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Kugabanya hydrogels ya fibrous kuri capillaries ntoya bifite akamaro kanini muri sisitemu y'ibinyabuzima na biomedical.Impagarara no guhagarika unixial hydrogels fibrous byakozweho ubushakashatsi bwimbitse, ariko igisubizo cyabo cyo kugumana biaxial muri capillaries ntikiramenyekana.Hano, turerekana mubigeragezo no mubitekerezo ko geles ya filamile isubiza muburyo butandukanye kuburizamo kuruta geles zoroshye zoroshye bitewe na asimmetrie mumiterere yubukanishi bwimiterere ya filaments, yoroshye mugusenyuka kandi ikaze mubibazo.Mugukomeza cyane, gel ya fibrous igaragaza uburebure buke no kugabanuka kwa asimptotike kugabanuka kwa biaxial Poisson igereranije na zeru, bikaviramo gukomera kwa gel hamwe no gutembera nabi kwamazi binyuze muri gel.Ibi bisubizo byerekana kurwanya trombi irambuye ya lysis na mitiweli ivura kandi igatera imbere gukura neza kwa embolisation endovasculaire ituruka kuri geles fibrous kugirango ihagarike kuva amaraso cyangwa guhagarika amaraso yibibyimba.
Imiyoboro ya fibrous nuburyo bwibanze bwubaka kandi bukora ibice byubaka ingirabuzimafatizo.Actin nigice cyingenzi cya cytoskeleton1;fibrin nikintu cyingenzi mugukiza ibikomere no gushiraho trombus2, na kolagen, elastine na fibronectine nibigize matrice idasanzwe mubwami bwinyamaswa3.Imiyoboro yagaruwe ya fibrous biopolymers yahindutse ibikoresho hamwe nibikorwa byinshi mubikorwa bya tissue4.
Imiyoboro ya filime yerekana icyiciro cyihariye cyibinyabuzima byoroshye bifite imiterere yubukanishi butandukanye nuyoboro woroshye wa molekile5.Bimwe muri ibyo bintu byahindutse mugihe cyubwihindurize kugirango bigenzure igisubizo cyibinyabuzima bihindagurika 6.Kurugero, imiyoboro ya fibrous yerekana umurongo wa elastique kumurongo muto 7.8 mugihe kumurongo munini bagaragaza kwiyongera gukomera 9,10, bityo bikagumana ubusugire bwimitsi.Ingaruka zindi miterere yubukorikori bwa geles fibrous, nkumunaniro mubi usanzwe usubiza ikibazo cyogosha11,12, ntikiramenyekana.
Imiterere yubukorikori bwa hydrogels ya fibrous fibrous yoroheje yakozwe mubushakashatsi bwikibazo cya uniaxial13,14 hamwe no kwikuramo 8,15, ariko kwikuramo biaxial kwidegemvya kwabo muri capillaries cyangwa tebes ntibyigeze byigwa.Hano twatanze ibisubizo byubushakashatsi kandi tunatanga ibitekerezo byerekana uburyo bwimyitwarire ya hydrogels fibrous munsi yo kugumana biaxial mumiyoboro ya microfluidic.
Fibrin microcels ifite ibipimo bitandukanye bya fibrinogen na trombine hamwe na diameter ya D0 iri hagati ya 150 na 220 µm byakozwe hakoreshejwe uburyo bwa microfluidic (Ishusho yinyongera 1).Ku mutini.1a yerekana amashusho ya fluorochrome yanditseho microgels yabonetse ukoresheje microscopi ya fluorescence (CFM).Microgels ni serefegitura, ifite polydispersity iri munsi ya 5%, kandi ihuriweho mumiterere kurwego rwapimwe na CFM (Amakuru yinyongera na firime S1 na S2).Impuzandengo ya pore ya microcels (igenwa no gupima Darcy permeability16) yagabanutse kuva kuri 2280 igera kuri 60 nm, fibrine yariyongereye kuva kuri 5.25 igera kuri 37.9 mg / mL, naho trombine yagabanutse kuva kuri 2,56 igera kuri 0.27 / mL.(Amakuru yinyongera).Umuceri.2), 3 n'imbonerahamwe y'inyongera 1).Ubukomezi bujyanye na microgel bwiyongera kuva kuri 0,85 bugera kuri 3,6 kPa (Ishusho yinyongera).Nkurugero rwa geles ikozwe muminyururu yoroheje, microgels ya agarose ikoreshwa muburyo butandukanye.
Fluorescence microscopi ishusho ya fluorescein isothiocyanate (FITC) yanditseho PM yahagaritswe muri TBS.Igipimo cyumurongo ni 500 µm.b SEM amashusho ya SM (hejuru) na RM (hepfo).Umwanya munini 500 nm.c Igishushanyo mbonera cyumuyoboro wa microfluidic ugizwe numuyoboro munini (diameter dl) nakarere kagufi kameze nka cone ifite inguni yinjira α ya 15 ° na diameter ya dc = 65 µm.d Uhereye ibumoso ugana iburyo: Amashusho ya microscope ya optique ya RM (diameter D0) mumiyoboro minini, zone ya conique no kugabanuka (kugabanya uburebure bwa gel Dz).Igipimo cyumurongo ni 100 µm.e, f TEM amashusho ya RM (e) idakozwe na RM (f) idashyizwe ahagaragara, yashyizweho kumasaha imwe hamwe nimbogamizi 1 / λr = 2.7, hakurikiraho kurekurwa no gukosora 5% bya misa.glutaraldehyde muri TBS.Diameter ya CO idakozwe ni 176 mm.Igipimo cyibipimo ni 100 nm.
Twibanze kuri microcels ya fibrin ifite ubukana bwa 0,85, 1.87 na 3,6 kPa (aha twavuga nka microgels yoroshye (SM), microcels ikomeye (MM) na microcels ikomeye (RM)).Uru rutonde rwa fibrin gel stiffness ni murwego rumwe rwubunini nko kumaraso 18,19 bityo rero geles ya fibrin yize mubikorwa byacu ifitanye isano itaziguye na sisitemu y'ibinyabuzima nyayo.Ku mutini.1b yerekana amashusho yo hejuru no hepfo yububiko bwa SM na RM bwabonetse hakoreshejwe scanning electron microscope (SEM).Ugereranije nuburyo bwa RM, imiyoboro ya SM ikorwa na fibre nini hamwe nuduce duto twamashami, bihuye na raporo zabanje 20, 21 (Ishusho yinyongera 5).Itandukaniro mu miterere ya hydrogel rifitanye isano nuburyo ibintu byifashe: ubwikorezi bwa gel bugabanuka hamwe no kugabanya ingano ya pore kuva kuri SM ikagera kuri MM na RM (Imbonerahamwe yinyongera 1), hamwe no gukomera kwa gel birahinduka.Nta gihinduka mumiterere ya microgel yagaragaye nyuma yo kubika kuri 4 ° C muminsi 30 (Ishusho yinyongera 6).
Ku mutini.1c yerekana igishushanyo cyumuyoboro wa microfluidic ufite igice cyambukiranya uruziga kirimo (uhereye ibumoso ugana iburyo): umuyoboro munini ufite diameter dl aho microgel ikomeza kuba idakora, igice kimeze nka cone kigabanya diameter dc
Ku mutini.1e, 1f yerekana kohereza microscopi ya electron (TEM) amashusho yimikorere ya RM idakozwe kandi igabanijwe.Nyuma yo kwikuramo RM, ingano ya microgel yagabanutse cyane kandi imiterere yabyo yabaye anisotropique ifite ubunini buto mu cyerekezo cyo kwikuramo, ibyo bikaba bihuye na raporo yabanjirije 23.
Kwiyunvira kwa Biaxial mugihe cyo kwikuramo bituma microgel irambura mu cyerekezo kitagira imipaka hamwe na coefficient λz = \ ({D} _ {{{{{{{\ rm {z}}}}}}} / \ ({D} _ { 0} \), aho \ ({D} _ {{{{({\ rm {z}}}}}}}} \) ni uburebure bwa microgel ifunze Ishusho 2a yerekana impinduka muri λzvs .1 / λr kuri microcels ya fibrin na agarose. Igitangaje ni uko mugukomeretsa gukomeye kwa 2.4 ≤ 1 / λr ≤ 4.2, mikorobe ya fibrin yerekana uburebure bwa 1.12 +/- 0.03 λz, bigira ingaruka nke gusa ku gaciro ka 1 / λr. imyitwarire ya microse ntoya ya agarose, igaragara no mugihe cyo kugabanuka guke 1 / λr = 2,6 kugeza kurambura nini λz = 1.3.
igeragezwa rya microgel ya Agarose hamwe na modulike itandukanye ya elastike (2,6 kPa, diyama yicyatsi kibisi; 8.3 kPa, uruziga rufunguye; 12.5 kPa, ikibanza cya orange gifunguye; inziga), MM (kare yumukara wirabura) na RM (inyabutatu yubururu ikomeye).Imirongo ihamye yerekana teoritiki yahanuwe λz kuri agarose (umurongo wicyatsi) na microcels ya fibrin (imirongo nibimenyetso byamabara amwe).b, c Umwanya wo hejuru: igishushanyo mbonera cyurunigi rwurusobe rwa agarose (b) na fibrin (c) mbere (ibumoso) na nyuma (iburyo) kwikuramo biaxial.Hasi: Imiterere y'urusobe rujyanye mbere na nyuma yo guhindura ibintu.Icyerekezo cya x na y cyerekanwa na magenta hamwe numwambi wijimye.Mu gishushanyo kiri hejuru, iminyururu y'urusobekerane rwerekejwe muri iyi x na y yerekanwe hamwe na magenta ijyanye n'imirongo yijimye, n'iminyururu yerekanwe mu cyerekezo cya z iterekanwa n'umurongo w'icyatsi.Muri fibrin gel (c), imirongo yumutuku nubururu mu cyerekezo cya x na y irunama cyane kuruta uko itigeze ihinduka, kandi imirongo yicyatsi mu cyerekezo cyerekezo cyunamye kandi kirambuye.Impagarara hagati yicyerekezo cyo kwikuramo no guhagarika umutima zinyuzwa mumutwe hamwe nicyerekezo hagati.Muri geles ya agarose, iminyururu mu byerekezo byose igena umuvuduko wa osmotic, ugira uruhare runini muguhindura gele.d Impinduka ziteganijwe mu kigereranyo cya biaxial Poisson,}} ^ {{{{{\ rm {eff}}}}}}} = - {{{{{\ rm {ln}}}}}} {\ lambda} _ { z} / {{{{{mInet yerekana biaxial deformation ya gel.e Guhindura umuvuduko wimpinduka ΔPtr, isanzwe kuri gel stiffness S, yateguwe nkigikorwa cyo kugereranya compression ya agarose na fibrin microgel.Ibimenyetso by'ibimenyetso bihuye n'amabara muri (a).Icyatsi kibisi n'umutuku byerekana isano iri hagati ya ΔPtr / S na 1 / λr kuri gela ya agarose na fibrin.Igice cyaciwe kumurongo utukura cyerekana kwiyongera kwa ΔPtr munsi yo kwikuramo gukomeye kubera imikoranire ya interiber.
Iri tandukaniro rifitanye isano nuburyo butandukanye bwo guhindura imiyoboro ya fibrin na agarose microgel, bigizwe na flex2424 na rigid25.Biaxial compression ya geles yoroheje itera kugabanuka kwijwi ryayo hamwe no kwiyongera kwingutu hamwe nigitutu cya osmotic, biganisha ku kurambura gel mu cyerekezo kitagira imipaka.Kurambura kwa nyuma kwa gele biterwa nuburinganire bwiyongera ryingufu za entropique yubusa yiminyururu irambuye no kugabanuka kwingufu zubusa za osmose bitewe nubushakashatsi bwa polymer buke muri gel irambuye.Mugihe cyo kwikuramo biaxial ikomeye, kurambura gel byiyongera hamwe na λz ≈ 0,6 \ ({{\ lambda} _ {{{\ rm {r}}}} {{- 2/3}} \) (reba Ishusho 2a muri igice cyo kuganira 5.3.3).Impinduka zijyanye n'iminyururu ihindagurika n'imiterere y'urusobe rujyanye mbere na nyuma yo kugumana biaxial bigaragara mu mashusho.2b.
Ibinyuranye, geles ya fibrous nka fibrin isanzwe isubiza muburyo butandukanye kugumana biaxial.Filaments yerekejwe cyane cyane nicyerekezo cyo guhuza flex (bityo bikagabanya intera iri hagati yambukiranya imipaka), mugihe filaments yiganjemo perpendicular yerekeza ku cyerekezo cyo kwikuramo igororotse kandi irambuye munsi yimbaraga za elastique, bigatuma gel irambura ( Igishushanyo 1).2c) Imiterere ya SM, MM na RM idakozwe neza yaranzwe no gusesengura amashusho yabo ya SEM na CFM (Ikiganiro cyinyongera Igice cya IV hamwe ninyongera Ishusho 9).Muguhitamo modulus ya elastike (E), diameter (d), uburebure bwumwirondoro (R0), intera iri hagati yimpera (L0 ≈ R0) nu mfuruka yo hagati (ψ0) yimigozi muri microgel ya fibrin idakozwe (Imbonerahamwe yinyongera 2) - 4), dusanga iyo nsanganyamatsiko igoramye modulus \ ({k} _ {{{{{{\ rm {b)))))))))} = \ frac {9 \ pi E {d} ^ {4}} {4 {\ psi} _ {0} ^ {2} {L} _ {0}} \) ni munsi cyane ugereranije na modulus yayo itoroshye \ ({k} _ {{{{{{ }}}} = E \ frac {\ pi {d} ^ {2} {R} _ {0}} {4} \), bityo kb / ks ≈ 0.1 (Imbonerahamwe 4).Rero, mubihe bya biaxial gel igumana, imigozi ya fibrin irunama byoroshye, ariko irwanya kurambura.Kurambura umuyoboro uteye ubwoba ukorerwa biaxial compression byerekanwe kumugereka wa 17.
Dutezimbere icyitegererezo cya affine (Ikiganiro Cyinyongera Igice cya V hamwe nishusho yinyongera 10-16) aho kurambura gel fibrous bigenwa kuburinganire bwaho bwimbaraga za elastique zikora muri gel kandi bugahanura ko muburyo bukomeye bwa biaxial λz - 1 munsi y'imbogamizi
Ikigereranyo (1) cyerekana ko no muri compression ikomeye (\ ({\ lambda} _ {{{\ mbox {r)))) \, \ to \, 0 \)) hariho kwaguka gake gake hamwe no guhindagurika kurambuye kuri kwiyuzuzamo λz - 1 = 0.15 ± 0.05.Iyi myitwarire ifitanye isano na (i) \ ({\ ibumoso ({k} _ {{{{({\ rm {b}}}}}}}} / {k} _ {{{{{\ rm) s s {3} \) kumurongo ukomeye wa biaxial. Ni ngombwa kumenya ko prefactor \ ({\ ibumoso ({k} _ {({\ mbox {b)))) / {k} _ {({\ mbox { s))). ψ0, bisa na SM, MM na RM (Imbonerahamwe y'inyongera 4).
Kugirango turusheho kwerekana itandukaniro riri hagati yubwisanzure buterwa nubwisanzure hagati ya geles yoroheje kandi yuzuye, turamenyekanisha igipimo cya biaxial Poisson \ ({\ nu} _ {{{({\ rm {b))))))} {{\ mbox { =}}} \, \ mathop {{\ lim}} \ imipaka _ {{\ lambda} _ {{{{({\ rm {r}}}}}}} \ to 1} \ frac {{\ lambda} _ {{{{{\ rm {z}}}}}} - 1} {1 - {\ lambda} _ {{({\ rm {r}}}}}}}}}, \) asobanura imbibi. Icyerekezo cya gel gisubiza ibibazo bingana mubyerekezo bibiri bya radiyo, kandi bigera no kumurongo munini umwe \ rm {b}}}}}}} ^ {{{{{\ rm {eff}}}}}}}} }} = - {{{{{\ rm {ln}}}}}}}} \ lambda} _ {z} / {{{({\ rm {ln)))))))) {{\ lambda } _ {{{({\ rm {r)))))))))} \).Ku mutini.2d yerekana \ ({{{{{{\ rm {\ nu}}}}}}} _ {{{({\ rm {b}}}}}} }}}}}}}) Kuri geles ya agarose mukumirwa gukomeye {\ rm {eff}}}}}}}} \) yiyongera ku gaciro ka asimptotic 2/3, naho kuri fibrin geles igabanuka kugeza kuri zeru, kuva lnλz / lnλr → 0, kuva λz yiyongera hamwe kwiyuzuza nkuko λr yiyongera.Menya ko mubigeragezo, microgels zifunze zifunze zidahinduka muburyo bumwe, kandi igice cyabo cyo hagati kigabanuka cyane;icyakora, extrapolation kumurongo munini wa 1 / λr ituma bishoboka kugereranya igeragezwa hamwe nigitekerezo cya geles yahinduwe kimwe.
Irindi tandukanyirizo ryimyitwarire ya geles yoroheje hamwe na geles ya filament yabonetse kubera kugenda kwabo kugabanuka.Umuvuduko wo guhinduranya ΔPtr, usanzwe kuri gel stiffness S, wiyongereye hamwe no kwiyongera (Ishusho 2e), ariko kuri 2.0 ≤ 1 / λr ≤ 3.5, microgel ya fibrin yerekanaga agaciro kari hasi ya ΔPtr / S hasi mugihe cyo kugabanuka.Kugumana microgel ya agarose biganisha ku kwiyongera k'umuvuduko wa osmotic, biganisha ku kurambura gel mu cyerekezo kirekire kuko molekile ya polymer irambuye (Ishusho 2b, ibumoso) no kwiyongera k'umuvuduko wo guhinduranya na ΔPtr / S ~ ( 1 / λr) 14/317.Ibinyuranye na byo, imiterere ya microgel ya fibrin ifunze igenwa nuburinganire bwingufu zurudodo rwo kwikuramo imirasire hamwe nimpagarara ndende, biganisha ku guhinduka kwinshi kwinshi λz ~ \ (\ sqrt {{k} _ {{{{{{ \ rm {b))))))))))))} / {k} _ {{{{{\Kuri 1 / λr ≫ 1, impinduka zumuvuduko wubuhinduzi zapimwe nka 1} {{{({\ rm {ln)))))) \ ibumoso ({{\ lambda}} _ {{{{{\ rm {}}}}}}}}Rero, ΔPtr ntigabanijwe cyane kuruta geles ya agarose.Kuri compression hamwe na 1 / λr> 3.5, kwiyongera gukabije kwijwi ryibice bya filaments hamwe nubusabane bwamafirime aturanye bigabanya ihinduka ryimiterere ya gel kandi biganisha ku gutandukana kubisubizo byubushakashatsi bivuye ku guhanura (umurongo utudomo utukura ku gishushanyo 2e).Twanzuye ko kuri kimwe 1 / λr na Δ \ ({P} _ {{{{{{{\ rm {tr}}}}}}}} _ {{{{\ rm {fibrin}}})) }}}} \) <ΔP <Δ \ ({P} _ {{{{{{\ rm {tr)))))))}}} _ {{{\ \ rm {agarose gel}}} \) gel ya agarose izafatwa na microchannel, kandi gel ya fibrin ifite ubukana bumwe izayinyuramo.Kuri ΔP <Δ \ ({P} _ {{{{{{\ rm {tr))))))))) _ {{{{{\ rm {fibrin)))))))))} \ ), Gele ebyiri zombi zizahagarika umuyoboro, ariko gel ya fibrin izasunika cyane kandi igabanye neza, ibuza amazi neza.Ibisubizo byerekanwe ku gishushanyo cya 2 byerekana ko gel fibrous ishobora kuba icyuma cyiza cyo kugabanya kuva amaraso cyangwa kubuza gutanga amaraso kubyimba.
Ku rundi ruhande, fibrin ikora scafold iganisha kuri tromboembolism, indwara y’indwara aho trombus ifata icyombo kuri ΔP <ΔPtr, nko mubwoko bumwe na bumwe bwubwonko bwa ischemic (Ishusho 3a).Kunanirwa gukabije guterwa no kurambura mikorobe ya fibrin byatumye habaho kwiyongera gukomeye kwa fibrin ya C / C fibrinogen ugereranije na geles zoroshye, aho C na C fibrinogen ibujijwe kandi mikorobe idakozwe neza.Polymer yibanze muri gel.Igishushanyo 3b cyerekana ko fibrinogen C / C muri SM, MM, na RM yiyongereyeho inshuro zirindwi kuri 1 / λr ≈ 4.0, biterwa no kubuzwa no kubura umwuma (Ishusho yinyongera 16).
Igishushanyo cyerekana uburyo bwo guhagarika imiyoboro yo mu bwonko yo hagati mu bwonko.b Kwibuza-guhuza ugereranije no kwiyongera kwa fibrin yibanda kuri SM (inziga zitukura zikomeye), MM (kare yumukara ukomeye), na RM (inyabutatu yubururu ikomeye).c Igishushanyo mbonera cyakoreshejwe mukwiga clavage ya fibrin yabujijwe.Igisubizo cya fluoresc cyanditseho tPA muri TBS cyatewe ku gipimo cya 5.6 × 107 µm3 / s hamwe n’igitutu cyiyongereyeho 0.7 Pa kumuyoboro uherereye kuri perpendikulari kuri axe ndende ya mikorobe nkuru.d Igicucu cyuzuye microscopique ishusho ya MM ibuza (D0 = 200 µm) kuri Xf = 28 µm, ΔP = 700 Pa no mugihe cyo gutandukana.Imirongo yerekana utudomo yerekana imyanya ibanza yinyuma yinyuma ninyuma ya MM kuri tlys = 0. Ibara ryatsi nijimye rihuye na FITC-dextran (70 kDa) na tPA yanditseho AlexaFluor633.e Igihe-gitandukanye ugereranije nubunini bwa RMs hamwe na D0 ya 174 µm (ubururu bwuguruye butatu butatu), 199 µm (inyabutatu yubururu ifunguye), na 218 µm (inyabutatu yubururu ifunguye), muburyo bumwe, muri microchannel hamwe na Xf = 28 ± 1 µm.ibice bifite ΔP 1200, 1800, na 3000 Pa, hamwe na Q = 1860 ± 70 µm3 / s.Inet yerekana RM (D0 = 218 µm) icomeka kuri microchannel.f Itandukanyirizo ryubunini bugereranije bwa SM, MM cyangwa RM yashyizwe kuri Xf = 32 ± 12 µm, kuri ΔP 400, 750 na 1800 Pa na ΔP 12300 Pa na Q 12300 mukarere ka conone ya microchannel, 2400 na 1860 µm3 / s.Xf yerekana umwanya wimbere wa microgel kandi ikagena intera yayo kuva itangiye kugabanuka.V (tlys) na V0 nubunini bwigihe gito bwa microgel ya lysed hamwe nubunini bwa microgel idahungabanye.Ibara ry'inyuguti rihuye n'amabara muri b.Imyambi yumukara kuri e, f ihuye nigihe cyanyuma cyigihe mbere yo kunyura mikorobe binyuze muri microchannel.Igipimo cyibipimo muri d, e ni 100 µm.
Kugira ngo dukore iperereza ku ngaruka zo kubuza kugabanuka kwamazi ya geles ya fibrine ibuza, twize lysis ya SM, MM, na RM yinjiye hamwe na trombolytic agent tissue plasminogen activateur (tPA).Igishushanyo 3c cyerekana igishushanyo mbonera cyakoreshejwe mubushakashatsi bwa lysis. Kuri ΔP = 700 Pa (<ΔPr karere. Kuri ΔP = 700 Pa (<ΔPr karere. При ΔP = 700 Па (<ΔPr Kuri ΔP = 700 Pa (<ΔPtr) nigipimo cyo gutemba, Q = 2400 µm3 / s, ya Tris buffered saline (TBS) ivanze na 0.1 mg / mL (fluorescein isothiocyanate) FITC-dextran, microgel yarimo microchannel ihuza.karere.Δ ΔP = 700 Pa (<ΔPtr) 和 流速 Q = 2400 μm3 / s 的 Tris 缓冲 盐水 (TBS) 与 0.1 mg / mL 的 （异 硫氰酸 荧光 素） FITC- 葡聚糖 混合 时 ， 微 凝胶 堵塞了 锥形 微 通道 地区。Δ ΔP = 700 Pa (<ΔPtr) 和 流速 Q = 2400 μm3 / s 了 锥形 微 通道 地区。 Микрогели Кану сививаются при смешивании трис-буферного солевого раствора (TBS) с 0,1 мг / К алов. Microgels yacometse mugihe Tris buffer saline (TBS) yavanze na 0.1mg / mL (fluorescein isothiocyanate) FITC-dextran kuri ΔP = 700 Pa (<ΔPtr) nigipimo cyogutemba Q = 2400 µm3 / s Uturere duhuza uturere twa mikorobe.Imyanya yimbere Xf ya microgel igena intera yayo kuva kugabanuka kwambere X0.Kugirango habeho lysis, igisubizo cya fluoresc yanditseho tPA muri TBS yatewe mumurongo uherereye hagati ya orthogonally kugera kumurongo muremure wa microchannel.
Igihe igisubizo cya tPA kigeze kuri MM idasanzwe, impande yinyuma ya microgel yahindutse urujijo, byerekana ko gukuramo fibrin byatangiye mugihe tlys = 0 (Ishusho 3d nigishushanyo cya 18).Mugihe cya fibrinolysis, irangi ryanditseho tPA ryirundarunda imbere muri MM kandi rihuza imigozi ya fibrin, ibyo bigatuma habaho kwiyongera buhoro buhoro ubukana bwibara ryijimye rya mikorobe.Kuri tlys = 60 min, MM isezerana kubera gusesa igice cyinyuma cyayo, hamwe numwanya wimbere Xf ihinduka bike.Nyuma yiminota 160, MM yasezeranye cyane yakomeje gusezerana, maze kuri tlys = 161 min, iragabanuka, bityo igarura amazi yatembye binyuze muri microchannel (Igishusho 3d nigishushanyo cya 18, inkingi iburyo).
Ku mutini.3e yerekana lysis-yunganirwa nigihe-biterwa no kugabanuka kwijwi V (tlys) risanzwe kugeza mububiko bwambere V0 bwa mikorobe itandukanye ya fibrin.CO hamwe na D0 174, 199, cyangwa 218 µm yashyizwe muri microchannel ifite ΔP 1200, 1800, cyangwa 3000 Pa, hamwe na Q = 1860 ± 70 µm3 / s kugirango bahagarike imiyoboro (Ishusho 3e, shyiramo).imirire.Microgel igenda igabanuka buhoro buhoro kugeza igihe ari gito bihagije kugirango inyure mumiyoboro.Kugabanuka k'ubunini bukomeye bwa CO hamwe na diameter nini ya mbere bisaba igihe kirekire cya lysis.Bitewe no gutembera bisa binyuze muri RM zitandukanye, clavage ibaho ku kigero kimwe, bigatuma igogorwa ryibice bito bya RM nini hamwe no gutinda kwabo.Ku mutini.3f yerekana kugabanuka ugereranije muri V (tlys) / V0 kubera gutandukana kwa SM, MM, na RM kuri D0 = 197 ± 3 µm byateguwe nkigikorwa cya tlys.Kuri SM, MM na RM, shyira buri microgel muri microchannel ifite ΔP 400, 750 cyangwa 1800 Pa na Q 12300, 2400 cyangwa 1860 µm3 / s.Nubwo igitutu cyakoreshwaga kuri SM cyikubye inshuro 4,5 ugereranije n’icya RM, umuvuduko wanyuze muri SM wikubye inshuro zirenga esheshatu kubera ubwinshi bw’imikorere ya SM, kandi kugabanuka kwa microgel byagabanutse kuva kuri SM kugera kuri MM na RM .Kurugero, kuri tlys = 78 min, SM ahanini yarashonze kandi irimurwa, mugihe MM na PM bakomeje gufunga imiyoboro mito, nubwo bagumana 16% na 20% gusa byubunini bwumwimerere.Ibisubizo byerekana akamaro ka lysis ya convection-medrated lysis ya fibrous geles igabanijwe kandi bifitanye isano na raporo zerekana igogorwa ryihuse ryibibyimba birimo fibrine yo hasi.
Rero, akazi kacu karerekana mubigeragezo no muburyo bwa tekinoroji uburyo geles ya filimile isubiza kwifungisha biaxial.Imyitwarire ya geles fibrous mumwanya muto igenwa na asimmetrie ikomeye yingufu zingutu za filaments (yoroshye muri compression kandi ikomeye muri tension) kandi nukugereranya gusa na ratio na curvature ya filaments.Iyi myitwarire itera kurambura geli ya fibrous ikubiye muri capillaries zifunganye, igipimo cya biaxial Poisson igabanuka hamwe no kwiyongera no kugabanuka gake.
Kubera ko biaxial igizwe nibice byoroshye byahinduwe bikoreshwa muburyo butandukanye bwikoranabuhanga, ibisubizo byacu bitera iterambere ryibikoresho bishya bya fibrous.By'umwihariko, kugumana biaxial gels ya filamile muri capillaries ntoya cyangwa tebes biganisha ku gukomera kwabo no kugabanuka gukabije.Kubuza cyane amazi gutembera binyuze muri geles fibrous idasanzwe bifite ibyiza iyo bikoreshejwe nk'amacomeka kugirango wirinde kuva amaraso cyangwa kugabanya amaraso kuri malignancies33,34,35.Ku rundi ruhande, igabanuka ry’amazi anyuze muri geli ya fibrine, bityo bikabuza lysis ya convective-medrated trombus lysis, bitanga ikimenyetso cyerekana lysis gahoro yimitsi idasanzwe [27, 36, 37].Sisitemu yacu yo kwerekana imiterere nintambwe yambere iganisha ku gusobanukirwa ningaruka zo gukanika imashini ya fibrous biopolymer hydrogels yo kugumana biaxial.Kwinjiza uturemangingo twamaraso cyangwa platine mumitsi ya fibrin ibuza bizagira ingaruka kumyitwarire yabo yo kubuza 38 kandi bizaba intambwe ikurikira muguhishura imyitwarire ya sisitemu igoye cyane yibinyabuzima.
Reagents zikoreshwa mugutegura microgel ya fibrin no guhimba ibikoresho bya MF byasobanuwe mumakuru yinyongera (Uburyo bwinyongera Igice cya 2 nicya 4).Fibrin microcels yateguwe muguhindura igisubizo kivanze cya fibrinogen, Tris buffer na trombine mumigezi yibanda kubikoresho bya MF, hanyuma bikurikirwa no gutonyanga.Igisubizo cya Bovine fibrinogen (60 mg / ml muri TBS), Tris buffer na bovine thrombin (5 U / ml muri 10 mM CaCl2 igisubizo) byatanzwe hakoreshejwe pompe ebyiri zigenga zigenga (PhD 200 Harvard Apparatus PHD 2000 Syring Pump).guhagarika MF, Amerika).Icyiciro cya F-amavuta gikomeza kirimo 1 wt.Ibitonyanga byakozwe mubikoresho bya MF bikusanyirizwa mumashanyarazi ya ml 15 ya centrifuge irimo amavuta ya F.Shira imiyoboro mu bwogero bwamazi kuri 37 ° C kuri 1 h kugirango urangize fibrine.FITC yanditseho microcels ya fibrin yateguwe kuvanga bovine fibrinogen na FITC yanditseho fibrinogen yabantu mubipimo bya 33: 1.Inzira nimwe yo gutegura mikorobe ya fibrin.
Hindura microgel ziva mumavuta F kuri TBS ukoresheje centrifuging ikwirakwiza kuri 185 g kuminota 2.Microgel yaguye yakwirakwijwe mu mavuta F ivanze na 20 wt.% Inzoga ya parfluorooctyl, hanyuma ikwirakwizwa muri hexane irimo 0.5 wt.% Span 80, hexane, 0.1 wt.% Triton X mumazi na TBS.Hanyuma, microgels yatatanye muri TBS irimo 0.01 wt% Hagati ya 20 hanyuma ibikwa kuri 4 ° C mugihe cibyumweru 1-2 mbere yubushakashatsi.
Guhimba ibikoresho bya MF byasobanuwe mumakuru yinyongera (Uburyo bwinyongera Igice cya 5).Mubigeragezo bisanzwe, agaciro keza ka ΔP kagenwa nuburebure bugereranije bwibigega byahujwe mbere na nyuma yigikoresho cya MF cyo kwinjiza mikorobe ifite diameter ya 150
Amakuru ashyigikira ibisubizo byubu bushakashatsi araboneka kubanditsi babisabwe.Amashusho ya SEM ya geli ya fibrin, TEM ishusho ya geli ya fibrin mbere na nyuma yo guterwa, hamwe namakuru yingenzi yinjiza kumashusho 1 na 2. 2 na 3 yatanzwe muri dosiye yibanze.Iyi ngingo itanga amakuru yumwimerere.
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Igihe cyo kohereza: Gashyantare-23-2023