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Ukopha okungalawuleki kungenye yezimbangela eziphambili zokufa.Ukuthola ukopha ngokushesha kuqinisekisa ukusinda kwesihloko njengosizo lokuqala ngesikhathi sokulwa, izingozi zomgwaqo, nemisebenzi yokunciphisa ukufa.I-Nanoporous fiber-reinforced composite scaffold (NFRCS) esuselwe ekwakhekeni kwefilimu ye-hemostatic elula (HFFC) njengesigaba esiqhubekayo singavusa futhi sithuthukise ukopha.Ukuthuthukiswa kwe-NFRCS kusekelwe ekwakhiweni kophiko lukajekamanzi.Isakhiwo sephiko likajekamanzi siqukethe amaphiko aguquguqukayo nabude, futhi ulwelwesi lwamaphiko luxhunywe komunye nomunye ukuze kugcinwe ubuqotho be-microstructure.I-HFFC imboza ngokulinganayo ingaphezulu lefayibha ngefilimu yogqinsi lwe-nanometer futhi ixhuma ugqinsi lukakotini olusatshalaliswe ngokungahleliwe (Ct) (isigaba esihlakazekile) ukuze yakhe isakhiwo esinama-nanoporous.Ukuhlanganiswa kwezigaba eziqhubekayo nezihlakazekile kunciphisa izindleko zomkhiqizo ngokuphindwe kashumi uma kuqhathaniswa nemikhiqizo etholakalayo kwezohwebo.I-NFRCS eguquliwe (ama-tampons noma amabhande esandla) angasetshenziswa ezinhlelweni ezihlukahlukene ze-biomedical.Ucwaningo lwe-in vivo luphethe ngokuthi i-Cp NFRCS ethuthukisiwe iqala futhi ithuthukise inqubo yokuhlangana endaweni yokufakwa kwesicelo.I-NFRCS ingakwazi ukushintsha i-microenvironment futhi isebenze ezingeni lamaselula ngenxa yesakhiwo sayo esinama-nanoporous okuholela ekuphulukisweni okungcono kwenxeba kumodeli yesilonda sokusikwa.
Ukopha okungalawuleki ngesikhathi sokulwa, ukuhlinzwa kanye nezimo eziphuthumayo kungaba yingozi enkulu empilweni yabalimele1.Lezi zimo ziphinde ziholele ekwandeni okuphelele kokungazweli kwemithambo ye-peripheral, okuholela ekushaqekeni kwe-hemorrhagic.Izinyathelo ezifanele zokulawula ukopha ngesikhathi nangemva kokuhlinzwa zibhekwa njengeziyingozi empilweni2,3.Ukulimala kwemikhumbi emikhulu kuholela ekulahlekelweni kwegazi okukhulu, okuholela ekufeni kwe-≤ 50% ekulweni kanye ne-31% ngesikhathi sokuhlinzwa1.Ukulahleka kwegazi okukhulu kuholela ekwehleni komthamo womzimba, okunciphisa ukuphuma kwenhliziyo.Ukwanda kokumelana okuphelele kwemithambo ye-peripheral kanye nokukhubazeka okuqhubekayo kwe-microcirculation kuholela ku-hypoxia ezithweni ezisekela ukuphila.Ukushaqeka kwe-hemorrhagic kungenzeka uma isimo siqhubeka ngaphandle kokungenelela okusebenzayo1,4,5.Ezinye izinkinga zihlanganisa ukuqhubekela phambili kwe-hypothermia kanye ne-metabolic acidosis, kanye nesifo sokuqina kwesibindi esikhinyabeza inqubo yokuhlangana.Ukushaqeka okukhulu ukopha kuhlotshaniswa nengozi enkulu yokufa6,7,8.Ebangeni lesi-III (okuqhubekayo) ukushaqeka, ukumpontshelwa igazi kubalulekile ukuze kuphile isiguli phakathi nokuhlinzwa kanye nokugula nokufa kwangemva kokuhlinzwa.Ukuze sinqobe zonke izimo ezingenhla ezisongela ukuphila, sisungule i-nanoporous fiber-reinforced composite scaffold (NFRCS) esebenzisa ukugxila okuncane kwe-polymer (0.5%) kusetshenziswa inhlanganisela yama-polymer e-hemostatic ancibilika emanzini.
Ngokusetshenziswa kwe-fiber reinforcement, imikhiqizo engabizi ingathuthukiswa.Imicu ehlelwe ngokungahleliwe ifana nokwakheka kwephiko likajekamanzi, elilinganiswe ngemigqa evundlile neqondile emaphikweni.I-transverse kanye ne-longitudinal veins yephiko ixhumana nolwelwesi lwephiko (Fig. 1).I-NFRCS iqukethe i-Ct eqinisiwe njengohlelo lwescaffold olunamandla angcono omzimba kanye nemishini (Umfanekiso 1).Ngenxa yokufinyeleleka nobungcweti, odokotela abahlinzayo bakhetha ukusebenzisa izikali zentambo kakotini (Ct) ngesikhathi sokusebenza nokugqoka. Ngakho-ke, uma kucatshangelwa izinzuzo zayo eziningi, okuhlanganisa > 90% i-crystalline cellulose (idlulisela ekuthuthukisweni komsebenzi we-hemostatic), i-Ct yasetshenziswa njengohlelo lwamathambo lwe-NFRCS9,10. Ngakho-ke, uma kucatshangelwa izinzuzo zayo eziningi, okuhlanganisa > 90% i-crystalline cellulose (idlulisela ekuthuthukisweni komsebenzi we-hemostatic), i-Ct yasetshenziswa njengohlelo lwamathambo lwe-NFRCS9,10. Следовательно, учитывая его многочисленные преимущества, в том числе > 90% кристаллической целюлозы (участвует в повыское высобега), пользовали в качестве скелетной системы NFRCS9,10. Ngakho-ke, uma kubhekwa izinzuzo zayo eziningi, okuhlanganisa> i-90% i-crystalline cellulose (ehileleke ekwandeni komsebenzi we-hemostatic), i-Ct isetshenziswe njenge-NFRCS skeletal system9,10.因此，考虑到它的多重益处，包括> 90% 的结晶纤维素（有助于增强止血活性）），Ct 被被,用结晶纤维素).因此，考虑到它的多重益处，包括> 90%Ngakho-ke, uma kubhekwa izinzuzo zayo eziningi, kuhlanganise ne-crystalline cellulose engaphezu kuka-90% (isiza ukuthuthukisa umsebenzi we-hemostatic), i-Ct isetshenziswe njengesikafula se-NFRCS9,10.I-Ct yayimbozwe phezulu (kwabonwa ukwakheka kwefilimu enogqinsi lwe-nano) futhi yaxhunyaniswa ne-hemostatic film-forming composition (HFFC).I-HFFC yenza njenge-matrigel, ibambe i-Ct ebekwe ndawonye ngokungahleliwe.Idizayini ethuthukisiwe idlulisela ukucindezeleka ngaphakathi kwesigaba esihlakazekile (imicu yokuqinisa).Kunzima ukuthola izakhiwo ze-nanoporous ezinamandla amahle wemishini usebenzisa ukugxila okuncane kwe-polymer.Ngaphezu kwalokho, akulula ukwenza ngendlela oyifisayo isikhunta sezinhlelo zokusebenza ezahlukene ze-biomedical.
Isibalo sibonisa umdwebo wedizayini ye-NFRCS esekelwe kuhlaka lwephiko likajekamanzi (A).Lesi sithombe sibonisa ukufaniswa okuqhathanisayo kwesakhiwo sephiko likajekamanzi (imithambo ephambanayo kanye nelongitudinal yephiko ixhumene) kanye nesithombe-micrograph esiphambanayo se-Cp NFRCS (B).Ukumelwa okuhleliwe kwe-NFRCS.
Ama-NFRC athuthukiswa kusetshenziswa i-HFFC njengesigaba esiqhubekayo sokubhekana nemikhawulo engenhla.I-HFFC yakhiwe ama-polymers e-hemostatic ahlukahlukene ahlanganisa ifilimu ehlanganisa i-chitosan (njenge-polymer eyinhloko ye-hemostatic) ene-methylcellulose (MC), i-hydroxypropyl methylcellulose (HPMC 50 cp) kanye ne-polyvinyl alcohol (PVA)) (125 kDa) njenge-polymer esekelayo ekhuthaza ukwakheka kwe-thrombus.ukwakheka.Ukwengezwa kwe-polyvinylpyrrolidine K30 (PVP K30) kuthuthukise umthamo wokumunca umswakama we-NFRCS.I-Polyethylene glycol 400 (PEG 400) yengezwe ukuze kuthuthukiswe i-polymer crosslinking emixubeni ye-polymer eboshiwe.Izingoma ezintathu ezihlukene ze-HFFC ze-hemostatic (Cm HFFC, Ch HFFC ne-Cp HFFC), okuyi-chitosan ene-MC (Cm), i-chitosan ene-HPMC (Ch), ne-chitosan ene-PVA (Cp), isetshenziswe ku-Ct.Ucwaningo oluhlukahlukene lwe-in vitro kanye ne-in vivo luqinisekisile umsebenzi wokuphulukisa we-hemostatic kanye nesilonda we-NFRCS.Izinto eziyinhlanganisela ezihlinzekwa yi-NFRCS zingasetshenziswa ukwenza ngendlela oyifisayo izinhlobo ezahlukene zescaffolding ukuhlangabezana nezidingo ezithile.
Ngaphezu kwalokho, i-NFRCS ingashintshwa njengebhandeshi noma umqulu ukuze imboze yonke indawo yokulimala kwamaphethelo aphansi nezinye izingxenye zomzimba.Ngokukhethekile ngokulimala kwemilenze yokulwa, idizayini eklanyelwe ye-NFRCS ingashintshwa ibe ingxenye yengalo noma umlenze ogcwele (Supplementary Figure S11).I-NFRCS ingenziwa ibhande esihlakaleni elineglue yethishu, engasetshenziswa ukumisa ukopha ekulimaleni okubi kakhulu kwesandla sokuzibulala.Umgomo wethu oyinhloko ukuthuthukisa i-NFRCS ene-polymer encane ngangokunokwenzeka engalethwa kubantu abaningi (ngaphansi komugqa wobumpofu) futhi engafakwa kukhithi yosizo lokuqala.Idizayini elula, ephumelelayo, futhi eyongayo, i-NFRCS izuzisa imiphakathi yasendaweni futhi ingaba nomthelela emhlabeni jikelele.
I-Chitosan (isisindo se-molecular 80 kDa) kanye ne-amaranth zathengwa eMerck, e-India.I-Hydroxypropyl methylcellulose 50 Cp, i-polyethylene glycol 400 ne-methylcellulose ithengwe kwa-Loba Chemie Pvt.LLC, Mumbai.I-Polyvinyl alcohol (isisindo se-molecular 125 kDa) (87-90% hydrolysed) yathengwa ku-National Chemicals, e-Gujarat.I-Polyvinylpyrrolidine K30 yathengwa e-Molychem, eMumbai, kwathengwa ama-swabs ayinyumba kwa-Ramaraju Surgery Cotton Mills Ltd., e-Tamil Nadu, ngamanzi e-Milli Q (uhlelo lokuhlanza amanzi oluqondile-Q3, i-Merck, e-India) njengesithwali.
I-NFRCS yasungulwa kusetshenziswa indlela ye-lyophilization11,12.Zonke izingoma ze-HFFC (Ithebula 1) zalungiswa kusetshenziswa isivunguvungu esingumshini.Lungiselela isixazululo esingu-0.5% se-chitosan usebenzisa i-acetic acid engu-1% emanzini ngokugqugquzela okuqhubekayo ku-800 rpm ku-stirrer engumshini.Isisindo esiqondile se-polymer elayishiwe ekhonjiswe kuThebula 1 sengezwe kwisisombululo se-chitosan futhi sinyakaziswa kuze kutholakale isisombululo se-polymer esicacile.I-PVP K30 kanye ne-PEG 400 zengezwe engxubeni ewumphumela ngamanani aboniswe kuThebula 1, futhi ukugoqa kwaqhutshekwa kwaze kwatholakala isisombululo se-polymer esicacile se-viscous.Ukugeza okuwumphumela wesixazululo se-polymer kwenziwa nge-sonication imizuzu engu-60 ukuze kukhishwe amabhamuza omoya avaleleke engxube ye-polymer.Njengoba kuboniswe Emfanekisweni Owengeziwe S1(b), i-Ct yasatshalaliswa ngokulinganayo emthonjeni ngamunye wepuleti elinemithombo engu-6 (isikhunta) esihlanganiswe no-5 ml we-HFFC.
Ipuleti elinemithombo eyisithupha lafakwa i-sonication for 60 min ukuze kuzuzwe ukumanziswa okufanayo nokusatshalaliswa kwe-HFFC kunethiwekhi ye-Ct.Bese ufaka ipuleti yemithombo eyisithupha ku-20 ° C amahora angu-8-12.Amapuleti okuqandisa afakwa lyophilized amahora angama-48 ukuze athole ukwakheka okuhlukahlukene kwe-NFRCS.Inqubo efanayo isetshenziselwa ukukhiqiza izimo nezakhiwo ezahlukene, njengama-tampons noma ama-cylindrical tampons, nanoma yimuphi omunye umumo wezinhlelo zokusebenza ezahlukene.
I-chitosan enesisindo esinembile (80 kDa) (3%) incibilika ku-1% we-acetic acid kusetshenziswa i-stirrer kazibuthe.Kusixazululo esiwumphumela se-chitosan sengezwe i-1% PEG 400 futhi sinyakaziswa imizuzu engama-30.Thela ikhambi eliwumphumela esitsheni esiyisikwele noma esingunxande bese ubamba iqhwa ku-80°C amahora angu-12.Amasampula aqandisiwe afakwa lyophilized amahora angu-48 ukuze athole i-Cs13 enezimbotshana.
I-NFRCS ethuthukisiwe yenziwa ukuhlolwa kusetshenziswa i-Fourier transform infrared spectroscopy (FTIR) (Shimadzu 8400 s FTIR, Tokyo, Japan) ukuze kuqinisekiswe ukuhambisana kwamakhemikhali kwe-chitosan namanye ama-polymers14,15.I-spectra ye-FTIR (ububanzi bebanga le-spectral kusuka ku-400 kuya ku-4000 cm-1) yawo wonke amasampula ahloliwe atholwe ngokwenza ama-scan angu-32.
Izinga lokumuncwa kwegazi (i-BAR) lakho konke ukwakheka lihlolwe kusetshenziswa indlela echazwe u-Chen et al.16 ngokulungiswa okuncane.Ama-NFRK athuthukisiwe azo zonke izingoma zomiswe kuhhavini wevacuum ku-105°C ngobusuku obunye ukuze kukhishwe isincibilikisi esisele.I-30 mg NFRCS (isisindo sesampula sokuqala - W0) kanye ne-30 mg Ct (ukulawula okuhle) zibekwe ezitsheni ezihlukene eziqukethe i-premix ye-3.8% ye-sodium citrate.Ngezikhawu zesikhathi ezinqunyiwe, okungukuthi 5, 10, 20, 30, 40 kanye namasekhondi angu-60, i-NFRCS yakhishwa futhi izindawo zayo zahlanzwa ngegazi elingamuncwanga ngokubeka amasampula ku-Ct imizuzwana engu-30.Isisindo sokugcina segazi elimuncwe yi-NFRCS 16 sacatshangelwa (W1) ngesikhathi ngasinye.Bala iphesenti le-BAR usebenzisa ifomula elandelayo:
Isikhathi sokujiya kwegazi (BCT) sanqunywa njengoba kubikwe uWang et al.17 .Isikhathi esidingekayo segazi eliphelele (igazi legundane elixutshwe kuqala ne-3.8% ye-sodium citrate) ukuze lihlule lapho kukhona i-NFRCS sibalwa njenge-BCT yesampula yokuhlola.Izingxenye ezihlukahlukene ze-NFRCS (30 mg) zafakwa ku-10 ml we-screw cap vials futhi zafakwa ku-37°C.Igazi (0.5 ml) lafakwa ku-vial futhi u-0.3 ml we-0.2 M CaCl2 wengezwa ukuze kusebenze ukujiya kwegazi.Ekugcineni, guqula i-vial njalo ngemizuzwana engu-15 (kufika ku-180°) kuze kube yilapho ihlule eliqinile libumba.I-BCT yesampula ilinganiselwa ngenani lama-flip vails17,18.Ngokusekelwe ku-BCT, izingoma ezimbili ezingcono kakhulu ezivela ku-NFRCS Cm, Ch kanye ne-Cp zikhethelwe izifundo eziqhubekayo zabalingiswa.
I-BCT ye-Ch NFRCS kanye ne-Cp NFRCS yokuqanjwa kwanqunywa ngokusebenzisa indlela echazwe ngu-Li et al.19 .Beka 15 x 15 mm2 Ch NFRCS, Cp NFRCS, kanye no-Cs (ukulawula okuhle) ezitsheni ezihlukene ze-Petri (37 °C).Igazi eliqukethe u-3.8% we-sodium citrate lixutshwe no-0.2 M CaCl2 ngesilinganiso sevolumu engu-10:1 ukuze kuqalwe inqubo yokujiyisa kwegazi.20 µl ka-0.2 M CaCl2 ingxube yegazi legundane isetshenziswe endaweni yesampula futhi yafakwa esitsheni se-Petri esingenalutho.Ukulawula kwakuyigazi elithelwa ezitsheni ezingenalutho zePetri ngaphandle kukaCt.Ngezikhathi ezinqunyiwe zemizuzu engu-0, 3, kanye ne-5, yeka ukujiyisa ngokungeza u-10 ml wamanzi e-deionized (DI) kusampula equkethe indishi ngaphandle kokuphazamisa ihlule.Ama-erythrocyte angaguquki (ama-erythrocyte) athola i-hemolysis lapho kukhona amanzi angenalutho futhi akhulule i-hemoglobin.I-Hemoglobin ngezikhathi ezihlukene (HA(t)) ikalwe ku-540 nm (λmax hemoglobin) kusetshenziswa i-UV-Vis spectrophotometer.Ukumuncwa ngokuphelele kwe-hemoglobin (AH(0)) kuminithi elingu-0 lika-20 µl wegazi ku-10 ml wamanzi akhishiwe kuthathwe njengezinga lenkomba.Ukutholwa kwe-hemoglobin (i-RHA) yegazi elijiyile kubalwe kusukela kusilinganiso HA(t)/HA(0) kusetshenziswa iqoqwana legazi elifanayo.
Kusetshenziswa i-analyzer yokuthungwa (Texture Pro CT V1.3 Build 15, Brookfield, USA), izici zokunamathela ze-NFRK ezithishuni ezilimele zanqunywa.Cindezela isitsha esiyicylindrical esivulekile esiphansi ngaphansi kwesikhumba sengulube (ngaphandle kongqimba lwamafutha).Amasampula (Ch NFRCS kanye ne-Cp NFRCS) asetshenziswe nge-cannula ekubunjweni okuyicylindrical ukudala ukunamathela esikhumbeni sengulube.Ngemva kokufakwa kwemizuzu emi-3 ekamelweni lokushisa (RT) (25° C.), amandla okunamathela e-NFRCS aqoshwa ngesilinganiso esingaguquki esingu-0.5 mm/sec.
Isici esiyinhloko sama-sealants okuhlinzwa ukwandisa ukujiya kwegazi ngenkathi kunciphisa ukulahleka kwegazi.I-coagulation engalahleki ku-NFRCS yahlolwa kusetshenziswa indlela eshicilelwe ngaphambilini enezinguquko ezincane 19.Yenza ithubhu le-microcentrifuge (2 ml) (ububanzi bangaphakathi obuyi-10 mm) nembobo engu-8 × 5 mm2 ohlangothini olulodwa lwethubhu ye-centrifuge (emele inxeba elivulekile).I-NFRCS isetshenziselwa ukuvala ukuvuleka futhi itheyiphu isetshenziselwa ukuvala imiphetho yangaphandle.Engeza u-20 µl ka-0.2 M CaCl2 kushubhu le-microcentrifuge eliqukethe u-3.8% we-sodium citrate premix.Ngemuva kwemizuzu engu-10, amashubhu e-microcentrifuge asusiwe ezitsheni futhi ukwanda kwenqwaba yezitsha kunqunywe ngenxa yokuphuma kwegazi ku-NFRK (n = 3).Ukulahleka kwegazi I-Ch NFRCS ne-Cp NFRCS ziqhathaniswe nama-Cs.
Ubuqotho obumanzi be-NFRCS banqunywa ngokusekelwe endleleni echazwe u-Mishra no-Chaudhary21 ngokulungiswa okuncane.Faka i-NFRCS eflaskini ye-Erlenmeyer engu-100 ml enamanzi angama-50 ml bese uzungeza amasekhondi angama-60 ngaphandle kokwenza phezulu.Ukuhlolwa okubonakalayo kanye nokubekwa phambili kwamasampuli obuqotho bomzimba ngokusekelwe ekuqoqweni.
Amandla abophezelayo e-HFFC ku-Ct ahlolisiswa kusetshenziswa izindlela ezishicilelwe ngaphambilini ezinezinguquko ezincane.Ubuqotho bokwemboza okungaphezulu buhlolwe ngokudalula i-NFRK kumagagasi e-acoustic (isisusa sangaphandle) ebukhoneni be-milliQ water (Ct).I-NFRCS Ch NFRCS ethuthukisiwe kanye ne-Cp NFRCS yafakwa kubhekha egcwele amanzi futhi yafakwa i-sonicated for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 30 min, ngokulandelana.Ngemva kokoma, umehluko wephesenti phakathi kwesisindo sokuqala nesokugcina se-NFRCS wasetshenziswa ukubala ukulahleka kwephesenti yezinto ezibonakalayo (HFFC).I-In vitro BCT iphinde yasekela amandla okubopha noma ukulahleka kwezinto ezingaphezulu.Ukusebenza kahle kokubophezela kwe-HFFC ku-Ct kunikeza ukuhlangana kwegazi kanye nokunwebeka okunwebekayo ebusweni be-Ct22.
I-homogeneity ye-NFRCS ethuthukisiwe yanqunywa i-BCT yamasampuli (30 mg) athathwe ezindaweni ezijwayelekile ezikhethwe ngokungahleliwe ze-NFRCS.Landela inqubo ye-BCT eshiwo ngaphambilini ukuze unqume ukuthobela i-NFRCS.Ukusondela phakathi kwawo wonke amasampula amahlanu kuqinisekisa ukumbozwa kwendawo efanayo kanye nokufakwa kwe-HFFC ku-Ct mesh.
Indawo yokuxhumana negazi (i-NBCA) yanqunywa njengoba kubikwe ngaphambilini ngokulungiswa okuthile.Hlanganisa igazi ngokucindezela u-20 µl wegazi phakathi kwezindawo ezimbili ze-Ct, Ch NFRCS, Cp NFRCS kanye ne-Cs.Ngemuva kwehora elingu-1, izingxenye ezimbili ze-stent zahlukaniswa futhi zikalwa mathupha indawo yehlule.Inani elimaphakathi lezimpinda ezintathu lithathwe njenge-NBCA NFRCS19.
Ukuhlaziywa kwe-Dynamic Vapor Sorption (DVS) kwasetshenziselwa ukuhlola ukusebenza kahle kwe-NFRCS ukumunca amanzi avela endaweni yangaphandle noma endaweni yokulimala enesibopho sokuqalisa ukuhlangana.I-DVS ihlola noma irekhoda ukukhuphuka komhwamuko kanye nokulahleka kwawo kusampula ngokwe-gravimetrically isebenzisa ibhalansi ezwela kakhulu ngokulungiswa kwesisindo esingu-±0.1 µg.Ingxenye yokucindezela komhwamuko (umswakama ohlobene) ikhiqizwa yisilawuli sokugeleza kwenqwaba ye-elekthronikhi ezungeze isampula ngokuxuba amagesi enethiwekhi agcwele kanye nawomile. Njengokweziqondiso ze-European Pharmacopeia, ngokusekelwe kumaphesenti okuthathwa komswakama ngamasampula, amasampula ahlukaniswe ngezigaba ezi-4 (0–0.012% w/w-non-hygroscopic, 0.2–2% w/w little hygroscopic, 2–15% moderately hygroscopic, kanye > 25 hygroscopic kakhulu) Njengokweziqondiso ze-European Pharmacopeia, ngokusekelwe kumaphesenti okuthathwa komswakama ngamasampula, amasampula ahlukaniswe ngezigaba ezi-4 (0–0.012% w/w-non-hygroscopic, 0.2–2% w/w kancane hygroscopic, 2–15 % ngokumaphakathi hygroscopic, kanye > 315 hygroscopic kakhulu) kakhulu.Ngokuvumelana nezincomo ze-European Pharmacopoeia, kuye ngephesenti lokumuncwa komswakama ngamasampula, amasampula ahlukaniswe abe yizigaba ezi-4 (0-0.012% w/w - non-hygroscopic, 0.2-2% w/w little hygroscopic, 2–teenfifteen%).% умеренно гигроскопичен и > 15% очень гигроскопичен)23. % i-hygroscopic emaphakathi futhi > 15% ene-hygroscopic kakhulu)23.根据欧洲药典指南，根据樣品吸收水分的百分比，样品分为4 类（0-0.012% w/w-澎家其公2.吸湿性, 2-15 % 适度吸湿，> 15% 非常吸湿）23.根据 欧洲 药典 指南 ， 根据 吸收 水分 的 百分比 样品 分為 分为 分为 类 %済/w-10 , , , 0.2-2% W/w 轻微, 2-15% 适度 吸湿 ，> 15 %非常吸湿）23.Ngokuvumelana nezincomo ze-European Pharmacopoeia, amasampula ahlukaniswe abe amakilasi ama-4 kuye ngephesenti lomswakama amuncwa isampula (0-0.012% ngesisindo - non-hygroscopic, 0.2-2% ngesisindo kancane hygroscopic, 2-15% ngesisindo).% умеренно гигроскопичен, > 15 % очень гигроскопичен) 23. % i-hygroscopic emaphakathi, > 15% ene-hygroscopic kakhulu) 23.Ukusebenza kahle kwe-hygroscopic kwe-NFCS X NFCS kanye ne-TsN NFCS kunqunywe ku-analyzer DVS TA TGA Q5000 SA.Phakathi nale nqubo, isikhathi sokugijima, umswakama ohlobene (RH), nesisindo sesampula sesikhathi sangempela ku-25 ° C24 sitholwe.Okuqukethwe komswakamo kubalwa ngokuhlaziywa kwenqwaba ye-NFRCS kusetshenziswa isibalo esilandelayo:
I-MC iwumswakama we-NFRCS.m1 - isisindo esomile sama-NSAID.i-m2 iyisisindo sesikhathi sangempela se-NFRCS ku-RH enikeziwe.
Indawo ephelele yalinganiselwa kusetshenziswa ukuhlolwa kwe-nitrogen adsorption nge-nitrogen ewuketshezi ngemva kokuthulula amasampula ku-25 °C amahora angu-10 (< 7 × 10–3 Torr). Indawo ephelele yalinganiselwa kusetshenziswa ukuhlolwa kwe-nitrogen adsorption nge-nitrogen ewuketshezi ngemva kokuthulula amasampula ku-25 °C amahora angu-10 (< 7 × 10–3 Torr). Общая площадь поверхности оценивалась с помощью эксперимента по адсорбции азота жидким азотом после опорожнения образцов при 12 × 12 × 12 × 3 Isiqephu). Indawo ephelele yalinganiselwa kusetshenziswa isilingo se-nitrogen adsorption nge-nitrogen ewuketshezi ngemva kokuba amasampula athululwa ku-25°C amahora angu-10 (< 7 × 10–3 Torr).在25°C 清空样品10 小时（< 7 × 10-3 Torr）后，使用液氮的氮吸附实验估计总表面积。Ku-25°C Общая площадь поверхности оценивалась с использованием экспериментов по адсорбции азота жидким азотом после опользованием экспериментов по адсорбции азотом азотом после опользованием экспериментов по адсорбции азотом оценивалась °C (< 7 × 10-3 тор). Indawo ephelele yalinganiselwa kusetshenziswa izivivinyo ze-nitrogen adsorption nge-nitrogen ewuketshezi ngemva kokuthi amasampuli athululwa amahora angu-10 ku-25°C (< 7 x 10-3 torr).Indawo ephelele, ivolumu yembotshana kanye nosayizi wembotshana we-NFRCS kunqunywe nge-Quantachrome evela ku-NOVA 1000e, e-Austria kusetshenziswa isofthiwe ye-RS 232.
Lungiselela ama-RBC angu-5% (asawoti njenge-diluent) egazini lonke.Bese udlulisela i-aliquot ye-HFFC (0.25 ml) epuleti lemithombo engu-96 kanye nesisindo esingu-5% se-RBC (0.1 ml).Fukamisa ingxube ku-37°C imizuzu engu-40.Ingxubevange yamangqamuzana egazi abomvu neserum kwakubhekwa njengokulawula okuhle, kanye nengxube kasawoti kanye namangqamuzana egazi abomvu njengokulawula okungalungile.I-Hemagglutination inqunywe ngokwesilinganiso se-Stajitzky.Izikali ezihlongozwayo zimi kanje: + + + + i-granular aggregate eminyene;+ + + amaphedi aphansi abushelelezi anemiphetho egobile;+ + amaphedi aphansi abushelelezi anemiphetho edabukile;+ izindandatho ezibomvu eziwumngcingo ezizungeze imiphetho yamaphedi abushelelezi;– (negative) inkinobho ebomvu discreter 12 phakathi nendawo ephansi umthombo.
I-hemocompatibility ye-NFRCS yafundwa ngendlela ye-International Organization for Standardization (ISO) (ISO10993-4, 1999)26,27.Indlela ye-gravimetric echazwe nguSingh et al.Ukuguqulwa okuncane kwenziwa ukuze kuhlolwe ukwakheka kwe-thrombus ebukhoneni noma ebusweni be-NFRCS.U-500 mg we-Cs, i-Ch NFRCS kanye ne-Cp NFRCS zafakwa ku-phosphate buffered saline (PBS) amahora angu-24 ku-37°C.Ngemva kwamahora angu-24, i-PBS yasuswa futhi i-NFRCS yaphathwa nge-2 ml yegazi eliqukethe i-3.8% ye-sodium citrate.Ebusweni be-NFRCS, engeza u-0.04 ml we-0.1 M CaCl2 kumasampuli afakiwe.Ngemuva kwemizuzu engama-45, u-5 ml wamanzi acwebile wengezwa ukumisa ukuhlangana.Igazi elihlanganisiwe ebusweni be-NFRK lelashwe ngesisombululo se-formaldehyde esingu-36-38%.Amahlule ahlanganiswe ne-formaldehyde ayomiswa futhi akalwe.Iphesenti le-thrombosis lalinganiselwa ngokubala isisindo sengilazi ngaphandle kwegazi nesampula (ukulawula okungalungile) kanye nengilazi enegazi (ukulawula okuhle).
Njengesiqiniseko sokuqala, amasampula abonwa ngaphansi kwesibonakhulu ukuze aqonde ikhono le-HFFC surface coating, Ct interconnected, kanye nenethiwekhi ye-Ct ukuze enze izimbotshana.Izingxenye ezizacile ze-Ch ne-Cp ezivela ku-NFRCS zasikwa nge-scalpel blade.Isigaba esiphumela sibekwe kwisilayidi seglasi, simbozwe nge-coverlip, futhi imiphetho ihlanganiswe neglue.Amaslayidi alungisiwe abukwe ngesibonakhulu esibonakalayo futhi izithombe zathathwa ngokukhuliswa okuhlukene.
Ukufakwa kwe-polymer kumanethiwekhi e-Ct kuboniswe kusetshenziswa i-fluorescence microscopy ngokusekelwe endleleni echazwe nguRice et al.29. Ukwakheka kwe-HFFC okusetshenziswe ekwenziweni kwahlanganiswa nodayi we-fluorescent (i-amaranth), futhi i-NFRCS (Ch & Cp) yalungiswa ngendlela eshiwo ngaphambilini. Ukwakheka kwe-HFFC okusetshenziswe ekwenziweni kwahlanganiswa nodayi we-fluorescent (i-amaranth), futhi i-NFRCS (Ch & Cp) yalungiswa ngendlela eshiwo ngaphambilini.Ukwakheka kwe-HFFC okusetshenziselwa ukwakheka kwaxutshwa nodayi we-fluorescent (i-amaranth) kanye ne-NFRCS (Ch ne-Cp) yatholwa ngokuya ngendlela eshiwo ngaphambilini.将用于配方的HFFC 组合物与荧光染料（苋菜）混合，并按照前面提到的方法制备NFRCSp））将用于配方的HFFC 组合物与荧光染料（苋菜）混合，并按照前面提到的方法制备NFRCSp））Ukwakheka kwe-HFFC okusetshenziswe ekwenziweni kuhlanganiswe nodayi we-fluorescent (i-Amaranth) futhi kwathola i-NCFRCS (Ch ne-Cp), njengoba kushiwo ngaphambili.Izingxenye ezizacile ze-NFRK zasikwa kumasampula atholiwe, zabekwa kumaslayidi engilazi, futhi zambozwa ngeziliphu zekhava.Bheka amaslayidi alungisiwe ngaphansi kwesibonakhulu se-fluorescent usebenzisa isihlungi esiluhlaza (310-380 nm).Izithombe zithathwe ku-4x magnification ukuze kuqondwe ubudlelwano be-Ct kanye nokufakwa kwe-polymer ngokweqile kunethiwekhi ye-Ct.
I-topography ye-NFRCS Ch ne-Cp yanqunywa kusetshenziswa i-atomic force microscope (AFM) ene-ultra-sharp TESP cantilever kumodi yokuthepha: 42 N/m, 320 kHz, ROC 2-5 nm, Bruker, Taiwan.Ubulukhuni bobuso bunqunywe yi-root mean square (RMS) kusetshenziswa isofthiwe (I-Scanning Probe Image Processor).Izindawo ezahlukahlukene ze-NFRCS zinikezwe ezithombeni ze-3D ukuze kuhlolwe ukufana kwendawo.Ukuchezuka okujwayelekile kwesikolo sendawo ethile kuchazwa njengokumahhadla kwendawo.Isibalo se-RMS sisetshenziselwe ukulinganisa ubulukhuni obungaphezulu be-NFRCS31.
Ucwaningo olusekelwe ku-FESEM lwenziwe kusetshenziswa i-FESEM, SU8000, HI-0876-0003, Hitachi, Tokyo, ukuze kuqondwe i-morphology ye-Ch NFRCS ne-Cp NFRCS, ebonise i-BCT engcono kune-Cm NFRCS.Ucwaningo lwe-FESEM lwenziwa ngokwendlela echazwe nguZhao et al.32 ngezinguquko ezincane.I-NFRCS 20 kuya ku-30 ​​mg I-Ch NFRCS ne-Cp NFRCS zixutshwe kuqala ne-20 µl ka-3.8% ye-sodium citrate ehlanganiswe negazi legundane.I-20 μl ye-0.2 M CaCl2 yengezwe kumasampuli eselashwe igazi ukuze kuqalwe ukuhlangana futhi amasampula afakwe ekamelweni lokushisa imizuzu engu-10.Ngaphezu kwalokho, ama-erythrocyte amaningi asusiwe ebusweni be-NFRCS ngokuhlanjululwa ngosawoti.
Amasampula alandelayo aphathwe nge-0.1% glutaraldehyde bese omiswa kuhhavini oshisayo ongama-37 ° C ukuze asuse umswakama.Amasampula omisiwe agcotshwe futhi ahlaziywa 32.Ezinye izithombe ezitholwe ngesikhathi sokuhlaziywa kwakuwukwakheka kwehlule ebusweni bemicu yekotini ngayinye, ukubekelwa phansi kwe-polymer phakathi kwe-Ct, i-erythrocyte morphology (umumo), ubuqotho behlule, kanye ne-erythrocyte morphology phambi kwe-NFRCS.Izindawo ze-NFRCS ezingaphenduliwe kanye nezindawo ze-NFRCS eziphathwe nge-Ch ne-Cp ezifakwe ngegazi zahlolwa ukuze kutholakale ama-ion ayisisekelo (i-sodium, i-potassium, i-nitrogen, i-calcium, i-magnesium, i-zinc, ithusi ne-selenium)33.Qhathanisa amaphesenti e-ion ayisisekelo phakathi kwamasampula alashiwe nanganakiwe ukuze uqonde ukunqwabelana kwe-ion eyisisekelo ngesikhathi sokwakheka kwehlule kanye ne-clot homogeneity.
Ubukhulu be-Cp HFFC surface coating endaweni ye-Ct bunqunywe kusetshenziswa i-FESEM.Izingxenye eziphambanayo ze-Cp NFRCS zasikwa kuhlaka futhi zambozwa nge-sputter.Amasampula okugcoba ama-sputter atholwe abonwa yi-FESEM futhi ubukhulu be-surface coating bulinganiswa 34 , 35 , 36 .
I-X-ray micro-CT ihlinzeka ngesithombe se-3D esinokulungiswa okuphezulu okungabhubhi futhi ikuvumela ukuthi ufunde ukuhlelwa kwesakhiwo kwangaphakathi kwe-NFRK.I-Micro-CT isebenzisa i-X-ray edlula isampula ukuze irekhode i-coefficient yasendaweni yomugqa wokuncisha ama-X-ray kusampula, esiza ukuthola ulwazi lwe-morphological.Indawo yangaphakathi ye-Ct ku-Cp NFRCS kanye ne-Cp NFRCS ephathwe igazi yahlolwa yi-micro-CT ukuze kuqondwe ukusebenza kahle kokumuncwa kanye nokujiyisa kwegazi phambi kwe-NFRCS37,38,39.Izakhiwo ze-3D zamasampuli e-Cp NFRCS alashwe igazi nanganakiwe aphinde akhiwa kusetshenziswa i-micro-CT (V|tome|x S240, Phoenix, Germany).Kusetshenziswa isoftware ye-VG STUDIO-MAX inguqulo engu-2.2, izithombe ze-X-ray ezimbalwa zathathwa ngama-engeli ahlukene (okungcono ukumbozwa okungu-360°) ukuze kwakhiwe izithombe ze-3D ze-NFRCS.Idatha yokuqagela eqoqiwe yakhiwa kabusha yaba izithombe zevolumu ye-3D kusetshenziswa isofthiwe efanayo ye-3D ScanIP Academic.
Ukwengeza, ukuze kuqondwe ukusatshalaliswa kwehlule, u-20 µl wegazi elixutshwe ngaphambili kanye no-20 µl ka-0.2 M CaCl2 kwengezwe ku-NFRCS ukuqalisa ukujiyisa kwegazi.Amasampula alungisiwe ashiywe ukuze aqine.Indawo ye-NFRK yaphathwa nge-0.5% glutaraldehyde futhi yomiswa kuhhavini lomoya oshisayo ku-30-40 ° C imizuzu engu-30.Ihlule legazi elakhiwe ku-NFRCS laskeniwa, lakhiwa kabusha, futhi kwabonwa ngeso lengqondo isithombe se-3D sehlule legazi.
Ukuhlolwa kwe-antibacterial kwenziwa ku-Cp NFRCS (okungcono kakhulu uma kuqhathaniswa ne-Ch NFRCS) kusetshenziswa indlela echazwe ngaphambilini enezinguquko ezincane.Umsebenzi wokulwa namagciwane we-Cp NFRCS kanye ne-Cp HFFC wanqunywa kusetshenziswa amagciwane okuhlola amathathu ahlukene [i-S.aureus (ibhaktheriya e-gram-positive), i-E.coli (i-gram-negative bacteria) kanye ne-Candida emhlophe (C.albicans)] ekhula ku-agar ezitsheni ze-Petri ku-incubator.Gcoba ngokulinganayo u-50 ml wokumiswa kwesiko lebhaktheriya elihlanjululwe ekuhlanganiseni kwe-105-106 CFU ml-1 endaweni ye-agar.Thela okuphakathi esitsheni sePetri futhi usivumele siqine.Imithombo yenziwa ebusweni bepuleti le-agar ukuze igcwaliswe nge-HFFC (imithombo emi-3 ye-HFFC no-1 yokulawula okungekuhle).Engeza u-200 µl HFFC emithonjeni engu-3 no-200 µl pH 7.4 PBS kumthombo wesi-4.Ngakolunye uhlangothi lwesitsha se-petri, beka idiski engu-12 mm Cp NFRCS ku-agar eqinisiwe bese umanzisa nge-PBS (pH 7.4).I-Ciprofloxacin, i-ampicillin kanye namaphilisi e-fluconazole abhekwa njengamazinga okubhekisela e-Staphylococcus aureus, i-Escherichia coli ne-Candida albicans.Linganisa indawo yokuvimbela mathupha futhi uthathe isithombe sedijithali sezoni yokuvimbela.
Ngemva kokugunyazwa kokuziphatha kwesikhungo, ucwaningo lwenziwa eKasturba Medical College of Education and Research eManipal, Karnataka, eningizimu yeNdiya.Iphrothokholi yokuhlola ye-in vitro ye-TEG iye yabuyekezwa futhi yavunyelwa yiKomidi Lezimiso Zokuziphatha le-Kasturba Medical College, Manipal, Karnataka (IEC: 674/2020).Izifundo zabuthwa kumavolontiya anikela ngegazi (abaneminyaka engu-18 kuya kwengu-55) ebhange legazi lasesibhedlela.Ukwengeza, ifomu lemvume enolwazi latholwa kumavolontiya ukuze kuqoqwe amasampula egazi.I-Teg yomdabu (N-TEG) ​​​​yasetshenziselwa ukutadisha umphumela wokwakheka kwe-Cp HFFC egazini lonke elixutshwe ne-sodium citrate.I-N-TEG yaziwa kabanzi ngendima yayo ekuvuseleleni ukunakekelwa kwamaphuzu, okudala izinkinga kubahlengikazi ngenxa yamathuba okubambezeleka okubalulekile emiphumeleni (ukuhlolwa kwe-coagulation evamile).Ukuhlaziywa kwe-N-TEG kwenziwa kusetshenziswa igazi lonke.Imvume enolwazi kanye nomlando wezokwelapha onemininingwane kutholwe kubo bonke ababambiqhaza.Ucwaningo aluzange lufake ababambiqhaza abanezinkinga ze-hemostatic noma i-thrombotic njengokukhulelwa/ngemuva kokubeletha noma isifo sesibindi.Izifundo ezithatha izidakamizwa ezithinta i-coagulation cascade nazo azifakwanga ocwaningweni.Ukuhlolwa kwaselabhorethri okuyisisekelo (i-hemoglobin, isikhathi se-prothrombin, i-thromboplastin ecushiwe nokubalwa kweplatelet) kwenziwa kubo bonke ababambiqhaza ngokwezinqubo ezijwayelekile.I-N-TEG inquma i-blood clot viscoelasticity, isakhiwo sokuqala se-clot, ukusebenzisana kwezinhlayiyana, ukuqinisa amahlule, kanye ne-clot lysis.Ukuhlaziywa kwe-N-TEG kunikeza idatha yezithombe nezinombolo emiphumeleni eqoqiwe yezinto ezimbalwa zamaselula kanye ne-plasma.Ukuhlaziywa kwe-N-TEG kwenziwa kumavolumu amabili ahlukene we-Cp HFFC (10 µl no-50 µl).Ngenxa yalokho, u-1 ml wegazi eliphelele eline-citric acid yengezwe ku-10 μl we-Cp HFFC.Engeza u-1 ml (Cp HFFC + igazi elicijile), 340 µl igazi elixutshwe ku-20 µl 0.2 M CaCl2 equkethe i-TEG dish.Ngemva kwalokho, izitsha ze-TEG zalayishwa ku-TEG® 5000, US ukukala u-R, K, i-engeli ye-alpha, MA, G, CI, TPI, EPL, LY 30% wamasampuli egazi phambi kwe-Cp HFFC41.
Iphrothokholi yocwaningo lwe-in vivo yabuyekezwa futhi yavunyelwa yiKomidi Lokuziphatha Kwezilwane Lesikhungo (IAEC), Isikole Sezokwelapha saseKasturba, Isikhungo Semfundo Ephakeme Manipal, Manipal (IAEC/KMC/69/2020).Zonke izivivinyo zezilwane zenziwe ngokuhambisana nezincomo zeKomidi Lokulawula Nokulawula Ukuhlolwa Kwezilwane (CPCSEA).Zonke izifundo ze-NFRCS ze-vivo (2 × 2 cm2) zenziwa kumagundane ama-Wistar wesifazane (anesisindo esingu-200 kuya ku-250 g).Zonke izilwane zazijwayele izinga lokushisa lika-24-26 ° C, izilwane zazikwazi ukufinyelela mahhala ekudleni okujwayelekile namanzi ad libitum.Zonke izilwane zahlukaniswa ngokungahleliwe zaba amaqembu ahlukene, iqembu ngalinye lalinezilwane ezintathu.Zonke izifundo zenziwe ngokuhambisana ne-Animal Studies: Report of In Vivo Experiments 43.Ngaphambi kocwaningo, izilwane zaxoshwa nge-anesthetized by intraperitoneal (ip) ukuphathwa kwengxube ye-20-50 mg ye-ketamine (nge-1 kg yesisindo somzimba) kanye ne-2-10 mg ye-xylazine (nge-1 kg yesisindo somzimba).Ngemva kocwaningo, ivolumu yokopha ibalwa ngokuhlola umehluko phakathi kwesisindo sokuqala nesokugcina samasampula, inani elilinganiselwe elitholwe ekuhlolweni okuthathu lithathwe njengevolumu yokopha kwesampula.
Imodeli yokunqunywa komsila wegundane yasetshenziswa ukuze kuqondwe amandla e-NFRCS okulinganisa ukopha ekuhlukumezekeni, ukulwa, noma engozini yomgwaqo (imodeli yokulimala).Sika u-50% womsila nge-scalpel blade futhi ubeke emoyeni imizuzwana engu-15 ukuqinisekisa ukopha okuvamile.Ngaphezu kwalokho, amasampula okuhlola abekwe emsila wegundane ngokufaka ingcindezi (Ct, Cs, Ch NFRCS kanye ne-Cp NFRCS).Ukopha kanye ne-PCT kwabikwa ngezifanekiso zokuhlola (n = 3) 17,45.
Ukusebenza kokulawula ingcindezi ye-NFRCS ekulweni kuye kwaphenywa ngemodeli yomthambo we-femoral ongaphezulu.Umthambo wesifazane uyavezwa, ubhobozwe nge-24G trocar, futhi wophe phakathi nemizuzwana engu-15.Ngemuva kokubona ukopha okungalawuleki, isampula yokuhlola ifakwa endaweni yokubhoboza ngengcindezi.Ngokushesha ngemva kokusetshenziswa kwesampula yokuhlola, isikhathi sokujiyisa sarekhodwa futhi ukusebenza kahle kwe-hemostatic kwabonwa imizuzu emi-5 elandelayo.Inqubo efanayo iphindwe nge-Cs ne-CT46.
UDowling et al.I-47 ihlongoze imodeli yokulimala kwesibindi ukuze kuhlolwe amandla we-hemostatic wezinto ze-hemostatic kumongo wokuphuma kwegazi ngaphakathi kokuhlinzwa.I-BCT yarekhodelwa amasampula e-Ct (ukulawula okungekuhle), uhlaka lwe-Cs (ukulawula okuhle), amasampuli e-Ch NFRCS, namasampuli e-Cp NFRCS.I-suprahepatic vena cava yegundane yavezwa ngokwenza i-laparotomy ephakathi.Ngemva kwalokho, ingxenye ekude ye-lobe yesokunxele yasikwa ngesikelo.Sika isibindi nge-scalpel blade futhi usivumele ukuthi sophe imizuzwana embalwa.Isisindo esinembile se-Ch NFRCS kanye ne-Cp NFRCS amasampula abekwe endaweni eyonakele ngaphandle kwengcindezi enhle futhi i-BCT yarekhodwa.Iqembu lokulawula (Ct) libe selisebenzisa ingcindezi elandelwa yi-Cs 30 s47 ngaphandle kokuphula ukulimala.
Ukuhlolwa kokuphola kwesilonda kwe-vivo kwenziwa kusetshenziswa imodeli yesilonda esikhethekile ukuze kuhlolwe izakhiwo zokupholisa amanxeba ze-NCFRCS esekelwe ku-polymer ethuthukisiwe.Amamodeli amanxeba okukhishwa akhethiwe futhi enziwa ngokwezindlela ezishicilelwe ngaphambilini ngokuguqulwa okuncane19,32,48.Zonke izilwane zalaliswa izinzwa njengoba kuchaziwe ngaphambili.Sebenzisa i-biopsy punch (12 mm) ukwenza isiyingi esijulile esikhumbeni somhlane.Izingosi zamanxeba ezilungisiwe zagqokiswa ngama-Cs (ukulawula okuhle), i-Ct (eqaphela ukuthi amaphedi kakotini aphazamisa ukuphulukiswa), i-Ch NFRCS ne-Cp NFRCS (iqembu lokuhlola) kanye nokulawula okungalungile ngaphandle kwanoma yikuphi ukwelashwa.Ngosuku ngalunye locwaningo, indawo yenxeba yayilinganiswa ngawo wonke amagundane.Sebenzisa ikhamera yedijithali ukuthatha isithombe sendawo yenxeba bese ufaka ingubo entsha.Iphesenti lokuvaleka kwenxeba likalwe ngefomula elandelayo:
Ngokusekelwe kumaphesenti okuvalwa kwenxeba ngosuku lwe-12 locwaningo, isikhumba samagundane seqembu elingcono kakhulu sasikwa ((Cp NFRCS) kanye neqembu lokulawula) futhi safundwa yi-H&E staining kanye ne-trichrome staining ye-Masson. Ngokusekelwe kumaphesenti okuvalwa kwenxeba ngosuku lwe-12 locwaningo, isikhumba samagundane seqembu elingcono kakhulu sasikwa ((Cp NFRCS) kanye neqembu lokulawula) futhi safundwa yi-H&E staining kanye ne-trichrome staining ye-Masson.Ngokusekelwe kumaphesenti okuvalwa kwamanxeba ngosuku lwe-12 locwaningo, isikhumba samagundane eqembu elingcono kakhulu ((Cp NFRCS) neqembu lokulawula) sasikwa futhi sahlolwa ngokungcoliswa nge-hematoxylin-eosin kanye ne-Masson's trichrome.根据研究第12天的伤口闭合百分比，切除最佳组((Cp NFRCS)和对照组)的大鼠皮肤，进海行H&E手究.根据研究第12天的伤口闭合百分比，切除最佳组((Cp NFRCS)和对照组)的大鼠皮肤，进行H&E扄)Amagundane aseqenjini elingcono kakhulu ((Cp NFRCS) kanye namaqembu okulawula) ahlinzwa ukuze athole ibala le-hematoxylin-eosin kanye ne-Masson's trichrome staining ngokusekelwe ekuvaleni kwenxeba okungamaphesenti ngosuku lwe-12 locwaningo.Inqubo yokufaka amabala esetshenzisiwe yenziwa ngokwezindlela ezichazwe ngaphambilini49,50.Kafushane, ngemva kokulungiswa ku-10% we-formalin, amasampuli aphelelwa amanzi kusetshenziswa uchungechunge lotshwala obuhleliwe.Sebenzisa i-microtome ukuze uthole izingxenye ezincane (ugqinsi olu-5 µm) lwezicubu ezisikiwe.Izingxenye ze-serial ezacile zokulawula kanye ne-Cp NFRCS zelashwe nge-hematoxylin ne-eosin ukuze kufundwe izinguquko ze-histopathological.Ibala likaMasson's trichrome lasetshenziswa ukuthola ukwakheka kwama-collagen fibrils.Imiphumela etholakele yacwaningwa ngokungaboni odokotela bezifo.
Ukuzinza kwamasampuli e-Cp NFRCS kwacwaningwa ezingeni lokushisa legumbi (25°C ± 2°C/60% RH ± 5%) izinyanga eziyi-1251.I-Cp NFRCS (ukuguquka kombala nokukhula kwe-microbial) yahlolwa ngokubukeka futhi yahlolelwa ukumelana nokugoqeka kanye ne-BCT ngokwezindlela ezingenhla ezishiwo esigabeni Sezinto kanye Nezindlela.
Ukukhula nokukhiqizwa kabusha kwe-Cp NFRCS kwahlolwa ngokulungiselela i-Cp NFRCS enosayizi ongu-15×15 cm2.Ukwengeza, amasampula angu-30 mg (n = 5) akhishwe kumafrakshini ahlukahlukene e-Cp NFRCS futhi i-BCT yamasampuli ahlolisisiwe yahlolwa njengoba kuchazwe ekuqaleni kwesigaba Sezindlela.
Sizame ukuthuthukisa umumo nezakhiwo ezahlukahlukene sisebenzisa izingoma ze-Cp NFRCS zezinhlelo zokusebenza ezahlukahlukene ze-biomedical.Umumo onjalo noma ukucupha kufaka phakathi ama-swabs ayi-conical okopha ekhaleni, izinqubo zamazinyo, nama-cylindrical swabs okopha kwesitho sangasese sowesifazane.
Wonke amasethi edatha avezwa njengokuchezuka okujwayelekile ± okujwayelekile futhi ahlaziywa i-ANOVA kusetshenziswa i-Prism 5.03 (GraphPad, San Diego, CA, USA) elandelwa ukuhlolwa kokuqhathanisa okuningi kwe-Bonferroni (*p<0.05).
Zonke izinqubo ezenziwa ezifundweni zabantu zazihambisana namazinga e-Institute kanye noMkhandlu Kazwelonke Wocwaningo, kanye neSimemezelo se-Helsinki 1964 kanye nezichibiyelo zaso ezalandela, noma izindinganiso zokuziphatha ezifanayo.Bonke ababambiqhaza baziswe ngezici zocwaningo kanye nesimo salo sokuzithandela.Idatha yombambi qhaza ihlala iyimfihlo uma isiqoqiwe.Iphrothokholi yokuhlola ye-in vitro ye-TEG iye yabuyekezwa futhi yavunyelwa yiKomidi Lezimiso Zokuziphatha le-Kasturba Medical College, Manipal, Karnataka (IEC: 674/2020).Amavolontiya asayine imvume enolwazi yokuqoqa amasampula egazi.
Zonke izinqubo ezenziwa ezifundweni zezilwane zenziwa ngokuhambisana ne-Kastuba Faculty of Medicine, i-Manipal Institute of Higher Education, Manipal (IAEC/KMC/69/2020).Zonke izivivinyo zezilwane eziklanyelwe zenziwe ngokuhambisana neziqondiso zeKomidi Lokulawula Nokulawula Ukuhlolwa Kwezilwane (CPCSEA).Bonke ababhali balandela imihlahlandlela ethi FAKA.
I-spectra ye-FTIR yazo zonke i-NFRCS yahlaziywa futhi yaqhathaniswa ne-chitosan spectrum eboniswe kuMfanekiso 2A.Iziqongo ze-spectral ze-chitosan (ezirekhodiwe) ku-3437 cm-1 (OH ne-NH ukwelula, ukuwela), 2945 kanye no-2897 cm-1 (CH stretching), 1660 cm-1 (NH2 strain), 1589 cm-1 (N-H bending ), 10 cm-106 cm-stretch (106 cm - 107 cm-stretch, 1157 cm-7 i-hydroxyl), 993 cm-1 (yelula CO, Bo-OH) 52.53.54.Ithebula le-Supplementary S1 libonisa amanani we-FTIR NFRCS wokumunca we-chitosan (intatheli), i-chitosan emsulwa, i-Cm, i-Ch, ne-Cp.I-spectra ye-FTIR yazo zonke i-NFRCS (Cm, Ch kanye ne-Cp) ibonise amabhande okumunca anesici esifanayo njenge-chitosan emsulwa ngaphandle kwezinguquko ezibalulekile (Umfanekiso 2A).Imiphumela ye-FTIR iqinisekise ukungabikho kokusebenzelana kwamakhemikhali noma ngokomzimba phakathi kwama-polymer asetshenziswa ukuthuthukisa i-NFRCS, okubonisa ukuthi ama-polymer asetshenzisiwe awasebenzi.
Ukulingiswa kwe-in vitro kwe-Cm NFRCS, i-Ch NFRCS, i-Cp NFRCS ne-Cs.(A) imele i-spectra ye-FTIR ehlanganisiwe ye-chitosan ne-Cm NFRCS, i-Ch NFRCS ne-Cp NFRCS ngaphansi kokucindezelwa.(B) a) Izinga lokuthatha igazi lonke le-NFRCS Cm, Ch, Cp, ne-Cg (n = 3);Amasampula e-Ct abonise i-BAR ephakeme ngenxa yokuthi i-swab kakotini inezinga eliphezulu lokumunca kahle;b) Igazi ngemva kokumuncwa kwegazi Umdwebo wesampula elimunnyiwe.Ukumelwa okunemifanekiso kwe-BCT yesampula yokuhlola C (i-Cp NFRCS ibe ne-BCT ehamba phambili (15 s, n = 3)). Idatha ku-C, D, E, kanye no-G iboniswe njengesho ukuthi ± SD, futhi amabha wephutha amele i-SD, ***p <0.0001. Idatha ku-C, D, E, kanye no-G iboniswe njengesho ukuthi ± SD, futhi amabha wephutha amele i-SD, ***p <0.0001. Данные в C, D, E and G представлены как среднее ± стандартное отклонение, а планки погрешностей представляют стандартное отклонение,0,0 ***p. Idatha ku-C, D, E, kanye no-G yethulwa njengencazelo ± ukuchezuka okujwayelekile, futhi amabha wamaphutha amelela ukuchezuka okujwayelekile, ***p<0.0001. C, D, E 和G 中的数据显示為平均值± SD，误差线代表SD,***p <0.0001. C, D, E 和G 中的数据显示為平均值± SD，误差线代表SD,***p <0.0001. Данные в C, D, E and G показаны как среднее значение ± стандартное отклонение, планки погрешностей представляют стандартное отклонение,0,010, ***p. Idatha ku-C, D, E, kanye no-G iboniswa njengesho ukuchezuka okujwayelekile ±, amabha wamaphutha amelela ukuchezuka okujwayelekile, ***p<0.0001.