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Insuline nanoparticules (NPs) minnu doni hakɛ ka ca, olu ye baarakɛcogo wɛrɛw sɔrɔ u tata hakɛ danfaralenw na.Nin baara in kun ye ka jalen ni fiyɛli kɛcogo nɔ jateminɛ inisɔndiyalen nanoparticules (chitosan nanoparticules) jɔcogo la, ni manitol ye walima ni mannitol tɛ a la i n’a fɔ cryoprotectant.An ye nin nanoparticules ninnu jogo fana jateminɛ ni u wulicogo kura ye.Sanni.Sanni jibɔ, nanofɛn minnu bɛ ɲɔgɔn cɛ kitosan/sodiyɔmu/insuline cross-linked nanoparticules hakɛ la, o kɛra 318 nm ye, PDI kɛra 0,18 ye, encapsulation efficiency kɛra 99,4% ye, ani doni hakɛ kɛra 25,01% ye.Reconstitution kɔfɛ, nanoparticules bɛɛ, fo minnu dilannen don ni freeze-drying fɛɛrɛ ye k’a sɔrɔ u ma kɛ of mannitol, maintained their spherical particle structure.Ni an y’a suma ni mannitol bɛ nanoparticules minnu na, minnu farikolojidɛsɛra ni fiyɛli kelen ye, mannitol tɛ minnu na ni fiyɛli ye, olu fana ye fɛnɲɛnamafagalanw hakɛ danmadɔ jira (376 nm) ani doni hakɛ cayalenba (25,02%) ni encapsulation hakɛ kelen ye (98,7%) ani PDI (0,20) ni u jalen don walima freeze-drying techniques.Nanofɛn jalenw ni fiyɛli jalen ye ni manitol tɛ, o fana kɛra sababu ye ka inisɔndiya bɔli teliya la ani ka selilɛriw tali nafa bonya.Nin baara in b’a jira ko fiyɛli jalen bɛ se ka inisɔndiya nanofɛnw farikolojidɛsɛ k’a sɔrɔ a ma kɛ ni cryoprotectants ye ni i y’a suma ni konfirɛzi jalen fɛɛrɛ kɔrɔw ye, o bɛ doni tali seko caman da, ka farali wajibiyalenw dɔgɔya ani ka baara kɛ musakaw nafa ka bon kosɛbɛ.
Kabini a sɔrɔla san 19221,2,3, inisɔndiya ni a furakɛli labɛnw ye sukarodunbana suguya fɔlɔ (T1DM) ni sukarodunbana suguya filanan (T1DM) banabagatɔw ka ɲɛnamaya kisi.Nka, k’a sababu kɛ a ka baarakɛcogo ye i n’a fɔ farikolojɔli dumuni min girinya ka bon, inisɔndiya bɛ fara ɲɔgɔn kan nɔgɔya la, ka tila ni farikolojɔlifɛnw ye, wa a bɛ ban ni tɛmɛsira fɔlɔ ye nɔ.Sukarodunbana suguya fɔlɔ bɛ mɔgɔ minnu na, olu mago bɛ inisɔndiya pikiri la u si bɛɛ kɔnɔ.Banabagatɔ caman minnu sɔrɔla sukarodunbana suguya 2 la fɔlɔ, olu fana ka kan ka inisɔndiya pikiri kuntaalajan ta.Don o don inisɔndiya pikiri ye don o don dimi ni dusukasi sababuba ye nin mɔgɔ ninnu bolo, ni nɔ jugu bila hakili kɛnɛya la.O de kosɔn, inisɔndiya suguya wɛrɛw bɛ dusukasi dɔgɔya, i n’a fɔ i n’a fɔ inisɔndiya da la, olu bɛ ka sɛgɛsɛgɛ kosɛbɛ5 bawo u bɛ se ka sukarodunbanabagatɔ miliyari 5 ɲɔgɔn ka ɲɛnamaya kɛcogo lasegin diɲɛ kɔnɔ.
Nanoparticules (nanoparticules) fɛɛrɛ ye ɲɛtaa caman di a ɲininiw ma ka inisɔndiya da la4,6,7.Min bɛ insuline (insuline) minɛ ka ɲɛ ani ka a tanga tiɲɛni ma walasa ka a lase farikolo yɔrɔ kɛrɛnkɛrɛnnenw na.Nka, nanoparticules (nanoparticules) furakɛli kɛcogo bɛ dan caman na, kɛrɛnkɛrɛnnenya la, ka da fɛnɲɛnamafagalanw (particules suspensions) sabatili koɲɛw kan.Jɛkulu dɔw bɛ se ka kɛ a maratuma na, o min bɛ dɔ bɔ biyoavailability of insuline-loaded nanoparticles8.Ka fara o kan, nanoparticules ni insuline ka polimɛri matirili sabatili kemikaliya fana ka kan ka jateminɛ walasa ka insuline nanoparticules (NPs) sabatili sabati.Sisan, freeze-drying technology ye sanu sariya ye min bɛ NP sabatilenw dilan ka sɔrɔ ka fɛn caman bali ka fɛn caman wuli u marali waati
Nka, nɛnɛ-jalen bɛ a ɲini ka fɛnɲɛnamafagalanw fara a kan walasa ka NPw ka sɛrɛkili jɔcogo bali ka nɔ bila jikuruw ka masinko degun fɛ.O bɛ dɔ bɔ kosɛbɛ inisɔndiya nanofɛnw doni na liyofili kɔfɛ, bawo kiriyɔprotɛki bɛ girinya hakɛ fanba ta.O la, a ka c’a la, inisɔndiyalan NP minnu bɛ bɔ, olu tɛ bɛn mugu jalenw dilanni, i n’a fɔ furakisɛw ni dawolo filimuw, ka da a kan a ka kan ka nanofɛn jalenw hakɛ caman sɔrɔ walasa ka inisɔndiya furakɛli finɛtiri sɔrɔ.
Fiyɛli jalen ye fɛɛrɛ ye min lakodɔnnen don kosɛbɛ, wa a sɔngɔ man gɛlɛn iziniw kɔnɔ, min bɛ kɛ ka mugu jalenw dilan ka bɔ jilamaw la furakɛli baarakɛyɔrɔw la10,11.Kɔlɔsili kɛli fɛnɲɛnamafagalanw dilanni taabolo kan, o bɛ kɛ sababu ye ka fɛnɲɛnamafagalan caman 12, 13 .Ka fara o kan, a kɛra fɛɛrɛ ɲuman ye min bɛ kɛ ka farikolojɔli dumuniw labɛn ka ɲɛ da kɔnɔ.Durring fiyɛli jalen, ji bɛ wuli joona kosɛbɛ, o bɛ dɛmɛ ka fɛnɲɛnamafagalanw kɔnɔna funteni dɔgɔya11,14, o b’a to a kɛli ka se ka funteni-sɛgɛsɛgɛli yɔrɔw datugu.Sanni fiyɛli ka ja, finimugu ja ka kan ka kɛ kelen ye ka ɲɛ ni furaji ye min kɔnɔfɛnw bɛ sɔrɔ a kɔnɔ11, encapsulation in spray-drying improves encapsulation efficiency during dehydration.Ikomi fiyɛli-jalen encapsulation kɛcogo tɛ cryoprotectants wajibiya, fiyɛli-jalen bɛ se ka kɛ ka NP jalenw dilan minnu doni hakɛ ka ca.
Nin kalan in bɛ inisɔndiyalen NPw dilanni fɔ ni kitosan ni sodiyɔmu tripolifosifati ɲɔgɔndan ye ni ion jeli fɛɛrɛ ye.Iyɔn jeli ye labɛnni fɛɛrɛ ye min bɛ sira di nanofɛnw dilanni ma ni kuran jɛɲɔgɔnyaw ye ion suguya fila walima u caman cɛ cogoya dɔw la.Fɛɛrɛ minnu bɛ ja ni fiyɛli ye, olu fila bɛɛ kɛra ka kitosan/sodiyɔmu min kɛra ka ɲɛ tripolyphosphate/insulin cross-linked nanoparticles.Jijibɔ kɔfɛ, u cogoya sɛgɛsɛgɛra ni SEM ye.U ka se ka ɲɔgɔn sɔrɔ kokura, o jateminɛ kɛra ni u bonya tilali ye, u sanfɛla charge, PDI, encapsulation efficiency, ani loading content sumani.Nanoparticules resolubilisées minnu bɛ sɔrɔ inisɔndiyacogo wɛrɛw fɛ, olu jogo fana jateminɛna ni u ka inisɔn lakanani, u bɔli kɛcogo, u bɔli kɛcogo sumani ye ani selilɛriw tali nafa.
Fura ɲagaminen pH ni kitosan ni inisɔndiya hakɛ ye fɛnba fila ye minnu bɛ nɔ bila NP labanw ka fɛnɲɛnamafagalanw hakɛ ni u ka fɛnɲɛnamafagalanw tali nafa (EE) la, bawo u bɛ nɔ bila ionotropique gelation processus la.A jirala ko fura ɲagaminen pH bɛ tali kɛ kosɛbɛ fɛnɲɛnamafagalanw hakɛ ni encapsulation nafama na (Ja 1a).I n’a fɔ a jiralen bɛ cogo min na ja 1a kɔnɔ, iko pH bɛ bonya ka bɔ 4,0 ka se 6,0 ma, fɛnɲɛnamafagalanw hakɛ dantɛmɛnen (nm) dɔgɔyara ani EE cayara kosɛbɛ, k’a sɔrɔ ni pH cayara ka se 6,5 ma, fɛnɲɛnamafagalanw hakɛ danmadɔ y’a daminɛ ka bonya ani EE ma Changé.Ni kitosan ni inisɔndiya hakɛ bɛ bonya, fɛnɲɛnamafagalanw hakɛ danmadɔ fana bɛ bonya.Ka fara o kan, fɛn si ma ye EE la ni nanofɛnw labɛnna ni u girinya hakɛ ye kitosan/insuline min ka ca ni 2,5:1 ye (k/kg) (Ja 1b).O la, labɛncogo ɲuman minnu bɛ nin kalan in kɔnɔ (pH 6,0, kitosan/insuline mass ratio of 2,5:1) olu kɛra ka insuline doni nanoparticules labɛn walasa ka kalan wɛrɛw kɛ.O labɛncogo in kɔnɔ, nanoparticules fɛnɲɛnamafagalanw hakɛ danmadɔ kɛra ka ɲɛ 318 nm (Ja 1c), PDI tun ye 0,18 ye, embedding efficiency tun ye 99,4% ye, zeta potentiel tun ye 9,8 mv ye, ani insuline doni tun ye 25,01% (m/m) ye.Ka da transmission electron microscopy (TEM) jaabiw kan, nanoparticules optimisés tun ye spherical ani discrete ye ni u bonya tun ye kelen ye (Ja 1d).
Insuline nanoparticules ka paramètre optimisation : (a) pH nɔ min bɛ inisɔndi nanoparticules (minnu labɛnna ni chitosan ni insuline mass ratio ye 5:1 ye) cɛmancɛ cɛmancɛ ni u encapsulation efficiency (EE) la (b) kitosan ni Inisɔndiya hakɛ hakɛ nɔ min bɛ inisɔndiya NPw cɛmancɛ cɛmancɛ ni u ka u kɔnɔfɛnw (EE) la (minnu labɛnna pH 6 la); (c) insuline nanoparticules optimisées ka fɛnɲɛnamafagalanw hakɛ tilatilali; (d) TEM mikrografu min bɛ inisɔndiya NP ɲumanw jira .
A dɔnna kosɛbɛ ko kitosan ye polielektroliti barikama ye min ka pKa ye 6,5 ye.A bɛ charge positive sɔrɔ asidifɛnw na bawo a ka amino kuluba bɛ kɛ proton ye ni ion idɔrɔzi ye15.O la, a ka ca a la, a bɛ kɛ tabaga ye walasa ka makɔrɔmolekyulu charge négativelenw datugu.Nin kalan in na, kitosan kɛra ka insuline encapsulate ni isoélectric point of 5.3.Since chitosan is used as a coating material, with the increase of its proportion, the thickness of the outer layer of the nanoparticles increases correspondingly, resulting in a larger average particle size.In addition, higher levels of chitosan can encapsulate more insulin.In our case, EE was highest when the ratio of chitosan and insulin reached 2.5:1, and there was no significant change in EE tuma min na o hakɛ tora ka caya.
Ka fara kitosan ni inisɔndiya hakɛ kan, pH fana jɔyɔrɔ ka bon NPw labɛnni na.Gan et al. 17 ye pH nɔ sɛgɛsɛgɛ kitosan nanofɛnw hakɛ la.U y’a ye ko fɛnɲɛnamafagalanw hakɛ bɛ dɔgɔya ka taa a fɛ fo pH ka se 6,0 ma, wa fɛnɲɛnamafagalanw hakɛ cayara kosɛbɛ pH > 6,0 na, o bɛ bɛn an ka kɔlɔsiliw ma.O ko in bɛ bɔ a la ko ni pH cayara, inisɔndiyalan bɛ surface charge jugu sɔrɔ, o la, a bɛ a fɛ kuran jɛɲɔgɔnyaw ni kitosan/sodium tripolyphosphate (TPP) complexe ye, o bɛ na ni fɛnɲɛnamafagalanw hakɛ fitinin ye ani EE caman .Nka, ni pH ladilanna ka se 6,5 ma, amino kulu minnu bɛ kitosan kan, olu bɔra porotɔni na, o kɛra sababu ye ka kitosan foli kɛ.O cogo la, pH caman bɛ na ni amino ionw dɔgɔyali ye TPP ni inisɔndiya la, o bɛ na ni ɲɔgɔndan dɔgɔyali ye, a laban cɛmancɛ ka bon fɛnɲɛnamafagalanw hakɛ ani EE dɔgɔyali.
NP minnu jalen don nɛnɛ na ani minnu jalen don ni fiyɛli ye, olu cogoya sɛgɛsɛgɛli bɛ se ka jibɔ ni mugu dilanni fɛɛrɛ ɲumanw sugandili ɲɛminɛ.Fɛɛrɛ min ka fisa, o ka kan ka fura sabatili di, ka fɛnɲɛnamafagalanw cogoya kelen, fura doni ka ca ani ka wuli ka ɲɛ furaji fɔlɔ la.Nin kalan in kɔnɔ, walasa ka fɛɛrɛ fila ninnu suma ɲɔgɔn na ka ɲɛ, inisɔndiya NP minnu bɛ ni manitɔli 1% ye walima ni u tɛ u la, olu kɛra jibɔ waati la.Mannitol bɛ kɛ i n’a fɔ a bulking agent or cryoprotectant in various dry powder formulations for freeze drying and spray drying.Ka ɲɛsin insuline nanoparticules lyophilisées ma ni mannitol tɛ, i n’a fɔ a jiralen bɛ cogo min na ja 2a kɔnɔ, mugu jɔcogo dɔ kɔlɔsira min kɔnɔna ka bon, a tɛ cogo la ani a ka gɛlɛn, o kɔlɔsira scanning electron microscopy (SEM) fɛ.Few discrete particles were detected in the powder after dehydration (Ja 2e).Nin jaabiw y’a jira ko NP fanba tolilen don nɛnɛ waati la, k’a sɔrɔ u ma kɛ ni kiriyɔprotectant si tɛ u la.Inisɔn nanoparticules jalen ni fiyɛli la, mannitol 1% bɛ minnu na, nanoparticules spheriques minnu ɲɛw bɛ nɔgɔ, olu kɔlɔsira (Ja 2b,d,f,h).Insuline nanoparticules fiyɛli ni fiyɛli ye ni mannitol tɛ, olu tora spherical nka a kurulen don a sanfɛla la (Ja 2c).Sɛrɛkili ni kurukurulenw bɛ kuma ka taa a fɛ bɔli kɛcogo ni selilɛriw tali sɛgɛsɛgɛliw la jukɔrɔ.Ka da NP jalenw cogoya yelen kan, NP jalenw ni fiyɛli fɛ minnu tɛ mannitɔli ye ani NP minnu jalen don ni fiyɛli ye ni u jalen don ni nɛnɛ ye ani minnu jalen don ni fiyɛli ye ni manitɔli ye, olu fila bɛɛ ye NPs mugu finmanw sɔrɔ (Jaa 2c). 2f,g,h).Ni yɔrɔ min bɛ fɛnɲɛnamafagalanw ni ɲɔgɔn cɛ, o bɛ kɛ sababu ye ka wuli ka caya, o la, a bɔli hakɛ bɛ caya.
Inisɔn NP jilamaw danfaralenw cogoya : (a) Inisɔn NP minnu bɛ liyofili la, olu ja SEM la, ni manitol tɛ u la; (b) SEM ja min bɛ inisɔndiya NPw kan minnu bɛ liyofili ni manitɔli ye; (c) inisɔndiya NP minnu jalen don ni fiyɛli ye ni manitol SEM ja tɛ u la ; (d) Inisɔn NPw ja SEM minnu jalen don ni manitol ye ni fiyɛli ye; (e) inisɔn NPs mugu liyofililen ja min tɛ manitɔli ye; (f) inisɔndiya NP minnu bɛ liyofili la ni manitɔli ye, olu ja; ( g) Inisɔn NPs mugu min jalen don ni fiyɛli ye, o ja min tɛ manitɔli ye; (h) inisɔn NPs mugu min jalen don ni fiyɛli ye ni manitol ye, o ja.
Jalen waati la, manitɔli bɛ kɛ i n’a fɔ sɔgɔsɔgɔninjɛlan, ka NPw mara cogo la min tɛ fɛn ye ani ka tiɲɛni bali ni jikuruw ye19.O ni jikuruw jalen tɛ kelen ye.O de kama, mannitɔli man kan ka kɛ nin fɛɛrɛ in na.Tiɲɛ na, NP minnu jalen don ni fiyɛli ye ni mannitɔli tɛ, olu ye NP finmanw di i n’a fɔ a ɲɛfɔra cogo min ka tɛmɛ.Nka, a bɛ kɛ cogo min na, o tɛ kelen ye, nɛnɛ kɛcogo tɛ kɛ ni jisuma jalen ye. manitɔli bɛ se ka kɛ hali bi fasa ye fiyɛli-jalen kɛcogo la walasa ka NPw di u ma ka kɛ sɛrɛkili ye20 (Ja 2d), o bɛ dɛmɛ ka o NP suguw bɔli kɛcogo kelen sɔrɔ.Ka fara o kan, a jɛlen don ko fɛnɲɛnamafagalan belebele dɔw bɛ se ka sɔrɔ inisɔndiya NP minnu jalen don nɛnɛ na ani minnu jalen don ni fiyɛli ye, mannitol bɛ minnu na (Ja 2b,d), o bɛ se ka kɛ sababu ye ka u dalajɛ manitol min bɛ fɛnɲɛnamafagalanw kɔnɔ ka fara insuline min bɛ a kɔnɔ. To.Chitosan layer.A ka kan ka kɔlɔsi ko nin kalan in kɔnɔ, walasa ka a jira ko sɛrɛkili jɔcogo ka to a cogo la farikolojidɛsɛ kɔfɛ, manitɔli ni kitosan hakɛ bɛ to 5:1 na, walasa fasa hakɛ caman fana ka se ka NP jalenw ka fɛnɲɛnamafagalanw hakɛ bonya.
Fourier transform infrared attenuated total reflection (FTIR-ATR) spectroscopie ye inisɔndiya fu, kitosan, chitosan, TPP ani insuline ɲagaminen farikolo tacogo jira.NP farikolojidɛsɛlenw bɛɛ kɛra ni FTIR-ATR spectroscopy ye.Kɛrɛnkɛrɛnnenya la, band intensities 1641, 1543 ani 1412 cm-1 kɔlɔsira NP encapsulé kɔnɔ freeze-dried with mannitol and in NPs spray-dried with and without mannitol (Ja 3).I n’a fɔ a fɔra cogo min na ka tɛmɛ, nin fanga bonya ninnu tun bɛ tali kɛ ɲɔgɔndan na kitosan, TPP ani insuline cɛ.Sɛgɛsɛgɛli min kɛra kitosan ni insuline cɛ jɛɲɔgɔnya kan, o y’a jira ko FTIR spectres kɔnɔ inisɔn doni kitosan nanoparticules kɔnɔ, kitosan band bɛ ɲɔgɔn sɔrɔ ni insuline ta ye, ka dɔ fara a kan karɔbɔli fanga (1641 cm-1) ani amine (1543 cm-1) cɛsirilan.TPP ka tripolifosifati kuluw bɛ tali kɛ amoniyɔmu kuluw la kitosan kɔnɔ, ka kɛ bandi ye santimɛtɛrɛ 1412 la.
FTIR-ATR spectres de insuline gratuite, chitosan, chitosan/TPP/insuline ani NPs farikolo ɲagaminenw farikolojidɛsɛ furakɛcogo wɛrɛw fɛ.
Ka fara o kan, nin jaabi ninnu bɛ bɛn ni SEM kɔnɔ jateminɛw ye, o min y’a jira ko NP minnu bɛ kɛ kaburu kɔnɔ, olu tora u cogo la ni u fiyɛra ani ni u jalen don ni mannitɔli ye, nka ni manitɔli tɛ yen, fiyɛli jalen dɔrɔn de bɛ fɛnɲɛnamafagalanw bɔ minnu sirilen don ka chitosan, TPP, ani insuline.Nin jaabi b’a jira ko chitosan, TPP ani insuline cɛsiraw tɛ sɔrɔ tugun NPw la minnu jalen don ni konfirɛzi ye ni mannitol tɛ.NPw jɔcogo tiɲɛna frize-drying waati la ni cryoprotectant tɛ, o bɛ se ka ye SEM jaabiw la (Ja 2a).Ka da inisɔndiyalan NPw cogoya ni FTIR jaabiw kan, dɔrɔn NP minnu kɛra ni liyofili ye, minnu jalen don ni fiyɛli ye, ani minnu tɛ manitɔli ye, olu kɛra ka kɛɲɛ ni sigicogo kura kɔlɔsili ye ani NP minnu tɛ mannitɔli ye, ka da NP minnu tɛ mannitɔli ye, olu tiɲɛni ye farikolojidɛsɛ waati. ka jɛkafɔ kɛ.
Farikolojidɛsɛ bɛ kɛ ka u mara waati jan kɔnɔ ani ka u labɛn kokura ka kɛ fura wɛrɛw ye.NP jalenw seko ka se ka u labɛn kokura u maralen kɔfɛ, o nafa ka bon kosɛbɛ u kɛli la furakɛlicogo wɛrɛw la i n’a fɔ furakisɛw ni filimuw.An y’a kɔlɔsi ko inisɔndiya NP minnu jalen don ni fiyɛli ye, olu ka fɛnɲɛnamafagalan hakɛ danmadɔ bɛ caya dɔɔni dɔrɔn u labɛncogo kura kɔfɛ.Fan wɛrɛ fɛ, fiyɛli kɛlen kɔfɛ ani insuline nanoparticules congelées avec mannitol, o cayara kosɛbɛ (Taabolo 1).PDI ni EE ma Changé kosɛbɛ (p > 0,05) NP bɛɛ lajɛlen kɔfɛ nin kalan in kɔnɔ (Taabolo 1).Nin jaabi in b’a jira ko particule fanba tora u cogo la u wulilen kɔfɛ kokura.Nka, manitol farali kɛra sababu ye ka insuline doni dɔgɔya kosɛbɛ lyophilized ani manitol nanoparticules jalen ni fiyɛli ye (Taabolo 1).O ni NP minnu jalen don ni fiyɛli ye ni mannitol tɛ u la, olu ka insuline doni hakɛ tora a cogo kɔrɔ la (Taabolo 1).
A dɔnna kosɛbɛ ko nanofɛnw doni nafa ka bon kosɛbɛ ni u bɛ kɛ fura dicogo la.Ni doni ka dɔgɔn NPw la, fɛn camanba de ka kan ka kɛ walasa ka se furakɛli dakun na.Nka, o NP hakɛ camanba suguw ka sɔgɔsɔgɔli ka bon, o bɛ na ni gɛlɛya ni gɛlɛya ye da la ani pikiri kɛli la, o cogo kelen na 22 .Ka fara o kan, NPw bɛ se ka kɛ fana bɛ kɛ ka furakisɛw ni biyofilimu sɔgɔlenw dilan23, 24, o min bɛ NP hakɛ caman kɛli wajibiya ni doni hakɛ dɔgɔyara, o bɛ na ni furakisɛ belebelebaw ni biyofilimu belebelebaw ye minnu tɛ bɛn da la.O la, NP minnu farikolojidɛsɛ don, ni inisɔndiya doni ka ca, olu ka ɲi kosɛbɛ.An ka jaabiw b’a jira ko NP minnu jalen don ni fiyɛli ye mannitɔli tɛ minnu na, olu ka inisɔndiya doni caman bɛ se ka caman di nafa minnu bɛ mɔgɔ sama ka taa o lasecogo wɛrɛw la.
NP farikolojidɛsɛlenw bɛɛ bilala friji kɔnɔ kalo saba kɔnɔ.SEM jaabiw y’a jira ko NP jilamaw bɛɛ cogoya ma Changé kosɛbɛ kalo saba marali kɔnɔ (Ja 4).Ni u labɛnna kokura ji la, NP bɛɛ ye EE dɔgɔya dɔɔni jira ani ka inisɔndiya hakɛ dɔɔni (~5%) bɔ kalo saba mara waati kɔnɔ (Taabolo 2).Nka, nanofɛnw bɛɛ ka fɛnɲɛnamafagalanw hakɛ danmadɔ cayara.NP minnu jalen don ni fiyɛli ye ni mannitɔli tɛ u la, olu hakɛ cayara ka se 525 nm ma, ka sɔrɔ NP minnu jalen don ni fiyɛli ye ani minnu jalen don nɛnɛ na ni mannitɔli ye, olu hakɛ cayara ka se 872 ani 921 nm ma (Taabolo 2).
Inisɔn NP jilama suguya minnu maralen don kalo saba kɔnɔ, olu cogoya: (a) Inisɔn NP minnu bɛ liyofili ni manitɔli ye, olu ja SEM; (b) SEM ja min bɛ inisɔndiya nanoparticules jalen fiyɛli la ni mannitol tɛ u la; (c) ni manitol SEM ja tɛ inisɔndiya NPw jalenw na ni fiyɛli ye.
Ka fara o kan, sanjiw yelen don inisɔndiya nanofɛnw na minnu labɛnna kokura, minnu jalen don ni manitoli ye ni fiyɛli ye ani minnu jalen don ni nɛnɛ ye (Ja S2).O bɛ se ka sɔrɔ fɛnɲɛnamafagalan belebelebaw ma minnu ma jɔ ji la ka ɲɛ.Nin jateminɛ minnu kɛra sanfɛ, olu bɛɛ b’a jira ko fiyɛli jalen fɛɛrɛ bɛ se ka inisɔndiya nanofɛnw tanga farikolojidɛsɛ ma ani ko inisɔndiya nanofɛnw doni caman bɛ se ka sɔrɔ ni fiyɛri si tɛ u la walima ka u ja cryoprotectants (banakisɛfagalanw).
Insuline marali sɛgɛsɛgɛra pH = 2,5 fɛnɲɛnamafagalan na ni pepsin, tripsin ani α-chymotrypsin ye walasa ka NPw tangacogo jira ka bɔ anzimu dumunikɛbaliya la farikolojidɛsɛ kɔfɛ.NPw farikolojidɛsɛlenw ka inisɔndiya marali ni NP labɛnna kura ta kɛra ɲɔgɔn fɛ, ani inisɔndiyalan gansan kɛra kɔrɔsili jugu ye.Nin sɛgɛsɛgɛli in na, inisɔndiya fu ye inisɔndiyalan bɔli teliya jira 4 kɔnɔ h enzyme furakɛli saba bɛɛ la (Ja 5a–c).O ni ɲɔgɔn cɛ, NP minnu jalen don ni mannitol ye ni nɛnɛ ye ani NP minnu jalen don ni fiyɛli ye ni mannitol ye walima ni u tɛ u la, olu ka inisɔndiya bɔli sɛgɛsɛgɛli y’a jira ko nin NP ninnu lakanani ka bon kosɛbɛ anzimu dumunikɛbaliya la, o ni inisɔndiya NP labɛnna kura ta bɔlen don ( ja 1).5a-c).Ni nanoparticules dɛmɛ ye pepsin kɔnɔ, tripsin, ani α-chymotrypsin, insuline ka ca ni 50%, 60% ani 75% ye, o tun bɛ se ka lakana lɛrɛ 4 kɔnɔ, o cogo kelen na (Ja 5a–c).O inisɔndiya lakanani se bɛ se ka dɔ fara inisɔndiya caman sɔrɔli kan joli la25.Nin jaabi ninnu b’a jira ko ni fiyɛli jalen don ni manitol ye walima ni mannitol tɛ a la ani ka jalen kɛ ni mannitol ye NPw ka se ka inisɔndiya tanga farikolojidɛsɛ kɔfɛ.
Inisɔn NPw lakanani ni u bɔli kɛcogo : (a) inisɔndiya lakanani pepisini jilama la; (b) inisɔndiya lakanani tripsin jilama la; (c) inisɔndiya lakanani ni α-kimotiripisini jilama ye; ( d) NP jilamaw bɔli kɛcogo pH = 2,5 jilama kɔnɔ; (e) NP jilamaw bɔli kɛcogo pH = 6,6 jilama kɔnɔ; (f) NP jilamaw bɔli kɛcogo pH = 7,0 jilama la.
Inisɔn NP jalen minnu labɛnna kokura ani minnu labɛnna kokura, olu bilala tampon suguya caman kɔnɔ (pH = 2,5, 6,6, 7,0) 37 °C la, ka kɔnɔbara, kɔnɔbara basigilen ani banakɔtaa sanfɛla pH sigida ladege, walasa ka inisɔndiya nɔ sɛgɛsɛgɛ inisɔndiya kɛlɛli la. Labɔli kɛcogo sigida wɛrɛw la.Kɔnɔbaralabana yɔrɔ yɔrɔ dɔ.Ni pH = 2,5 ye, NP minnu bɛ inisɔndiya la ani inisɔndiyalan NP jalenw minnu bɛ wuli ka bɔ u nɔ na, ​​olu ye bɔli fɔlɔ jira ka bɔ lɛrɛ kelen fɔlɔ kɔnɔ, o kɔfɛ, bɔli teliya la lɛri 5 nataw kɔnɔ (Ja 5d).O bɔli teliya la daminɛ na, a ka c’a la, o bɛ bɔ farikolojɔli dumuniw bɔli teliya la, minnu tɛ dafalen ye immobilized in the internal structure of the particle.Ni pH = 6,5 ye, NP minnu bɛ inisɔndiya la ani insuline NP jalenw minnu labɛnna kokura, olu ye bɔli nɔgɔya ni dɔɔni dɔɔni jira lɛrɛ 6 kɔnɔ, barisa sɛgɛsɛgɛli furaji pH tun bɛ tali kɛ NPw labɛnna furaji la (Ja 5e).Ni pH = 7 ye, NPw tun tɛ sabati ani u tun bɛ ɲini ka tiɲɛ pewu sanga fila fɔlɔ kɔnɔ (Ja 5f).O bɛ kɛ barisa kitosan deprotonation bɛ kɛ pH caman na, o bɛ na ni polimɛri rezo ye min tɛ ɲɔgɔn ta kosɛbɛ ani inisɔndiya donilen bɔli.
Ka fara o kan, inisɔndiya NP minnu jalen don ni fiyɛli ye ni mannitɔli tɛ, olu ye bɔli teliya jira ka tɛmɛ NP farikolojidɛsɛlen tɔw kan (Ja 5d–f).I n’a fɔ a ɲɛfɔra cogo min ka tɛmɛ, inisɔndiya NP minnu jalen don kokura ni manitɔli tɛ, olu ye fɛnɲɛnamafagalan hakɛ fitinin jira.Fura misɛnninw bɛ yɔrɔba di, o la fura min bɛ tali kɛ o la, o fanba bɛna kɛ fɛnɲɛnamafagalanw sanfɛla la walima a kɛrɛfɛ, o bɛ na ni fura bɔli teliya ye
NPw ka sitotokisisi sɛgɛsɛgɛra ni MTT sɛgɛsɛgɛli ye.I n’a fɔ a jiralen bɛ cogo min na ja S4 kɔnɔ, a jirala ko NP minnu farikolojidɛsɛlen don, olu bɛɛ tɛ nɔba bila seliluw ka ɲɛnamaya la ni u hakɛ ye 50–500 μg/ml ye, o b’a jira ko NP farikolojidɛsɛlenw bɛɛ bɛ se ka baara kɛ ni u ye lafiya la walasa ka se furakɛli finɛtiri la.
Sugunɛ ye farikolo yɔrɔba ye, inisɔndiya bɛ a farikolo baara kɛ min fɛ.HepG2 selilɛriw ye hadamadenw ka sugunɛbilenni selilɛri ye min bɛ kɛ ka caya i n’a fɔ sugunɛbilenni tali misali in vitro.Yan, HepG2 selilɛriw kɛra ka NPw minɛcogo jateminɛ selilɛriw la minnu farikolojidɛsɛlen don ni frize-drying ani spray-drying fɛɛrɛw ye.Cellular uptake by confocal laser scanning using flow sitometri ni yeli kɔfɛ lɛri caman kɔnɔ ni FITC inisɔndiyalan fu ye min hakɛ ye 25 μg/mL ye, FITC inisɔndiyalan NP minnu labɛnna kura ye ani FITC inisɔndiyalen NP minnu farikolojidɛsɛ ye inisɔndiya hakɛ kelen na, olu kɔlɔsili kɛra test.NP minnu labɛnna kura inisɔndiyalen don, NP minnu dilannen don ni manitol ye, ani NP minnu jalen don ni fiyɛli ye ni mannitol ye ani minnu tɛ u la (Ja 6a) olu ka selilu kɔnɔ fluorescence fanga tun ka ca ni 4,3, 2,6, 2,4 ani 4,1 ye ka tɛmɛ hɔrɔnyalenw kan.FITC-insuline kulu, o cogo kelen na (Ja 6b).Nin jaabiw b’a jira ko inisɔndiya min bɛ don a kɔnɔ, o fanga ka bon selilɛriw tali la ka tɛmɛ inisɔndiyalan gansan kan, kɛrɛnkɛrɛnnenya la, o bɛ bɔ inisɔndiyalan nanofɛnw hakɛ dɔgɔyali la minnu bɛ bɔ kalan in na.
HepG2 selilɛriw tali kɔfɛ lɛrɛ 4 kɔnɔ ni NP labɛnnen kuraw ni NPw ye minnu farikolojidɛsɛlen don: (a) FITC-insuline tali tilatilali HepG2 selilɛriw fɛ.(b) Fluorescence fanga hakɛ jateminɛlen jateminɛlen ni flow cytometry ye (n = 3), *P < 0,05 ni inisɔndiya gansan ta kɛra ɲɔgɔn fɛ.
O cogo kelen na, CLSM jaw y’a jira ko FITC-insuline-loaded NPs labɛnna kuraw ni FITC-insuline-loaded spray-dried NPs (mannitol tɛ minnu na) olu ka FITC fluorescence fanga fanga tun ka bon kosɛbɛ ka tɛmɛ sample tɔw ta kan (Ja 6a).Ka fara o kan, ni mannitol farala a kan, ni furaji in sɔgɔli ka bon, o bɛ dɔ fara u ka se ka bana kɛlɛ ka se selilɛriw tali ma, o bɛ na ni inisɔndiya caya dɔgɔyali ye.Nin jaabi ninnu b’a jira ko NP minnu jalen don ni fiyɛli ye, ni mannitɔli tɛ minnu na, olu ye selilɛriw tacogo ɲuman jira bawo u fɛnɲɛnamafagalanw hakɛ tun ka dɔgɔn ni NP jalenw ta ye minnu bɛ kɛ nɛnɛ na, u wulilen kɔfɛ.
Kitosan (a girinya hakɛ cayalenba ye 100 KDa, 75–85% deacetylated) sanna Sigma-Aldrich fɛ.(Oakville, Ontario, Canada).Sodium tripolyphosphate (TPP) sanna VWR fɛ (Radnor, Pennsylvania, USA).Hadamaden ka insuline recombinant min kɛra nin ɲinini in na, o bɔra Fisher Scientific fɛ (Waltham, MA, USA).Fluorescein isothiocyanate (FITC)-labeled human insulin and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) sanna Sigma-Aldrich fɛ.(Oakville, Ontario, Canada).HepG2 selilɛri sɔrɔla ATCC fɛ (Manassas, Virginia, USA).Reactif tɔw bɛɛ tun ye analytique walima chromatographie grade ye.
Aw bɛ CS jilama miligaramu 1/ml labɛn ni aw ye a wuli ji fila la (DD ji) la, asidi asetiki 0,1% bɛ min na.Aw bɛ TPP ni inisɔndiya miligaramu 1/ml labɛn ni aw ye u wuli DD ji la ani asidi asetiki 0,1% la.A pre-emulsion labɛnna ni politrɔni PCU-2-110 teliyaba homojenizari ye (Brinkmann Ind. Westbury, NY, USA).A labɛncogo ye nin ye: a fɔlɔ, TPP jilama mililitiri 2 bɛ fara inisɔndiya mililitiri 4 kan, ka sɔrɔ ka o ɲagaminen wuli miniti 30 kɔnɔ ka a ɲagami ka ɲɛ.O kɔfɛ, ji ɲagaminen bɛ fara CS jilama kan dɔɔni dɔɔni pikirijikɛlan fɛ ni wuli teliya la (10.000 rpm).A ɲagaminenw bilala teliya la ka u lamaga (15.000 rpm) jisuma na miniti 30 kɔnɔ, ka u ladilan ka se pH dɔ ma walasa ka inisɔndi NPw sɔrɔ minnu bɛ ɲɔgɔn cɛ.Walisa ka inisɔndiya NPw kɛ kelen ye ka taa a fɛ ani ka u fɛnɲɛnamafagalanw hakɛ dɔgɔya, u ye sonikɛ miniti 30 wɛrɛ kɔnɔ jisuma na ni sonikatɛri suguya ye (UP 200ST, Hielscher Ultrasonics, Teltow, Alimanjamana).
Insuline NPS sɛgɛsɛgɛra Z-average diameter, polydispersity index (PDI) ani zeta potentiel (zeta potentiel) la ni dynamic light scattering (DLS) sumanikɛlanw ye ni Litesizer 500 (Anton Paar, Graz, Austria) ye ni u dilancogo ye DD ji la 25°C la.Morphology ni bonya tilatilali kɛra Hitachi H7600 transmission electron microscope (TEM) fɛ (Hitachi, Tokyo, Japon), ani jaw sɛgɛsɛgɛra o kɔfɛ ni Hitachi ja porogaramu ye (Hitachi, Tokyo, Japon).Walisa ka insuline NPw ka encapsulation efficiency (EE) ani loading capacity (LC) jateminɛ, NPw kɛra pipet ye ka don ultrafiltration tubes kɔnɔ ni molecular weight cut-off ye 100 kDa ye, ka centrifuged kɛ 500 xg la fo 30 min.Insuline min ma kɛ filtrate kɔnɔ, o hakɛ jateminɛna ni Agilent 1100 Series HPLC system (Agilent, Santa Clara, California, USA) ye min bɛ kɛ ni pɔnpe naani ye, autosampler, column heater, ani DAD detector.Insuline sɛgɛsɛgɛra ni C18 koloma ye (Zorbax, 3,5 μm, 4,6 mm × 150 mm, Agilent, Etazuni) ani a kɔlɔsira ni 214 nm ye.Faza mobili tun ye asetonitirili ni ji ye, TFA 0,1% bɛ min na, gradient ratios bɛ bɔ 10/90 na ka se 100/0 ma, ani ka boli miniti 10 kɔnɔ.Fase mobile tun bɛ pɔnpe ni jibɔ hakɛ ye 1,0 ml/min.Colon funteni bilala 20 °C la.Aw bɛ EE ni LC kɛmɛsarada la jate ni equations ye.(1) ani Eq.(2) ye.
CS/insuline hakɛ suguya caman minnu bɛ daminɛ 2,0 na ka se 4,0 ma, olu sɛgɛsɛgɛra walasa ka insuline NP kɛcogo ɲuman sɔrɔ.CS jilama hakɛ danfaralenw farala a kan labɛnni senfɛ, ka sɔrɔ insuline/TPP ɲagaminen tora a cogo la.Insuline NPw labɛnna pH hakɛ la min bɛ se 4,0 fo 6,5 ma ni ɲagaminen pH kɔlɔsili ye ka ɲɛ furaji bɛɛ faralen kɔfɛ (insuline, TPP ani CS).Insuline nanoparticules ka EE ni particules hakɛ jateminɛna pH hakɛw ni CS/insuline mass ratios danfaralenw na walasa ka insuline NPw dilancogo ɲɛ.
Insuline NPs optimisés bilala aliminiyɔmu minɛn kan, ka u datugu ni tissue ye min sirilen don ni kase dɔw ye.O kɔfɛ, minɛn minnu sirilen don, olu bilala Labconco FreeZone freeze dryer kɔnɔ (Labconco, Kansas City, MO, USA) min bɛ ni tray dryer ye.Future ni vacuum tansiyɔn bilala -10 °C, 0,350 Torr la lɛrɛ 2 fɔlɔ kɔnɔ, ani 0 °C ani 0,120 Torr lɛrɛ 22 tɔ la lɛrɛ 24 kɔnɔ walasa ka inisɔndiya NP jalenw sɔrɔ.
Buchi Mini Spray Dryer B-290 (BÜCHI, Flawil, Suwisi) kɛra ka insuline encapsulé (insuline encapsulé) dilan.Jalen kɛcogo sugandilenw tun ye : funteni 100 °C, balo bɔli 3 L/min, ani gazi bɔli 4 L/min.
Insuline NPs ka kɔn farikolojidɛsɛ ɲɛ ani a kɔfɛ, olu taamasiɲɛw kɛra ni FTIR-ATR spectroscopie ye.Nanoparticules déhydrates ani insuline libres ani chitosan, olu sɛgɛsɛgɛra ni Spectrum 100 FTIR spectrophotometer (PerkinElmer, Waltham, Massachusetts, USA) ye min bɛ ni universel ATR sampling accessoire (PerkinElmer, Waltham, Massachusetts, USA) ye.Signal cɛmancɛw sɔrɔla ka bɔ sɛgɛsɛgɛli 16 la ni janya ye santimɛtɛrɛ 4 ye santimɛtɛrɛ 4000-600 hakɛ la.
Insuline NP jalenw cogoya jateminɛna ni SEM ja ye minnu bɛ inisɔndiya NP jalenw ni fiyɛli la minnu minɛna Helios NanoLab 650 Focused Ion Beam-Scanning Electron Microscope (FIB-SEM) fɛ (FEI, Hillsboro, Oregon, Etazuni). Paramɛtiri kunba min kɛra, o tun ye tension 5 keV ye ani current 30 mA.
Insuline NP minnu farikolojidɛsɛlen don, olu bɛɛ wulila kokura ji dd kɔnɔ.A fɛnɲɛnamafagalanw hakɛ, PDI, EE ani LC sɛgɛsɛgɛra kokura ni fɛɛrɛ kelen ye min kofɔra ka tɛmɛ walasa k’u jogo jateminɛ farikolojidɛsɛ kɔfɛ.Anhydroinsuline NPw sabatili fana jateminɛna ni NPw jogo sɛgɛsɛgɛli ye u marali waati jan kɔfɛ.Nin kalan in kɔnɔ, NP bɛɛ lajɛlen jibɔlen kɔfɛ, olu maralen don friji kɔnɔ fo saba kalo.Kalo saba maralen kɔfɛ, NPw sɛgɛsɛgɛra ka ɲɛsin u cogoya ma, PDI, EE ani LC.
Aw bɛ NPw labɛn kokura mililitiri 5 kɛ mililitiri 45 la, kɔnɔbaraji simulatigɛlen (pH 1,2, pepsin 1% bɛ min na), banakɔtaa ji (pH 6,8, tripsin 1% bɛ min na) walima kimotiripisini jilama (100 g/mL, fositɛri tampon na, pH 7,8) bɛ min na walasa ka inisɔndiya nafa jateminɛ NPw lakanani na o kɔfɛ farikolojidɛsɛ.U bilala 37°C la ni wuli teliya ye 100 rpm.500 μL furaji lajɛra waati danfaralenw na ani inisɔndiya hakɛ jateminɛna ni HPLC ye.
Insuline NP minnu labɛnna kokura ani minnu ji ma bɔ u la, olu bɔli kɛcogo in vitro sɛgɛsɛgɛra ni dialyse bag fɛɛrɛ ye (molecules weight cut-off 100 kDa, Spectra Por Inc.).NP jalenw labɛnna kokura ani minnu labɛnna kokura, olu diyalizera ji la minnu pH ye pH 2,5, pH 6,6, ani pH 7,0 (0,1 M phosphate-buffered saline, 0,1 M phosphate-buffered saline PBS) walasa ka kɔnɔbara, kɔnɔbara basigilen ani sanfɛla banakɔtaa pH sigida ladege.Aw bɛɛ lajɛlen tun bɛ tobi 37 °C la ni u lamagacogo basigilen ye ni 200 ye miniti kɔnɔ.Aw bɛ ji sama ka bɔ mililitiri 5 diyalizi foroko kɔfɛ nin waati ninnu na: 0,5, 1, 2, 3, 4, ani 6 h, ka sɔrɔ ka a hakɛ fa ni kura ye o yɔrɔnin bɛɛ dialysate.Insuline nɔgɔli ji la, o sɛgɛsɛgɛra ni HPLC ye, ani inisɔndiya bɔli hakɛ min bɛ bɔ nanofɛnw na, o jate kɛra ka bɔ inisɔndiya gansan bɔlen ni inisɔndiya bɛɛ lajɛlen hakɛ la min bɛ nanofɛnw kɔnɔ (Equation 3).
Adamadenw ka sugunɛbilenni kansɛri selilɛri HepG2 seliluw falenna minɛnw kɔnɔ minnu janya ye milimɛtɛrɛ 60 ye ni Dulbecco’s Modified Eagle’s Medium (DMEM) ye min kɔnɔ, den kɔnɔbara basigilen 10%, penisilini 100 IU/mL, ani 100 μg/mL streptomycine29.Sɛnɛfɛnw maralen don 37°C la, 95% danfaralen nɛnɛ, ani 5% CO2.Ka ɲɛsin minɛni sɛgɛsɛgɛliw ma, HepG2 seliluw tun bɛ kisɛ kɛ ni selilu 1 × 105/ml ye Nunc Lab-Tek chamber slide system kan kɔlɔn 8 kɔnɔ (Thermo Fisher, NY, Etazuni).Ka kɛɲɛ ni sitotokisi sɛgɛsɛgɛli ye, u tun bɛ kisɛ kɛ kɔlɔn 96 tabali kɔnɔ (Corning, NY, Etazuni) ni u hakɛ ye selilu 5 × 104 ye/ml.
MTT sɛgɛsɛgɛli kɛra ka inisɔndiya NPs30 labɛn kuraw ni farikolojidɛsɛlenw ka sitotokisi jateminɛ.HepG2 seliluw kɛra kɔlɔn 96 kɔnɔ, u hakɛ ye 5 × 104 selilu/mL ye, ka u falen tile 7 kɔnɔ ka sɔrɔ ka sɛgɛsɛgɛli kɛ.Insuline NPw kɛra ka u kɛ hakɛ caman na (50 fo 500 μg/mL) sɛnɛfɛnw na, o kɔfɛ a bɛ di seliluw ma.Lɛrɛ 24 kɔfɛ, seliluw ko siɲɛ 3 ni PBS ye ka u tobi ni fɛnɲɛnamafagalan ye min kɔnɔ MTT miligaramu 0,5/ml bɛ sɔrɔ fo lɛri 4 wɛrɛ.Sitotokisi jateminɛ kɛra ni tetarazoliyɔmu MTT jɛman ka dɔgɔyali anzimatiki sumani ye ka se fɔrmazan bulama ma 570 nm la ni Tecan infinite M200 pro spectrophotometer ye tabali kalanbaga (Tecan, Männedorf, Suwisi).
NPw ka selilɛri tacogo ɲuman sɛgɛsɛgɛra ni confocal laser scanning microscopy ani flow cytometry analysis ye.Nunc Lab-Tek chamber slide system kɔlɔn kelen-kelen bɛɛ furakɛra ni FITC-insuline hɔrɔnyalen ye, FITC-insuline-loaded NPs, ka 25 μg/mL FITC-insulin NPs jilamaw labɛn kokura u hakɛ kelen na ka u bila ka u bila Lɛrɛ 4.Kɔnɔw ko siɲɛ 3 ni PBS ye ka u jɔ ni paraformaldehyde 4% ye.Nukiliw nɔgɔlen don ni 4′,6-diamidino-2-phenylindole (DAPI) ye.Insuline localisation kɔlɔsira ni Olympus FV1000 laser scanning/two-photon confocal microscope ye (Olympus, Shinjuku City, Tokyo, Japan).For flow sitomɛtiri sɛgɛsɛgɛli la, FITC-insuline hɔrɔnyalen 10 μg/mL, FITC-insuline-loaded NPs, ani FITC-insuline NPs jilama minnu bɛ wuli ka bɔ a nɔ na, ​​olu hakɛ kelenw farala kɔlɔn 96-platiw kan minnu kisɛra ni HepG2 selilɛriw ye ka u bila sanga 4 kɔnɔ .Lɛrɛ 4 kɛlen kɔfɛ, seliluw bɔra ka u ko siɲɛ 3 ni FBS.5 × 104 selilɛriw sɛgɛsɛgɛli kɛra ni BD LSR II flow cytometer ye (BD, Franklin Lakes, New Jersey, Etazuni).
Nafa bɛɛ b’a jira i n’a fɔ cɛmancɛ ± danfaralan.Jɛkulu bɛɛ ni ɲɔgɔn cɛ danfara minnu kɛra, olu jateminɛna ni sira kelen ANOVA walima t-test ye IBM SPSS Statistics 26 fɛ Mac fɛ (IBM, Endicott, New York, Etazuni) ani p < 0,05 jatera jateminɛ nafama ye.
Nin kalan in b’a jira ko fiyɛli jalen bɛ se ka kɛ ka jibɔ ni kitosan/TPP/insuline nanoparticules cross-linked ye ni u labɛncogo ɲuman ye ni i y’a suma ni freeze-drying methods standards ye minnu bɛ kɛ ni bulking agents walima cryoprotectants capacity ye ani ka doni hakɛ caman sɔrɔ.The optimized insulin nanoparticles yielded an average particle size of 318 nm and an encapsulation efficiency of 99.4%.SEM and FTIR results after dehydration showed that the spherical structure was maintained only in spray-dried NPs with and without mannitol and lyophilized with mannitol, but lyophilized NPs without mannitol were decomposed during dehydration.In the reconstitution ability test, insulin nanoparticles spray-dried without mannitol showed the smallest mean particle size and the highest loading upon reconstitution.Nin NP farikolojidɛsɛlenw bɛɛ bɔli kɛcogo y’a jira ko u bɛ bɔ teliya la jilamaw la minnu ye pH = 2,5 ani pH = 7 ye, wa u bɛ sabati kosɛbɛ jilamaw la minnu pH ye pH = 6,5 ye.Ni an y’a suma ni NP jilamalen wɛrɛw ye minnu wulilen don kokura, NP minnu jalen don ni fiyɛli ye ni manitol tɛ, olu ye bɔli teliya jira.O jaabi bɛ bɛn ni min kɔlɔsira selilɛri wulicogo sɛgɛsɛgɛli la, i n’a fɔ NP minnu jalen don ni fiyɛli ye ni mannitɔli tɛ yen, olu bɛ se ka NP labɛnnen kuraw ka selilɛri tali nafa sabati pewu.Nin jaabi ninnu b’a jira ko inisɔndiya nanofɛn jalenw minnu labɛnna ni fiyɛli jalen ye ni mannitɔli tɛ minnu na, olu bɛnnen don kosɛbɛ ka baara kɛ ka taa a fɛ ka kɛ fura tata hakɛ wɛrɛw ye minnu tɛ ji sɔrɔ, i n’a fɔ furakisɛ minnu bɛ ta da la walima filimu minnu bɛ kɛ ni biyoadhesive ye.
K’a sababu kɛ hakililata koɲɛw ye, kunnafonidilan minnu kɛra ani/walima minnu sɛgɛsɛgɛra nin kalan in senfɛ, olu tɛ sɔrɔ foroba la, nka u bɛ sɔrɔ sɛbɛnnikɛla kelen-kelen bɛɛ fɛ ni ɲinini kɛra cogo bɛnnen na.
Kagan, A. Sukarodunbana suguya 2: a bɔyɔrɔ sigida ni dɔnniya la, furakɛli gɛlɛyaw, ani a nɔfɛkow banabagatɔw ni mɔgɔ wɛrɛw la.(McFarlane, 2009).
Singh, AP, Guo, Y., Singh, A., Xie, W. & Jiang, P. Insuline encapsulation yiriwali: yala a tali da la bɛ se ka kɛ sisan wa?J. Farmasi.bio-farmasi.jidagayɔrɔ.1, 74–92 (2019).
Wong, CY, Al-Salami, H. & Dass, CR Kɔsa in na, ɲɛtaa minnu kɛra liposomew dicogo la minnu bɛ inisɔndiya da la, sukarodunbana furakɛli kama.Fasiri.J. Furakɛcogo.549, 201–217 (2018).