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Ubonisa ikharawusi yezilayidi ezithathu ngaxeshanye.Sebenzisa amaqhosha angaphambili nalandelayo ukutyhutyha izilayidi ezithathu ngexesha, okanye sebenzisa amaqhosha esilayidi ekupheleni ukuya kwizilayidi ezithathu ngexesha.
Ukuthintelwa kwee-fibrous hydrogels kwi-capillaries emxinwa kubaluleke kakhulu kwiinkqubo zebhayoloji kunye ne-biomedical.Uxinzelelo kunye nokunyanzeliswa kwe-uniaxial ye-fibrous hydrogels ifundwe ngokubanzi, kodwa impendulo yabo ekugcinweni kwe-biaxial kwii-capillaries ihlala ingahlolisiswa.Apha, sibonisa ngovavanyo kunye nethiyori ukuba iigels ze-filamentous ziphendula ngokusemgangathweni ngokwahlukileyo ukunyanzeliswa kuneejeli eziguquguqukayo zekhonkco ngenxa ye-asymmetry kwiipropati zomatshini we-filaments edibeneyo, ethambileyo ekunyanzelweni kunye nokuqina koxinzelelo.Ngaphantsi kogcino oluluqilima, ijeli yefibrous ibonisa ubude obuncinci kunye nokuncipha kwe-asymptotic kumlinganiselo we-biaxial Poisson ukuya kwi-zero, okukhokelela ekuhlanganiseni okuqinileyo kwejeli kunye nokungenwa kakuhle kolwelo ngejeli.Ezi ziphumo zibonisa ukuxhathisa kwe-thrombi e-occlusive eyoluliweyo kwi-lysis ngamayeza onyango kwaye ivuselela uphuhliso lwe-endovascular embolization esebenzayo ukusuka kwi-fibrous gels ukumisa ukopha kwe-vascular okanye ukuvimbela ukunikezelwa kwegazi kwamathumba.
Uthungelwano lweFibrous lusisiseko sesakhiwo kunye neebhloko zokwakha ezisebenzayo zezicubu kunye neeseli eziphilayo.I-Actin yinxalenye enkulu ye-cytoskeleton1;I-fibrin yinto ephambili ekuphulukisweni kwesilonda kunye ne-thrombus formation2, kunye ne-collagen, i-elastin kunye ne-fibronectin ngamalungu e-matrix e-extracellular kwizilwanyana3.Uthungelwano olufunyenweyo lwee-fibrous biopolymers ziye zaba zizixhobo ezinosetyenziso olubanzi kubunjineli beethishu4.
Uthungelwano lweFilamentous lumele udidi olwahlukileyo lwento ethambileyo yebhayoloji eneempawu zomatshini ezahlukileyo kuthungelwano lweemolekyuli eziguquguqukayo5.Ezinye zezi zakhiwo ziye zavela ngexesha lokuzivelela ukuze kulawuleke ukuphendulwa kwezinto zebhayoloji kwi-deformation6.Umzekelo, uthungelwano lwe-fibrous lubonisa i-elasticity yomgca kwii-strains ezincinci7,8 ngelixa kwiintlobo ezinkulu zibonakalisa ukuqina okwenyuka kwe-9,10, ngaloo ndlela igcina ingqibelelo yezicubu.Iimpembelelo zezinye iipropathi zemishini yeegel ze-fibrous, ezifana noxinzelelo olubi oluqhelekileyo ekuphenduleni i-shear strain11,12, ayikafunyanwa.
Iimpawu zemishini ye-semi-flexible fibrous hydrogels ziye zafundwa phantsi kwe-uniaxial tension13,14 kunye ne-compression8,15, kodwa ukunyanzeliswa kwe-biaxial yenkululeko kwi-capillaries encinci okanye iityhubhu ayizange ifundwe.Apha sichaza iziphumo zovavanyo kwaye siphakamisa ngokwethiyori indlela yokuziphatha kwee-fibrous hydrogels phantsi kokugcinwa kwe-biaxial kumajelo e-microfluidic.
Ii-microgels ze-Fibrin ezinemilinganiselo eyahlukeneyo ye-fibrinogen kunye ne-thrombin kunye nobukhulu be-D0 ukusuka kwi-150 ukuya kwi-220 µm zenziwa kusetyenziswa indlela ye-microfluidic (umzobo owongezelelweyo 1).Kwikhiwane.I-1a ibonisa imifanekiso ye-fluorochrome ebhalwe nge-microgels efunyenwe kusetyenziswa i-confocal fluorescence microscopy (CFM).Ii-microgels ziyi-spherical, zine-polydispersity engaphantsi kwe-5%, kwaye zifanayo kwisakhiwo kuzo zonke izikali ezihlolwe yi-CFM (uLwazi olongezelelweyo kunye ne-Movies S1 kunye ne-S2).Umyinge we-pore ye-microgels (enqunywe ngokulinganisa i-Darcy permeability16) yehla ukusuka kwi-2280 ukuya kwi-60 nm, umxholo we-fibrin wanda ukusuka kwi-5.25 ukuya kwi-37.9 mg / mL, kwaye i-concentration ye-thrombin yehla ukusuka kwi-2.56 ukuya kwi-0.27 iiyunithi / mL, ngokulandelanayo.(Iinkcukacha ezongezelelweyo).Irayisi.2), 3 kunye netheyibhile eyongezelelweyo 1).Ukuqina okuhambelanayo kwe-microgel kwanda ukusuka kwi-0.85 ukuya kwi-3.6 kPa (i-Supplementary Fig. 4).Njengemizekelo yeegels ezenziwe kwiintambo eziguquguqukayo, i-agarose microgels yobunzima obuhlukeneyo busetyenziswa.
Umfanekiso we-Fluorescence microscopy we-fluorescein isothiocyanate (FITC) obhalwe ukuba PM unqunyanyisiwe kwi-TBS.Isikali sebha ngama-500 µm.b imifanekiso ye-SEM ye-SM (phezulu) kunye ne-RM (ezantsi).Isikali bar 500 nm.c Umzobo weSchematic wetshaneli yemicrofluidic equka umjelo omkhulu (idiameter dl) kunye nommandla ocutheneyo omile okwekhowuni ene-engile yokungena α ye-15 ° kunye nedayamitha ye-dc = 65 µm.d Ukusuka ekhohlo ukuya ekunene: Imifanekiso yemakroskopu ye-RM (i-diameter D0) kwimijelo emikhulu, ummandla we-conical kunye noxinzelelo (ukunciphisa ubude bejeli ye-Dz).Isikali sebha li-100 µm.e, f imifanekiso ye-TEM ye-RM engaguqukiyo (e) kunye ne-RM evaliweyo (f), ehleliweyo iyure enye kunye ne-constriction 1 / λr = 2.7, ilandelwa kukukhululwa kunye nokulungiswa kwe-5% yobunzima.glutaraldehyde kwi-TBS.Ububanzi be-CO engaguqukiyo yi-176 μm.Ibar yesikali yi-100 nm.
Sigxininise kwi-fibrin microgels enobunzima be-0.85, 1.87 kunye ne-3.6 kPa (emva koku kuthiwa yi-microgels ethambileyo (SM), i-microgels ephakathi (MM) kunye ne-microgels enzima (RM), ngokulandelanayo).Olu luhlu lwe-fibrin gel stiffness lunomyalelo ofanayo wobukhulu kunye ne-blood clots18,19 kwaye ngoko ke iigels ze-fibrin ezifundwe kumsebenzi wethu zihambelana ngokuthe ngqo kwiinkqubo ze-biological zangempela.Kwikhiwane.I-1b ibonisa imifanekiso ephezulu kunye nesezantsi yezakhiwo ze-SM kunye ne-RM ezifunyenwe ngokusebenzisa i-microscope ye-electron yokuskena (SEM), ngokulandelanayo.Xa kuthelekiswa nezakhiwo ze-RM, uthungelwano lwe-SM lwenziwa ngamacandelo amaninzi kunye namanqaku amancinci amasebe, ahambelana neengxelo zangaphambili ze-20, i-21 (i-Supplementary Fig. 5).Umahluko kwisakhiwo se-hydrogel sihambelana ne-trend yeempawu zayo: ukunyanzeliswa kwe-gel kunciphisa kunye nokunciphisa ubungakanani be-pore ukusuka kwi-SM ukuya kwi-MM kunye ne-RM (iThebhile eyoNgezelelweyo 1), kunye nokuqina kwe-gel ukuguqula.Akukho tshintsho kwisakhiwo se-microgel saphawulwa emva kokugcinwa kwi-4 ° C kwiintsuku ze-30 (i-Supplementary Fig. 6).
Kwikhiwane.I-1c ibonisa umzobo we-microfluidic channel kunye necandelo lomnqamlezo ojikelezayo oqulethe (ukusuka kwesobunxele ukuya ekunene): umjelo omkhulu onobubanzi be-dl apho i-microgel ihlala ingenakuguqulwa, icandelo le-cone-shaped kunye nokunciphisa ububanzi be-dc
Kwikhiwane.I-1e, i-1f ibonisa ukuhanjiswa kwe-electron microscopy (TEM) imifanekiso ye-RM engaguqukiyo kunye ne-biaxially limited RM izakhiwo.Emva kokunyanzeliswa kwe-RM, ubukhulu be-pore ye-microgel bunciphe kakhulu kwaye imilo yabo yaba yi-anisotropic kunye nobukhulu obuncinci kwicala loxinzelelo, oluhambelana nengxelo yangaphambili ye-23.
Uxinzelelo lwe-Biaxial ngexesha lokufinyela lubangela ukuba i-microgel ibe yobude kwicala elingasikelwanga mda kunye ne-coefficient λz = \({D}_{{{{{{\rm{z}}}}}}}}/\({D }_ { 0}\) , apho \({D}_{{{({\rm{z}}}}}}}\) bubude be microgel evaliweyo Umfanekiso 2a ubonisa utshintsho kwi λzvs .1/ λr Okumangalisayo kukuba, phantsi koxinzelelo olunamandla lwe-2.4 ≤ 1/λr ≤ 4.2, i-fibrin microgels ibonisa ubude obungathandabuzekiyo be-1.12 +/- 0.03 λz, echatshazelwa kancinci kuphela ngexabiso le-1 / λr. i-agarose microgels encinci, ebonwa nakwi-compression ebuthakathaka 1/λr = 2.6 ukuya kubude obukhulu λz = 1.3.
imifuniselo ye-Agarose microgel enemodyuli elastiki eyahlukileyo (2.6 kPa, idayimani eluhlaza evulekileyo; 8.3 kPa, isangqa esimdaka ngebala; 12.5 kPa, isikwere esiorenji esivulekileyo; 20.2 kPa, imagenta evulekileyo unxantathu ojikiweyo) kunye neSM (obomvu oqinileyo) Guqula kulinganiso lobude λz ( izangqa), MM (izikwere ezimnyama eziqinileyo) kunye neRM (oonxantathu obhlowu oqinileyo).Imigca eqinileyo ibonisa ingqikelelo yethiyori λz ye-agarose (umgca oluhlaza) kunye ne-fibrin microgels (imigca kunye neempawu zombala ofanayo).b, c Iphaneli ephezulu: umzobo weskim semixokelelwane yenethiwekhi ye-agarose (b) kunye nefibrin (c) ngaphambi (ngasekhohlo) nasemva (ekunene) yoxinzelelo lwe-biaxial.Ezantsi: Ubume bothungelwano oluhambelanayo ngaphambi nasemva kokuguqulwa.Izalathiso zoxinzelelo zika-x kunye no-y ziboniswa ngama-magenta kunye neentolo ezimdaka ngebala, ngokulandelelanayo.Kulo mfanekiso ungentla, amatyathanga othungelwano olujoliswe kula x kunye no-y ulwalathiso luboniswa kunye nemigca yemagenta ehambelanayo kunye ne-brown, kunye namatyathanga ajoliswe kwi-arbitrary z isalathiso imelwe yimigca eluhlaza.Kwi-fibrin gel (c), imigca emfusa kunye nemdaka ku-x kunye no-y izalathiso igoba ngaphezu kwemeko engaguqukiyo, kunye nemigca eluhlaza kwicala lika-z igobe kwaye yolule.Uxinzelelo phakathi kwezalathiso zoxinzelelo kunye noxinzelelo luhanjiswa ngemisonto enezalathiso eziphakathi.Kwiigels ze-agarose, amatyathanga kuwo onke amacala amisela uxinzelelo lwe-osmotic, okwenza igalelo elibalulekileyo kwi-deformation ye-gel.d Utshintsho oluqikelelweyo kumlinganiselo we-biaxial Poisson, } }^{{{{{\rm{eff}}}}}}}} =-{{{{\rm{ln}}}}}}}{\lambda }_{ z}/{{{{{{ \rm{ln}}}}}} {\ lambda }_{r}\ ), ukwenzela ukunyanzeliswa kwe-equibiaxial ye-agarose (umgca oluhlaza) kunye ne-fibrin (umgca obomvu) iigels.I-inset ibonisa ukuguqulwa kwe-biaxial ye-gel.e Ukutshintsha koxinzelelo lwe-Translocation ΔPtr, i-normalized to gel stiffness S, icwangciswe njengomsebenzi we-compression ratio ye-agarose kunye ne-fibrin microgels.Imibala yesimboli ihambelana nemibala eku-(a).Imigca eluhlaza kunye nebomvu ibonisa ubudlelwane bethiyori phakathi kwe-ΔPtr/S kunye ne-1/λr ye-agarose kunye nejeli ye-fibrin, ngokulandelelanayo.Inxalenye edayiweyo yomgca obomvu ibonisa ukunyuka kwe-ΔPtr phantsi koxinzelelo olunamandla ngenxa yokusebenzisana kwe-interfiber.
Lo mahluko unxulumene neendlela ezahlukeneyo zokuguqulwa kwe-fibrin kunye ne-agarose microgel networks, equkethe i-flexible24 kunye ne-rigid25 threads, ngokulandelanayo.Ukunyanzeliswa kwe-Biaxial yeegels eziguquguqukayo kukhokelela ekunciphiseni umthamo wabo kunye nokunyuka okuhambelana noxinzelelo kunye noxinzelelo lwe-osmotic, olukhokelela ekwandiseni ijeli kwicala elingenamkhawulo.Ukwandiswa kokugqibela kwejeli kuxhomekeke kwibhalansi yokunyuka kwamandla e-entropic asimahla amatyathanga awoluliweyo kunye nokuncipha kwamandla asimahla e-osmosis ngenxa yoxinzelelo oluphantsi lwepolymer kwijeli eyoluliweyo.Ngaphantsi koxinzelelo olunamandla lwe-biaxial, ukwandiswa kwejeli kuyanda nge-λz ≈ 0.6 \({{\lambda}_{{{\rm{r}}}}^{-2/3}}\) (jonga umzobo 2a icandelo lengxoxo 5.3.3).Utshintsho oluhambelanayo kwiityathanga eziguquguqukayo kunye nokuma kweenethiwekhi ezihambelanayo ngaphambi nangemva kokugcinwa kwe-biaxial kuboniswe kwiMifanekiso.2b.
Ngokwahlukileyo, iijeli zefibrous ezifana nefibrin ngokwendalo ziphendula ngokwahlukileyo kugcino lwe-biaxial.I-filaments ijolise ikakhulu ngokuhambelana nolwalathiso lwe-flexion flex (ngaloo ndlela inciphisa umgama phakathi kwe-cross-links), ngelixa i-filaments ubukhulu becala i-perpendicular ukuya kwicala lokunyanzeliswa kolula kwaye yolulele phantsi kwesenzo samandla e-elastic, ebangela ukuba ijeli yolule ( Umzobo 1).I-2c) Izakhiwo ze-SM, i-MM kunye ne-RM engatshintshiyo zibonakaliswe ngokuhlalutya imifanekiso yazo ye-SEM kunye ne-CFM (iCandelo leNgxoxo eyoNgezelelweyo ye-IV kunye ne-9).Ngokumisela i-modulus elastic (E), ububanzi (d), ubude beprofayili (R0), umgama phakathi kweziphelo (L0 ≈ R0) kunye ne-angle ephakathi (ψ0) yeentambo kwi-fibrin microgels engatshintshiyo (iThebhile eyoNgezelelweyo 2) - 4), sifumanisa ukuba imodyuli yokugoba intambo \({k}_{{{{{\rm{b))))))))))}=\frac{9\pi E{d}^{4}} {4 {\psi } _{0}^{2}{L}_{0}}\) ingaphantsi kakhulu kunemodyuli yayo yokuqina\({k}_{{{{{\rm{s}}}}} }} }}=E\frac{\pi {d}^{2}{R}_{0}}{4}\), ngoko kb/ks ≈ 0.1 (Itheyibhile eyoNgezelelweyo 4).Ngaloo ndlela, phantsi kweemeko zokugcinwa kwejeli ye-biaxial, iintambo ze-fibrin zigoba ngokulula, kodwa zixhathise ukolula.Ukwandiswa kwenethiwekhi ye-filamentous ephantsi koxinzelelo lwe-biaxial kuboniswe kwi-Supplementary Fig. 17.
Siphuhlisa imodeli yonxulumano lwethiyori (Ingxoxo eyoNgezelelweyo yeCandelo V kunye neMizobo eyoNgezelelweyo 10–16) apho ukwandiswa kwejeli yefibrous kugqitywe ukusuka kwi-equilibrium yasekuhlaleni ye-elastic forces esebenza kwijeli kwaye iqikelela ukuba kwi-biaxial strain eyomeleleyo λz - 1 phantsi kwesithintelo
Inxaki (1) ibonisa ukuba naphantsi koxinzelelo olunamandla (\({\lambda }_{{{\mbox{r))))\,\ukuya \,0\)) kukho ukwandiswa kwejeli okuncinci kunye nokuguqulwa kobude obulandelayo ukuzaliseka λz–1 = 0.15 ± 0.05.Le ndlela yokuziphatha inxulumene (i) \({\ left({k}_{{{({\rm{b}}}}}}}}}}/{k}_{{{{\rm} {s }}}}}}}\ ekunene)}^{1/2}\) ≈ 0.15−0.4 kunye (ii) igama kwizibiyeli ezisisikwere ngokungafaniyo uqikelelo \(1{{\mbox{/}}} \ sqrt 3 s))))\ ekunene)}^{1/ 2 }\) ayinanto yakwenza nokuqina komsonto E, kodwa imiselwa kuphela ngumyinge wembonakalo yomsonto d/L0 kunye ne-engile esembindini ye-arc. ψ0, efana ne-SM, MM kunye neRM (iThebhile eyoNgezelelweyo ye-4).
Ukugxininisa ngakumbi umahluko kuxinzelelo olubangelwa inkululeko phakathi kweegels eziguquguqukayo kunye ne-filamentous, sazisa umlinganiselo we-biaxial Poisson \({\nu }_{{{({\rm{b)))))))} {{\ mbox { =}}\,\mathop{{\lim}}\imida_{{\lambda}_{{{{({\rm{r}}}}}}}}\ukuya 1}\ frac{{\ lambda } _{{{{\rm{z}}}}}-1}{1-{\lambda }_{{({\rm{r}}}}}}}}}}, \) ichaza into engenamida Ukuqhelaniswa koxinzelelo lwejeli ekuphenduleni ubunzima obulinganayo kumacala amabini eradial, kwaye yandise oku kwiintlobo ezinkulu ezifanayo \ rm{b}}}}}}}}}}^{{{{\rm{eff}}}}}}}} }}=-{{{{{\rm{ln}}}}}}}} { \lambda } _{z} /{{{({\rm{ln)))))))}} {\lambda }_{{({\rm{r))))))))}\) .Kwikhiwane.2d ibonisa \({{{{\rm{\nu}}}}}}}}_{{({{\rm{b}}}}}}}}}}^{{{{\rm { eff }}}}}}}}\) ukunyanzeliswa kwe-biaxial efanayo ye-flexible (efana ne-agarose) kunye ne-rigid (efana ne-fibrin) iigels (Ingxoxo eyongezelelweyo, iCandelo 5.3.4), kwaye igxininisa ubudlelwane phakathi kokungafani okunamandla kwiimpendulo zokuvalelwa. Kwijeli ze-agarose phantsi kwezithintelo ezinamandla {\rm{eff}}}}}}}}\) yonyuka kwixabiso le-asymptotic 2/3, kunye nejeli ye-fibrin yehla iye ku-zero, ekubeni lnλz/lnλr → 0, ekubeni i-λz inyuka nge Ukuzaliseka njengoko i-λr isanda.Qaphela ukuba kwimifuniselo, i-microgels evaliweyo engqukuva iguquka ngokungafaniyo, kwaye indawo yazo esembindini ifumana uxinzelelo olunamandla;kunjalo, extrapolation kwixabiso elikhulu 1/λr kwenza kube nokwenzeka ukuthelekisa umfuniselo kunye ithiyori yeejeli ngokufanayo deformed.
Olunye ulwahlulo ekuziphatheni kweegels eziguquguqukayo kunye neegels ze-filamentous zafunyanwa ngenxa yokuhamba kwazo phezu kokunciphisa.Uxinzelelo lwe-translocation ΔPtr, oluqhelekileyo kwi-gel ukuqina kwe-S, kwanda ngokunyuka koxinzelelo (Umfanekiso 2e), kodwa kwi-2.0 ≤ 1 / λr ≤ 3.5, i-fibrin microgels ibonise amaxabiso aphantsi kakhulu e-ΔPtr / S phantsi ngexesha lokunciphisa.Ukugcinwa kwe-agarose microgel kukhokelela ekunyukeni koxinzelelo lwe-osmotic, okukhokelela ekwaluleni kwejeli kwicala lobude njengoko iimolekyuli ze-polymer zoluliwe (umzobo 2b, ngasekhohlo) kunye nokunyuka koxinzelelo lwe-translocation ngu-ΔPtr / S ~( 1/λr)14/317.Ngokuchasene noko, imilo ye-fibrin evaliweyo ye-microgels imiselwa yibhalansi yamandla kwintambo yoxinzelelo lwe-radial kunye noxinzelelo lwe-longitudinal, ekhokelela kwideformation ephezulu ye-longitudinal λz ~\(\sqrt{{k}_{{{{{{{{ \rm{ b))))))))} /{k}_{{{{{{\rm{s}}}}}}}}}}\).Kwi-1/λr ≫ 1, utshintsho kuxinzelelo lokufuduswa kukalwa njenge-1}{{({\rm{ln)))))))\left({{\lambda }}_{{{{{\rm} {r} }}}}}}} ^ {{-} 1} \ ekunene) \) (Ingxoxo eyoNgezelelweyo, iCandelo 5.4), njengoko kubonisiwe ngomgca obomvu oqinileyo kwiSazobe 2e.Ke, i-ΔPtr ayinyanzelwanga kangako kuneejeli ze-agarose.Ukunyanzeliswa nge-1 / λr> 3.5, ukunyuka okubalulekileyo kwi-volume fraction of filaments kunye nokusebenzisana kwe-filaments engabamelwane kunciphisa ukuguqulwa kwejel kwaye kukhokelela ekuphambukeni kweziphumo zovavanyo ukusuka kuqikelelo (umgca wamachaphaza obomvu kwi-Fig. 2e).Siphetha ngelithi nge-1/λr efanayo kunye ne-Δ\({P}_{{{{{{\rm{tr}}}}}}}}}}_{{{\rm{fibrin}}})) }}}}\) < ΔP < Δ\({P}_{{{{{{\rm{tr)))))))}}}_{{{{\rm{agarose}}}}}} } } }}\) ijeli ye-agarose iya kubanjwa yi-microchannel, kwaye i-fibrin gel enobunzima obufanayo iya kudlula kuyo.Ye ΔP < Δ\({P}_{{{{{\rm{tr))))))))))_{{{{\rm{fibrin)))))))))}\\ ), Iigel ezimbini zombini ziya kuvala umjelo, kodwa i-fibrin gel iya kutyhala inzulu kwaye icinezele ngokufanelekileyo, ivimbele ukuhamba kwamanzi ngokufanelekileyo.Iziphumo eziboniswe kuMzobo 2 zibonisa ukuba ijeli yefibrous inokusebenza njengeplagi esebenzayo yokunciphisa ukopha okanye inqanda unikezelo lwegazi kumathumba.
Ngakolunye uhlangothi, i-fibrin yenza i-clot scaffold ekhokelela kwi-thromboembolism, imeko ye-pathological apho i-thrombus ivala isitya kwi-ΔP <ΔPtr, njengezinye iintlobo ze-ischemic stroke (Umfanekiso 3a).Ukwandiswa okubuthathaka okubangelwa kukuthintelwa kwe-fibrin microgels kubangele ukwanda okunamandla kwi-fibrin concentration ye-C / C fibrinogen xa kuthelekiswa neegels zekhonkco eziguquguqukayo, apho i-C kunye ne-C fibrinogen zithintelwe kunye nee-microgels ezingatshintshiyo, ngokulandelanayo.Ugxininiso lwepolymer kwijeli.Umzobo we-3b ubonisa ukuba i-fibrinogen C / C kwi-SM, MM, kunye ne-RM yanda ngaphezu kwezihlandlo ezisixhenxe kwi-1 / λr ≈ 4.0, eqhutywa kukuthintelwa kunye nokuchithwa kwamanzi (i-Supplementary Fig. 16).
Umzobo weskimu wokuvaleka komthambo ophakathi wobuchopho ebuchotsheni.b Ukwandiswa kwesalamane esithintelwe sisithintelo kwi-fibrin yoxinaniso kwi-SM ephazamisayo (izangqa ezibomvu eziqinileyo), i-MM (izikwere ezimnyama eziqinileyo), kunye ne-RM (onxantathu oluhlaza okwesibhakabhaka).c Uyilo lovavanyo olusetyenziselwa uphononongo lokucandeka kweejeli zefibrin ezithintelweyo.Isisombululo se-tPA ene-fluorescent ebhalwe nge-tPA kwi-TBS yatofwa ngesantya sokuhamba kwe-5.6 × 107 µm3/s kunye nokuhla koxinzelelo olongezelelweyo lwe-0.7 Pa kwiitshaneli ezibekwe kwi-perpendicular kwi-axis ende ye-microchannel engundoqo.d Umfanekiso odityanisiweyo we-multichannel microscopic ye-MM ephazamisayo (D0 = 200 µm) e-Xf = 28 µm, ΔP = 700 Pa kunye nangexesha lokwahlula.Imigca enamachaphaza ethe nkqo ibonisa izikhundla zokuqala ze-posterior and anterior edges of the MM at tlys = 0. Imibala eluhlaza kunye nepinki ihambelana neFITC-dextran (70 kDa) kunye ne-tPA ebhalwe nge-AlexaFluor633, ngokulandelanayo.e Umthamo wokutshintsha kwexesha ohambelanayo we-RMs ezivaliweyo kunye ne-D0 ye-174 µm (unxantathu oluhlaza ovulekileyo oguqulweyo), 199 µm (unxantathu ovulekileyo obhlowu), kunye ne-218 µm (unxantathu oblowu ovulekileyo), ngokulandelelanayo, kwi-microchannel ekhonkoni ene-Xf = 28 ± 1 µm.amacandelo ane-ΔP 1200, 1800, kunye ne-3000 Pa, ngokulandelelana, kunye ne-Q = 1860 ± 70 µm3/s.I-inset ibonisa i-RM (D0 = 218 µm) iplaga i-microchannel.f Ukwahluka kwexesha lomthamo ozalanayo we-SM, MM okanye i-RM ebekwe kwi-Xf = 32 ± 12 µm, kwi-ΔP 400, 750 kunye ne-1800 Pa kunye ne-ΔP 12300 Pa kunye ne-Q 12300 kwingingqi ye-conical ye-microchannel, ngokulandelelana kwe-23600 kunye ne-1m800 µ /s.I-Xf imele indawo yangaphambili ye-microgel kwaye imisela umgama wayo ukusuka ekuqaleni kokuncipha.I-V (tlys) kunye ne-V0 yivolumu yesikhashana ye-lysed microgel kunye nomthamo we-microgel engaphazamisekiyo, ngokulandelanayo.Imibala yeempawu ihambelana nemibala eku-b.Iintolo ezimnyama kwi-e, f zihambelana nomzuzu wokugqibela wexesha phambi kokudlula kwe-microgels kwi-microchannel.Ibar yesikali ku-d, e li-100 µm.
Ukuphanda isiphumo sothintelo ekunciphiseni ukuhamba kwamanzi kwiijeli ze-fibrin ezithintelayo, sifunde i-lysis ye-SM, MM, kunye ne-RM yangeniswa nge-thrombolytic agent tissue plasminogen activator (tPA).Umzobo 3c ubonisa uyilo lokulinga olusetyenziselwa imifuniselo ye-lysis. Kwi-ΔP = 700 Pa (<ΔPtr) kunye nesantya sokuhamba, Q = 2400 μm3 / s, ye-Tris-buffered saline (TBS) exutywe ne-0.1 mg/mL ye (fluorescein isothiocyanate) FITC-Dextran, i-microgel ivalele i-microchannel tapered ummandla. Kwi-ΔP = 700 Pa (<ΔPtr) kunye nesantya sokuhamba, Q = 2400 μm3 / s, ye-Tris-buffered saline (TBS) exutywe ne-0.1 mg/mL ye (fluorescein isothiocyanate) FITC-Dextran, i-microgel ivalele i-microchannel tapered ummandla. При ΔP = 700 Па (<ΔPtr) kunye скорости потока, Q = 2400 мкм3/с, трис-буферного солевого раствора (TBS), смешанного с 0,1 мг/млл (флицизренский) микрогель перекрывал сужающийся микроканал. Kwi-ΔP = 700 Pa (<ΔPtr) kunye nesantya sokuhamba, Q = 2400 µm3 / s, ye-Tris buffered saline (TBS) exutywe ne-0.1 mg / mL (fluorescein isothiocyanate) FITC-dextran, i-microgel ivalele i-microchannel eguqukayo.ummandla.在ΔP = 700 Pa (<ΔPtr) 和流速Q = 2400 μm3/s 的Tris 缓冲盐水(TBS) 与0.1 mg/mL的（异硫氰酸荧光素光红荧光素荣路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 (флуоресцеинизотиоцианат) FITC-декстра FITC-декстра сти потока Q = 2400 мкм3/с Конические области микроканалов. I-Microgels iplagwe xa i-Tris buffered saline (TBS) yaxutywa ne-0.1mg/mL (fluorescein isothiocyanate) FITC-dextran e-ΔP = 700 Pa (<ΔPtr) kunye nesantya sokuhamba Q = 2400 µm3/s Imimandla yeConical ye-microchannels.Isikhundla sangaphambili i-Xf ye-microgel imisela umgama wayo ukusuka kwindawo yokuqala yokunciphisa i-X0.Ukuphembelela i-lysis, isisombululo se-fluorescently ebhalwe i-tPA kwi-TBS yatofwa ukusuka kumjelo obekwe nge-orthogonally ukuya kwi-axis ende ye-microchannel engundoqo.
Xa isisombululo se-tPA sifikelele kwi-occlusal MM, i-posterior edge ye-microgel yaba yinto engabonakaliyo, ebonisa ukuba i-fibrin cleavage iqalile ngexesha le-tlys = 0 (Umfanekiso we-3d kunye ne-Supplementary Fig. 18).Ngexesha le-fibrinolysis, i-tPA ebhalwe ngedayi iqokelela ngaphakathi kwe-MM kwaye ibophelele kwimicu ye-fibrin, ekhokelela ekunyukeni kancinci kancinci kombala opinki we-microgels.Kwi-tlys = i-60 min, iikhontrakthi ze-MM ngenxa yokuchithwa kwenxalenye yayo yangasemva, kunye nesikhundla somda wayo ohamba phambili we-Xf utshintsha kancinci.Emva kwemizuzu eyi-160, i-MM enesivumelwano esinamandla yaqhubeka ikhontrakthi, kwaye kwi-tlys = i-161 min, yafumana i-contraction, ngaloo ndlela ibuyisela ukuhamba kwamanzi kwi-microchannel (Umfanekiso we-3d kunye ne-Supplementary Fig. 18, ikholamu yasekunene).
Kwikhiwane.I-3e ibonisa ukuncipha kwexesha elixhomekeke kwi-lysis-mediated kwi-volume V (tlys) eqhelekileyo kumthamo wokuqala we-V0 we-fibrin microgels ezahlukeneyo.I-CO ene-D0 174, 199, okanye i-218 µm ifakwe kwi-microchannel kunye ne-ΔP 1200, 1800, okanye i-3000 Pa, ngokulandelanayo, kunye ne-Q = 1860 ± 70 µm3 / s ukuvala i-microchannel (Umfanekiso 3e, i-inset).isondlo.I-microgels iyancipha ngokuthe ngcembe de ibe yincinci ngokwaneleyo ukuba idlule kwiitshaneli.Ukuncipha komthamo obalulekileyo we-CO kunye nobubanzi obukhulu bokuqala kufuna ixesha elide le-lysis.Ngenxa yokuqukuqela okufanayo kwii-RM ezinobungakanani obahlukeneyo, ukuqhekeka kwenzeka ngesantya esifanayo, okukhokelela ekwetyisweni kwamaqhezu amancinane ee-RM ezinkulu kunye nokulibaziseka kokufuduswa kwazo.Kwikhiwane.I-3f ibonisa unciphiso olunxulumeneyo kwi-V (tlys)/V0 ngenxa yokwahlulwa kwe-SM, MM, kunye ne-RM kwi-D0 = 197 ± 3 µm ecetywe njengomsebenzi we-tlys.Kwi-SM, MM kunye ne-RM, faka i-microgel nganye kwi-microchannel ene-ΔP 400, 750 okanye i-1800 Pa kunye ne-Q 12300, i-2400 okanye i-1860 µm3 / s, ngokulandelanayo.Nangona uxinzelelo olusetyenzisiweyo kwi-SM lwaluyi-4.5 ngamaxesha angaphantsi kwe-RM, ukuhamba nge-SM kwakunamandla angaphezu kwamathandathu ngenxa yokuphakama okuphezulu kwe-SM, kwaye i-shrinkage ye-microgel iyancipha ukusuka kwi-SM ukuya kwi-MM kunye ne-RM. .Ngokomzekelo, kwi-tlys = i-78 min, i-SM ininzi yachithwa kwaye yachithwa, ngelixa i-MM kunye ne-PM baqhubeka bevala i-microchannels, nangona begcina kuphela i-16% kunye ne-20% yomthamo wabo wokuqala, ngokulandelanayo.Ezi ziphumo zibonisa ukubaluleka kwe-lysis ye-convection-mediated lysis ye-gels ye-fibrous constricted kwaye ihambelana neengxelo zokugaya ngokukhawuleza kwamahlwili kunye nomxholo ophantsi we-fibrin.
Ke, umsebenzi wethu ubonisa ngokulinga kunye nethiyori indlela apho iigels ze-filamentous ziphendula ukuvalelwa kwe-biaxial.Ukuziphatha kweegel ze-fibrous kwisithuba esilinganiselweyo kunqunywe yi-asymmetry eyomeleleyo ye-strain energy ye-filaments (i-soft in compression and hard in tension) kwaye kuphela nge-aspect ratio kunye ne-curvature ye-filaments.Oku kusabela kukhokelela kubude obuncinci beejeli zefibrous eziqulethwe kwiicapillaries ezimxinwa, umlinganiselo wabo we-biaxial Poisson wehla ngoxinzelelo olukhulayo kunye noxinzelelo oluncinci lokukhanya.
Ekubeni isiqulatho se-biaxial se-soft deformable particles sisetyenziswa kuluhlu olubanzi lwetekhnoloji, iziphumo zethu zikhuthaza ukuphuhliswa kwezinto ezintsha ze-fibrous.Ngokukodwa, ukugcinwa kwe-biaxial yeegels ze-filamentous kwii-capillaries ezincinci okanye iibhubhu zikhokelela ekuhlanganiseni kwazo okunamandla kunye nokuncipha okubukhali kwi-permeability.Ukuthintela okunamandla kokuhamba kwamanzi ngokusebenzisa i-occlusive fibrous gels kunenzuzo xa isetyenziswe njengeeplagi zokuthintela ukuphuma kwegazi okanye ukunciphisa ukunikezelwa kwegazi kwi-malignancies33,34,35.Ngakolunye uhlangothi, ukuncipha kokuhamba kwamanzi ngejeli ye-occlusal fibrin, ngaloo ndlela inqanda i-convective-mediated thrombus lysis, inika isalathisi sokucotha kwe-lysis ye-occlusal clots [27, 36, 37].Inkqubo yethu yokwenza imodeli linyathelo lokuqala ekuqondeni iziphumo zempendulo yoomatshini ye-fibrous biopolymer hydrogels kugcino lwe-biaxial.Ukudibanisa iiseli zegazi okanye iiplatelet kwiigels ze-fibrin ezithintelayo ziya kuchaphazela ukuziphatha kwazo okuthintelweyo 38 kwaye iya kuba linyathelo elilandelayo ekutyhileni ukuziphatha kweenkqubo ezibaluleke kakhulu zebhayoloji.
Ii-reagents ezisetyenziselwa ukulungisa i-fibrin microgels kunye nokwenza izixhobo ze-MF zichazwe kwiiNkcukacha ezongezelelweyo (Iindlela ezongezelelweyo zeCandelo 2 kunye ne-4).I-microgels ye-Fibrin yalungiswa ngokufaka i-emulsifying isisombululo esixubileyo se-fibrinogen, i-Tris buffer kunye ne-thrombin ekuhambeni okugxininise kwisixhobo se-MF, kulandelwa yi-droplet gelation.Isisombululo se-bovine fibrinogen (60 mg / ml kwi-TBS), isisombululo se-Tris buffer kunye ne-bovine thrombin isisombululo (i-5 U / ml kwi-10 mM isisombululo se-CaCl2) salawulwa kusetyenziswa iimpompo zesirinji ezimbini ezizimeleyo (PhD 200 Harvard Apparatus PHD 2000 Syring Pump).ukubhloka MF, USA).Isigaba esiqhubekayo se-oyile esiqulethe i-1 wt.% ibhloko ye-copolymer PFPE-P(EO-PO)-PFPE, yaziswa kwiyunithi ye-MF kusetyenziswa impompo yesirinji yesithathu.Amaconsi akhiwe kwisixhobo se-MF aqokelelwa kwi-tube ye-centrifuge ye-15 ml equkethe i-F-oyile.Beka iityhubhu kwindawo yokuhlambela amanzi kwi-37 °C ngeyure enye ukugqiba i-fibrin gelation.I-FITC ebhalwe i-fibrin microgels yalungiswa ngokuxuba i-bovine fibrinogen kunye ne-FITC ebhalwe i-fibrinogen yabantu kumlinganiselo we-33: 1 wobunzima, ngokulandelelanayo.Inkqubo iyafana nokulungiswa kwe-fibrin microgels.
Dlulisa i-microgels ukusuka kwioli F ukuya kwi-TBS ngokugxilisa i-dispersion kwi-185 g ye-2 min.Ii-microgels ezincitshisiweyo zasasazwa kwi-oyile F exutywe ne-20 wt.% ye-perfluorooctyl alcohol, emva koko yasasazwa kwi-hexane equlethe i-0.5 wt.% I-Span 80, i-hexane, i-0.1 wt.% i-Triton X emanzini kunye ne-TBS.Ekugqibeleni, i-microgels yahlakazwa kwi-TBS equkethe i-0.01 wt% Phakathi kwe-20 kwaye igcinwe kwi-4 ° C malunga neeveki ze-1-2 ngaphambi kovavanyo.
Ukwenziwa kwesixhobo seMF kuchazwe kwiiNkcukacha ezoNgezelelweyo (Iindlela ezongezelelweyo zeCandelo lesi-5).Kuvavanyo oluqhelekileyo, ixabiso elilungileyo le-ΔP lichongwa kubude obuhambelanayo beendawo zokugcina amanzi ezixhunywe ngaphambi nangemva kwesixhobo se-MF sokwazisa i-microgels kunye nobubanzi be-150
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Ixesha lokuposa: Feb-23-2023