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Ukopha okungalawulwa kungenye yezimbangela eziphambili zokufa. Ukufeza ukuphuma kwegazi okusheshayo kuqinisekisa ukusinda komuntu njengosizo lokuqala ngesikhathi sokulwa, izingozi zomgwaqo, kanye nemisebenzi yokunciphisa ukufa. I-scaffold ye-fibre-reinforced composite scaffold (NFRCS) etholakala ku-composition elula ye-hemostatic film-forming (HFFC) njengesigaba esiqhubekayo ingabangela futhi ithuthukise ukuphuma kwegazi. Ukuthuthukiswa kwe-NFRCS kusekelwe ekwakhiweni kwephiko le-dragonfly. Isakhiwo sephiko le-dragonfly sakhiwe ngamaphiko ajikelezayo nawade, futhi ama-membrane ephiko axhumene ukuze kugcinwe ubuqotho besakhiwo esincane. I-HFFC imboza ngokulinganayo ubuso be-fiber ngefilimu yobukhulu be-nanometer futhi ixhumanisa ubukhulu bekotoni obusakazwa ngokungahleliwe (Ct) (isigaba esihlakazekile) ukuze kwakhiwe isakhiwo se-nanoporous. Ukuhlanganiswa kwezigaba eziqhubekayo nezihlakazekile kunciphisa izindleko zomkhiqizo ngokuphindwe kayishumi uma kuqhathaniswa nemikhiqizo etholakala kwezentengiselwano. I-NFRCS eguquliwe (ama-tampons noma ama-wristbands) ingasetshenziswa ezinhlotsheni ezahlukene ze-biomedical. Izifundo ze-in vivo ziphethe ngokuthi i-Cp NFRCS ethuthukisiwe ivuselela futhi ithuthukise inqubo yokujiya endaweni yokusetshenziswa. I-NFRCS ingaguqula indawo encane futhi isebenze ezingeni lamaseli ngenxa yesakhiwo sayo se-nanoporous okuholela ekuphulukisweni okungcono kwenxeba kumodeli yenxeba lokususwa.
Ukopha okungalawulwa ngesikhathi sokulwa, izimo zangaphakathi kokuhlinzwa kanye nezimo eziphuthumayo kungaba usongo olukhulu empilweni yabalimele1. Lezi zimo ziholela ekwandeni okuphelele kokumelana kwemithambo yegazi, okuholela ekushaqekeni kwegazi. Izinyathelo ezifanele zokulawula ukopha ngesikhathi nangemva kokuhlinzwa zibhekwa njengezingasongela impilo2,3. Ukulimala kwemithambo emikhulu kuholela ekulahlekelweni kwegazi okukhulu, okuholela ezingeni lokufa elingu-≤ 50% empini kanye no-31% ngesikhathi sokuhlinzwa1. Ukulahlekelwa igazi okukhulu kuholela ekwehleni komthamo womzimba, okunciphisa ukuphuma kwegazi kwenhliziyo. Ukwanda kokumelana okuphelele kwemithambo yegazi kanye nokuphazamiseka okuqhubekayo kwe-microcirculation kuholela ku-hypoxia ezithweni ezisekela impilo. Ukushaqeka kwegazi kungase kwenzeke uma isimo siqhubeka ngaphandle kokungenelela okusebenzayo1,4,5. Ezinye izinkinga zifaka phakathi ukuqhubeka kwe-hypothermia kanye ne-metabolic acidosis, kanye nesifo sokujiya kwegazi esivimbela inqubo yokujiya kwegazi. Ukushaqeka kwegazi okukhulu kuhlotshaniswa nengozi ephezulu yokufa6,7,8. Ku-grade III (progressive) shock, ukumpontshelwa igazi kubalulekile ukuze isiguli sisinde ngesikhathi sokugula nokufa kwangaphakathi kokuhlinzwa kanye nangemuva kokuhlinzwa. Ukuze sinqobe zonke lezi zimo ezisongela ukuphila ezingenhla, sakhe i-nanoporous fiber-reinforced composite scaffold (NFRCS) esebenzisa ukuhlushwa okuncane kwe-polymer (0.5%) sisebenzisa inhlanganisela yama-polymer e-hemostatic ancibilikayo emanzini.
Ngokusebenzisa ukuqinisa ifayibha, imikhiqizo engabizi kakhulu ingathuthukiswa. Imicu ehlelwe ngokungahleliwe ifana nesakhiwo sephiko le-dragonfly, ilinganiswe ngemigqa evundlile neqondile emaphikweni. Imithambo evundlile nemide yephiko ixhumana ne-membrane yephiko (Isithombe 1). I-NFRCS yakhiwe yi-Ct eqinisiwe njengesistimu ye-scaffold enamandla angcono ngokomzimba nangokwemishini (Isithombe 1). Ngenxa yokuthengeka kwayo kanye nobuciko bayo, odokotela abahlinzayo bakhetha ukusebenzisa ama-gauge entambo yekotoni (Ct) ngesikhathi sokuhlinzwa kanye nokugqoka. Ngakho-ke, uma sicabangela izinzuzo zayo eziningi, okuhlanganisa i-cellulose ekristalu engu-> 90% (enikeza ukuthuthukiswa komsebenzi we-hemostatic), i-Ct yasetshenziswa njengohlelo lwamathambo lwe-NFRCS9,10. Ngakho-ke, uma sicabangela izinzuzo zayo eziningi, okuhlanganisa i-cellulose ekristalu engu-> 90% (enikeza ukuthuthukiswa komsebenzi we-hemostatic), i-Ct yasetshenziswa njengohlelo lwamathambo lwe-NFRCS9,10. Следовательно, учитывая его многочисленные преимущества, в том числе > 90% кристаллической целюлозы (участвует в повыской активности), Ct использовали в качестве скелетной системы NFRCS9,10. Ngakho-ke, uma sibheka izinzuzo zayo eziningi, okuhlanganisa ne-cellulose ekristalu engaphezulu kuka-90% (ehilelekile emsebenzini owengeziwe we-hemostatic), i-Ct yasetshenziswa njengohlelo lwamathambo lwe-NFRCS9,10.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,作的骨架系统。因此，考虑到它的多重益处，包括> 90%Ngakho-ke, uma ubheka izinzuzo zayo eziningi, okuhlanganisa i-cellulose ekristalu engaphezu kuka-90% (kusiza ukuthuthukisa umsebenzi we-hemostatic), i-Ct yasetshenziswa njenge-scaffold ye-NFRCS9,10.I-Ct yayimbozwe ngokungaphezulu (kwabonakala ukwakheka kwefilimu enobukhulu obuncane) futhi ixhunywe nokwakheka kwefilimu okwakha i-hemostatic (HFFC). I-HFC isebenza njenge-matrigel, ibambe i-Ct ebekwe ngokungahleliwe ndawonye. Umklamo othuthukisiwe udlulisa ukucindezeleka ngaphakathi kwesigaba esihlakazekile (imicu yokuqinisa). Kunzima ukuthola izakhiwo ezinama-nanoporous ezinamandla amahle omshini kusetshenziswa amazinga amancane e-polymer. Ngaphezu kwalokho, akulula ukwenza ngezifiso izikhunta ezahlukene zezinhlelo zokusebenza ezahlukene zezokwelapha.
Isithombe sibonisa umdwebo womklamo we-NFRCS osekelwe esakhiweni sephiko le-dragonfly (A). Lesi sithombe sibonisa ukufaniswa okuqhathanisayo kwesakhiwo sephiko le-dragonfly (imithambo ehlanganayo kanye nemithambo emide yephiko ixhumene) kanye ne-photomicrograph ye-Cp NFRCS (B). Ukumelwa kwe-NFRCS ngokwesikimu.
Ama-NFRC athuthukiswe kusetshenziswa i-HFC njengesigaba esiqhubekayo sokubhekana nemikhawulo engenhla. I-HFC yakhiwe ngama-polymer ahlukahlukene e-hemostatic akha ifilimu afaka phakathi 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. Ukwengezwa kwe-polyvinylpyrrolidine K30 (PVP K30) kuthuthukise amandla okumunca umswakama we-NFRCS. I-Polyethylene glycol 400 (PEG 400) yengezwe ukuthuthukisa ukuxhumanisa kwe-polymer kuma-polymer ahlanganisiwe. Izinhlanganisela ezintathu ezahlukene ze-HFCC hemostatic (Cm HFCC, Ch HFCC kanye ne-Cp HFCC), okuyi-chitosan ene-MC (Cm), i-chitosan ene-HPMC (Ch), kanye ne-chitosan ene-PVA (Cp), zasetshenziswa ku-Ct. Izifundo ezahlukahlukene zokuchaza i-in vitro kanye ne-in vivo ziqinisekisile umsebenzi wokupholisa amanxeba kanye nowokuphola kwe-NFRCS. Izinto ezihlanganisiwe ezinikezwa yi-NFRCS zingasetshenziswa ukwenza ngezifiso izinhlobo ezahlukene ze-scaffolding ukuze kuhlangatshezwane nezidingo ezithile.
Ngaphezu kwalokho, i-NFRCS ingashintshwa njengebhandeji noma i-roll ukuze imboze yonke indawo yokulimala yemilenze engezansi nezinye izingxenye zomzimba. Ngokukhethekile ngokulimala kwemilenze yokulwa, umklamo we-NFRCS oklanyelwe ungashintshwa ube yingalo eyodwa noma umlenze ogcwele (Isithombe Esingeziwe S11). I-NFRCS ingenziwa ibe yibhande lesihlakala elineglue yezicubu, engasetshenziswa ukuvimba ukuphuma kwegazi ngenxa yokulimala okukhulu kwesihlakala. Umgomo wethu oyinhloko ukuthuthukisa i-NFRCS ene-polymer encane ngangokunokwenzeka engalethwa kubantu abaningi (ngaphansi komugqa wobumpofu) futhi engafakwa ekhithini yosizo lokuqala. Idizayini elula, ephumelelayo, futhi engabizi, i-NFRCS izuzisa imiphakathi yendawo futhi ingaba nomthelela emhlabeni jikelele.
I-Chitosan (isisindo sama-molecule angu-80 kDa) kanye ne-amaranth kuthengwe eMerck, eNdiya. I-Hydroxypropyl methylcellulose 50 Cp, i-polyethylene glycol 400 kanye ne-methylcellulose kuthengwe kwa-Loba Chemie Pvt. LLC, eMumbai. I-Polyvinyl alcohol (isisindo sama-molecule esingu-125 kDa) (87-90% i-hydrolysed) ithengwe kwa-National Chemicals, eGujarat. I-Polyvinylpyrrolidine K30 ithengwe kwa-Molychem, eMumbai, ama-swabs angenamagciwane athengwe kwa-Ramaraju Surgery Cotton Mills Ltd., eTamil Nadu, ngamanzi e-Milli Q (uhlelo lokuhlanza amanzi lwe-Direct-Q3, eMerck, eNdiya) njengomthwali.
I-NFRCS yathuthukiswa kusetshenziswa indlela ye-lyophilization11,12. Zonke izakhi ze-HFCC (Ithebula 1) zalungiswa kusetshenziswa i-mechanical stirrer. Lungisa isixazululo esingu-0.5% se-chitosan usebenzisa i-acetic acid engu-1% emanzini ngokuxubha okuqhubekayo ku-800 rpm ku-mechanical stirrer. Isisindo esiqondile se-polymer elayishiwe esiboniswe kuThebula 1 sengezwe kusisombululo se-chitosan futhi savuselelwa kwaze kwatholakala isixazululo se-polymer esicacile. I-PVP K30 kanye ne-PEG 400 zengezwe kungxube ephumayo ngamanani aboniswe kuThebula 1, futhi kwavuselelwa kwaqhubeka kwaze kwatholakala isixazululo se-polymer esicacile esiqinile. Ukugeza okubangelwayo kwesisombululo se-polymer kwafakwa i-sonicated imizuzu engama-60 ukuze kususwe amabhamuza omoya abanjwe engxubeni ye-polymer. Njengoba kuboniswe ku-Supplementary Figure S1(b), i-Ct yasatshalaliswa ngokulinganayo emthonjeni ngamunye wepuleti le-6-well (mold) elihlanganiswe ne-5 ml ye-HFCC.
Ipuleti lemithombo eyisithupha lafakwa i-sonic imizuzu engama-60 ukuze kutholakale ukumanzisa kanye nokusatshalaliswa okufanayo kwe-HFC kunethiwekhi ye-Ct. Bese kubanda ipuleti lemithombo eyisithupha ku--20°C amahora angu-8-12. Amapuleti okuqandisa afakwa i-lyophilized amahora angama-48 ukuze kutholakale izinhlobo ezahlukene ze-NFRCS. Inqubo efanayo isetshenziselwa ukukhiqiza izimo nezakhiwo ezahlukene, njenge-tampon noma i-cylindrical tampons, noma olunye uhlobo lwezinhlelo zokusebenza ezahlukene.
I-chitosan esikaliwe kahle (80 kDa) (3%) incibilikiswa ku-1% acetic acid kusetshenziswa i-magnetic stirrer. Esixazululweni esiphumela se-chitosan kwenezelwe i-1% PEG 400 futhi kwavuselelwa imizuzu engama-30. Thela isixazululo esiphumelayo esitsheni esiyisikwele noma esingunxande bese uqandisa ku--80°C amahora ayi-12. Amasampula aqandisiwe ahlungiwe amahora angama-48 ukuze kutholakale i-Cs13 enezimbobo.
I-NFRCS ethuthukisiwe yahlolwa kusetshenziswa i-Fourier transform infrared spectroscopy (FTIR) (Shimadzu 8400 s FTIR, Tokyo, Japan) ukuqinisekisa ukuhambisana kwamakhemikhali kwe-chitosan namanye ama-polymer14,15. Ama-spectra e-FTIR (ububanzi bebanga le-spectral kusukela ku-400 kuya ku-4000 cm-1) awo wonke amasampula ahlolwe atholwe ngokwenza amaskeni angu-32.
Izinga lokumuncwa kwegazi (i-BAR) lazo zonke izinhlobo lihlolwe kusetshenziswa indlela echazwe nguChen et al. 16 ngokuguqulwa okuncane. Ama-NFRK athuthukisiwe azo zonke izithako omisiwe kuhhavini we-vacuum ku-105°C ubusuku bonke ukuze kususwe i-solvent esele. Ama-30 mg e-NFRCS (isisindo sesampula sokuqala - i-W0) kanye nama-30 mg e-Ct (ukulawula okuhle) abekwe ezitsheni ezihlukene eziqukethe i-premix ye-3.8% sodium citrate. Ngezikhathi ezinqunyiwe, okungukuthi imizuzwana emi-5, 10, 20, 30, 40 kanye ne-60, ama-NFRCS asuswa futhi izindawo zawo zahlanzwa igazi elingamuncwanga ngokubeka amasampula ku-Ct imizuzwana engama-30. Isisindo sokugcina segazi elimuncwa yi-NFRCS 16 sacatshangelwa (i-W1) ngesikhathi ngasinye. Bala iphesenti le-BAR usebenzisa ifomula elandelayo:
Isikhathi sokujiya kwegazi (i-BCT) sinqunywe njengoba kubikwe nguWang et al. 17. Isikhathi esidingekayo ukuze igazi eliphelele (igazi legundane elixutshwe ne-3.8% sodium citrate) lijiye phambi kwe-NFRCS sibalwa njenge-BCT yesampula yokuhlola. Izingxenye ezahlukene ze-NFRCS (30 mg) zifakwe emabhodleleni e-screw cap angu-10 ml futhi zafakwa ku-37°C. Igazi (0.5 ml) lafakwa ebhodleleni kwathi u-0.3 ml we-0.2 M CaCl2 wafakwa ukuze kusebenze ukujiya kwegazi. Ekugcineni, guqula ibhodlela njalo ngemizuzwana eyi-15 (kufika ku-180°) kuze kube yilapho kwakheka i-clot eqinile. I-BCT yesampula ilinganiswa ngenani lama-flips vails17,18. Ngokusekelwe ku-BCT, kukhethwe ukwakheka okubili okuhle kakhulu okuvela ku-NFRCS Cm, Ch kanye ne-Cp ukuze kuqhutshekwe nokuhlolwa kwezimpawu.
I-BCT ye-Ch NFRCS kanye ne-Cp NFRCS compositions inqunywe ngokusebenzisa indlela echazwe nguLi et al. 19. Beka i-15 x 15 mm2 Ch NFRCS, i-Cp NFRCS, kanye ne-Cs (ukulawula okuhle) ezitsheni ezihlukene ze-Petri (37 °C). Igazi eliqukethe i-3.8% sodium citrate lixutshwe ne-0.2 M CaCl2 ngesilinganiso sevolumu esingu-10:1 ukuqala inqubo yokujiya kwegazi. I-20 µl yengxube yegazi legundane le-0.2 M CaCl2 yafakwa ebusweni besampula yafakwa esitsheni se-Petri esingenalutho. Ukulawula kwakuthiwa igazi lithululelwe ezitsheni ze-Petri ezingenalutho ngaphandle kwe-Ct. Ngezikhawu ezimisiwe zemizuzu engu-0, 3, kanye ne-5, yeka ukujiya ngokungeza i-10 ml yamanzi a-deionized (DI) esitsheni esiqukethe isitsha ngaphandle kokuphazamisa i-clot. Ama-erythrocyte angagoqekile (ama-erythrocyte) angena e-hemolysis lapho kukhona amanzi a-deionized futhi akhiphe i-hemoglobin. I-hemoglobin ngezikhathi ezahlukene (HA(t)) ilinganiswe ku-540 nm (i-hemoglobin ye-λmax) kusetshenziswa i-UV-Vis spectrophotometer. Ukumuncwa okuphelele kwe-hemoglobin (AH(0)) ku-0 min ye-20 µl yegazi ku-10 ml yamanzi angenawo i-ion kwathathwa njengendinganiso yokubhekisela. I-relative hemoglobin uptake (RHA) yegazi elihlanganisiwe yabalwa kusukela ku-HA(t)/HA(0) kusetshenziswa i-batch efanayo yegazi.
Kusetshenziswa i-texture analyzer (Texture Pro CT V1.3 Build 15, Brookfield, USA), kwatholakala ukuthi izimfanelo zokunamathela ze-NFRK ezicutshini ezonakele. Cindezela isitsha esiyisilinda esivulekile phansi ngaphakathi kwesikhumba sengulube (ngaphandle kongqimba lwamafutha). Amasampula (Ch NFRCS kanye ne-Cp NFRCS) afakwa nge-cannula ezikhumbeni zesilinda ukuze kudalwe ukunamathela esikhumbeni sengulube. Ngemva kokufukamela imizuzu emi-3 ekushiseni kwegumbi (RT) (25° C.), amandla okunamathela e-NFRCS aqoshwa ngesivinini esingaguquki esingu-0.5 mm/sec.
Isici esiyinhloko sezinto zokuvala igazi ngokuhlinzwa ukwandisa ukujiya kwegazi ngenkathi kunciphisa ukulahleka kwegazi. Ukujiya okungenakulahlekelwa ku-NFRCS kuhlolwe kusetshenziswa indlela eyanyatheliswa ngaphambilini enezinguquko ezincane 19. Yenza ithubhu le-microcentrifuge (2 ml) (ububanzi bangaphakathi obuyi-10 mm) elinembobo engu-8 × 5 mm2 ohlangothini olulodwa lwethubhu le-centrifuge (elimele inxeba elivulekile). I-NFRCS isetshenziselwa ukuvala ukuvulwa bese kusetshenziswa itheyiphu ukuvala imiphetho yangaphandle. Engeza u-20 µl we-0.2 M CaCl2 ethubhu le-microcentrifuge eliqukethe i-3.8% sodium citrate premix. Ngemuva kwemizuzu eyi-10, amathubhu e-microcentrifuge asuswa ezitsheni futhi ukwanda kwesisindo sezitsha kwanqunywa ngenxa yokuphuma kwegazi kusuka ku-NFRK (n = 3). Ukulahlekelwa yigazi i-Ch NFRCS kanye ne-Cp NFRCS kwaqhathaniswa ne-Cs.
Ubuqotho obumanzi be-NFRCS bunqunywe ngokusekelwe endleleni echazwe nguMishra noChaudhary21 ngezinguquko ezincane. Beka i-NFRCS ebhodleleni lika-Erlenmeyer elingu-100 ml elinamanzi angu-50 ml bese ujikeleza imizuzwana engama-60 ngaphandle kokwenza ingaphezulu. Ukuhlolwa okubonakalayo kanye nokubekwa phambili kwamasampula ukuze kube nobuqotho obungokomzimba ngokusekelwe ekuqoqweni.
Amandla okubopha e-HFCC ku-Ct afundwe kusetshenziswa izindlela ezishicilelwe ngaphambilini ngezinguquko ezincane. Ubuqotho bokumboza ubuso buhlolwe ngokudalula i-NFRK kumaza e-acoustic (isikhuthazo sangaphandle) lapho kukhona amanzi e-milliQ (Ct). Ama-NFRCS Ch NFRCS kanye ne-Cp NFRCS athuthukisiwe afakwa ebhekheni eligcwele amanzi futhi afakwa i-sonicated imizuzu eyi-1, 2, 3, 4, 5, 6, 7, 8, 9, 10, kanye ne-30 ngokulandelana. Ngemva kokomisa, umehluko wephesenti phakathi kwesisindo sokuqala nesokugcina se-NFRCS wasetshenziswa ukubala ukulahleka kwephesenti lezinto (HFCC). I-BCT ye-in vitro isekele amandla okubopha noma ukulahleka kwezinto ezingaphezulu. Ukusebenza kahle kokubopha kwe-HFCC ku-Ct kunikeza ukujiya kwegazi kanye nokwemboza okunwebekayo ebusweni be-Ct22.
Ukulingana kwe-NFRCS ethuthukisiwe kunqunywe yi-BCT yamasampula (30 mg) athathwe ezindaweni ezijwayelekile ezikhethiwe ngokungahleliwe ze-NFRCS. Landela inqubo ye-BCT eshiwo ngaphambilini ukuze unqume ukuhambisana ne-NFRCS. Ukusondelana phakathi kwawo wonke amasampula amahlanu kuqinisekisa ukumbozwa okufanayo kwendawo kanye nokufakwa kwe-HFC ku-Ct mesh.
Indawo yokuxhumana kwegazi (i-NBCA) inqunywe njengoba kubikwe ngaphambilini ngokushintsha okuthile. Hlanganisa igazi ngokulibamba ngama-20 µl egazi phakathi kwezindawo ezimbili ze-Ct, i-Ch NFRCS, i-Cp NFRCS kanye ne-Cs. Ngemva kwehora eli-1, izingxenye ezimbili ze-stent zahlukaniswa futhi kwalinganiswa ngesandla indawo ye-clot. Inani elimaphakathi lokuphindaphinda kathathu labhekwa njenge-NBCA NFRCS19.
Ukuhlaziywa kwe-Dynamic Vapor Sorption (DVS) kwasetshenziswa ukuhlola ukusebenza kahle kwe-NFRCS ekumunceni amanzi endaweni yangaphandle noma endaweni yokulimala ebangela ukuqala ukujiya. I-DVS ihlola noma iqopha ukumuncwa nokulahleka komhwamuko kusampula ngokusebenzisa ibhalansi ezwela kakhulu enesisombululo sesisindo esingu-±0.1 µg. Umfutho womhwamuko oyingxenye (umswakama ohlobene) ukhiqizwa yisilawuli sokugeleza kobuningi be-elekthronikhi esizungeze isampula ngokuxuba amagesi agcwele kanye nawomile. Ngokweziqondiso ze-European Pharmacopeia, ngokusekelwe kumaphesenti okumuncwa komswakama ngamasampula, amasampula ahlukaniswe ngezigaba ezine (0–0.012% w/w− non-hygroscopic, 0.2–2% w/w hygroscopic kancane, 2–15% hygroscopic ephakathi nendawo, kanye >15% hygroscopic kakhulu)23. Ngokweziqondiso ze-European Pharmacopeia, ngokusekelwe kumaphesenti okumuncwa komswakama ngamasampula, amasampula ahlukaniswe ngezigaba ezine (0–0.012% w/w− non-hygroscopic, 0.2–2% w/w hygroscopic kancane, 2–15% hygroscopic ephakathi nendawo, kanye >15% hygroscopic kakhulu)23.Ngokuhambisana nezincomo ze-European Pharmacopoeia, kuye ngephesenti lokumuncwa komswakama ngamasampula, amasampula ahlukaniswe ngezigaba ezine (0–0.012% w/w – okungeyona i-hygroscopic, 0.2–2% w/w kancane i-hygroscopic, 2–15%).% умеренно гигроскопичен и > 15% очень гигроскопичен)23. % i-hygroscopic ephakathi nendawo kanye no-> 15% i-hygroscopic kakhulu)23.根据欧洲药典指南，根据樣品吸收水分的百分比，样品分為4 类（0-0.012% w/w-水分的百分比，样品分為4 类（0-0.012% w/w-搸/w2.轻微吸湿性、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 （0（w-10 类 （0（w-10 .性 , , , 0.2-2% W/w 轻微 , 2-15% 适度 吸湿 ，> 15 % 非常吸湿）23.Ngokuhambisana nezincomo ze-European Pharmacopoeia, amasampula ahlukaniswe ngezigaba ezine kuye ngephesenti lomswakama omuncwa yisampula (0-0.012% ngesisindo - okungeyona i-hygroscopic, 0.2-2% ngesisindo esi-hygroscopic kancane, 2-15% ngesisindo).% умеренно гигроскопичен, > 15 % очень гигроскопичен) 23. % hygroscopic ephakathi nendawo, > 15% hygroscopic kakhulu) 23.Ukusebenza kahle kwe-NFCS X NFCS kanye ne-TsN NFCS okune-hygroscopic kunqunywe ku-analyzer DVS TA TGA Q5000 SA. Phakathi nale nqubo, isikhathi sokusebenza, umswakama ohlobene (RH), kanye nesisindo sesampula sesikhathi sangempela ku-25°C24 kutholakale. Okuqukethwe komswakama kubalwa ngokuhlaziywa kwesisindo se-NFRCS okunembile kusetshenziswa i-equation elandelayo:
I-MC iwumswakama we-NFRCS. m1 - isisindo esomile sama-NSAID. m2 iyisisindo se-NFRCS sangempela ku-RH ethile.
Indawo yonke yobuso ilinganiswe kusetshenziswa ukuhlolwa kokumuncwa kwe-nitrogen nge-nitrogen ewuketshezi ngemuva kokuthulula amasampula ku-25 °C amahora ayi-10 (< 7 × 10–3 Torr). Indawo yonke yobuso ilinganiswe kusetshenziswa ukuhlolwa kokumuncwa kwe-nitrogen nge-nitrogen ewuketshezi ngemuva kokuthulula amasampula ku-25 °C amahora ayi-10 (< 7 × 10–3 Torr). Общая площадь поверхности оценивалась с помощью эксперимента по адсорбции азота жидким азотом после опорожнения образцов при 12 C. × 10–3 Торр). Indawo yonke yobuso ilinganiswe kusetshenziswa ukuhlolwa kokumuncwa kwe-nitrogen nge-nitrogen ewuketshezi ngemuva kokuba amasampula ekhishwe ku-25°C amahora ayi-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 азотом после опользованием экспериментов азота жидким азотом после опользованием экспериментов по адсорбции при 25°C (< 7 × 10-3 тор). Indawo yonke engaphezulu ilinganiswe kusetshenziswa ukuhlolwa kokumuncwa kwe-nitrogen nge-nitrogen ewuketshezi ngemuva kokuba amasampula ekhishwe amahora ayi-10 ku-25°C (< 7 x 10-3 torr).Indawo yonke yobuso, ivolumu yembobo kanye nosayizi wembobo ye-NFRCS kunqunywe nge-Quantachrome evela ku-NOVA 1000e, e-Austria kusetshenziswa isofthiwe ye-RS 232.
Lungisa ama-RBC angu-5% (usawoti njengesihlanzi) egazini lonke. Bese udlulisela i-aliquot ye-HFCC (0.25 ml) epuletini elinamanzi angu-96 kanye nesisindo sama-RBC angu-5% (0.1 ml). Faka ingxube ku-37°C imizuzu engu-40. Ingxube yamangqamuzana abomvu egazi kanye ne-serum yabhekwa njengokulawula okuhle, kanye nengxube yamangqamuzana egazi abomvu kanye namangqamuzana egazi abomvu njengokulawula okungalungile. I-Hemagglutination yanqunywa ngokwesilinganiso se-Stajitzky. Izikali eziphakanyisiwe zimi kanje: + + + + ama-aggregates aqinile e-granular; + + + ama-bottom pads abushelelezi anemiphetho egobile; + + ama-bottom pads abushelelezi anemiphetho edabukile; + amasongo abomvu amancane azungeze imiphetho yama-bottom pads abushelelezi; – (negative) inkinobho ebomvu ehlukanisiwe 12 maphakathi nomthombo ongezansi.
Ukuhambisana kwegazi kwe-NFRCS kwafundwa ngokwendlela ye-International Organisation for Standardization (ISO) (ISO10993-4, 1999)26,27. Indlela ye-gravimetric echazwe nguSingh et al. Kwenziwe izinguquko ezincane ukuhlola ukwakheka kwe-thrombus lapho kukhona noma phezu kwe-NFRCS. Ama-500 mg e-Cs, i-Ch NFRCS kanye ne-Cp NFRCS afakwe ku-phosphate buffered saline (PBS) amahora angama-24 ku-37°C. Ngemuva kwamahora angama-24, i-PBS yasuswa kwathi i-NFRCS yaphathwa nge-2 ml yegazi equkethe i-3.8% sodium citrate. Ebusweni be-NFRCS, engeza u-0.04 ml we-0.1 M CaCl2 kumasampula afakwe ku-incubated. Ngemuva kwemizuzu engama-45, kwanezelwa ama-5 ml amanzi acwengekile ukuze kumiswe ukujiya. Igazi elihlanganisiwe ebusweni be-NFRK laphathwa ngesisombululo se-formaldehyde esingu-36-38%. Amahlule ahlanganiswe ne-formaldehyde omisiwe futhi alinganiswa. Iphesenti le-thrombosis lalinganiselwa ngokubala isisindo sengilazi ngaphandle kwegazi kanye nesampula (ukulawula okungekuhle) kanye nengilazi enegazi (ukulawula okuhle).
Njengokuqinisekiswa kokuqala, amasampula abonwe ngeso lengqondo ngaphansi kwe-microscope ebonakalayo ukuze kuqondwe ikhono le-HFCC surface coating, i-Ct exhunyiwe, kanye nenethiwekhi ye-Ct yokwakha ama-pores. Izingxenye ezincane ze-Ch kanye ne-Cp ezivela ku-NFRCS zanqunywa nge-scalpel blade. Ingxenye eyalandela yabekwa ku-slide yengilazi, yambozwa nge-coverlip, kanti imiphetho yaqiniswa nge-glue. Ama-slide alungisiwe abhekwa ngaphansi kwe-microscope ebonakalayo futhi izithombe zathathwa ngokukhulisa okuhlukile.
Ukufakwa kwe-polymer kumanethiwekhi e-Ct kwabonakala kusetshenziswa i-fluorescence microscopy ngokusekelwe endleleni echazwe nguRice et al.29. Ukwakheka kwe-HFC okusetshenziswe kule nhlanganisela kwaxutshwa nodayi we-fluorescent (amaranth), kwathi i-NFRCS (Ch & Cp) yalungiswa ngokwendlela eshiwo ngaphambilini. Ukwakheka kwe-HFC okusetshenziswe kule nhlanganisela kwaxutshwa nodayi we-fluorescent (amaranth), kwathi i-NFRCS (Ch & Cp) yalungiswa ngokwendlela eshiwo ngaphambilini.Ukwakheka kwe-HFC okusetshenziselwa ukwakheka kwahlanganiswa nodayi we-fluorescent (amaranth) kwathi i-NFRCS (Ch kanye ne-Cp) yatholakala ngokwendlela 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-HFCC okusetshenziswe kule ndlela kwahlanganiswa nodayi we-fluorescent (Amaranth) futhi kwatholakala i-NFRCS (Ch kanye ne-Cp), njengoba kushiwo ngaphambili.Izingxenye ezincane ze-NFRK zasikwa kumasampula atholiwe, zabekwa kumaslayidi engilazi, zambozwa ngamaslayidi okumboza. Bheka amaslayidi alungisiwe ngaphansi kwe-microscope ye-fluorescent usebenzisa isihlungi esiluhlaza (310-380 nm). Izithombe zathathwa ngokukhulisa okungu-4x ukuze kuqondwe ubudlelwano be-Ct kanye nokufakwa kwe-polymer okweqile kunethiwekhi ye-Ct.
I-topography yobuso be-NFRCS Ch kanye ne-Cp inqunywe 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 (Scanning Probe Image Processor). Izindawo ezahlukene ze-NFRCS zenziwe ezithombeni ze-3D ukuhlola ukufana kobuso. Ukuphambuka okujwayelekile kwesilinganiso sendawo ethile kuchazwa njengobunzima bobuso. I-RMS equation isetshenziswe ukulinganisa ubulukhuni bobuso be-NFRCS31.
Izifundo ezisekelwe ku-FESEM zenziwe kusetshenziswa i-FESEM, i-SU8000, i-HI-0876-0003, eHitachi, eTokyo, ukuze kuqondwe isimo sobuso be-Ch NFRCS kanye ne-Cp NFRCS, ebonise i-BCT engcono kune-Cm NFRCS. Ucwaningo lwe-FESEM lwenziwe ngokwendlela echazwe nguZhao et al. 32 ngokuguqulwa okuncane. I-NFRCS 20 kuya ku-30 ​​mg ye-Ch NFRCS kanye ne-Cp NFRCS zaxutshwa kusengaphambili ne-20 µl ye-3.8% sodium citrate exutshwe kusengaphambili negazi legundane. I-20 μl ye-0.2 M CaCl2 yanezelwa kumasampula aphathwe ngegazi ukuze kuqalwe ukujiya futhi amasampula afakwa ekushiseni kwegumbi imizuzu eyi-10. Ngaphezu kwalokho, ama-erythrocyte amaningi asuswa ebusweni be-NFRCS ngokuhlanzwa nge-saline.
Amasampula alandelayo aphathwe nge-0.1% glutaraldehyde abese omiswa kuhhavini oshisayo ku-37°C ukuze kususwe umswakama. Amasampula omisiwe ambozwa futhi ahlaziywa 32. Ezinye izithombe ezitholwe ngesikhathi sokuhlaziywa kwakuwukwakheka kwe-clot ebusweni bezintambo zikakotini ngazinye, ukufakwa kwe-polymer phakathi kwe-Ct, i-erythrocyte morphology (ukuma), ubuqotho be-clot, kanye ne-erythrocyte morphology lapho kukhona i-NFRCS. Izindawo ze-NFRCS ezingalashwanga kanye nezindawo ze-NFRCS eziphathwe nge-Ch kanye ne-Cp ezifakwe igazi ziskeniwe ukuthola ama-ion ayisisekelo (i-sodium, i-potassium, i-nitrogen, i-calcium, i-magnesium, i-zinc, ithusi kanye ne-selenium)33. Qhathanisa amaphesenti ama-ion ayisisekelo phakathi kwamasampula aphathwe kanye nalawo angalashwanga ukuze uqonde ukuqongelela kwama-ion ayisisekelo ngesikhathi sokwakheka kwe-clot kanye ne-homogeneity ye-clot.
Ubukhulu bokumbozwa kobuso be-Cp HFFC ebusweni be-Ct bunqunywe kusetshenziswa i-FESEM. Izingxenye eziphambene ze-Cp NFRCS zisikwe ohlakeni futhi zambozwa nge-sputter. Amasampula okumbozwa kwe-sputter aphume abonwa yi-FESEM kanti ubukhulu bokumbozwa kobuso bulinganiswe ngama-34, 35, 36.
I-X-ray micro-CT inikeza isithombe se-3D esingabhubhisi esine-resolution ephezulu futhi ikuvumela ukuthi utadishe ukuhlelwa kwesakhiwo sangaphakathi se-NFRK. I-Micro-CT isebenzisa umsebe we-X-ray odlula esampula ukuze urekhode i-coefficient yendawo yokunciphisa umugqa we-X-ray esampula, okusiza ukuthola ulwazi lwe-morphological. Indawo yangaphakathi ye-Ct ku-Cp NFRCS kanye ne-Cp NFRCS elashwe ngegazi kuhlolwe yi-micro-CT ukuze kuqondwe ukusebenza kahle kokumuncwa kanye nokujiya kwegazi lapho kukhona i-NFRCS37,38,39. Izakhiwo ze-3D zamasampula e-Cp NFRCS alashwe ngegazi futhi angalashwa zakhiwa kabusha kusetshenziswa i-micro-CT (V|tome|x S240, Phoenix, Germany). Kusetshenziswa inguqulo yesofthiwe ye-VG STUDIO-MAX 2.2, izithombe eziningana ze-X-ray zathathwa ngezindlela ezahlukene (okungcono kakhulu ukumbozwa okungu-360°) ukuthuthukisa izithombe ze-3D ze-NFRCS. Idatha yokuqoqwa eqoqwe yakhiwa kabusha yaba izithombe ze-3D volumetric kusetshenziswa isofthiwe elula ye-3D ScanIP Academic.
Ngaphezu kwalokho, ukuze kuqondwe ukusatshalaliswa kwe-clot, ama-20 µl egazi elixutshwe kusengaphambili kanye nama-20 µl e-0.2 M CaCl2 afakwe ku-NFRCS ukuze kuqalwe ukujiya kwegazi. Amasampula alungisiwe ashiywa ukuze aqine. Ubuso be-NFRK buphathwe nge-0.5% glutaraldehyde futhi bomiswa kuhhavini womoya oshisayo ku-30–40°C imizuzu engama-30. I-clot yegazi eyakheka ku-NFRCS yaskenwa, yakhiwa kabusha, futhi kwabonakala isithombe se-3D se-clot yegazi.
Ukuhlolwa kwe-antibacterial kwenziwa ku-Cp NFRCS (uma kuqhathaniswa ne-Ch NFRCS) kusetshenziswa indlela echazwe ngaphambilini enezinguquko ezincane. Umsebenzi we-antibacterial we-Cp NFRCS kanye ne-Cp HFCC unqunywe kusetshenziswa ama-microorganism amathathu ahlukene okuhlola [i-S.aureus (i-gram-positive bacteria), i-E.coli (i-gram-negative bacteria) kanye ne-Candida emhlophe (i-C.albicans)] ekhula ku-agar ezitsheni ze-Petri ku-incubator. Faka ngokulinganayo i-50 ml ye-bacterial culture suspension exutshwe kahle ku-concentration ye-105-106 CFU ml-1 ku-agar medium. Thela i-medium esitsheni se-Petri bese uyivumela iqine. Kwenziwe imigodi ebusweni bepuleti le-agar ukuze kugcwaliswe nge-HFCC (imigodi emi-3 ye-HFCC kanye ne-1 yokulawula okungekuhle). Engeza i-200 µl HFCC emigxotsheni emi-3 kanye ne-200 µl pH 7.4 PBS emgqonyeni wesine. Ngakolunye uhlangothi lwesitsha se-petri, beka idiski engu-12 mm Cp NFRCS ku-agar eqinile bese umanzisa nge-PBS (pH 7.4). Amaphilisi e-Ciprofloxacin, ampicillin kanye ne-fluconazole abhekwa njengezindinganiso zokubhekisela ze-Staphylococcus aureus, i-Escherichia coli kanye ne-Candida albicans. Kala indawo yokuvinjelwa ngesandla bese uthatha isithombe sedijithali sendawo yokuvinjelwa.
Ngemva kokuvunyelwa kokuziphatha kwesikhungo, ucwaningo lwenziwe eKasturba Medical College of Education and Research eManipal, eKarnataka, eningizimu yeNdiya. Iphrothokholi yokuhlola ye-TEG ye-in vitro ibuyekezwe futhi yavunyelwa yiKomidi Lokuziphatha Lezikhungo laseKasturba Medical College, eManipal, eKarnataka (IEC: 674/2020). Izihloko zaqashwa kubanikeli begazi abazithandelayo (abaneminyaka engu-18 kuya ku-55) ebhange legazi lesibhedlela. Ngaphezu kwalokho, ifomu lemvume enolwazi latholakala kubavolontiya ukuze kuqoqwe amasampula egazi. I-Native TEG (N-TEG) ​​​​yasetshenziswa ukutadisha umphumela wokwakheka kwe-Cp HFCC egazini eliphelele elixutshwe ne-sodium citrate. I-N-TEG yaziwa kabanzi ngendima yayo ekuvuseleleni indawo yokunakekelwa, okudala izinkinga kodokotela ngenxa yokuthi kungenzeka kube nokubambezeleka okukhulu emiphumeleni (ukuhlolwa kokuqina kwegazi okuvamile). Ukuhlaziywa kwe-N-TEG kwenziwa kusetshenziswa igazi eliphelele. Imvume enolwazi kanye nomlando wezokwelapha onemininingwane kwatholakala kubo bonke abahlanganyeli. Ucwaningo aluzange lubandakanye abahlanganyeli abanezinkinga ze-hemostatic noma ze-thrombotic njengokukhulelwa/ngemuva kokubeletha noma isifo sesibindi. Abantu abathatha izidakamizwa ezithinta i-coagulation cascade nabo bakhishwe ocwaningweni. Ukuhlolwa okuyisisekelo kwelabhorethri (i-hemoglobin, isikhathi se-prothrombin, i-thromboplastin esebenzayo kanye nokubalwa kwamaplatelet) kwenziwa kubo bonke abahlanganyeli ngokwezinqubo ezijwayelekile. I-N-TEG inquma i-blood clot viscoelasticity, isakhiwo sokuqala se-clot, ukusebenzisana kwezinhlayiya, ukuqinisa i-clot, kanye ne-clot lysis. Ukuhlaziywa kwe-N-TEG kunikeza idatha yesithombe nezinombolo ngemiphumela ehlangene yezakhi eziningana zamaseli kanye ne-plasma. Ukuhlaziywa kwe-N-TEG kwenziwa kumavolumu amabili ahlukene e-Cp HFCC (10 µl kanye no-50 µl). Ngenxa yalokho, i-1 ml yegazi eliphelele eline-citric acid yanezelwa ku-10 μl ye-Cp HFCC. Engeza i-1 ml (Cp HFCC + igazi elicitrated), i-340 µl yegazi elixubile ku-20 µl 0.2 M CaCl2 equkethe isitsha se-TEG. Ngemva kwalokho, izitsha ze-TEG zafakwa ku-TEG® 5000, US ukuze kulinganiswe i-R, K, i-alpha angle, i-MA, i-G, i-CI, i-TPI, i-EPL, i-LY ama-30% amasampula egazi lapho kukhona i-Cp HFFC41.
Iphrothokholi yokufunda ngaphakathi kwe-vivo ibuyekezwe futhi yavunywa yiKomidi Lokuziphatha Kwezilwane Lesikhungo (IAEC), iKasturba School of Medicine, iManipal Institute of Higher Education, iManipal (IAEC/KMC/69/2020). Zonke izivivinyo zezilwane zenziwe ngokuhambisana nezincomo zeKomidi Lokulawula Nokugada Ukuhlolwa Kwezilwane (CPCSEA). Zonke izifundo ze-NFRCS ngaphakathi kwe-vivo (2 × 2 cm2) zenziwe kumagundane e-Wistar wesifazane (anesisindo esingu-200 kuya ku-250 g). Zonke izilwane zajwayelana nokushisa okungu-24-26°C, izilwane zazinokufinyelela kwamahhala kokudla okujwayelekile namanzi ngokushesha. Zonke izilwane zahlukaniswa ngokungahleliwe ngamaqembu ahlukene, iqembu ngalinye lalakhiwe yizilwane ezintathu. Zonke izifundo zenziwa ngokuhambisana ne-Animal Studies: Umbiko we-In Vivo Experiments 43. Ngaphambi kocwaningo, izilwane zalaliswa i-anesthesia ngokuphathwa kwe-intraperitoneal (ip) 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 yabalwa ngokuhlola umehluko phakathi kwesisindo sokuqala nesokugcina samasampula, inani elimaphakathi elitholwe ezivivinyweni ezintathu lathathwa njengevolumu yokopha yesampula.
Imodeli yokunqunywa komsila wegundane yasetshenziswa ukuze kuqondwe amandla e-NFRCS okulawula ukopha kwegazi ekulimaleni, empini, noma engozini yemoto (imodeli yokulimala). Sika u-50% womsila nge-scalpel blade bese uwubeka emoyeni imizuzwana eyi-15 ukuqinisekisa ukopha okuvamile. Ngaphezu kwalokho, amasampula okuhlola abekwa emsileni wegundane ngokusebenzisa ingcindezi (Ct, Cs, Ch NFRCS kanye ne-Cp NFRCS). Ukopha kanye ne-PCT kubikwe kumasampula okuhlola (n ​​= 3)17,45.
Ukusebenza kahle kokulawulwa kwengcindezi ye-NFRCS empini kuhlolwe kumodeli yomthambo we-femoral ongaphezulu. Umthambo we-femoral uyavezwa, ubhotshozwe nge-trocar engu-24G, bese wopha zingakapheli imizuzwana eyi-15. Ngemva kokubonwa ukopha okungalawuleki, isampula yokuhlola ibekwa endaweni yokubhotshozwa ngengcindezi esetshenziswayo. Ngokushesha ngemva kokusetshenziswa kwesampula yokuhlola, isikhathi sokujiya kwaqoshwa futhi ukusebenza kahle kwe-hemostatic kwabonwa imizuzu emi-5 elandelayo. Inqubo efanayo yaphindwa nge-Cs kanye ne-Ct46.
UDowling nabanye abangu-47 baphakamise imodeli yokulimala kwesibindi ukuze kuhlolwe amandla okugcoba kwezinto ezikhipha igazi emzimbeni uma kuziwa ekuphumeni kwegazi ngesikhathi sokuhlinzwa. I-BCT yaqoshwa kumasampula e-Ct (ukulawula okungekuhle), uhlaka lwe-Cs (ukulawula okuhle), amasampula e-Ch NFRCS, kanye namasampula e-Cp NFRCS. I-vena cava yegundane engenhla yadalulwa ngokwenza i-laparotomy ephakathi. Ngemuva kwalokho, ingxenye ekude ye-lobe yesobunxele yasikwa ngesikele. Yenza isike esibindini nge-scalpel blade bese uyivumela ukuthi yophe imizuzwana embalwa. Amasampula okuhlola e-Ch NFRCS kanye ne-Cp NFRCS alinganiswe kahle abekwa endaweni eyonakele ngaphandle kokucindezela okuhle futhi i-BCT yaqoshwa. Iqembu lokulawula (Ct) labe selisebenzisa ingcindezi elandelwa yi-Cs 30 s47 ngaphandle kokuphula ukulimala.
Ukuhlolwa kokuphulukiswa kwenxeba emzimbeni kwenziwa kusetshenziswa imodeli yenxeba elikhipha izicubu ukuze kuhlolwe izakhiwo zokuphulukiswa kwenxeba zama-NFRCS asekelwe ku-polymer athuthukisiwe. Amamodeli amanxeba akhipha izicubu akhethiwe futhi enziwa ngokwezindlela ezishicilelwe ngaphambilini ngezinguquko ezincane19,32,48. Zonke izilwane zahlanzwa izicubu njengoba kuchaziwe ngaphambilini. Sebenzisa i-biopsy punch (12 mm) ukwenza ukusika okujulile okujikelezayo esikhumbeni somhlane. Izindawo zamanxeba ezilungisiwe zagqokwa nge-Cs (ukulawula okuhle), i-Ct (ukubona ukuthi amaphedi kakotini aphazamisa ukuphulukiswa), i-Ch NFRCS kanye ne-Cp NFRCS (iqembu lokuhlola) kanye nokulawula okungekuhle ngaphandle kokwelashwa. Ngosuku ngalunye locwaningo, indawo yenxeba yayilinganiswa kuwo wonke amagundane. Sebenzisa ikhamera yedijithali ukuthatha isithombe sendawo yenxeba bese ugqoka ibhandeji elisha. Iphesenti lokuvalwa kwenxeba lalinganiswa ngale ndlela elandelayo:
Ngokusekelwe kumaphesenti okuvalwa kwenxeba ngosuku lwe-12 locwaningo, isikhumba segundane seqembu elihle kakhulu sakhishwa ((Cp NFRCS) kanye neqembu lokulawula) futhi safundwa yi-H&E staining kanye ne-Masson's trichrome staining. Ngokusekelwe kumaphesenti okuvalwa kwenxeba ngosuku lwe-12 locwaningo, isikhumba segundane seqembu elihle kakhulu sakhishwa ((Cp NFRCS) kanye neqembu lokulawula) futhi safundwa yi-H&E staining kanye ne-Masson's trichrome staining.Ngokusekelwe kumaphesenti okuvalwa kwenxeba ngosuku lwe-12 locwaningo, isikhumba samagundane eqembu elihle kakhulu ((Cp NFRCS) kanye neqembu lokulawula) sasuswa futhi sahlolwa ngokufaka i-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，进行H&E染色 kanye noMasson三色染色研究。根据研第第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 eqenjini elihle kakhulu ((Cp NFRCS) kanye namaqembu okulawula) akhishwa ukuze kufakwe i-hematoxylin-eosin staining kanye ne-Masson's trichrome staining ngokusekelwe ekuvalekeni kwenxeba ngamaphesenti ngosuku lwe-12 locwaningo.Inqubo yokufaka umbala esetshenzisiwe yenziwa ngokwezindlela ezichazwe ngaphambilini49,50. Kafushane, ngemva kokufakwa ku-formalin engu-10%, amasampula akhishwa amanzi kusetshenziswa uchungechunge lwe-alcohols ezilinganisiwe. Sebenzisa i-microtome ukuthola izingxenye ezincane (ubukhulu obungu-5 µm) zezicubu ezikhishwe. Izingxenye ezincane zokulawula kanye ne-Cp NFRCS zelashwe nge-hematoxylin kanye ne-eosin ukuze kufundwe izinguquko ze-histopathological. I-Masson's trichrome stain yasetshenziswa ukuthola ukwakheka kwe-collagen fibrils. Imiphumela etholakele yafundwa ngokungazi lutho ngososayensi bezifo.
Ukuqina kwamasampula e-Cp NFRCS kwafundwa ekushiseni kwegumbi (25°C ± 2°C/60% RH ± 5%) izinyanga eziyi-1251. I-Cp NFRCS (ukushintsha kombala womphezulu kanye nokukhula kwamagciwane) yahlolwa ngokubonakalayo futhi yahlolwa ukumelana nokuguguleka kokugoba kanye ne-BCT ngokwezindlela ezingenhla ezichazwe esigabeni sezinto zokwakha kanye nezindlela.
Ukukwazi ukukhulisa kanye nokuphindaphinda kwe-Cp NFRCS kuhlolwe ngokulungiselela i-Cp NFRCS enosayizi ongu-15×15 cm2. Ngaphezu kwalokho, amasampula angu-30 mg (n = 5) akhishwe ezingxenyaneni ezahlukene ze-Cp NFRCS kanti i-BCT yamasampula afundwe yahlolwa njengoba kuchaziwe ngaphambili esigabeni sezindlela.
Sizame ukuthuthukisa izimo nezakhiwo ezahlukahlukene sisebenzisa ukwakheka kwe-Cp NFRCS kwezinhlelo ezahlukene zezokwelapha. Lezi zakhiwo noma ukwakheka kufaka phakathi ama-swab aqinile okopha ekhaleni, izinqubo zamazinyo, kanye nama-swab asilinda okopha esithweni sangasese sowesifazane.
Wonke amasethi wedatha avezwa njengokwehluka okujwayelekile okungu-± futhi ahlaziywe yi-ANOVA kusetshenziswa i-Prism 5.03 (GraphPad, San Diego, CA, USA) kulandelwe ukuhlolwa kokuqhathanisa okuningi kukaBonferroni (*p<0.05).
Zonke izinqubo ezenziwa ezifundweni zabantu zazihambisana nezindinganiso ze-Institute kanye ne-National Research Council, kanye neSimemezelo saseHelsinki sika-1964 kanye nezichibiyelo zaso ezalandela, noma izindinganiso zokuziphatha ezifanayo. Bonke abahlanganyeli baziswa ngezici zocwaningo kanye nohlobo lwalo lokuzithandela. Idatha yabahlanganyeli ihlala iyimfihlo uma isiqoqiwe. Iphrothokholi yokuhlola ye-TEG ye-in vitro ibuyekezwe futhi yavunyelwa yiKomidi Lokuziphatha Lezikhungo laseKasturba Medical College, eManipal, eKarnataka (IEC: 674/2020). Abavolontiya basayine imvume enolwazi yokuqoqa amasampula egazi.
Zonke izinqubo ezenziwa ezifundweni zezilwane zenziwe ngokuhambisana neKastuba Faculty of Medicine, Manipal Institute of Higher Education, Manipal (IAEC/KMC/69/2020). Zonke izivivinyo zezilwane eziklanywe zenziwa ngokuhambisana neziqondiso zeKomidi Lokulawula Nokugada Ukuhlolwa Kwezilwane (CPCSEA). Bonke ababhali balandela iziqondiso ze-ARRIVE.
Ama-spectra e-FTIR awo wonke ama-NFRCS ahlaziywe futhi aqhathaniswa ne-spectrum ye-chitosan eboniswe ku-Figure 2A. Iziqongo ze-spectral ze-chitosan (eziqoshiwe) ku-3437 cm-1 (ukwelula kwe-OH kanye ne-NH, ukugqagqana), 2945 kanye no-2897 cm-1 (ukwelula kwe-CH), 1660 cm-1 (uhlobo lwe-NH2), 1589 cm-1 (ukugoba kwe-N–H), 1157 cm-1 (ukwelula kwebhuloho O-), 1067 cm-1 (ukwelula kwe-C–O, i-hydroxyl yesibili), 993 cm-1 (ukwelula kwe-CO, i-Bo-OH) 52.53.54. Ithebula Elingeziwe S1 libonisa amanani e-spectrum yokumuncwa kwe-FTIR NFRCS ye-chitosan (intatheli), i-chitosan emsulwa, i-Cm, i-Ch, kanye ne-Cp. Ama-spectra e-FTIR awo wonke ama-NFRCS (Cm, Ch kanye ne-Cp) abonise amabhande okumunca afanayo njenge-chitosan emsulwa ngaphandle kwezinguquko ezibalulekile (Isithombe 2A). Imiphumela ye-FTIR iqinisekisile ukungabikho kokuxhumana kwamakhemikhali noma ngokomzimba phakathi kwama-polymer asetshenziswa ukuthuthukisa i-NFRCS, okubonisa ukuthi ama-polymer asetshenzisiwe awasebenzi.
Ukucaciswa kwe-in vitro kwe-Cm NFRCS, i-Ch NFRCS, i-Cp NFRCS kanye ne-Cs. (A) imele ama-spectra e-FTIR ahlanganisiwe ezingxenyeni ze-chitosan kanye ne-Cm NFRCS, i-Ch NFRCS kanye ne-Cp NFRCS ngaphansi kokucindezelwa. (B) a) Izinga lokungenisa igazi lonke le-NFRCS Cm, Ch, Cp, kanye ne-Cg (n = 3); Amasampula e-Ct abonise i-BAR ephezulu ngoba i-cotton swab inokusebenza kahle kokumunca okuphezulu; b) Igazi ngemva kokumunca igazi Umfanekiso wesampula emuncwe. Ukumelwa kwesithombe se-BCT yesampula yokuhlola i-C (i-Cp NFRCS yayine-BCT engcono kakhulu (15 s, n = 3)). Idatha ku-C, D, E, kanye no-G iboniswe njenge-± SD ephakathi, kanti amabha amaphutha amele i-SD, ***p < 0.0001. Idatha ku-C, D, E, kanye no-G iboniswe njenge-± SD ephakathi, kanti amabha amaphutha 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 njengokuphambuka okujwayelekile okungu-±, kanti amabha amaphutha amelela ukuphambuka 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 njengokuphambuka okujwayelekile okungu-±, amabha amaphutha amelela ukuphambuka okujwayelekile, ***p<0.0001.