavelength yog 745 nm, thiab qhov emission wavelength yog 815 nm. Lub Living Image 4.5.5 software (PerkinElmer Inc.) tau siv los ntsuas qhov muaj zog fluorescence ntawm cov qhov dej uas muaj conjugate.
Qhov muaj zog fluorescence thiab kev sib sau ua ke ntawm MNP-PLA-EDA-ICG conjugate tau ntsuas hauv cov nas qauv qog nqaij hlav hauv vivo, tsis muaj qhov muaj thiab siv lub zog sib nqus ntawm qhov chaw xav tau. Cov nas tau raug tshuaj loog nrog isoflurane, thiab tom qab ntawd 0.1 mL ntawm MNP-PLA-EDA-ICG conjugate tau txhaj los ntawm cov leeg ntawm tus Tsov tus tw. Cov nas uas tsis tau kho tau siv ua kev tswj tsis zoo kom tau txais keeb kwm yav dhau los fluorescent. Tom qab muab cov conjugate rau hauv cov hlab ntsha, muab tus tsiaj tso rau ntawm theem cua sov (37 ° C) hauv chav ntawm IVIS Lumina LT series III fluorescence imager (PerkinElmer Inc.) thaum tswj kev ua pa nrog 2% isoflurane anesthetization. Siv ICG lub lim dej ua ke (745–815 nm) rau kev ntes teeb liab 1 feeb thiab 15 feeb tom qab kev qhia txog MNP.
Txhawm rau ntsuam xyuas qhov sib sau ua ke ntawm conjugate hauv qog, thaj chaw peritoneal ntawm tus tsiaj tau npog nrog ntawv, uas ua rau nws muaj peev xwm tshem tawm qhov ci ntsa iab uas cuam tshuam nrog kev sib sau ua ke ntawm cov khoom me me hauv lub siab. Tom qab kawm txog kev faib tawm ntawm MNP-PLA-EDA-ICG, cov tsiaj tau raug tua los ntawm kev siv tshuaj isoflurane anesthesia ntau dhau rau kev sib cais tom qab ntawm thaj chaw qog thiab kev ntsuas ntau ntawm cov hluav taws xob fluorescence. Siv Living Image 4.5.5 software (PerkinElmer Inc.) los ua cov txheej txheem tshuaj xyuas teeb liab los ntawm thaj chaw xaiv. Peb qhov kev ntsuas tau coj mus rau txhua tus tsiaj (n = 9).
Hauv kev tshawb fawb no, peb tsis tau ntsuas qhov kev thauj khoom ntawm ICG rau ntawm MNPs-ICG. Tsis tas li ntawd xwb, peb tsis tau piv rau qhov kev ua haujlwm ntawm cov nanoparticles nyob rau hauv kev cuam tshuam ntawm cov hlau nplaum tas mus li ntawm ntau yam duab. Tsis tas li ntawd xwb, peb tsis tau soj ntsuam qhov cuam tshuam mus sij hawm ntev ntawm lub zog sib nqus ntawm kev khaws cia cov nanoparticles hauv cov ntaub so ntswg qog.
Cov nanoparticles tseem ceeb, nrog qhov nruab nrab loj ntawm 195.4 nm. Tsis tas li ntawd, cov kua nplaum muaj cov agglomerates nrog qhov nruab nrab loj ntawm 1176.0 nm (Daim Duab 5A). Tom qab ntawd, qhov feem tau lim los ntawm lub lim centrifugal. Lub zeta potential ntawm cov khoom me me yog -15.69 mV (Daim Duab 5B).
Daim Duab 5 Cov khoom siv lub cev ntawm cov khoom ncua: (A) kev faib tawm ntawm qhov loj me ntawm cov khoom me me; (B) kev faib tawm ntawm cov khoom me me ntawm zeta lub peev xwm; (C) TEM daim duab ntawm cov nanoparticles.
Qhov loj ntawm cov khoom me me yog 200 nm (Daim Duab 5C), muaj ib qho MNP nrog qhov loj ntawm 20 nm, thiab PLA-EDA-ICG conjugated organic plhaub nrog qhov ceev ntawm electron qis dua. Kev tsim cov agglomerates hauv cov kua dej tuaj yeem piav qhia los ntawm qhov modulus qis ntawm lub zog electromotive ntawm cov nanoparticles ib leeg.
Rau cov hlau nplaum tas mus li, thaum lub magnetization yog concentrated nyob rau hauv lub ntim V, qhov kev qhia tawm integral yog muab faib ua ob integrals, uas yog lub ntim thiab qhov chaw:
Yog tias muaj cov qauv uas muaj magnetization tas li, qhov ceev tam sim no yog xoom. Tom qab ntawd, qhov kev qhia tawm ntawm lub magnetic induction vector yuav siv daim ntawv hauv qab no:
Siv cov kev pab cuam MATLAB (MathWorks, Inc., USA) rau kev suav lej, ETU "LETI" daim ntawv tso cai kawm lej 40502181.
Raws li pom hauv Daim Duab 7 Daim Duab 8 Daim Duab 9 Daim Duab-10, lub zog sib nqus muaj zog tshaj plaws yog tsim los ntawm lub hlau nplaum uas taw qhia axially los ntawm qhov kawg ntawm lub tog raj kheej. Lub vojvoog ua haujlwm zoo yog sib npaug rau geometry ntawm lub hlau nplaum. Hauv cov hlau nplaum cylindrical nrog lub tog raj kheej uas nws ntev dua nws txoj kab uas hla, lub zog sib nqus muaj zog tshaj plaws yog pom nyob rau hauv qhov kev taw qhia axial-radial (rau cov khoom sib xws); yog li ntawd, ib khub ntawm lub tog raj kheej nrog qhov sib piv loj dua (txoj kab uas hla thiab ntev) MNP adsorption yog qhov zoo tshaj plaws.
Daim Duab 7 Cov khoom ntawm lub zog sib nqus Bz raws li Oz axis ntawm lub hlau nplaum; qhov loj me ntawm lub hlau nplaum: kab dub 0.5 × 2 hli, kab xiav 2 × 2 hli, kab ntsuab 3 × 2 hli, kab liab 5 × 2 hli.
Daim Duab 8 Lub khoom siv hluav taws xob sib nqus Br yog perpendicular rau lub hlau nplaum axis Oz; qhov loj me ntawm cov hlau nplaum: kab dub 0.5 × 2 hli, kab xiav 2 × 2 hli, kab ntsuab 3 × 2 hli, kab liab 5 × 2 hli.
Daim Duab 9 Qhov sib nqus induction zog Bz Cheebtsam ntawm qhov deb r ntawm qhov kawg axis ntawm lub hlau nplaum (z = 0); qhov loj me ntawm lub hlau nplaum: kab dub 0.5 × 2 hli, kab xiav 2 × 2 hli, kab ntsuab 3 × 2 hli, kab liab 5 × 2 hli.
Daim Duab 10 Cov khoom siv sib nqus induction raws li qhov kev taw qhia radial; qhov loj me ntawm cov hlau nplaum txheem: kab dub 0.5 × 2 hli, kab xiav 2 × 2 hli, kab ntsuab 3 × 2 hli, kab liab 5 × 2 hli.
Cov qauv hydrodynamic tshwj xeeb siv tau los kawm txog txoj kev xa MNP mus rau cov nqaij mob qog, ua kom cov nanoparticles sib sau ua ke hauv thaj chaw tsom, thiab txiav txim siab tus cwj pwm ntawm cov nanoparticles nyob rau hauv cov xwm txheej hydrodynamic hauv lub cev ncig. Cov hlau nplaum tas mus li siv tau ua cov teb sib nqus sab nraud. Yog tias peb tsis quav ntsej txog kev sib cuam tshuam magnetostatic ntawm cov nanoparticles thiab tsis xav txog cov qauv kua sib nqus, nws txaus los kwv yees qhov kev sib cuam tshuam ntawm cov hlau nplaum thiab ib qho nanoparticle nrog kev kwv yees dipole-dipole.
Qhov twg m yog lub zog sib nqus ntawm lub hlau nplaum, r yog lub vojvoog vector ntawm qhov chaw uas lub nanoparticle nyob, thiab k yog qhov system factor. Hauv qhov kev kwv yees dipole, lub teb ntawm lub hlau nplaum muaj qhov teeb tsa zoo sib xws (Daim Duab 11).
Hauv ib lub teb sib nqus sib npaug, cov nanoparticles tsuas yog tig raws cov kab ntawm lub zog. Hauv ib lub teb sib nqus tsis sib npaug, lub zog ua rau nws:
Qhov twg yog qhov derivative ntawm ib qho kev taw qhia l. Tsis tas li ntawd, lub zog rub cov nanoparticles mus rau hauv cov cheeb tsam tsis sib npaug tshaj plaws ntawm lub teb, uas yog, qhov curvature thiab density ntawm cov kab ntawm lub zog nce.
Yog li ntawd, nws yog qhov xav tau siv lub hlau nplaum muaj zog txaus (lossis cov saw hlau nplaum) nrog cov axial anisotropy pom tseeb hauv thaj chaw uas cov khoom me me nyob.
Rooj 1 qhia txog lub peev xwm ntawm ib lub hlau nplaum ua lub hauv paus hlau nplaum txaus los ntes thiab khaws cia MNP hauv lub txaj vascular ntawm daim teb thov.