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Cov nanoparticles insulin (NPs) uas muaj cov khoom thauj ntau tau pom ntau daim ntawv thov hauv ntau hom tshuaj sib txawv. Daim ntawv no lub hom phiaj yog los soj ntsuam qhov cuam tshuam ntawm cov txheej txheem khov-qhuav thiab cov txheej txheem ziab-tsuag rau cov qauv ntawm cov nanoparticles chitosan uas muaj insulin, nrog lossis tsis muaj mannitol ua tus tiv thaiv cryoprotectant. Peb kuj tau soj ntsuam qhov zoo ntawm cov nanoparticles no los ntawm kev rov yaj lawv. Ua ntej dehydration, qhov loj me ntawm cov nanoparticles chitosan/sodium tripolyphosphate/insulin cross-linked nanoparticles tau ua kom zoo tshaj plaws kom yog 318 nm, PDI yog 0.18, qhov ua tau zoo ntawm encapsulation yog 99.4%, thiab qhov loading yog 25.01%. Tom qab rov ua dua, txhua cov nanoparticles, tshwj tsis yog cov uas tsim los ntawm txoj kev khov-qhuav yam tsis siv mannitol, tswj lawv cov qauv spherical particle. Piv rau cov nanoparticles uas muaj mannitol uas qhuav los ntawm kev tsuag, cov nanoparticles uas tsis muaj mannitol kuj qhia tau tias qhov me tshaj plaws nruab nrab qhov loj me (376 nm) thiab qhov loading siab tshaj plaws. cov ntsiab lus (25.02%) nrog rau qhov sib xws encapsulation tus nqi (98.7%) thiab PDI (0.20) los ntawm kev ziab lossis khov-qhuav cov txheej txheem. Cov nanoparticles qhuav los ntawm kev tsuag ziab tsis muaj mannitol kuj ua rau muaj kev tso tawm sai tshaj plaws ntawm insulin thiab qhov ua tau zoo tshaj plaws ntawm kev nqus ntawm tes. Cov haujlwm no qhia tau hais tias kev tsuag ziab tuaj yeem ua rau cov insulin nanoparticles qhuav yam tsis tas yuav siv cryoprotectants piv rau cov txheej txheem khov ziab ib txwm muaj, tsim kom muaj peev xwm thauj khoom ntau dua, txo qis qhov xav tau ntxiv thiab cov nqi ua haujlwm zoo dua.
Txij li thaum nws tau tshawb pom hauv xyoo 1922, insulin thiab nws cov tshuaj tau cawm txoj sia ntawm cov neeg mob uas muaj ntshav qab zib hom 1 (T1DM) thiab ntshav qab zib hom 2 (T1DM). Txawm li cas los xij, vim nws cov khoom ua ib qho protein hnyav molecular siab, insulin yooj yim sib sau ua ke, tawg los ntawm cov enzymes proteolytic, thiab tshem tawm los ntawm thawj-dhau nyhuv. Cov neeg uas kuaj pom tias muaj ntshav qab zib hom 1 xav tau kev txhaj tshuaj insulin rau tas lawv lub neej. Ntau tus neeg mob uas kuaj pom tias muaj ntshav qab zib hom 2 kuj xav tau kev txhaj tshuaj insulin ntev. Kev txhaj tshuaj insulin txhua hnub yog qhov ua rau mob thiab tsis xis nyob txhua hnub rau cov neeg no, nrog rau cov teebmeem tsis zoo rau kev noj qab haus huv ntawm lub hlwb. Yog li ntawd, lwm hom kev txhaj tshuaj insulin uas ua rau tsis xis nyob tsawg dua, xws li kev txhaj tshuaj insulin hauv qhov ncauj, tab tom raug kawm ntau yam5 vim lawv muaj peev xwm rov qab kho lub neej zoo ntawm kwv yees li 5 billion tus neeg uas muaj ntshav qab zib thoob ntiaj teb.
Kev siv tshuab nanoparticle tau muab kev nce qib tseem ceeb hauv kev sim noj insulin4,6,7. Ib qho uas zoo encapsulates thiab tiv thaiv insulin los ntawm kev puas tsuaj rau kev xa mus rau qhov chaw tshwj xeeb ntawm lub cev. Txawm li cas los xij, kev siv cov nanoparticle formulations muaj ntau qhov kev txwv, feem ntau yog vim muaj teeb meem kev ruaj khov ntawm cov khoom me me. Qee qhov kev sib sau ua ke yuav tshwm sim thaum lub sijhawm khaws cia, uas txo qhov bioavailability ntawm insulin-loaded nanoparticles8. Tsis tas li ntawd, qhov kev ruaj khov tshuaj lom neeg ntawm cov polymer matrix ntawm nanoparticles thiab insulin kuj yuav tsum tau xav txog kom ntseeg tau tias muaj kev ruaj khov ntawm insulin nanoparticles (NPs). Tam sim no, kev siv tshuab freeze-drying yog tus qauv kub rau kev tsim cov NPs ruaj khov thaum tiv thaiv kev hloov pauv tsis xav tau thaum lub sijhawm khaws cia9.
Txawm li cas los xij, kev khov-qhuav yuav tsum tau ntxiv cov cryoprotectants kom tiv thaiv cov qauv kheej kheej ntawm NPs los ntawm kev cuam tshuam los ntawm kev ntxhov siab ntawm cov dej khov. Qhov no txo ​​​​​​qhov kev thauj khoom ntawm cov nanoparticles insulin tom qab lyophilization, vim tias cryoprotectant nyob feem ntau ntawm qhov hnyav piv. Yog li ntawd, cov insulin NPs tsim tawm feem ntau pom tias tsis haum rau kev tsim cov hmoov qhuav, xws li cov ntsiav tshuaj noj thiab cov yeeb yaj kiab noj, vim tias xav tau ntau cov nanoparticles qhuav kom ua tiav lub qhov rais kho mob ntawm insulin.
Kev ziab tshuaj tsuag yog ib qho txheej txheem lag luam uas paub zoo thiab pheej yig rau kev tsim cov hmoov qhuav los ntawm cov kua hauv kev lag luam tshuaj10,11. Kev tswj hwm cov txheej txheem tsim cov khoom me me tso cai rau kev encapsulation zoo ntawm ntau cov tshuaj bioactive 12, 13. Ntxiv mus, nws tau dhau los ua ib txoj hauv kev zoo rau kev npaj cov protein encapsulated rau kev tswj hwm qhov ncauj. Thaum lub sijhawm ziab tshuaj tsuag, dej ua pa sai heev, uas pab ua kom qhov kub ntawm lub hauv paus khoom me me qis11,14, ua rau nws daim ntawv thov los encapsulate cov khoom kub-rhiab. Ua ntej ziab tshuaj tsuag, cov khoom siv txheej yuav tsum tau sib xyaw ua ke nrog cov tshuaj uas muaj cov khoom xyaw encapsulated11,14. Tsis zoo li kev ziab khov, homogenization ua ntej encapsulation hauv kev ziab tshuaj tsuag txhim kho kev ua haujlwm encapsulation thaum lub sijhawm dehydration. Txij li thaum cov txheej txheem ziab tshuaj tsuag tsis xav tau cryoprotectants, ziab tshuaj tsuag tuaj yeem siv los tsim cov NPs qhuav nrog cov ntsiab lus thauj khoom siab.
Daim ntawv tshawb fawb no qhia txog kev tsim cov NPs uas muaj insulin los ntawm kev sib txuas ntawm chitosan thiab sodium tripolyphosphate siv txoj kev ion gel. Ion gelation yog ib txoj kev npaj uas tso cai rau kev tsim cov nanoparticles los ntawm kev sib cuam tshuam electrostatic ntawm ob lossis ntau hom ionic nyob rau hauv qee qhov xwm txheej. Ob qho tib si freeze-drying thiab spray-drying techniques tau siv los dehydrate cov chitosan / sodium tripolyphosphate / insulin cross-linked nanoparticles zoo tshaj plaws. Tom qab dehydration, lawv cov morphology tau soj ntsuam los ntawm SEM. Lawv lub peev xwm recombination tau soj ntsuam los ntawm kev ntsuas lawv qhov kev faib tawm loj, qhov them ntawm qhov chaw, PDI, encapsulation efficiency, thiab loading content. Qhov zoo ntawm resolubilized nanoparticles tsim los ntawm ntau txoj kev dehydration kuj tau soj ntsuam los ntawm kev sib piv lawv cov insulin tiv thaiv, kev tso tawm tus cwj pwm, thiab cellular uptake efficiency.
Lub pH ntawm cov tshuaj sib xyaw thiab qhov piv ntawm chitosan thiab insulin yog ob qho tseem ceeb uas cuam tshuam rau qhov loj me ntawm cov khoom me me thiab kev ua haujlwm ntawm encapsulation (EE) ntawm cov NPs kawg, vim lawv cuam tshuam ncaj qha rau cov txheej txheem ionotropic gelation. Lub pH ntawm cov tshuaj sib xyaw tau pom tias muaj feem cuam tshuam nrog qhov loj me ntawm cov khoom me me thiab kev ua haujlwm ntawm encapsulation (Daim Duab 1a). Raws li pom hauv Daim Duab 1a, thaum pH nce ntawm 4.0 txog 6.0, qhov nruab nrab ntawm cov khoom me me (nm) txo qis thiab EE nce ntxiv, thaum thaum pH nce mus txog 6.5, qhov nruab nrab ntawm cov khoom me me pib nce thiab EE tseem tsis hloov pauv. Raws li qhov piv ntawm chitosan rau insulin nce, qhov nruab nrab ntawm cov khoom me me kuj nce ntxiv. Ntxiv mus, tsis muaj kev hloov pauv hauv EE tau pom thaum nanoparticles tau npaj ntawm qhov sib piv ntawm chitosan / insulin siab dua 2.5: 1 (w / w) (Daim Duab 1b). Yog li ntawd, cov xwm txheej npaj zoo tshaj plaws hauv kev tshawb fawb no (pH 6.0, chitosan / insulin mass ratio ntawm 2.5: 1) tau siv. los npaj cov nanoparticles uas muaj insulin rau kev kawm ntxiv. Hauv qab qhov kev npaj no, qhov nruab nrab ntawm cov khoom me me ntawm cov nanoparticles insulin tau raug kho kom zoo kom yog 318 nm (Daim Duab 1c), PDI yog 0.18, qhov ua tau zoo ntawm kev ntxig yog 99.4%, lub peev xwm zeta yog 9.8 mv, thiab qhov kev thauj khoom insulin yog 25.01% (m/m). Raws li cov txiaj ntsig ntawm kev siv tshuab hluav taws xob kis (TEM), cov nanoparticles uas tau kho kom zoo yog cov kheej kheej thiab sib cais nrog qhov loj sib xws (Daim Duab 1d).
Kev ua kom zoo dua ntawm cov khoom siv insulin nanoparticles: (a) qhov cuam tshuam ntawm pH ntawm qhov nruab nrab txoj kab uas hla thiab kev ua haujlwm encapsulation (EE) ntawm cov khoom siv insulin nanoparticles (npaj ntawm 5: 1 pawg ntawm chitosan thiab insulin); (b) chitosan thiab Kev cuam tshuam ntawm pawg piv ntawm insulin ntawm qhov nruab nrab txoj kab uas hla thiab kev ua haujlwm encapsulation (EE) ntawm insulin NPs (npaj ntawm pH 6); (c) kev faib tawm qhov loj me ntawm cov khoom siv insulin nanoparticles zoo tshaj plaws; (d) TEM micrograph ntawm cov khoom siv insulin NPs zoo tshaj plaws.
Nws paub zoo tias chitosan yog ib qho polyelectrolyte tsis muaj zog nrog pKa ntawm 6.5. Nws muaj qhov them zoo hauv cov kua qaub vim tias nws cov amino pawg tseem ceeb yog protonated los ntawm hydrogen ions15. Yog li ntawd, nws feem ntau siv ua tus nqa khoom los encapsulate cov macromolecules uas muaj qhov tsis zoo. Hauv kev tshawb fawb no, chitosan tau siv los encapsulate insulin nrog qhov isoelectric point ntawm 5.3. Txij li thaum chitosan yog siv ua cov khoom siv txheej, nrog rau kev nce ntawm nws qhov sib piv, qhov tuab ntawm txheej sab nraud ntawm cov nanoparticles nce ntxiv, ua rau qhov nruab nrab loj dua. Tsis tas li ntawd, cov qib siab dua ntawm chitosan tuaj yeem encapsulate ntau insulin. Hauv peb qhov teeb meem, EE yog qhov siab tshaj plaws thaum qhov piv ntawm chitosan thiab insulin ncav cuag 2.5: 1, thiab tsis muaj kev hloov pauv tseem ceeb hauv EE thaum qhov piv txuas ntxiv nce.
Dhau li ntawm qhov sib piv ntawm chitosan thiab insulin, pH kuj tau ua lub luag haujlwm tseem ceeb hauv kev npaj NPs. Gan et al. 17 tau kawm txog qhov cuam tshuam ntawm pH rau qhov loj me ntawm cov khoom me me ntawm chitosan nanoparticles. Lawv pom tias qhov txo qis ntawm qhov loj me ntawm cov khoom me me kom txog thaum pH mus txog 6.0, thiab qhov nce ntxiv ntawm qhov loj me ntawm cov khoom me me tau pom ntawm pH > 6.0, uas yog sib xws nrog peb cov kev soj ntsuam. Qhov xwm txheej no yog vim qhov tseeb tias nrog kev nce pH, cov tshuaj insulin molecule tau txais qhov them tsis zoo, yog li, nyiam kev sib cuam tshuam electrostatic nrog chitosan / sodium tripolyphosphate (TPP) complex, ua rau qhov loj me ntawm cov khoom me me thiab EE siab. Txawm li cas los xij, thaum pH raug kho rau 6.5, cov amino pawg ntawm chitosan tau deprotonated, ua rau chitosan folding. Yog li, pH siab ua rau muaj kev cuam tshuam tsawg dua ntawm amino ions rau TPP thiab insulin, ua rau kev sib txuas qis dua, qhov nruab nrab loj dua thiab EE qis dua.
Kev tshuaj xyuas cov khoom morphological ntawm cov NPs khov-qhuav thiab cov tshuaj tsuag-qhuav tuaj yeem coj kev xaiv cov txheej txheem ua kom qhuav thiab hmoov zoo dua. Txoj kev nyiam yuav tsum muab cov tshuaj ruaj khov, cov duab sib npaug, cov tshuaj ntau thiab solubility zoo hauv cov tshuaj qub. Hauv kev tshawb fawb no, los sib piv ob txoj kev zoo dua, insulin NPs nrog lossis tsis muaj 1% mannitol tau siv thaum lub sijhawm qhuav. Mannitol yog siv ua tus neeg sawv cev bulking lossis cryoprotectant hauv ntau yam hmoov qhuav rau kev ziab khov thiab ziab tsuag. Rau cov lyophilized insulin nanoparticles tsis muaj mannitol, raws li pom hauv Daim Duab 2a, cov qauv hmoov porous heev nrog cov qhov loj, tsis sib xws thiab ntxhib tau pom nyob rau hauv scanning electron microscopy (SEM). Ob peb qhov sib cais tau pom hauv cov hmoov tom qab ziab (Daim Duab 2e). Cov txiaj ntsig no qhia tau tias feem ntau NPs tau decomposed thaum lub sijhawm ziab khov tsis muaj cryoprotectant. Rau cov khoom qhuav khov thiab cov tshuaj tsuag-qhuav insulin nanoparticles uas muaj 1% mannitol, cov nanoparticles spherical nrog cov qhov du tau pom (Daim Duab. 2b, d, f, h). Cov nanoparticles insulin uas txau-qhuav tsis muaj mannitol tseem yog lub voos xwmfab tab sis ntsws ntawm qhov chaw (Daim Duab 2c). Cov chaw kheej kheej thiab ntsws tau tham ntxiv hauv kev tso tawm tus cwj pwm thiab kev ntsuam xyuas cellular uptake hauv qab no. Raws li qhov pom ntawm cov NPs qhuav, ob qho tib si txau-qhuav NPs tsis muaj mannitol thiab NPs khov-qhuav thiab txau-qhuav nrog mannitol tau tsim cov hmoov NPs zoo (Daim Duab 2f, g, h). Qhov loj dua thaj chaw ntawm cov khoom me me, qhov siab dua qhov solubility thiab yog li ntawd qhov siab dua qhov tso tawm.
Daim duab ntawm cov insulin NPs uas qhuav dej sib txawv: (a) Daim duab SEM ntawm cov insulin NPs uas lyophilized tsis muaj mannitol; (b) Daim duab SEM ntawm cov insulin NPs uas lyophilized nrog mannitol; (c) cov insulin NPs uas qhuav tshuaj tsuag tsis muaj mannitol Daim duab SEM ntawm; (d) Daim duab SEM ntawm cov insulin NPs uas qhuav tshuaj tsuag nrog mannitol; (e) daim duab ntawm cov insulin NPs hmoov uas lyophilized tsis muaj mannitol; (f) daim duab ntawm cov insulin NPs uas qhuav tshuaj tsuag nrog mannitol; (g) Daim duab ntawm cov insulin NPs hmoov uas qhuav tshuaj tsuag tsis muaj mannitol; (h) daim duab ntawm cov insulin NPs hmoov uas qhuav tshuaj tsuag nrog mannitol.
Thaum lub sijhawm ua kom qhuav, mannitol ua haujlwm ua tus tiv thaiv kev khov, khaws cov NPs hauv daim ntawv amorphous thiab tiv thaiv kev puas tsuaj los ntawm cov dej khov crystals19. Qhov sib txawv, tsis muaj kauj ruam khov thaum lub sijhawm ziab tshuaj tsuag. Yog li ntawd, mannitol tsis tas yuav tsum muaj nyob rau hauv txoj kev no. Qhov tseeb, cov NPs qhuav tshuaj tsuag uas tsis muaj mannitol tau tsim cov NPs zoo dua li tau piav qhia ua ntej. Txawm li cas los xij, mannitol tseem tuaj yeem ua haujlwm ua tus sau hauv cov txheej txheem ziab tshuaj tsuag kom muab NPs ib qho qauv kheej kheej ntau dua20 (Daim Duab 2d), uas pab kom tau txais kev tso tawm sib xws ntawm cov NPs encapsulated. Tsis tas li ntawd, nws yog qhov tseeb tias qee cov khoom loj tuaj yeem pom hauv ob qho tib si khov-qhuav thiab tshuaj tsuag-qhuav insulin NPs uas muaj mannitol (Daim Duab 2b, d), uas tej zaum yuav yog vim muaj kev sib sau ua ke ntawm mannitol hauv lub hauv paus ntawm cov khoom me me ua ke nrog cov insulin encapsulated. Rau.Chitosan txheej. Nws yog ib qho tsim nyog sau tseg tias nyob rau hauv txoj kev tshawb no, txhawm rau kom ntseeg tau tias cov qauv kheej kheej tseem nyob ruaj khov tom qab dehydration, qhov piv ntawm mannitol thiab chitosan yog khaws cia ntawm 5: 1, yog li ntawd ntau ntawm cov khoom sau kuj tseem tuaj yeem ua rau qhov loj me ntawm cov NPs qhuav loj dua.
Fourier transform infrared attenuated total reflection (FTIR-ATR) spectroscopy tau piav qhia txog qhov sib xyaw ntawm cov tshuaj insulin dawb, chitosan, chitosan, TPP thiab insulin. Tag nrho cov dehydrated NPs tau piav qhia los ntawm kev siv FTIR-ATR spectroscopy. Qhov tseem ceeb, cov kab sib zog ntawm 1641, 1543 thiab 1412 cm-1 tau pom hauv cov encapsulated NPs khov-qhuav nrog mannitol thiab hauv NPs tsuag-qhuav nrog thiab tsis muaj mannitol (Daim Duab 3). Raws li tau tshaj tawm ua ntej, cov kev nce ntxiv ntawm lub zog no tau cuam tshuam nrog kev sib txuas ntawm chitosan, TPP thiab insulin. Kev tshawb nrhiav ntawm kev sib cuam tshuam ntawm chitosan thiab insulin tau qhia tias hauv FTIR spectra ntawm insulin-loaded chitosan nanoparticles, cov kab chitosan overlapped nrog cov insulin, ua rau cov carbonyl intensity (1641 cm-1) thiab amine (1543 cm-1) siv. Cov pawg tripolyphosphate ntawm TPP txuas nrog cov pawg ammonium hauv chitosan, tsim ib txoj kab ntawm 1412 cm-1.
FTIR-ATR spectra ntawm cov insulin dawb, chitosan, cov khoom sib xyaw ntawm chitosan/TPP/insulin thiab NPs uas tau qhuav los ntawm ntau txoj kev sib txawv.
Ntxiv mus, cov txiaj ntsig no zoo ib yam li cov uas tau pom hauv SEM, uas qhia tau tias cov NPs uas tau muab kaw cia tseem zoo li qub thaum txau thiab khov-qhuav nrog mannitol, tab sis yog tsis muaj mannitol, tsuas yog txau-qhuav tsim cov khoom me me uas tau muab kaw cia. Qhov sib txawv, cov txiaj ntsig FTIR-ATR spectral ntawm NPs khov-qhuav tsis muaj mannitol zoo ib yam li cov khoom sib xyaw ntawm chitosan, TPP, thiab insulin. Cov txiaj ntsig no qhia tau tias cov kev sib txuas ntawm chitosan, TPP thiab insulin tsis muaj nyob hauv NPs khov-qhuav tsis muaj mannitol lawm. Cov qauv NPs raug rhuav tshem thaum lub sijhawm khov-qhuav tsis muaj cryoprotectant, uas tuaj yeem pom hauv cov txiaj ntsig SEM (Daim Duab 2a). Raws li cov qauv morphology thiab FTIR ntawm cov insulin NPs uas qhuav dej, tsuas yog lyophilized, tsuag-qhuav, thiab mannitol-free NPs tau siv rau kev sim rov ua dua thiab mannitol-free NPs vim yog kev rhuav tshem ntawm mannitol-free NPs thaum lub sijhawm qhuav dej. tham txog.
Kev ziab yog siv rau kev khaws cia ntev thiab rov ua dua rau lwm cov qauv. Lub peev xwm ntawm cov NPs qhuav kom rov ua dua tom qab khaws cia yog qhov tseem ceeb rau lawv siv rau hauv cov qauv sib txawv xws li cov ntsiav tshuaj thiab cov yeeb yaj kiab. Peb pom tias qhov nruab nrab ntawm cov khoom me me ntawm cov tshuaj insulin NPs uas qhuav tshuaj tsuag thaum tsis muaj mannitol nce me ntsis tom qab rov ua dua. Ntawm qhov tod tes, qhov loj me ntawm cov tshuaj insulin nanoparticles uas qhuav tshuaj tsuag thiab khov tshuaj tsuag nrog mannitol nce ntxiv ntau heev (Rooj 1). PDI thiab EE tsis tau hloov pauv ntau heev (p > 0.05) tom qab rov ua ke ntawm txhua NPs hauv kev tshawb fawb no (Rooj 1). Qhov tshwm sim no qhia tau tias feem ntau ntawm cov khoom me me tseem nyob ruaj khov tom qab yaj dua. Txawm li cas los xij, kev ntxiv mannitol ua rau kev thauj khoom insulin ntawm lyophilized thiab qhuav tshuaj tsuag mannitol nanoparticles (Rooj 1). Hauv kev sib piv, cov ntsiab lus ntawm insulin ntawm NPs uas qhuav tshuaj tsuag yam tsis muaj mannitol tseem zoo ib yam li yav dhau los (Rooj 1).
Nws paub zoo tias qhov kev thauj khoom ntawm cov nanoparticles yog qhov tseem ceeb thaum siv rau lub hom phiaj xa tshuaj. Rau NPs uas muaj kev thauj khoom tsawg, cov khoom siv ntau heev yuav tsum tau mus txog qhov chaw kho mob. Txawm li cas los xij, qhov viscosity siab ntawm cov NP concentration siab ua rau muaj kev tsis yooj yim thiab nyuaj rau kev tswj hwm qhov ncauj thiab cov tshuaj txhaj, raws li 22. Tsis tas li ntawd, insulin NPs kuj tseem siv tau los ua cov ntsiav tshuaj thiab cov biofilms viscous23, 24, uas yuav tsum tau siv cov NPs ntau ntawm cov qib thauj khoom qis, ua rau cov ntsiav tshuaj loj thiab cov biofilms tuab uas tsis haum rau kev siv qhov ncauj. Yog li ntawd, dehydrated NPs nrog cov insulin load siab yog qhov xav tau heev. Peb cov txiaj ntsig qhia tias qhov insulin load siab ntawm mannitol-free spray-dried NPs tuaj yeem muab ntau qhov zoo rau cov txheej txheem xa khoom lwm txoj hauv kev no.
Tag nrho cov NPs uas qhuav lawm tau khaws cia rau hauv lub tub yees rau peb lub hlis. Cov txiaj ntsig SEM qhia tau hais tias cov morphology ntawm tag nrho cov NPs uas qhuav lawm tsis hloov pauv ntau thaum lub sijhawm khaws cia peb lub hlis (Daim Duab 4). Tom qab rov ua dua hauv dej, tag nrho cov NPs tau qhia txog qhov txo qis me ntsis hauv EE thiab tso tawm kwv yees li me me (~ 5%) ntawm insulin thaum lub sijhawm khaws cia peb lub hlis (Rooj 2). Txawm li cas los xij, qhov nruab nrab ntawm cov khoom me me ntawm txhua cov nanoparticles tau nce ntxiv. Qhov loj me ntawm cov NPs uas tsuag-qhuav tsis muaj mannitol nce mus txog 525 nm, thaum cov tsuag-qhuav thiab khov-qhuav NPs nrog mannitol nce mus txog 872 thiab 921 nm, raws li (Rooj 2).
Cov duab ntawm cov tshuaj insulin NPs sib txawv uas tau qhuav thiab khaws cia rau peb lub hlis: (a) Daim duab SEM ntawm cov tshuaj insulin NPs uas tau lyophilized nrog mannitol; (b) Daim duab SEM ntawm cov tshuaj insulin nanoparticles uas tau txau-qhuav uas tsis muaj mannitol; (c) tsis muaj mannitol Daim duab SEM ntawm cov tshuaj insulin NPs uas tau txau-qhuav.
Tsis tas li ntawd xwb, cov av noo tau pom nyob rau hauv cov nanoparticles insulin uas tau rov ua dua tshiab uas tau txau-qhuav nrog mannitol thiab khov-qhuav (Daim Duab S2). Qhov no yuav yog vim cov khoom loj tsis tau dai kom zoo hauv dej. Tag nrho cov txiaj ntsig saum toj no qhia tau hais tias cov txheej txheem ziab tshuaj tsuag tuaj yeem tiv thaiv cov nanoparticles insulin los ntawm kev qhuav dej thiab tias cov khoom ntim ntau ntawm cov nanoparticles insulin tuaj yeem tau txais yam tsis muaj cov khoom sau lossis cryoprotectants.
Kev khaws cia insulin tau sim hauv pH = 2.5 nruab nrab nrog pepsin, trypsin, thiab α-chymotrypsin los qhia txog lub peev xwm tiv thaiv ntawm NPs tiv thaiv kev zom zaub mov enzymatic tom qab dehydration. Kev khaws cia insulin ntawm cov dehydrated NPs tau piv nrog cov NPs tshiab npaj, thiab insulin dawb tau siv ua kev tswj tsis zoo. Hauv kev tshawb fawb no, insulin dawb tau qhia txog kev tshem tawm insulin sai hauv 4 teev hauv tag nrho peb txoj kev kho enzymatic (Daim duab 5a-c). Qhov sib piv, kev sim tshem tawm insulin ntawm NPs khov-qhuav nrog mannitol thiab NPs tsuag-qhuav nrog lossis tsis muaj mannitol tau qhia txog kev tiv thaiv siab dua ntawm cov NPs no tiv thaiv kev zom zaub mov enzymatic, uas zoo ib yam li cov insulin NPs tshiab npaj (Daim duab 1).5a-c). Nrog kev pab los ntawm nanoparticles hauv pepsin, trypsin, thiab α-chymotrypsin, ntau dua 50%, 60%, thiab 75% ntawm insulin tuaj yeem tiv thaiv hauv 4 teev, feem (Daim duab 5a-c). Lub peev xwm tiv thaiv insulin no yuav ua rau muaj feem ntau ntawm kev nqus insulin ntau dua. mus rau hauv cov ntshav25. Cov txiaj ntsig no qhia tau tias kev ziab tshuaj tsuag nrog lossis tsis muaj mannitol thiab kev ziab khov nrog mannitol tuaj yeem khaws lub peev xwm tiv thaiv insulin ntawm NPs tom qab lub cev qhuav dej.
Kev tiv thaiv thiab kev tso tawm ntawm cov insulin NPs uas qhuav dej: (a) kev tiv thaiv insulin hauv cov kua pepsin; (b) kev tiv thaiv insulin hauv cov kua trypsin; (c) kev tiv thaiv insulin los ntawm α-chymotrypsin kua; (d) Tus cwj pwm tso tawm ntawm cov NPs uas qhuav dej hauv pH = 2.5 kua; (e) tus cwj pwm tso tawm ntawm cov NPs uas qhuav dej hauv pH = 6.6 kua; (f) tus cwj pwm tso tawm ntawm cov NPs uas qhuav dej hauv pH = 7.0 kua.
Cov tshuaj insulin qhuav NPs uas tau npaj tshiab thiab rov ua dua tshiab tau muab tso rau hauv ntau yam buffers (pH = 2.5, 6.6, 7.0) ntawm 37 ° C, ua qauv pH ib puag ncig ntawm lub plab, duodenum, thiab txoj hnyuv me sab saud, los tshuaj xyuas qhov cuam tshuam ntawm insulin rau kev tiv thaiv insulin. Kev tso tawm tus cwj pwm hauv ntau qhov chaw sib txawv. Ib feem ntawm txoj hnyuv plab. Ntawm pH = 2.5, cov NPs uas muaj insulin thiab cov NPs qhuav uas tau yaj tau qhia txog kev tso tawm thawj zaug hauv ib teev thawj zaug, tom qab ntawd tso tawm qeeb qeeb hauv 5 teev tom ntej (Daim Duab 5d). Qhov kev tso tawm sai sai no thaum pib feem ntau yog qhov tshwm sim ntawm kev tso tawm sai ntawm cov protein molecules uas tsis tau immobilized tag nrho hauv cov qauv sab hauv ntawm cov khoom me me. Ntawm pH = 6.5, cov NPs uas muaj insulin thiab cov NPs qhuav uas tau tsim dua tshiab tau qhia txog kev tso tawm du thiab qeeb qeeb hauv 6 teev, vim tias pH ntawm cov kua kuaj zoo ib yam li cov kua NPs uas tau npaj (Daim Duab 5e). Ntawm pH = 7, cov NPs tsis ruaj khov thiab yuav luag tag nrho decomposed hauv ob teev thawj zaug (Daim Duab 5f). Qhov no yog vim tias deprotonation ntawm chitosan tshwm sim ntawm pH siab dua, uas ua rau lub network polymer tsawg dua thiab tso tawm ntawm cov insulin uas tau thauj khoom.
Ntxiv mus, cov insulin NPs uas txau-qhuav tsis muaj mannitol tau qhia txog qhov tso tawm sai dua li lwm cov NPs uas qhuav (Daim duab 5d-f). Raws li tau piav qhia ua ntej, cov insulin NPs uas tau rov ua dua tshiab uas qhuav tsis muaj mannitol tau qhia txog qhov me me ntawm cov khoom me me. Cov khoom me me muab thaj chaw loj dua, yog li feem ntau ntawm cov tshuaj cuam tshuam yuav nyob ntawm lossis ze ntawm qhov chaw ntawm cov khoom me me, ua rau cov tshuaj tso tawm sai26.
Qhov cytotoxicity ntawm NPs tau raug tshawb xyuas los ntawm MTT assay.Raws li pom hauv Daim Duab S4, txhua qhov dehydrated NPs tau pom tias tsis muaj kev cuam tshuam loj rau lub cell muaj sia nyob ntawm cov concentration ntawm 50–500 μg / ml, qhia tias txhua qhov dehydrated NPs tuaj yeem siv tau zoo kom mus txog lub qhov rais kho mob.
Lub siab yog lub cev tseem ceeb uas insulin ua haujlwm rau nws lub cev. Cov hlwb HepG2 yog cov kab hlwb hepatoma tib neeg feem ntau siv ua qauv hauv vitro hepatocyte uptake. Ntawm no, cov hlwb HepG2 tau siv los ntsuas kev nqus ntawm cov hlwb NPs uas qhuav dej siv cov txheej txheem khov-qhuav thiab tshuaj tsuag-qhuav. Kev nqus ntawm cov hlwb los ntawm kev ntsuas laser confocal siv cov cytometry ntws thiab kev pom tom qab ob peb teev ntawm kev sib xyaw nrog FITC insulin dawb ntawm qhov concentration ntawm 25 μg / mL, FITC insulin-loaded NPs tshiab npaj thiab dehydrated FITC insulin-loaded NPs ntawm cov insulin sib npaug zos Quantitative microscopy (CLSM) kev soj ntsuam tau ua. Lyophilized NPs tsis muaj mannitol tau raug rhuav tshem thaum lub sijhawm dehydration thiab tsis tau soj ntsuam hauv qhov kev xeem no. Qhov intracellular fluorescence intensities ntawm cov hlwb NPs tshiab npaj insulin-loaded, lyophilized NPs nrog mannitol, thiab tshuaj tsuag-qhuav NPs nrog thiab tsis muaj mannitol (Daim duab 6a) yog 4.3, 2.6, 2.4, thiab 4.1-npaug siab dua li cov dawb.FITC-insulin pawg, raws li (Daim Duab 6b). Cov txiaj ntsig no qhia tau hais tias encapsulated insulin muaj zog dua hauv kev nqus ntawm tes dua li cov insulin dawb, feem ntau yog vim qhov me me ntawm cov nanoparticles uas muaj insulin tsim tawm hauv kev tshawb fawb.
Kev nqus ntawm cov hlwb HepG2 tom qab 4 teev kev sib xyaw nrog cov NPs tshiab thiab cov NPs qhuav: (a) Kev faib tawm ntawm FITC-insulin kev nqus los ntawm cov hlwb HepG2. (b) Geometric nruab nrab ntawm qhov sib zog fluorescence uas tau soj ntsuam los ntawm kev ntws cytometry (n = 3), *P < 0.05 piv nrog cov insulin dawb.
Ib yam li ntawd, cov duab CLSM qhia tau hais tias qhov FITC fluorescence intensities ntawm cov FITC-insulin-loaded NPs tshiab thiab FITC-insulin-loaded spray-dried NPs (tsis muaj mannitol) muaj zog dua li lwm cov qauv (Daim Duab 6a). Ntxiv mus, nrog rau kev ntxiv mannitol, qhov viscosity siab dua ntawm cov tshuaj ua rau muaj kev tiv thaiv rau kev nqus ntawm cov hlwb, ua rau muaj kev loj hlob ntawm insulin tsawg dua. Cov txiaj ntsig no qhia tau hais tias cov mannitol-free spray-dried NPs tau pom tias muaj kev nqus ntawm cov hlwb siab tshaj plaws vim tias lawv qhov loj me me dua li cov freeze-dried NPs tom qab rov yaj.
Chitosan (qhov nruab nrab molecular hnyav 100 KDa, 75–85% deacetylated) tau yuav los ntawm Sigma-Aldrich. (Oakville, Ontario, Canada). Sodium tripolyphosphate (TPP) tau yuav los ntawm VWR (Radnor, Pennsylvania, USA). Recombinant human insulin siv hauv kev tshawb fawb no yog los ntawm Fisher Scientific (Waltham, MA, USA). Fluorescein isothiocyanate (FITC)-labeled human insulin thiab 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) tau yuav los ntawm Sigma-Aldrich. (Oakville, Ontario, Canada). Cov kab HepG2 cell tau txais los ntawm ATCC (Manassas, Virginia, USA). Tag nrho lwm cov reagents yog analytical lossis chromatographic qib.
Npaj cov tshuaj CS 1 mg/ml los ntawm kev yaj nws hauv dej ob zaug (DD dej) uas muaj 0.1% acetic acid. Npaj cov tshuaj TPP thiab insulin 1 mg/ml los ntawm kev yaj lawv hauv dej DD thiab 0.1% acetic acid, raws li. Cov tshuaj pre-emulsion tau npaj nrog lub tshuab homogenizer ceev ceev polytron PCU-2-110 (Brinkmann Ind. Westbury, NY, USA). Cov txheej txheem npaj yog raws li nram no: ua ntej, ntxiv 2ml ntawm cov tshuaj TPP rau hauv 4ml ntawm cov tshuaj insulin, thiab cov khoom sib xyaw raug do rau 30 feeb thiab sib xyaw tag nrho. Tom qab ntawd, cov tshuaj sib xyaw tau ntxiv rau hauv cov tshuaj CS los ntawm lub koob txhaj tshuaj hauv qab kev sib xyaw ceev ceev (10,000 rpm). Cov khoom sib xyaw tau khaws cia rau hauv kev sib xyaw ceev ceev (15,000 rpm) hauv lub dab dej khov rau 30 feeb, thiab lawv tau kho rau qee qhov pH kom tau txais cov insulin NPs sib txuas. Txhawm rau kom sib xyaw ntxiv thiab txo qhov loj me ntawm insulin NPs, lawv tau sonicated rau ntxiv 30 feeb hauv lub dab dej khov siv lub tshuab ntsuas sonicator (UP 200ST, Hielscher Ultrasonics, Teltow, Lub Tebchaws Yelemees).
Cov Insulin NPS raug sim rau Z-qhov nruab nrab txoj kab uas hla, polydispersity index (PDI) thiab zeta muaj peev xwm siv cov kev ntsuas dynamic light scattering (DLS) siv Litesizer 500 (Anton Paar, Graz, Austria) los ntawm kev diluting lawv hauv DD dej ntawm 25 ° C. Morphology thiab qhov loj me faib tau piav qhia los ntawm Hitachi H7600 transmission electron microscope (TEM) (Hitachi, Tokyo, Nyiv), thiab cov duab tau raug soj ntsuam tom qab siv Hitachi imaging software (Hitachi, Tokyo, Nyiv). Txhawm rau ntsuas qhov encapsulation efficiency (EE) thiab loading capacity (LC) ntawm insulin NPs, cov NPs tau pipetted rau hauv ultrafiltration raj nrog molecular weight cut-off ntawm 100 kDa thiab centrifuged ntawm 500 xg rau 30 feeb. Unencapsulated insulin hauv filtrate tau ntsuas siv Agilent 1100 Series HPLC system (Agilent, Santa Clara, California, USA) uas muaj lub twj tso kua mis quaternary, autosampler, column heater, thiab DAD. lub tshuab ntes. Insulin tau raug tshuaj xyuas los ntawm C18 kem (Zorbax, 3.5 μm, 4.6 mm × 150 mm, Agilent, Tebchaws USA) thiab tau kuaj pom ntawm 214 nm. Lub theem txav tau yog acetonitrile thiab dej, muaj 0.1% TFA, qhov sib piv ntawm 10/90 txog 100/0, thiab khiav rau 10 feeb. Lub theem txav tau raug pumped ntawm tus nqi ntws ntawm 1.0 ml / min. Lub kem kub tau teeb tsa rau 20 ° C. Xam cov feem pua ​​​​​​ntawm EE thiab LC siv cov qauv. (1) thiab Eq. (2).
Ntau yam CS/insulin piv txij li 2.0 txog 4.0 tau raug sim los ua kom zoo dua insulin NP. Cov tshuaj CS sib txawv tau ntxiv thaum lub sijhawm npaj, thaum cov tshuaj insulin/TPP sib xyaw tau khaws cia tas li. Insulin NPs tau npaj rau hauv pH ntawm 4.0 txog 6.5 los ntawm kev tswj hwm pH ntawm cov tshuaj tom qab ntxiv tag nrho cov tshuaj (insulin, TPP thiab CS). EE thiab qhov loj me ntawm cov insulin nanoparticles tau raug soj ntsuam ntawm cov nqi pH sib txawv thiab CS/insulin pawg sib piv kom zoo dua kev tsim cov insulin NPs.
Cov insulin NPs uas zoo tshaj plaws tau muab tso rau ntawm lub thawv txhuas thiab npog nrog cov ntaub so ntswg uas nruj nrog qee daim kab xev. Tom qab ntawd, cov thawv ntsia hlau tau muab tso rau hauv lub tshuab ziab khov Labconco FreeZone (Labconco, Kansas City, MO, USA) uas muaj lub tshuab ziab tais. Qhov kub thiab lub zog nqus tau teeb tsa ntawm -10 ° C, 0.350 Torr rau 2 teev thawj zaug, thiab 0 ° C thiab 0.120 Torr rau 22 teev seem ntawm 24 teev kom tau txais cov insulin NPs qhuav.
Lub Tshuab Ziab Khaub Ncaws Buchi Mini Spray B-290 (BÜCHI, Flawil, Switzerland) tau siv los tsim cov tshuaj insulin uas tau muab kaw cia. Cov kev ntsuas ziab uas tau xaiv yog: kub 100 °C, ntws pub 3 L/min, thiab ntws roj 4 L/min.
Cov Insulin NPs ua ntej thiab tom qab dehydration tau txheeb xyuas siv FTIR-ATR spectroscopy. Cov nanoparticles dehydrated nrog rau cov insulin dawb thiab chitosan tau soj ntsuam siv Spectrum 100 FTIR spectrophotometer (PerkinElmer, Waltham, Massachusetts, USA) uas muaj cov khoom siv universal ATR sampling accessory (PerkinElmer, Waltham, Massachusetts, USA). Cov cim nruab nrab tau txais los ntawm 16 scans ntawm qhov kev daws teeb meem ntawm 4 cm2 hauv qhov ntau zaus ntawm 4000-600 cm2.
Kev soj ntsuam cov qauv ntawm cov insulin NPs qhuav tau raug soj ntsuam los ntawm cov duab SEM ntawm cov insulin NPs uas tau khov thiab qhuav tshuaj tsuag uas tau ntes los ntawm Helios NanoLab 650 Focused Ion Beam-Scanning Electron Microscope (FIB-SEM) (FEI, Hillsboro, Oregon, USA). Cov yam ntxwv tseem ceeb uas siv yog voltage 5 keV thiab tam sim no 30 mA.
Tag nrho cov insulin NPs uas qhuav lawm tau yaj dua hauv dej dd. Qhov loj ntawm cov khoom me me, PDI, EE thiab LC tau raug sim dua siv tib txoj kev uas tau hais ua ntej los ntsuas lawv qhov zoo tom qab qhuav dej. Qhov ruaj khov ntawm anhydroinsulin NPs kuj tau ntsuas los ntawm kev sim cov khoom ntawm NPs tom qab khaws cia ntev. Hauv kev tshawb fawb no, tag nrho cov NPs tom qab qhuav dej tau khaws cia rau hauv lub tub yees rau peb lub hlis. Tom qab peb lub hlis ntawm kev khaws cia, NPs tau raug sim rau qhov loj ntawm cov khoom me me, PDI, EE thiab LC.
Yaj 5 mL ntawm cov NPs uas tau rov ua dua tshiab rau hauv 45 mL uas muaj cov kua dej hauv plab (pH 1.2, muaj 1% pepsin), cov kua hauv plab (pH 6.8, muaj 1% trypsin) lossis cov kua chymotrypsin (100 g/mL, hauv phosphate buffer, pH 7.8) los ntsuas qhov ua tau zoo ntawm insulin hauv kev tiv thaiv NPs tom qab dehydration. Lawv tau incubated ntawm 37°C nrog qhov ceev ntawm 100 rpm. 500 μL ntawm cov kua tau sau ntawm ntau lub sijhawm sib txawv thiab qhov concentration ntawm insulin tau txiav txim siab los ntawm HPLC.
Tus cwj pwm tso tawm hauv vitro ntawm cov insulin NPs tshiab uas tau npaj thiab qhuav tau raug sim los ntawm txoj kev dialysis hnab (molecular weight cut-off 100 kDa, Spectra Por Inc.). Cov NPs qhuav uas tau npaj thiab rov ua dua tshiab tau dialyzed hauv cov kua ntawm pH 2.5, pH 6.6, thiab pH 7.0 (0.1 M phosphate-buffered saline, PBS) los ua qauv pH ib puag ncig ntawm lub plab, duodenum, thiab txoj hnyuv me sab saud, raws li. Tag nrho cov qauv tau incubated ntawm 37 ° C nrog kev co tas li ntawm 200 rpm. Nqus cov kua sab nraum lub hnab dialysis 5 mL ntawm cov sijhawm hauv qab no: 0.5, 1, 2, 3, 4, thiab 6 teev, thiab tam sim ntawd rov ua kom tiav qhov ntim nrog dialysate tshiab. Kev ua qias tuaj ntawm insulin hauv cov kua tau raug soj ntsuam los ntawm HPLC, thiab tus nqi ntawm insulin tso tawm los ntawm cov nanoparticles tau suav los ntawm qhov piv ntawm cov insulin dawb tso tawm rau tag nrho cov insulin encapsulated hauv cov nanoparticles (Equation 3).
Cov hlwb Hepatocellular carcinoma ntawm tib neeg HepG2 tau cog rau hauv cov tais diav uas muaj txoj kab uas hla 60 hli siv Dulbecco's Modified Eagle's Medium (DMEM) uas muaj 10% fetal bovine serum, 100 IU/mL penicillin, thiab 100 μg/mL streptomycin29. Cov kab mob tau khaws cia ntawm 37°C, 95% qhov av noo, thiab 5% CO2. Rau kev ntsuas kev nqus, cov hlwb HepG2 tau cog rau ntawm 1 × 105 hlwb/ml rau ntawm 8-qhov Nunc Lab-Tek chamber slide system (Thermo Fisher, NY, USA). Rau kev ntsuas cytotoxicity, lawv tau cog rau hauv 96-qhov phaj (Corning, NY, USA) ntawm qhov ceev ntawm 5 × 104 hlwb/ml.
Qhov kev sim MTT tau siv los ntsuas qhov cytotoxicity ntawm cov insulin NPs30 uas tau npaj tshiab thiab qhuav dej. Cov hlwb HepG2 tau muab noob rau hauv cov phaj 96-qhov ntawm qhov ceev ntawm 5 × 104 hlwb / mL thiab cog qoob loo rau 7 hnub ua ntej kev sim. Insulin NPs tau diluted rau ntau yam concentration (50 txog 500 μg / mL) hauv cov kab lis kev cai thiab tom qab ntawd muab rau cov hlwb. Tom qab 24 teev ntawm kev cog qoob loo, cov hlwb tau ntxuav 3 zaug nrog PBS thiab cog qoob loo nrog cov kab lis kev cai uas muaj 0.5 mg / ml MTT rau ntxiv 4 teev. Cytotoxicity tau soj ntsuam los ntawm kev ntsuas qhov kev txo qis enzymatic ntawm daj tetrazolium MTT rau xim av formazan ntawm 570 nm siv Tecan infinite M200 pro spectrophotometer phaj nyeem ntawv (Tecan, Männedorf, Switzerland).
Kev siv lub tshuab ntsuas confocal laser scanning microscopy thiab kev ntsuas flow cytometry. Txhua lub qhov ntawm Nunc Lab-Tek chamber slide system tau kho nrog FITC-insulin dawb, FITC-insulin-loaded NPs, thiab rov ua dua 25 μg/mL ntawm cov FITC-insulin NPs uas qhuav dej ntawm tib qhov concentration thiab incubated rau 4 teev. Cov hlwb tau ntxuav 3 zaug nrog PBS thiab kho nrog 4% paraformaldehyde. Nuclei tau stained nrog 4′,6-diamidino-2-phenylindole (DAPI). Kev tshawb pom qhov chaw ntawm insulin tau pom siv Olympus FV1000 laser scanning/two-photon confocal microscope (Olympus, Shinjuku City, Tokyo, Nyiv). Rau kev ntsuas flow cytometry, tib qhov concentration ntawm 10 μg/mL dawb FITC-insulin, FITC-insulin-loaded NPs, thiab resolubilized dehydrated FITC-insulin NPs tau ntxiv rau 96-well plates. noob nrog HepG2 hlwb thiab incubated rau 4 teev. Tom qab 4 teev ntawm incubation, cov hlwb raug tshem tawm thiab ntxuav 3 zaug nrog FBS. 5 × 104 hlwb ib qho qauv tau soj ntsuam los ntawm BD LSR II flow cytometer (BD, Franklin Lakes, New Jersey, Tebchaws Meskas).
Txhua tus nqi raug qhia ua qhov nruab nrab ± tus qauv sib txawv. Kev sib piv ntawm txhua pawg tau raug soj ntsuam siv ib txoj kev ANOVA lossis t-test los ntawm IBM SPSS Statistics 26 rau Mac (IBM, Endicott, New York, USA) thiab p < 0.05 tau suav hais tias yog qhov tseem ceeb ntawm kev suav lej.
Daim ntawv tshawb fawb no qhia txog kev yooj ywm thiab lub peev xwm ntawm kev ziab tshuaj tsuag kom qhuav cov chitosan/TPP/insulin nanoparticles uas txuas nrog cross-linked nrog kev rov ua dua zoo dua piv rau cov txheej txheem khov-qhuav siv cov neeg sawv cev bulking lossis cryoprotectants muaj peev xwm thiab muaj peev xwm thauj khoom ntau dua. Cov nanoparticles insulin zoo tshaj plaws tau muab qhov nruab nrab ntawm cov khoom me me ntawm 318 nm thiab kev ua haujlwm encapsulation ntawm 99.4%. Cov txiaj ntsig SEM thiab FTIR tom qab dehydration qhia tau hais tias cov qauv spherical tsuas yog khaws cia hauv cov NPs qhuav tshuaj tsuag nrog thiab tsis muaj mannitol thiab lyophilized nrog mannitol, tab sis lyophilized NPs tsis muaj mannitol tau decomposed thaum lub sijhawm dehydration. Hauv kev sim peev xwm rov ua dua, insulin nanoparticles qhuav tshuaj tsuag tsis muaj mannitol qhia qhov nruab nrab me tshaj plaws thiab qhov thauj khoom siab tshaj plaws thaum rov ua dua. Cov cwj pwm tso tawm ntawm tag nrho cov dehydrated NPs no qhia tau hais tias lawv tau tso tawm sai sai hauv cov tshuaj ntawm pH = 2.5 thiab pH = 7, thiab ruaj khov heev hauv cov tshuaj ntawm pH = 6.5. Piv rau lwm cov dehydrated NPs uas yaj dua, cov NPs cov tshuaj tsuag qhuav uas tsis muaj mannitol qhia tau tias muaj kev tso tawm sai tshaj plaws. Qhov tshwm sim no yog sib xws nrog qhov pom hauv kev ntsuas kev nqus ntawm tes, vim tias cov tshuaj tsuag qhuav NPs uas tsis muaj mannitol yuav luag tswj tau qhov ua tau zoo ntawm tes ntawm cov NPs uas tau npaj tshiab. Cov txiaj ntsig no qhia tau tias cov nanoparticles insulin qhuav uas tau npaj los ntawm kev ziab tshuaj tsuag tsis muaj mannitol yog qhov tsim nyog tshaj plaws rau kev ua ntxiv rau lwm cov ntawv tshuaj anhydrous, xws li cov ntsiav tshuaj noj lossis cov yeeb yaj kiab bioadhesive.
Vim muaj teeb meem txog kev txawj ntse, cov ntaub ntawv uas tau tsim thiab/lossis tshuaj xyuas thaum lub sijhawm kawm tam sim no tsis muaj rau pej xeem, tab sis muaj los ntawm cov kws sau ntawv raws li qhov kev thov tsim nyog.
Kagan, A. Kab mob ntshav qab zib hom 2: keeb kwm hauv zej zog thiab kev tshawb fawb, teeb meem kev kho mob, thiab cov teeb meem rau cov neeg mob thiab lwm tus. (McFarlane, 2009).
Singh, AP, Guo, Y., Singh, A., Xie, W. & Jiang, P. Kev txhim kho ntawm insulin encapsulation: puas yog kev tswj hwm qhov ncauj tam sim no ua tau? J. Pharmacy.bio-pharmacy.reservoir.1, 74–92 (2019).
Wong, CY, Al-Salami, H. & Dass, CR Cov kev nce qib tsis ntev los no hauv cov txheej txheem xa khoom liposome uas muaj insulin rau kev kho mob ntshav qab zib. Kev Txhais Lus.J. Pharmacy.549, 201–217 (2018).