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Ukukhawulelwa kwama-hydrogel ane-fibrous ukuthi anciphise ama-capillaries kubaluleke kakhulu ezinhlelweni zebhayoloji nezama-biomedical.I-tension kanye nokucindezelwa kwe-uniaxial kwama-fibrous hydrogels kuye kwacwaningwa kabanzi, kodwa impendulo yawo ekugcinweni kwe-biaxial kuma-capillaries ayikahlolisiswa.Lapha, sibonisa ngokuhlola nangethiyori ukuthi amajeli ane-filamentous aphendula ngendlela ehlukile evimbelweni kunamaketanga aguquguqukayo ngenxa ye-asymmetry kuzakhiwo zemishini zemicu ehlanganisiwe, ethambile ekuminyanisweni futhi eqinile ekucindezelekeni.Ngaphansi kokugcinwa okuqinile, ijeli ene-fibrous ibonisa ukunwebeka okuncane kanye nokwehla okungabonakali kwe-asymptotic kwesilinganiso se-biaxial Poisson kuya ku-zero, okuholela ekuhlanganeni kwejeli okuqinile nokungenwa kahle koketshezi ejeli.Le miphumela ikhombisa ukumelana kwe-thrombi e-occlusive enwetshiwe ku-lysis ngama-ejenti okwelapha futhi ikhuthaze ukuthuthukiswa kwe-endovascular embolization ephumelelayo kusuka kuma-fibrous gels ukumisa ukopha kwemithambo noma ukuvimbela ukunikezwa kwegazi kwezimila.
Amanethiwekhi ama-Fibrous ayizakhiwo eziyisisekelo nezisebenzayo zezicubu namaseli aphilayo.I-Actin iyingxenye enkulu ye-cytoskeleton1;I-fibrin iyisici esiyinhloko ekwelapheni kwesilonda kanye nokwakheka kwe-thrombus2, futhi i-collagen, i-elastin ne-fibronectin yizingxenye ze-matrix ye-extracellular embusweni wezilwane3.Amanethiwekhi atholiwe ama-fibrous biopolymers asephenduke izinto ezisetshenziswa kabanzi kubunjiniyela bezicubu4.
Amanethiwekhi e-filamentous amele isigaba esihlukile sezinto ezithambile zebhayoloji ezinezakhiwo ezisebenza ngomshini ezihlukile kumanethiwekhi amangqamuzana aguquguqukayo5.Ezinye zalezi zakhiwo ziye zavela ngesikhathi sokuziphendukela kwemvelo ukuze kulawulwe ukusabela kwezinto eziphilayo ekuwohlokeni6.Isibonelo, amanethiwekhi ama-fibrous abonisa ukunwebeka komugqa ezinkingeni ezincane7,8 kuyilapho ezinhlobonhlobo ezinkulu abonisa ukuqina okwenyuka9,10, ngaleyo ndlela agcine ubuqotho bezicubu.Imithelela yezinye izici zemishini zamajeli ane-fibrous, njengokucindezeleka okuvamile okungekuhle ekuphenduleni i-shear strain11,12, ayikatholakali.
Izakhiwo zemishini ye-semi-flexible fibrous hydrogels ziye zafundwa ngaphansi kwe-uniaxial tension13,14 kanye nokucindezela8,15, kodwa ukucindezelwa kwe-biaxial okubangelwa inkululeko kuma-capillaries amancane noma amashubhu akuzange kufundwe.Lapha sibika imiphumela yokuhlolwa futhi siphakamisa ngokunengqondo indlela yokuziphatha kwama-fibrous hydrogel ngaphansi kokugcinwa kwe-biaxial eziteshini ezincane ze-microfluidic.
Ama-microgel e-Fibrin anezilinganiso ezihlukahlukene ze-fibrinogen nokugxilisa kwe-thrombin kanye nobubanzi be-D0 obusuka ku-150 kuya ku-220 µm akhiqizwa kusetshenziswa indlela ye-microfluidic (Umfanekiso Owengeziwe 1).Emkhiwaneni.I-1a ibonisa izithombe ze-fluorochrome abhalwe ukuthi ama-microgel atholwe kusetshenziswa i-confocal fluorescence microscopy (CFM).Ama-microgel ayindilinga, ane-polydispersity engaphansi kuka-5%, futhi ayafana ngesakhiwo kuzo zonke izikali ezihlolwe yi-CFM (Ulwazi Olwengeziwe nama-Movie S1 kanye ne-S2).Isilinganiso sosayizi we-pore wama-microgels (okunqunywe ngokulinganisa i-Darcy permeability16) yehla ukusuka ku-2280 kuya ku-60 nm, okuqukethwe kwe-fibrin kukhuphuke kusuka ku-5.25 kuya ku-37.9 mg / mL, futhi ukuhlushwa kwe-thrombin kwehla kusuka ku-2.56 kuya ku-0.27 amayunithi / mL, ngokulandelana.(Ulwazi Olwengeziwe).Ilayisi.2), 3 kanye nethebula elingeziwe 1).Ukuqina okuhambisanayo kwe-microgel kukhuphuka kusuka ku-0.85 kuya ku-3.6 kPa (I-Supplementary Fig. 4).Njengezibonelo zamajeli akhiwe ngamaketanga aguquguqukayo, kusetshenziswa ama-agarose microgel okuqina okuhlukahlukene.
Isithombe se-fluorescence microscopy se-fluorescein isothiocyanate (FITC) esibhalwe ukuthi PM simisiwe ku-TBS.Isikali sebha singu-500 µm.b izithombe ze-SEM ze-SM (phezulu) ne-RM (phansi).Ibha yesikali 500 nm.c Umdwebo wohlelo weshaneli encane ye-microfluidic ehlanganisa ishaneli enkulu (ububanzi dl) kanye nesifunda esimise okwekhoni ewumngcingo ene-engeli yokungena engu-α engu-15° kanye nobubanzi buka-dc = 65 µm.d Kwesobunxele kuye kwesokudla: Izithombe ze-Optical microscope ye-RM (ububanzi D0) eziteshini ezinkulu, indawo eyi-conical kanye nokuminyana (umkhawulo wejeli Dz).Isikali sebha singu-100 µm.e, f izithombe ze-TEM ze-RM engalungiswanga (e) kanye ne-RM evalekile (f), ezimiswe ihora elilodwa ngokuminya 1/λr = 2.7, okulandelwa ukukhululwa nokulungiswa kuka-5% wesisindo.i-glutaraldehyde ku-TBS.Ububanzi be-CO engaguquki bungu-176 μm.Ibha yesikali ingu-100 nm.
Sigxile kuma-fibrin microgel anobulukhuni obungu-0.85, 1.87 kanye no-3.6 kPa (ngemuva kwalokhu okubizwa ngokuthi ama-microgel athambile (SM), ama-microgel aqinile aphakathi nendawo (MM) nama-microgel aqinile (RM), ngokulandelana).Lolu hlu lokuqina kwejeli ye-fibrin luhlelo olufanayo lobukhulu njengamahlule egazi18,19 futhi yingakho amajeli e-fibrin afundwa emsebenzini wethu ahlobene ngokuqondile nezinhlelo zebhayoloji zangempela.Emkhiwaneni.I-1b ibonisa izithombe ezingaphezulu neziphansi zezakhiwo ze-SM ne-RM ezitholwe kusetshenziswa isibonakhulu se-electron yokuskena (SEM), ngokulandelanayo.Uma kuqhathaniswa nezakhiwo ze-RM, amanethiwekhi e-SM akhiwa imicu emikhulu kanye namaphoyinti amagatsha ambalwa, ahambisana nemibiko yangaphambilini ye-20, i-21 (I-Supplementary Fig. 5).Umehluko ekwakhiweni kwe-hydrogel uhambisana nokuthambekela kwezakhiwo zayo: ukungena kwejeli kuncipha ngokuncipha kosayizi we-pore ukusuka ku-SM kuye ku-MM kanye ne-RM (Ithebula Lokwengeza 1), futhi ukuqina kwejeli kuyahlehla.Azikho izinguquko esakhiweni se-microgel eziphawulwe ngemva kokugcinwa ku-4 ° C izinsuku ezingu-30 (I-Fig. 6 eyengeziwe).
Emkhiwaneni.I-1c ibonisa umdwebo wesiteshi esincane se-microfluidic esinesigaba esiphambano esiyindilinga esiqukethe (kusuka kwesobunxele kuye kwesokudla): umzila omkhulu onobubanzi obungu-dl lapho i-microgel ihlala ingashintshile, ingxenye emise okwekhoni enobubanzi obuncane obungu-dc
Emkhiwaneni.I-1e, i-1f ibonisa i-electron microscopy (TEM) yokudlulisela izithombe zezakhiwo ze-RM ezingaguquki futhi ezinomkhawulo we-biaxially.Ngemuva kokucindezelwa kwe-RM, usayizi we-microgel pore wehla kakhulu futhi umumo wabo waba yi-anisotropic enobukhulu obuncane ekuqondeni kokucindezela, okuhambisana nombiko wangaphambili we-23.
Ukucindezelwa kwe-Biaxial phakathi nokufinyela kubangela ukuthi i-microgel ikhule ngendlela engenamkhawulo nge-coefficient λz = \({D}_{{{{{{\rm{z}}}}}}}}/\({D }_ { 0}\) , lapho \({D}_{{{{(({\rm{z}}}}}}}}\) kuwubude be-microgel evaliwe Umfanekiso 2a ubonisa ushintsho ku-λzvs .1/ λr I-fibrin kanye ne-agarose microgels. Kuyamangaza ukuthi ngaphansi kokucindezelwa okunamandla okungu-2.4 ≤ 1/λr ≤ 4.2, ama-fibrin microgel abonisa ubude obuncane obungu-1.12 +/- 0.03 λz, obuthintwa kancane kuphela inani lika-1/λr. ama-agarose microgels alinganiselwe, abonwa ngisho nasekucindezelweni okubuthakathaka 1/λr = 2.6 kuya kubude obukhudlwana λz = 1.3.
ukuhlolwa kwe-Agarose microgel enemoduli enwebekayo ehlukile (2.6 kPa, idayimane eliluhlaza elivulekile; 8.3 kPa, indilinga evulekile ensundu; 12.5 kPa, isikwele esisawolintshi esivulekile; 20.2 kPa, unxantathu ohlanekezelwe wemagenta ovulekile) kanye ne-SM (okubomvu okuqinile) Guqula ubude obulinganisiwe λz ( imibuthano), MM (izikwele ezimnyama eziqinile) kanye ne-RM (onxantathu abaluhlaza okwesibhakabhaka okuqinile).Imigqa eqinile ibonisa u-λz obikezelwe ngokwethiyori we-agarose (umugqa oluhlaza) kanye nama-fibrin microgels (imigqa nezimpawu zombala ofanayo).b, c Iphaneli ephezulu: umdwebo womdwebo wamaketanga enethiwekhi ye-agarose (b) ne-fibrin (c) ngaphambi (kwesobunxele) nangemuva (kwesokudla) ukucindezelwa kwe-biaxial.Ngezansi: Umumo wenethiwekhi ehambisanayo ngaphambi nangemuva kokuguqulwa.Izikhombisi-ndlela zokuminyanisa u-x kanye no-y ziboniswa ngemicibisholo e-magenta nensundu, ngokulandelana.Emfanekisweni ongenhla, amaketango amanethiwekhi aqondiswe kulezi zikhombisi-ndlela zika-x kanye no-y aboniswa nemigqa ehambisanayo ye-magenta kanye nensundu, futhi amaketango aqondiswe ku-z inkombandlela amelwe imigqa eluhlaza.Kujeli ye-fibrin (c), imigqa ensomi nensundu eziqondisweni zika-x kanye no-y igoba kakhulu kunesimo sokungaguquki, futhi imigqa eluhlaza ku-z iyagoba futhi yeluleke.Ukungezwani phakathi kwezikhombisi-ndlela zokuminyanisa nokushuba kudluliselwa ngemicu enezikhombisi-ndlela eziphakathi nendawo.Emajeli e-agarose, amaketanga kuzo zonke izinkomba anquma ukucindezela kwe-osmotic, okwenza kube negalelo elikhulu ekuguqulweni kwejeli.d Ushintsho olubikezelwe esilinganisweni se-biaxial Poisson, } }^{{{{{\rm{eff}}}}}}}} =-{{{{\rm{ln}}}}}} {\lambda }_{ z}/{{{{{ {{ \rm{ln}}}}}}{\lambda }_{r}\ ), ngokuminyanisa okulinganayo kwamajeli e-agarose (umugqa oluhlaza) kanye ne-fibrin (umugqa obomvu).I-inset ibonisa ukuguqulwa kwe-biaxial yejeli.e Ukushintsha kwengcindezi yokuthutha i-ΔPtr, ejwayele ukuqina kwejeli S, ihlelwe njengomsebenzi wesilinganiso sokucindezelwa kwe-agarose ne-fibrin microgel.Imibala yophawu ihambisana nemibala eku-(a).Imigqa eluhlaza nebomvu ibonisa ubudlelwano bethiyori phakathi kwe-ΔPtr/S ne-1/λr ye-agarose nejeli ye-fibrin, ngokulandelanayo.Ingxenye edeshisiwe yomugqa obomvu ibonisa ukukhuphuka kwe-ΔPtr ngaphansi kokucindezelwa okuqinile ngenxa yokusebenzisana kwe-interfiber.
Lo mehluko uhlotshaniswa nezindlela ezihlukene zokuguqulwa kwe-fibrin ne-agarose microgel networks, ehlanganisa izintambo eziguquguqukayo ezingama-24 kanye ne-rigid25, ngokulandelana.Ukucindezelwa kwe-Biaxial kwama-gel aguquguqukayo kuholela ekwehleni kwevolumu yawo kanye nokwenyuka okuhambisana nokugxila kanye nengcindezi ye-osmotic, okuholela ekwandiseni ijeli ngendlela engenamkhawulo.Ukwelulwa kokugcina kwejeli kuncike ebhalansini yokwanda kwamandla wamahhala we-entropic wamaketanga alulekile kanye nokuncipha kwamandla amahhala e-osmosis ngenxa yokugxila okuphansi kwe-polymer kujeli eyeluliwe.Ngaphansi kokucindezelwa okuqinile kwe-biaxial, ukwelulwa kwejeli kukhuphuka ngokuthi λz ≈ 0.6 \({{\lambda}_{{{\rm{r}}}}^{-2/3}}\) (bona Fig. 2a ku- isigaba sengxoxo 5.3.3).Izinguquko ezihambisanayo kumaketanga aguquguqukayo kanye nokuma kwamanethiwekhi ahambisanayo ngaphambi nangemva kokugcinwa kwe-biaxial kuboniswa ku-Fig.2b.
Ngokuphambene, amajeli ane-fibrous njenge-fibrin ngokwemvelo asabela ngendlela ehlukile ekugcinweni kwe-biaxial.Imicu eqondiswe ngokuyinhloko ngokuhambisana nesiqondiso se-compression flex (ngaleyo ndlela yehlisa ibanga phakathi kwezixhumanisi eziphambanayo), kuyilapho imicu egxile kakhulu ekuqondeni kokucindezelwa iqonde futhi yelule ngaphansi kwesenzo samandla okunwebeka, okwenza ijeli ibe yinde ( Umfanekiso 1).2c) Izakhiwo ze-SM, MM kanye ne-RM ezingashintshiwe ziphawulwe ngokuhlaziya izithombe zazo ze-SEM ne-CFM (Isigaba Sengxoxo Eyengeziwe IV kanye Nomfanekiso Owengeziwe 9).Ngokunquma i-elastic modulus (E), ububanzi (d), ubude bephrofayili (R0), ibanga phakathi kweziphetho (L0 ≈ R0) kanye ne-engeli emaphakathi (ψ0) yezintambo kuma-microgels we-fibrin angashintshiwe (Ithebula Lesengezo 2) - 4), sithola ukuthi intambo yokugoba imodulus \({k}_{{{{{{\rm{b))))))))))}=\frac{9\pi E{d}^{4} } {4 {\psi } _{0}^{2}{L}_{0}}\) incane kakhulu kune-tensile modulus\({k}_{{{{{{\rm{s}}}}} }} }}=E\frac{\pi {d}^{2}{R}_{0}}{4}\), ngakho-ke kb/ks ≈ 0.1 (Ithebula Lokwengeza 4).Ngakho-ke, ngaphansi kwezimo zokugcinwa kwejeli ye-biaxial, imicu ye-fibrin igoba kalula, kodwa imelana nokwelula.Ukwelulwa kwenethiwekhi ye-filamentous ngaphansi kokucindezelwa kwe-biaxial kuboniswa ku-Supplementary Fig. 17.
Sakha imodeli yethiyori ehambisanayo (Isigaba Sengxoxo Eyengeziwe V kanye Nezibalo Ezingeziwe 10–16) lapho ukunwetshwa kwejeli ene-fibrous kunqunywa kusukela ekulinganisweni kwendawo kwamandla okunwebeka asebenza kujeli futhi sibikezele ukuthi kuhlobo oluqinile lwe-biaxial λz - 1 ngaphansi kwesibopho
I-equation (1) ibonisa ukuthi nangaphansi kokucindezelwa okuqinile (\({\lambda }_{{{\mbox{r))))\,\kuya \,0\)) kukhona ukunwetshwa kwejeli kancane kanye nokuguqulwa kwe-elongation okulandelayo saturation λz–1 = 0.15 ± 0.05.Lokhu kuziphatha kuhlobene (i) \({\left({k}_{{{{({\rm{b}}}}}}}}}}/{k}_{{{{{{\rm} {s }}}}}}\kwesokudla)}^{1/2}\) ≈ 0.15−0.4 kanye (ii) negama elikubakaki abayisikwele lilinganisa ngokungafaniyo \(1{{\mbox{/}}} \sqrt { 3 }\) kumabhondi e-biaxial aqinile. Kubalulekile ukuqaphela ukuthi i-prefactor \({\left({k}_{({\mbox{b)))))/{k}_{({\mbox{ s))))\kwesokudla)}^{1/ 2 }\) akuhlangene nokuqina kocu E, kodwa kunqunywa kuphela yi-aspect ratio yochungechunge d/L0 kanye ne-engeli emaphakathi ye-arc. ψ0, efana ne-SM, MM kanye ne-RM (Ithebula Lokwengeza 4).
Ukuze siqhubeke sigqamisa umehluko wobunzima obubangelwa inkululeko phakathi kwamajeli aguquguqukayo kanye nama-filamentous, sethula isilinganiso se-biaxial Poisson \({\nu }_{{{({\rm{b)))))))) {{\ mbox { =}}}\,\mathop{{\lim}}\imikhawulo_{{\lambda}_{{{{({({\rm{r}}}}}}}}\kuya ku-1}\ frac{{\ lambda } _{ {{{{\rm{z}}}}}}-1}{1-{\lambda }_{{({\rm{r}}}}}}}}}, \) ichaza okungenamkhawulo ukuma kohlobo lwejeli ekuphenduleni uhlobo olulinganayo eziqondisweni ezimbili zama-radial, futhi inweba lokhu kuzinhlobo ezinkulu ezifanayo \ rm{b }}}}}}}}} ^ {{{{\ rm{eff}}}}}}} }}=-{{{{{\rm{ln}}}}}}}} { \lambda } _{z} /{{({({\rm{ln))))))))} {\lambda }_{{{({\rm{r))))))))}\) .Emkhiwaneni.2d imibukiso \({{{{{\rm{\nu }}}}}}}_{{{({\rm{b}}}}}}}}^{{{ {{\rm { eff }}}}}}}}\) yokuminyanisa okufanayo kwe-biaxial kwezimo eziguquguqukayo (njenge-agarose) namajeli aqinile (njenge-fibrin) (Ingxoxo engeziwe, Isigaba 5.3.4), futhi igqamisa ubudlelwane phakathi komehluko oqinile ezimpendulweni zokuvalelwa. Kumajeli e-agarose ngaphansi kwemikhawulo eqinile {\rm{eff}}}}}}}}\) ikhuphukela enanini le-asymptotic engu-2/3, futhi kumajeli e-fibrin yehla ibe ziro, njengoba lnλz/lnλr → 0, njengoba i-λz inyuka ngo saturation njengoba λr ikhula.Qaphela ukuthi ekuhlolweni, ama-microgel ayindilinga avaliwe ashintshashintsha ngendlela efanayo, futhi ingxenye yawo emaphakathi iba nokucindezelwa okunamandla;kodwa-ke, ukukhishwa kwenani elikhulu elingu-1/λr kwenza kube nokwenzeka ukuqhathanisa ukuhlolwa nethiyori yamajeli akhubazeke ngendlela efanayo.
Omunye umehluko ekuziphatheni kwama-gel e-flexible chain kanye nama-filamentous gels atholakala ngenxa yokunyakaza kwawo ekusikeni.Ingcindezi yokuthutha i-ΔPtr, eyenzelwe ukuqina kwejeli S, yanda ngokucindezelwa okwandayo (Fig. 2e), kodwa ku-2.0 ≤ 1/λr ≤ 3.5, ama-fibrin microgel abonise amanani aphansi kakhulu we-ΔPtr/S phansi ngesikhathi sokuncipha.Ukugcinwa kwe-agarose microgel kuholela ekwenyukeni kwengcindezi ye-osmotic, okuholela ekwelulweni kwejeli endaweni ye-longitudinal njengoba ama-molecule e-polymer enwetshwa (Umfanekiso 2b, kwesokunxele) kanye nokwanda kwengcindezi yokudluliselwa kwe-ΔPtr/S ~( 1/λr)14/317.Ngokuphambene nalokho, ukuma kwama-microgel e-fibrin avaliwe kunqunywa ibhalansi yamandla yezintambo zokucindezelwa kwe-radial kanye nokuqina kwe-longitudinal, okuholela ekuguqulweni okuphezulu kwe-longitudinal λz ~\(\sqrt{{k}_{{{{{{{ \rm{ b))))))))} /{k}_{{{{{{{\rm{s}}}}}}}}}\).Ku-1/λr ≫ 1, ukuguqulwa kwengcindezi yokuthutha kukalwa ngokuthi 1}{{({\rm{ln)))))))\left({{\lambda }}_{{{{{{\rm) {r} }}}}}}}^{{-} 1} \kwesokudla)\) (Ingxoxo Eyengeziwe, Isigaba 5.4), njengoba kuboniswa umugqa obomvu oqinile ku-Fig. 2e.Ngakho-ke, i-ΔPtr ibambeke kancane kunakumajeli e-agarose.Ngokucindezelwa nge-1 / λr> 3.5, ukwanda okuphawulekayo kwengxenye yevolumu ye-filaments kanye nokuxhumana kwe-filaments engumakhelwane kunciphisa ukuguqulwa okuqhubekayo kwejeli futhi kuholela ekuphambukeni kwemiphumela yokuhlola kusukela ekubikezelweni (umugqa wamachashazi abomvu ku-Fig. 2e).Siphetha ngokuthi ku-1/λr efanayo kanye ne-Δ\({P}_{{{{{{{\rm{tr}}}}}}}}}_{{{{\rm{fibrin}}}} )) } }}}\) < ΔP < Δ\({P}_{{{{{{{{\rm{tr))))))))}}_{{{{\rm{agarose}}}}}} } } }}\) ijeli ye-agarose izothathwa i-microchannel, futhi ijeli ye-fibrin enokuqina okufanayo izodlula kuyo.Okwe-ΔP < Δ\({P}_{{{{{{\rm{tr))))))))))_{{{{{\rm{fibrin))))))))))}\ ), Amabili Womabili ama-gel azovimbela isiteshi, kodwa ijeli ye-fibrin izophusha ijule futhi icindezele ngokuphumelelayo, ivimbele ukugeleza kwamanzi ngokuphumelelayo.Imiphumela eboniswe kuMfanekiso 2 ibonisa ukuthi ijeli ene-fibrous ingasebenza njengepulaki ephumelelayo yokunciphisa ukopha noma ivimbele ukutholakala kwegazi ezimila.
Ngakolunye uhlangothi, i-fibrin yenza i-clot scaffold eholela ku-thromboembolism, isimo se-pathological lapho i-thrombus ivala umkhumbi ku-ΔP < ΔPtr, njengezinye izinhlobo ze-ischemic stroke (Fig. 3a).Ukwelulwa okubuthakathaka okubangelwa ukuvinjelwa kwama-microgel e-fibrin kubangele ukwanda okunamandla kokugxilwa kwe-fibrin ye-C/C fibrinogen uma kuqhathaniswa namajeli eketango aguquguqukayo, lapho i-C ne-C fibrinogen ikhawulelwe futhi ama-microgels angashintshile, ngokulandelana.Ukugxila kwe-polymer kujeli.Umfanekiso we-3b ubonisa ukuthi i-fibrinogen C / C ku-SM, MM, ne-RM yanda ngaphezu kokuphindwe kasikhombisa ku-1 / λr ≈ 4.0, eqhutshwa ukuvinjelwa nokuphelelwa amandla kwamanzi (I-Supplementary Fig. 16).
Umfanekiso ohleliwe wokuvaleka komthambo wobuchopho ophakathi ebuchosheni.b Ukwenyuka okuhlobene okulinganiselwe okulinganiselwe ekugxilweni kwe-fibrin ku-SM evimbelayo (imibuthano ebomvu eqinile), MM (izikwele ezimnyama eziqinile), kanye ne-RM (onxantathu abaluhlaza okwesibhakabhaka okuqinile).c Idizayini yokuhlola esetshenziselwa ukutadisha ukuqhekeka kwamajeli e-fibrin anomkhawulo.Isixazululo se-tPA esine-fluorescently enelebula ku-TBS sajovwa ngesilinganiso sokugeleza esingu-5.6 × 107 µm3/s kanye nokwehla kwengcindezi eyengeziwe okungu-0.7 Pa kumashaneli abekwe eceleni kwe-eksisi ende ye-microchannel enkulu.d Isithombe esincane esihlanganisiwe samashaneli amaningi we-MM ovimbelayo (D0 = 200 µm) kokuthi Xf = 28 µm, ΔP = 700 Pa naphakathi nokuhlukaniswa.Imigqa enamachashazi aqondile ibonisa ukuma kokuqala konqenqema olungemuva nangaphambili lwe-MM ku-tlys = 0. Imibala eluhlaza nepinki ihambisana ne-FITC-dextran (70 kDa) kanye ne-tPA enelebula elithi AlexaFluor633, ngokulandelana.e Ivolumu yesihlobo esishintshayo isikhathi sama-RM avaliwe ano-D0 ka-174 µm (unxantathu ohlaziywe ohlanekezelwe ovulekile ohlaza okwesibhakabhaka), 199 µm (unxantathu ovulekile oluhlaza okwesibhakabhaka), kanye no-218 µm (unxantathu ovulekile oluhlaza okwesibhakabhaka), ngokulandelanayo, kumshaneli omncane oyikhone one-Xf = 28 ± 1 µm.izigaba zine-ΔP 1200, 1800, ne-3000 Pa, ngokulandelana, kanye ne-Q = 1860 ± 70 µm3/s.Isifakeli sibonisa i-RM (D0 = 218 µm) ixhuma i-microchannel.f Ukuhluka kwesikhathi kwevolumu ehlobene ye-SM, MM noma RM ebekwe ku-Xf = 32 ± 12 µm, ku-ΔP 400, 750 kanye no-1800 Pa kanye ne-ΔP 12300 Pa kanye ne-Q 12300 endaweni eyikhonikhi ye-microchannel, ngokulandelana ku-23600 no-1m800 µ /s.I-Xf imele indawo yangaphambili ye-microgel futhi inquma ibanga layo kusukela ekuqaleni kokuncipha.I-V (tlys) ne-V0 ivolumu yesikhashana ye-lysed microgel kanye nevolumu ye-microgel engaphazanyiswa, ngokulandelanayo.Imibala yezinhlamvu ihambisana nemibala eku-b.Imicibisholo emnyama ku-e, f ihambisana nesikhathi sokugcina ngaphambi kokudlula kwama-microgel ku-microchannel.Ibha yesikali ku-d, e ingu-100 µm.
Ukuze siphenye umthelela wokuvinjelwa ekuncishisweni kokugeleza koketshezi kuwo wonke amajeli e-fibrin avimbelayo, sifunde i-lysis ye-SM, MM, ne-RM engene ngesisebenzeli se-thrombolytic tissue plasminogen activator (tPA).Umfanekiso 3c ubonisa idizayini yokuhlola esetshenziselwa ukuhlola kwe-lysis. Ku-ΔP = 700 Pa (<ΔPtr) kanye nezinga lokugeleza, Q = 2400 μm3/s, ye-Tris-buffered saline (TBS) exutshwe no-0.1 mg/mL we (fluorescein isothiocyanate) FITC-Dextran, i-microgel ivalele umzila omncane ocijile. isifunda. Ku-ΔP = 700 Pa (<ΔPtr) kanye nezinga lokugeleza, Q = 2400 μm3/s, ye-Tris-buffered saline (TBS) exutshwe no-0.1 mg/mL we (fluorescein isothiocyanate) FITC-Dextran, i-microgel ivalele umzila omncane ocijile. isifunda. I-При ΔP = 700 Па (<ΔPtr) kanye ne- скорости потока, Q = 2400 мкм3/с, трис-буферного солевого раствора (TBS), смешанного с 0,1 мг/мл (флицанца), микрогель перекрывал сужающийся микроканал. Ku-ΔP = 700 Pa (<ΔPtr) kanye nezinga lokugeleza, Q = 2400 µm3/s, ye-Tris buffered saline (TBS) exutshwe no-0.1 mg/mL (fluorescein isothiocyanate) FITC-dextran, i-microgel ivalele umzila omncane oguqukayo.isifunda.在ΔP = 700 Pa (<ΔPtr) 和流速Q = 2400 μm3/s 的Tris 缓冲盐水(TBS) 与0.1 mg/mL微凝胶堵塞了锥形微通道地区。在ΔP = 700 Pa (<ΔPtr) 和流速Q = 2400 μm3/s了锥形微通道地区。 Микрогели закупориваются при смешивании трис-буферного солевого раствора (TBS) с 0,1 мг/мл (флуоресцеинизотиоцианианат) FITC-декстра Δправгей (7) FITC-декстран сти потока Q = 2400 мкм3/с Конические области микроканалов. Ama-Microgel axhunywa lapho i-Tris buffered saline (TBS) ixutshwa no-0.1mg/mL (fluorescein isothiocyanate) FITC-dextran kokuthi ΔP = 700 Pa (<ΔPtr) kanye nesilinganiso sokugeleza Q = 2400 µm3/s Izifunda ze-Conical zamashaneli amancane.Indawo eya phambili i-Xf ye-microgel inquma ibanga layo ukusuka endaweni yokuqala yokushwabana u-X0.Ukuze kusungulwe i-lysis, isixazululo se-tPA ebhalwe ngokukhanyayo ku-TBS sajovwa sisuka esiteshini esibekwe nge-orthogonally kuya ku-eksisi ende yomzila omncane omkhulu.
Lapho isixazululo se-tPA sifinyelela ku-occlusal MM, umkhawulo ongemuva we-microgel waba mnyama, okubonisa ukuthi i-fibrin cleavage iqale ngesikhathi i-tlys = 0 (Fig. 3d kanye ne-Supplementary Fig. 18).Ngesikhathi se-fibrinolysis, i-tPA ebhalwe udayi inqwabelana ngaphakathi kwe-MM futhi ibophe emicu ye-fibrin, okuholela ekwandeni kancane kancane kokuqina kombala opinki wama-microgels.Ku-tlys = 60 min, i-MM yenza izinkontileka ngenxa yokuhlakazwa kwengxenye yayo engemuva, futhi indawo yonqenqema oluhamba phambili lwe-Xf ishintsha kancane.Ngemva kwemizuzu engu-160, i-MM enenkontileka enamandla yaqhubeka inkontileka, futhi ku-tlys = 161 min, yadlula, ngaleyo ndlela ibuyisela ukugeleza kwamanzi nge-microchannel (Fig. 3d kanye ne-Supplementary Fig. 18, ikholomu yesokudla).
Emkhiwaneni.I-3e ibonisa ukwehla okuncike esikhathini se-lysis-mediated kuvolumu V (tlys) okujwayelekile kuvolumu yokuqala ye-V0 yama-microgels we-fibrin anosayizi ohlukile.I-CO ene-D0 174, 199, noma 218 µm yafakwa ku-microchannel ene-ΔP 1200, 1800, noma 3000 Pa, ngokulandelana, kanye ne-Q = 1860 ± 70 µm3/s ukuze kuvinjwe i-microchannel (Fig. 3e, inset).umsoco.Ama-microgels ancipha kancane kancane aze abe mancane ngokwanele ukudlula eziteshini.Ukuncipha kwevolumu ebalulekile ye-CO enobubanzi obukhulu bokuqala kudinga isikhathi eside se-lysis.Ngenxa yokugeleza okufanayo kuma-RM anosayizi abahlukene, ukuqhekeka kwenzeka ngenani elifanayo, okuholela ekugayeni kwezingxenye ezincane zama-RM amakhulu kanye nokudluliselwa kwawo okubambezelekile.Emkhiwaneni.I-3f ibonisa ukuncishiswa okuhlobene ku-V(tlys)/V0 ngenxa yokuhlukaniswa kwe-SM, MM, ne-RM kokuthi D0 = 197 ± 3 µm ehlelwe njengomsebenzi we-tlys.Ku-SM, MM ne-RM, beka i-microgel ngayinye ku-microchannel ene-ΔP 400, 750 noma 1800 Pa kanye ne-Q 12300, 2400 noma 1860 µm3/s, ngokulandelanayo.Nakuba ingcindezi esetshenziswa ku-SM yayingaphansi ngokuphindwe izikhathi ezingu-4.5 kunaleyo ye-RM, ukugeleza kwe-SM kwakunamandla izikhathi ezingaphezu kweziyisithupha ngenxa yokufinyeleleka okuphezulu kwe-SM, futhi ukushwabana kwe-microgel kwehle ukusuka ku-SM kuya ku-MM kanye ne-RM. .Isibonelo, ku-tlys = 78 min, i-SM yahlakazeka kakhulu futhi yasuswa, kuyilapho u-MM no-PM beqhubeka nokuvala iziteshi ezincane, naphezu kokugcina kuphela i-16% kanye ne-20% yevolumu yabo yasekuqaleni, ngokulandelana.Le miphumela iphakamisa ukubaluleka kwe-convection-mediated lysis yamajeli e-fibrous constricted futhi ihlobane nemibiko yokugaya ngokushesha kwamahlule anokuqukethwe kwe-fibrin ephansi.
Ngakho-ke, umsebenzi wethu ubonisa ngokuhlola kanye nethiyori indlela ama-gel ane-filamentous asabela ngayo ekuboshweni kwe-biaxial.Ukuziphatha kwama-gel ane-fibrous endaweni elinganiselwe kunqunywa i-asymmetry eqinile yamandla obunzima be-filaments (i-soft in compression kanye ne-tension eqinile) futhi kuphela nge-aspect ratio kanye nokugoba kwe-filaments.Lokhu kusabela kuholela ekwandiseni okuncane kwamajeli ane-fibrous aqukethwe kuma-capillaries amancane, isilinganiso sawo se-biaxial Poisson sehla ngokucindezelwa okwandayo kanye nokucindezela okuncane kokukhanya.
Njengoba ukuqukatha kwe-biaxial kwezinhlayiya ezithambile ezikhubazekile kusetshenziswa ezinhlobonhlobo zobuchwepheshe, imiphumela yethu ikhuthaza ukuthuthukiswa kwezinto ezintsha ze-fibrous.Ikakhulukazi, ukugcinwa kwe-biaxial kwama-gel e-filamentous kuma-capillaries amancane noma amashubhu kuholela ekuhlanganiseni kwawo okuqinile kanye nokwehla okubukhali kokungena.Ukuvinjelwa okunamandla kokugeleza koketshezi ngamajeli e-occlusive fibrous kunezinzuzo lapho kusetshenziswa njengamapulaki okuvimbela ukopha noma ukunciphisa ukunikezwa kwegazi kuma-malignancies33,34,35.Ngakolunye uhlangothi, ukwehla kokugeleza koketshezi ngejeli ye-occlusal fibrin, ngaleyo ndlela kuvimbela i-convective-mediated thrombus lysis, kunikeza inkomba yokuncipha kancane kwamahlule e-occlusal [27, 36, 37].Uhlelo lwethu lokumodela luyisinyathelo sokuqala sokuqonda imithelela yempendulo yemishini yama-fibrous biopolymer hydrogels ekugcinweni kwe-biaxial.Ukufaka amangqamuzana egazi noma ama-platelet kumajeli e-fibrin avimbelayo kuzothinta ukuziphatha kwawo okukhawulelwe 38 futhi kuzoba isinyathelo esilandelayo sokwembula ukuziphatha kwezinhlelo ezibaluleke kakhulu zebhayoloji.
Ama-reagents asetshenziselwa ukulungisa ama-microgel e-fibrin nokwenza amadivayisi e-MF achazwa kokuthi Ulwazi Olwengeziwe (Izindlela Ezingeziwe Izigaba 2 no-4).Ama-microgel e-Fibrin alungiswa ngokufaka isixazululo esixubile se-fibrinogen, i-Tris buffer kanye ne-thrombin ekugelezeni okugxile kudivayisi ye-MF, kulandele i-droplet gelation.Isixazululo se-bovine fibrinogen (60 mg/ml ku-TBS), isixazululo se-Tris buffer ne-bovine thrombin (5 U/ml kusixazululo se-10 mM CaCl2) sasetshenziswa kusetshenziswa amaphampu esirinji amabili alawulwa ngokuzimela (PhD 200 Harvard Apparatus PHD 2000 Syring Pump).ukuvimba MF, USA).Isigaba esiqhubekayo sikawoyela esiqukethe i-1 wt.% block copolymer PFPE-P(EO-PO)-PFPE, yethulwa kuyunithi ye-MF kusetshenziswa iphampu yesirinji yesithathu.Amaconsi akhiwe kudivayisi ye-MF aqoqwa kushubhu ye-centrifuge engu-15 ml equkethe uwoyela we-F.Faka amashubhu kubhavu wamanzi ongu-37 °C ihora elingu-1 ukuze ugcwalise i-fibrin gelation.I-FITC ebizwa ngokuthi ama-fibrin microgels alungiswa ngokuxuba i-bovine fibrinogen kanye ne-FITC ebhalwe ukuthi i-fibrinogen yomuntu ngesilinganiso sesisindo esingu-33:1, ngokulandelanayo.Inqubo iyafana nokulungiswa kwe-fibrin microgels.
Dlulisa ama-microgels ukusuka emafutheni F ukuya ku-TBS ngokufaka i-centrifuging ukuhlakazeka ku-185 g imizuzu emi-2.Ama-microgel anciphile ahlakazwa emafutheni F ahlanganiswe no-20 wt.% we-perfluorooctyl alcohol, wabe esehlakazwa ku-hexane equkethe u-0.5 wt.% Span 80, i-hexane, 0.1 wt.% i-Triton X emanzini ne-TBS.Ekugcineni, ama-microgels ahlakazwa ku-TBS equkethe i-0.01 wt% Phakathi kwe-20 futhi agcinwa ku-4 ° C cishe amasonto angu-1-2 ngaphambi kokuhlolwa.
Ukwenziwa kwedivayisi ye-MF kuchazwe Olwazini Olwengeziwe (Izindlela Zokwengeza Isigaba 5).Esivivinyweni esijwayelekile, inani eliphozithivu le-ΔP linqunywa ubude obuhlobene bamachibi axhunywe ngaphambi nangemuva kwedivayisi ye-MF yokwethula ama-microgel anobubanzi obungu-150
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