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Ukopha okungalawulekiyo ngomnye woonobangela abaphambili bokusweleka.Ukufezekisa i-hemostasis ngokukhawuleza kuqinisekisa ukusinda kwesifundo njengoncedo lokuqala ngexesha lokulwa, iingozi zendlela, kunye nemisebenzi yokunciphisa ukufa.I-Nanoporous fiber-reinforced composite scaffold (NFRCS) ephuma kwi-hemostatic film-forming composition (HFFC) njengesigaba esiqhubekayo sinokubangela kwaye sikhulise i-hemostasis.Uphuhliso lwe-NFRCS lusekwe kuyilo lwephiko ledragonfly.Isakhiwo sephiko le-dragonfly siquka amaphiko anqamlezayo kunye ne-longitudinal, kwaye iimbumba zamaphiko zixhunywe omnye komnye ukuze zigcine ingqibelelo ye-microstructure.I-HFFC inxibe ngokulinganayo umphezulu wefiber kunye nefilimu yobukhulu be-nanometer kwaye idibanisa ubuninzi bekotoni obusasazwa ngokungaqhelekanga (Ct) (isigaba esihlakaziweyo) ukwenza isakhiwo se-nanoporous.Ukudityaniswa kwezigaba eziqhubekayo kunye nezihlakazekileyo kunciphisa iindleko zemveliso ngamaxesha alishumi xa kuthelekiswa neemveliso ezithengiswayo.I-NFRCS elungisiweyo (iitamponi okanye i-wristbands) inokusetyenziswa kwiinkqubo ezahlukeneyo ze-biomedical.Izifundo ze-vivo ziye zagqiba ekubeni i-Cp NFRCS ephuhlisiwe ivuselela kwaye iphucula inkqubo yokudibanisa kwindawo yokusetyenziswa.I-NFRCS inokumodareyitha i-microenvironment kwaye isebenze kwinqanaba leselula ngenxa yesakhiwo sayo se-nanoporous esibangela ukuphulukiswa kwamanxeba okungcono kwimodeli yesilonda sokuqhawula.
Ukopha okungalawulekiyo ngexesha lokulwa, uqhaqho kunye neemeko zongxamiseko kunokubangela ingozi enkulu kubomi bonzakeleyo1.Ezi meko zikhokelela ngakumbi ekunyukeni ngokubanzi kwi-peripheral vascular resistance, ekhokelela kwi-hemorrhagic shock.Amanyathelo afanelekileyo okulawula ukopha ngexesha nangemva kotyando abonwa njengobungozi bokuphila2,3.Ukulimala kwiinqanawa ezinkulu kukhokelela ekulahlekeni kwegazi elikhulu, okubangelwa izinga lokufa kwe-≤ 50% ekulweni kunye ne-31% ngexesha lotyando1.Ukulahleka kwegazi okukhulu kukhokelela ekunciphiseni umthamo womzimba, okunciphisa ukuphuma kwentliziyo.Ukunyuka kwe-peripheral vascular resistance kunye nokuphazamiseka okuqhubekayo kwe-microcirculation kukhokelela kwi-hypoxia kwizitho zokuxhasa ubomi.Ukutshatyalaliswa kwe-Hemorrhagic kungenzeka ukuba imeko iyaqhubeka ngaphandle kokungenelela okusebenzayo1,4,5.Ezinye iingxaki ziquka ukuqhubela phambili kwe-hypothermia kunye ne-metabolic acidosis, kunye nokuphazamiseka kwe-coagulation ephazamisa inkqubo yokudibanisa.Umothuko owopha kakhulu unxulunyaniswa nomngcipheko omkhulu wokufa6,7,8.Kwinqanaba le-III (eliqhubekayo) ukutshitshiswa, ukufakelwa kwegazi kubalulekile ekusindeni kwesigulane ngexesha le-intraoperative kunye ne-postoperative morbidity kunye nokufa.Ukoyisa zonke ezi meko zingasentla ezisongela ubomi, siye saqulunqa i-nanoporous fiber-reinforced composite scaffold (NFRCS) esebenzisa i-concentration encinci ye-polymer (0.5%) usebenzisa i-polymers ye-hemostatic e-soluble yamanzi.
Ngokusetyenziswa kwe-fiber reinforcement, iimveliso ezingabizi kakhulu zinokuphuhliswa.Imicu elungelelaniswe ngokungacwangciswanga ifana nephiko likahlabamanzi, elilungelelaniswe ngemigca ethe tye nethe nkqo emaphikweni.Imithanjeni enqamlezayo kunye ne-longitudinal yephiko inxibelelana ne-membrane yephiko (umzobo 1).I-NFRCS iqukethe i-Ct eqinisiweyo njengenkqubo ye-scaffold enamandla angcono omzimba kunye noomatshini (Umfanekiso 1).Ngenxa yokufikeleleka kunye nobugcisa, oogqirha botyando bakhetha ukusebenzisa i-cotton thread gauges (Ct) ngexesha lokusebenza kunye nokugqoka. Ngenxa yoko, ngokuqwalasela iinzuzo zayo ezininzi, kubandakanywa> i-90% ye-crystalline cellulose (inika ukuphuculwa komsebenzi we-hemostatic), i-Ct isetyenziswe njengenkqubo yamathambo ye-NFRCS9,10. Ngenxa yoko, ngokuqwalasela iinzuzo zayo ezininzi, kubandakanywa> i-90% ye-crystalline cellulose (inika ukuphuculwa komsebenzi we-hemostatic), i-Ct isetyenziswe njengenkqubo yamathambo ye-NFRCS9,10. Следовательно, учитывая его многочисленные преимущества, в том числе > 90% кристаллической целюлозы (участвущет в повыское), пользовали в качестве скелетной системы NFRCS9,10. Ngoko ke, ngenxa yeenzuzo zayo ezininzi, kubandakanywa> i-90% ye-crystalline cellulose (ebandakanyekayo ekunyuseni komsebenzi we-hemostatic), i-Ct isetyenziswe 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（有助于强止血活性）），Ct 结晶纤维素).因此，考虑到它的多重益处，包括> 90%Ngoko ke, ngenxa yeenzuzo zayo ezininzi, kubandakanywa ngaphezu kwe-90% ye-crystalline cellulose (inceda ukunyusa umsebenzi we-hemostatic), i-Ct isetyenziswe njenge-scaffold ye-NFRCS9,10.I-Ct yayigqunywe phezulu (ukwenziwa kwefilimu ye-nano-thick yabonwa) kwaye idibaniswe ne-hemostatic film-forming composition (HFFC).I-HFFC isebenza njenge-matrigel, ibambe i-Ct ngokungacwangciswanga kunye.Uyilo oluphuhlisiweyo luhambisa uxinzelelo ngaphakathi kwesigaba esihlakaziweyo (ukuqinisa iifibers).Kunzima ukufumana izakhiwo ze-nanoporous ezinamandla afanelekileyo omatshini usebenzisa i-polymer concentrations encinci.Ukongeza, akukho lula ukwenza iimolds ezahlukeneyo kwizicelo ezahlukeneyo ze-biomedical.
Umzobo ubonisa umzobo woyilo lwe-NFRCS olusekwe kulwakhiwo lwephiko ledragonfly (A).Lo mfanekiso ubonisa uthelekiso oluthelekisayo lwesakhiwo sephiko ledragonfly (i-intersecting and longitudinal veins of the phiko idityanisiwe) kunye ne-cross-sectional photomicrograph ye-Cp NFRCS (B).Umelo olucwangcisiweyo lwe-NFRCS.
Ii-NFRC zaphuhliswa kusetyenziswa i-HFFC njengesigaba esiqhubekayo ukujongana nemida engentla.I-HFFC yenziwe ngeepolymers ze-hemostatic ezahlukeneyo zefilimu ezibandakanya i-chitosan (njenge-polymer hemostatic main) kunye ne-methylcellulose (MC), i-hydroxypropyl methylcellulose (HPMC 50 cp) kunye ne-polyvinyl alcohol (PVA)) (125 kDa) njengepolymer yenkxaso ekhuthaza ukubunjwa kwe-thrombus.ukwakheka.Ukongezwa kwepolyvinylpyrrolidine K30 (PVP K30) kuphuculwe amandla okufunxa ukufuma kwe-NFRCS.I-Polyethylene glycol 400 (PEG 400) yongezwa ukuphucula i-polymer crosslinking kwi-polymer bonded blends.Iingoma ezintathu ezahlukeneyo ze-HFFC ze-hemostatic (Cm HFFC, Ch HFFC kunye neCp HFFC), ezizezi i-chitosan ene-MC (Cm), i-chitosan ene-HPMC (Ch), kunye ne-chitosan ene-PVA (Cp), zisetyenziswe kwi-Ct.Izifundo ezahlukeneyo ze-in vitro kunye ne-vivo ziqinisekisile umsebenzi wokuphilisa i-hemostatic kunye nenxeba le-NFRCS.Imathiriyeli ehlanganisiweyo ebonelelwa yi-NFRCS inokusetyenziselwa ukulungelelanisa iindlela ezahlukeneyo zesikafula ukuhlangabezana neemfuno ezithile.
Ukongeza, i-NFRCS inokuguqulwa njenge-bandage okanye i-roll ukugubungela yonke indawo yokulimala yeendawo ezisezantsi kunye namanye amalungu omzimba.Ngokukodwa ukulwa nokulimala kwamalungu, idizayini ye-NFRCS eyiliweyo ingatshintshwa ibe yingalo yesiqingatha okanye umlenze ogcweleyo (Umfanekiso owongezelelweyo S11).I-NFRCS inokwenziwa ibe yintambo yesihlahla eneglue enokuthi isetyenziswe ukunqanda ukopha ngenxa yokwenzakala kakhulu esihlahleni sokuzibulala.Injongo yethu ephambili kukuphuhlisa i-NFRCS kunye ne-polymer encinci ngokunokwenzeka enokuthi ihanjiswe kubantu abaninzi (ngaphantsi kwentlupheko) kwaye inokufakwa kwikiti yoncedo lokuqala.Ilula, isebenzayo, kwaye inoqoqosho kuyilo, i-NFRCS inceda uluntu lwendawo kwaye inokuba nefuthe kwihlabathi.
I-Chitosan (i-molecular weight 80 kDa) kunye ne-amaranth zathengwa kwi-Merck, e-Indiya.I-Hydroxypropyl methylcellulose 50 Cp, i-polyethylene glycol 400 kunye ne-methylcellulose zathengwa kwi-Loba Chemie Pvt.LLC, eMumbai.I-Polyvinyl alcohol (i-molecular weight 125 kDa) (87-90% hydrolysed) yathengwa kwiMichiza yeSizwe, eGujarat.I-Polyvinylpyrrolidine K30 yathengwa kwi-Molychem, eMumbai, i-swabs eyinyumba yathengwa kwi-Ramaraju Surgery Cotton Mills Ltd., i-Tamil Nadu, kunye ne-Milli Q yamanzi (inkqubo yokucoca amanzi ngokuthe ngqo, i-Merck, e-Indiya) njengomthwali.
I-NFRCS yaphuhliswa kusetyenziswa indlela ye-lyophilization11,12.Zonke iingoma ze-HFFC (Itheyibhile 1) zalungiswa ngokusebenzisa i-stirrer mechanical.Lungiselela isisombululo se-0.5% se-chitosan usebenzisa i-1% ye-acetic acid emanzini ngokuvuselela ngokuqhubekayo kwi-800 rpm kwi-stirrer mechanical.Ubunzima obuchanekileyo bepolymer elayishiweyo eboniswe kwiThebhile 1 yongezwa kwisisombululo se-chitosan kwaye yaxutywa de kwafunyanwa isisombululo se-polymer esicacileyo.I-PVP K30 kunye ne-PEG 400 zongezwa kumxube obangelwayo kwiimali eziboniswe kwiThebhile 1, kwaye ukuvuselela kwaqhubeka de kwafunyanwa isisombululo esicacileyo se-polymer.Ukuhlamba okubangelwa yisisombululo se-polymer kwafakwa kwi-sonicated imizuzu engama-60 ukususa amaqamza omoya avaleleke kumxube wepolymer.Njengoko kubonisiwe kwi-Supplementary Figure S1 (b), i-Ct yasasazwa ngokulinganayo kwiqula ngalinye le-6-well plate (mold) eyongezwa nge-5 ml ye-HFFC.
I-plate ye-six-well plate yayifakwe kwi-sonication ye-60 min ukufezekisa ukumanzisa okufanayo kunye nokuhanjiswa kwe-HFFC kwinethiwekhi ye-Ct.Emva koko qhwaba ipleyiti emithandathu kwi-20 ° C kwiiyure eziyi-8-12.Iipleyiti zokukhenkceza ziye zahluzwa iiyure ezingama-48 ukufumana imixube eyahlukeneyo ye-NFRCS.Inkqubo efanayo isetyenziselwa ukuvelisa iimilo ezahlukeneyo kunye nezakhiwo, ezifana neetamponi okanye i-cylindrical tampons, okanye nayiphi na enye imilo kwiinkqubo ezahlukeneyo.
Ukulinganisa ngokuchanekileyo i-chitosan (80 kDa) (3%) ichithwa kwi-1% ye-acetic acid usebenzisa i-stirrer magnetic.Kwisisombululo esiphumela kwi-chitosan yongezwa i-1% ye-PEG 400 kwaye ixutywe imizuzu engama-30.Thela isisombululo esibangelwayo kwisikwere okanye kwisitya esixande kwaye umise kwi-80 ° C kwiiyure ze-12.Iisampulu emkhenkceni ziye lyophilized iiyure 48 ukufumana Cs13 porous.
I-NFRCS ephuhlisiwe yayiphantsi kovavanyo kusetyenziswa i-Fourier transform infrared spectroscopy (FTIR) (Shimadzu 8400 s FTIR, Tokyo, Japan) ukuqinisekisa ukuhambelana kweekhemikhali ze-chitosan kunye nezinye iipolymers14,15.I-spectra ye-FTIR (ububanzi be-spectral range ukusuka kwi-400 ukuya kwi-4000 cm-1) yazo zonke iisampuli ezivavanyiweyo zifunyenwe ngokwenza i-32 scans.
Izinga lokufunxa igazi (BAR) kuzo zonke iifomyula zavavanywa kusetyenziswa indlela echazwe nguChen et al.16 kunye nohlengahlengiso oluncinci.I-NFRKs ephuhlisiwe yazo zonke iingoma zomiswe kwi-oven vacuum kwi-105 ° C ngobusuku bonke ukususa i-solvent eseleyo.I-30 mg NFRCS (ubunzima bokuqala besampula - W0) kunye ne-30 mg Ct (ulawulo oluhle) zifakwe kwiitya ezihlukeneyo ezine-premix ye-3.8% ye-sodium citrate.Ngexesha elimiselweyo ngaphambili, oko kukuthi i-5, i-10, i-20, i-30, i-40 kunye ne-60 imizuzwana, i-NFRCS yasuswa kwaye iindawo zabo zahlanjululwa ngegazi elingenakunqandwa ngokubeka iisampuli kwi-Ct imizuzwana ye-30.Ubunzima bokugqibela begazi elixutywe yi-NFRCS 16 laliqwalaselwa (W1) ngexesha ngalinye.Bala ipesenti yeBAR usebenzisa le fomula ilandelayo:
Ixesha lokuvala igazi (BCT) linqunywe njengoko kuchazwe nguWang et al.17 .Ixesha elifunekayo kwigazi elipheleleyo (igazi legundane elixutywe kunye ne-3.8% ye-sodium citrate) ukuvala phambi kwe-NFRCS kubalwe njenge-BCT yesampula yovavanyo.Amacandelo ahlukeneyo e-NFRCS (30 mg) afakwe kwi-10 ml ye-screw cap vials kwaye yaqaliswa kwi-37°C.Igazi (i-0.5 ml) yongezwa kwi-vial kunye ne-0.3 ml ye-0.2 M CaCl2 yongezwa ukuze kusebenze i-coagulation yegazi.Ekugqibeleni, guqula i-vial rhoqo ngemizuzwana ye-15 (ukuya kwi-180 °) de ihlwili eliqinileyo lenze.I-BCT yesampuli iqikelelwa ngenani le-flip vails17,18.Ngokusekwe kwi-BCT, iingoma ezimbini ezizezona zilungileyo ezivela kwi-NFRCS Cm, i-Ch kunye ne-Cp ziye zakhethwa ukuba ziqhubele phambili izifundo zabalinganiswa.
I-BCT ye-Ch NFRCS kunye ne-Cp NFRCS yokuqulunqwa kwagqitywa ngokuphumeza indlela echazwe nguLi et al.19 .Beka i-15 x 15 mm2 I-Ch NFRCS, i-Cp NFRCS, kunye ne-Cs (ulawulo oluhle) kwiitya zePetri ezahlukileyo (37 °C).Igazi eliqukethe i-3.8% ye-sodium citrate yaxutywa kunye ne-0.2 M CaCl2 kwi-10: i-1 umthamo we-volume ratio ukuqala inkqubo yokuqhawula igazi.I-20 µl ye-0.2 M CaCl2 umxube wegazi lempuku igalelwe kumphezulu wesampulu yaze yafakwa kwisitya sePetri esingenanto.Ulawulo yayiligazi eligalelwe izitya ezingenanto zePetri ngaphandle kweCt.Ngexesha elimiselweyo le-0, i-3, kunye nemizuzu emi-5, yeka ukuvala ngokufaka i-10 ml yamanzi e-deionized (DI) kwisampuli equkethe isitya ngaphandle kokuphazamisa i-clot.I-erythrocytes engaguqukiyo (i-erythrocytes) idlula i-hemolysis phambi kwamanzi adibeneyo kwaye ikhulule i-hemoglobin.I-Hemoglobin ngamaxesha ahlukeneyo (HA (t)) ilinganiswe kwi-540 nm (λmax hemoglobin) isebenzisa i-UV-Vis spectrophotometer.Ukufunxwa ngokupheleleyo kwe-hemoglobin (AH(0)) kwi-0 min ye-20 µl yegazi kwi-10 ml yamanzi adibeneyo athathwe njengomgangatho wereferensi.Ukunyuka kwe-hemoglobin (RHA) yegazi elidibeneyo kubalwa kumlinganiselo we-HA (t)/HA (0) kusetyenziswa ibhetshi yegazi elifanayo.
Ukusebenzisa i-analyzer ye-texture (Texture Pro CT V1.3 Yakha i-15, i-Brookfield, i-USA), i-adhesive properties of NFRK kwi-tissue eyonakeleyo inqunywe.Cinezela isitya se-cylindrical esivulekileyo esisezantsi ngaphakathi kwesikhumba sehagu (ngaphandle kwe-fat fat).Iisampulu (i-Ch NFRCS kunye ne-Cp NFRCS) zisetyenziswe nge-cannula kwi-cylindrical molds ukudala ukunamathela kwisikhumba sehagu.Emva kwemizuzu ye-3 incubation kwiqondo lokushisa (RT) (25 ° C.), amandla okunamathela kwe-NFRCS abhalwe kwizinga eliqhubekayo le-0.5 mm / sec.
Olona phawu luphambili lwezitywini zotyando kukwandisa ukujika kwegazi ngelixa kunciphisa ukulahleka kwegazi.I-Coagulation engenakulahlekelwa kwi-NFRCS yavavanywa ngokusebenzisa indlela epapashwe ngaphambili kunye nokuguqulwa okuncinci kwe-19.Yenza ityhubhu ye-microcentrifuge (2 ml) (ububanzi bangaphakathi be-10 mm) kunye ne-8 × 5 mm2 umngxuma kwicala elinye lombhobho we-centrifuge (emele isilonda esivulekileyo).I-NFRCS isetyenziselwa ukuvala ukuvula kwaye i-tape isetyenziselwa ukutywina imida yangaphandle.Yongeza i-20 µl ye-0.2 M CaCl2 kwi-tube ye-microcentrifuge equkethe i-3.8% ye-sodium citrate premix.Emva kwemizuzu eyi-10, iibhubhu ze-microcentrifuge zisuswe izitya kwaye ukwanda kobunzima bezitya kunqunywe ngenxa yokuphuma kwegazi kwi-NFRK (n = 3).Ukulahleka kwegazi I-Ch NFRCS kunye neCp NFRCS yafaniswa neCs.
Ingqibelelo emanzi ye-NFRCS yamiselwa ngokusekelwe kwindlela echazwe nguMishra kunye ne-Chaudhary21 ngokuguqulwa okuncinci.Beka i-NFRCS kwi-flask ye-Erlenmeyer ye-100 ml kunye ne-50 ml yamanzi kwaye ujikeleze i-60 s ngaphandle kokwenza phezulu.Ukuhlolwa okubonakalayo kunye nokubekwa phambili kweesampulu zokuthembeka ngokomzimba ngokusekelwe ekuqokeleleni.
Amandla okubopha i-HFFC kwi-Ct afundwe kusetyenziswa iindlela ezipapashwe ngaphambili kunye nokuguqulwa okuncinci.Ukunyaniseka kwengubo yobuso kwavavanywa ngokutyhila i-NFRK kumaza e-acoustic (i-stimulus yangaphandle) phambi kwamanzi e-milliQ (Ct).I-NFRCS Ch NFRCS ephuhlisiwe kunye ne-Cp NFRCS zifakwe kwi-beaker ezaliswe ngamanzi kwaye i-sonicated kwi-1, 2, 3, 4, 5, 6, 7, 8, 9, 10, kunye ne-30 min, ngokulandelanayo.Emva kokumisa, umehluko wepesenti phakathi kobunzima bokuqala kunye nobokugqibela be-NFRCS yayisetyenziselwa ukubala ipesenti yokulahlekelwa kwezinto (HFFC).I-In vitro BCT ixhasa ngakumbi amandla okubopha okanye ukulahleka kwezinto ezingaphezulu.Ukusebenza kakuhle kwe-HFFC ebophelela kwi-Ct ibonelela ngokuhlangana kwegazi kunye ne-elastic coating kumphezulu we-Ct22.
I-homogeneity ye-NFRCS ephuhlisiwe igqitywe yi-BCT yeesampuli (30 mg) ezithathwe kwiindawo eziqhelekileyo ezikhethiweyo ze-NFRCS.Landela inkqubo ye-BCT ekhankanywe ngaphambili ukumisela ukuthotyelwa kwe-NFRCS.Ukusondela phakathi kwazo zonke iisampulu ezintlanu kuqinisekisa ukugqunywa komhlaba okufanayo kunye nokubekwa kwe-HFFC kwi-Ct mesh.
Indawo yoqhagamshelwano lwegazi ebizwa ngokuba yi-NBCA yamiselwa njengoko kwakuxelwe ngaphambili ngohlengahlengiso oluthile.Hlanganisa igazi ngokucinezela i-20 µl yegazi phakathi kwemiphezulu emibini ye-Ct, i-Ch NFRCS, i-Cp NFRCS kunye ne-Cs.Emva kweyure enye, iinxalenye ezimbini ze-stent zahlulwa kwaye zalinganisa ngesandla indawo yehlwili.Ixabiso elingumndilili lokuphindaphinda okuthathu lithathwe njenge-NBCA NFRCS19.
Uhlalutyo lwe-Dynamic Vapor Sorption (DVS) lusetyenziselwe ukuvavanya ukusebenza kakuhle kwe-NFRCS ukufunxa amanzi kwindawo yangaphandle okanye kwindawo yokwenzakala enoxanduva lokuqalisa i-coagulation.I-DVS ivavanya okanye irekhode ukufunxwa komphunga kunye nelahleko kwisampulu yegravimetrically usebenzisa ibhalansi ebuthathaka kakhulu kunye nesisombululo sobunzima esingu-±0.1 µg.Uxinzelelo lomphunga oluyinxenye (ukufuma okuhambelanayo) uveliswa sisilawuli sokuhamba kobunzima be-elektroniki malunga nesampulu ngokuxuba igesi egcweleyo neyomileyo. Ngokwezikhokelo ze-European Pharmacopeia, ngokusekwe kwipesenti yokufunxa ukufuma ngeesampulu, iisampulu zahlulwa zibe ziindidi ezi-4 (0–0.012% w/w-non-hygroscopic, 0.2–2% w/w little hygroscopic, 2–15% moderately hygroscopic, and 2 hygroscopic) kakhulu> 15 . Ngokwezikhokelo ze-European Pharmacopeia, ngokusekelwe kwipesenteji yokufuma okufuma kwiisampuli, iisampulu zahlulwa zibe ziindidi ezi-4 (0–0.012% w/w-non-hygroscopic, 0.2–2% w/w little hygroscopic, 2–15 % moderately hygroscopic, and> 315 hygroscopic) kakhulu.Ngokuhambelana neengcebiso ze-European Pharmacopoeia, kuxhomekeke kwipesenti yokufunxa umswakama ngamasampuli, iisampuli zahlulahlulwe kwiindidi ze-4 (0-0.012% w / w - non-hygroscopic, 0.2-2% w / w / w little hygroscopic, 2-15%).% умеренно гигроскопичен и > 15% очень гигроскопичен)23. % ngokuphakathi kwehygroscopic kunye > 15% kakhulu hygroscopic)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2 类（0-0.012% w/w-澎澅公富/w2.吸湿性、2-15 % 适度吸湿，> 15% 非常吸湿）23.9 , , , 0.2-2% W/w 轻微 、 2-15% 适度 吸湿 ，> 15 %非常吸湿）23.Ngokuhambelana neengcebiso ze-European Pharmacopoeia, iisampulu zihlulwe zibe ziiklasi ezi-4 ngokuxhomekeke kwipesenti yokufuma efunnwe yisampulu (0-0.012% ngobunzima - non-hygroscopic, 0.2-2% ngobunzima obuncinci, i-2-15% ubunzima).% умеренно гигроскопичен, > 15 % очень гигроскопичен) 23. I-% ngokwe-hygroscopic ephakathi, > 15% i-hygroscopic kakhulu) 23.Ukusebenza kakuhle kwe-hygroscopic ye-NFCS X NFCS kunye ne-TsN NFCS kumiselwe kwi-analyzer DVS TA TGA Q5000 SA.Ngethuba le nkqubo, ixesha lokugijima, umswakama ohambelanayo (RH), kunye nesisindo sesampula sangempela kwi-25 ° C24 yafunyanwa.Umxholo wokufuma ubalwa ngohlalutyo oluchanekileyo lwe-NFRCS kusetyenziswa le nxaki ilandelayo:
I-MC kukufuma kwe-NFRCS.m1 - ubunzima obomileyo bee-NSAID.m2 lolona xesha lokwenene ubunzima be-NFRCS kwi-RH enikiweyo.
Umhlaba uwonke uqikelelwe kusetyenziswa umfuniselo we-nitrogen adsorption kunye nenitrogen engamanzi emva kokukhupha iisampuli kwi-25 °C nge-10 h (<7 × 10–3 Torr). Umhlaba uwonke uqikelelwe kusetyenziswa umfuniselo we-nitrogen adsorption kunye nenitrogen engamanzi emva kokukhupha iisampuli kwi-25 °C nge-10 h (<7 × 10–3 Torr). Общая площадь поверхности оценивалась с помощью эксперимента по адсорбции азота жидким азотом после опорожнения образцов ° 17 × 17 × 3 Торр). Ummandla opheleleyo womhlaba uqikelelwe kusetyenziswa umfuniselo we-nitrogen adsorption kunye ne-nitrogen engamanzi emva kokuba iisampuli zithululwe kwi-25 ° C nge-10 h (< 7 × 10–3 Torr).在25°C 清空樣品10 小时（< 7 × 10-3 Torr）后，使用液氮的氮吸附实验估计总表面积。Kuma-25°C Общая площадь поверхности оценивалась с использованием экспериментов по азота жидким азотом после опорожнения образ20 °C (< 7 × 10-3 тор). Ummandla opheleleyo womhlaba waqikelelwa kusetyenziswa imifuniselo ye-nitrogen adsorption kunye ne-nitrogen engamanzi emva kokuba iisampulu zithululwe iiyure ezili-10 kwi-25 ° C (< 7 x 10-3 torr).Ubungakanani bomphezulu opheleleyo, umthamo wepore kunye ne-NFRCS ubungakanani bepore bugqitywe nge-Quantachrome evela kwi-NOVA 1000e, e-Austria usebenzisa i-RS 232 software.
Lungiselela i-5% ye-RBCs (ityuwa njengediluent) kwigazi elipheleleyo.Emva koko udlulisele i-aliquot ye-HFFC (0.25 ml) kwi-plate ye-96-well kunye ne-5% ye-RBC mass (0.1 ml).Faka umxube kwi-37 ° C imizuzu engama-40.Umxube weeseli ezibomvu zegazi kunye ne-serum yayithathwa njengolawulo oluhle, kunye nomxube we-saline kunye neeseli ezibomvu zegazi njengolawulo olubi.I-hemagglutination inqunywe ngokwezinga le-Stajitzky.Izikali ezicetywayo zimi ngolu hlobo lulandelayo: + + + + i-granular aggregates eshinyeneyo;+ + + iipads ezigudileyo ezisezantsi ezinemiphetho egobileyo;+ + iipads ezigudileyo ezisezantsi ezinemiphetho ekrazukileyo;+ amakhonkco abomvu acutheneyo ajikeleze imiphetho yeepads ezigudileyo;– (negative) discrete iqhosha elibomvu 12 embindini wequla elisezantsi.
I-hemocompatibility ye-NFRCS ifundwe ngokwendlela ye-International Organisation for Standardization (ISO) (ISO10993-4, 1999)26,27.Indlela yegravimetric echazwe nguSingh et al.Uhlengahlengiso oluncinci lwenziwa ukuvavanya ukubunjwa kwe-thrombus kubukho okanye kumphezulu we-NFRCS.I-500 mg ye-Cs, i-Ch NFRCS kunye ne-Cp NFRCS zifakwe kwi-phosphate buffered saline (PBS) kwiiyure ezingama-24 kwi-37 ° C.Emva kweeyure ze-24, i-PBS yasuswa kwaye i-NFRCS yaphathwa nge-2 ml yegazi equkethe i-3.8% ye-sodium citrate.Kumphezulu we-NFRCS, yongeza i-0.04 ml ye-0.1 M CaCl2 kwiisampuli ezifakelweyo.Emva kwemizuzu engama-45, i-5 ml yamanzi adibeneyo yongezwa ukumisa i-coagulation.Igazi elidibeneyo ebusweni be-NFRK liphathwe nge-36-38% yesisombululo se-formaldehyde.Amahlwili alungiswe nge-formaldehyde omisiwe kwaye alinganiswe.Ipesenti ye-thrombosis yaqikelelwa ngokubala ubunzima beglasi ngaphandle kwegazi kunye nesampuli (ulawulo olubi) kunye neglasi enegazi (ulawulo oluhle).
Njengobungqina bokuqala, iisampuli zabonwa phantsi kwe-microscope ye-optical ukuqonda amandla omgangatho we-HFFC, i-Ct edibeneyo, kunye ne-Ct network ukwenza iipores.Amacandelo amancinci e-Ch kunye ne-Cp esuka kwi-NFRCS aye alungiswa nge-scalpel blade.Icandelo elibangelwayo lifakwe kwisilayidi seglasi, lihlanganiswe ne-coverlip, kwaye imida yayigxininiswe ngeglue.Izilayidi ezilungisiweyo zajongwa phantsi kwe-microscope ebonakalayo kwaye iifoto zathathwa ngokwahlukileyo.
Ukufakwa kwepolymer kwiinethiwekhi ze-Ct kwabonwa kusetyenziswa i-fluorescence microscopy ngokusekelwe kwindlela echazwe yiRice et al.29. Ukuqulunqwa kwe-HFFC esetyenzisiweyo ekuqulunqweni kwaxutywa ngedayi ye-fluorescent (i-amaranth), kunye ne-NFRCS (Ch & Cp) yalungiswa ngokwendlela ekhankanywe ngaphambili. Ukuqulunqwa kwe-HFFC esetyenzisiweyo ekuqulunqweni kwaxutywa ngedayi ye-fluorescent (i-amaranth), kunye ne-NFRCS (Ch & Cp) yalungiswa ngokwendlela ekhankanywe ngaphambili.Ukuqulunqwa kwe-HFFC esetyenzisiweyo ekuqulunqweni kwaxutywa ngedayi ye-fluorescent (i-amaranth) kunye ne-NFRCS (i-Ch kunye ne-Cp) ifunyenwe ngokwendlela ekhankanywe ngaphambili.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）Ukuqulunqwa kwe-HFFC esetyenziswe ekuqulunqweni kwaxutywa ngedayi ye-fluorescent (i-Amaranth) kwaye yafumana i-NCFRCS (Ch kunye neCp), njengoko kukhankanyiwe ngaphambili.Amacandelo amancinci e-NFRK anqunywe kwiisampuli ezifunyenweyo, ezibekwe kwiislayidi zeglasi, kwaye zigqunywe ngeesilayidi zokugubungela.Jonga izilayidi ezilungisiweyo phantsi kwe-microscope ene-fluorescent usebenzisa isihluzo esiluhlaza (310-380 nm).Imifanekiso ithathwe kwi-4x yokukhulisa ukuqonda ubudlelwane be-Ct kunye nokugqithiswa kwepolymer kwi-network ye-Ct.
I-topography yomhlaba ye-NFRCS Ch kunye ne-Cp inqunywe ngokusebenzisa i-atomic force microscope (AFM) kunye ne-ultra-sharp TESP cantilever kwimodi yokucofa: 42 N / m, 320 kHz, ROC 2-5 nm, Bruker, Taiwan.Uburhabaxa bomphezulu bugqitywe yingcambu yentsingiselo yesikwere (RMS) kusetyenziswa isoftwe (Skena Probe Image Processor).Iindawo ezahlukeneyo ze-NFRCS zanikezelwa kwimifanekiso ye-3D ukujonga ukufana komphezulu.Ukutenxa okusemgangathweni kwamanqaku ommandla onikiweyo kuchazwa njengoburhabaxa bomphezulu.I-equation ye-RMS yasetyenziselwa ukulinganisa uburhabaxa bomphezulu be-NFRCS31.
Izifundo ezisekelwe kwi-FESEM zenziwa ngokusebenzisa i-FESEM, i-SU8000, i-HI-0876-0003, i-Hitachi, i-Tokyo, ukuqonda i-morphology yomhlaba we-Ch NFRCS kunye ne-Cp NFRCS, ebonisa i-BCT engcono kune-Cm NFRCS.Uphononongo lwe-FESEM lwenziwa ngokwendlela echazwe nguZhao et al.32 kunye nohlengahlengiso oluncinci.I-NFRCS 20 ukuya kwi-30 mg I-Ch NFRCS kunye ne-Cp NFRCS yayixutywe ngaphambili kunye ne-20 µl ye-3.8% ye-sodium citrate exutywe ngegazi le-rat.I-20 μl ye-0.2 M CaCl2 yongezwa kwiisampuli eziphathwe ngegazi ukuqalisa i-coagulation kwaye iisampulu zifakwe kwiqondo lokushisa le-10 imizuzu.Ukongezelela, i-erythrocytes engaphezulu isuswe kwi-NFRCS ebusweni ngokuhlanjululwa nge-saline.
Iisampuli ezilandelayo zaphathwa nge-0.1% glutaraldehyde kwaye zomiswa kwi-oven yomoya oshushu kwi-37 ° C ukususa umswakama.Iisampuli ezomileyo zifakwe kwaye zihlalutywe i-32.Eminye imifanekiso efunyenwe ngexesha lokuhlalutya yayiyi-clot ye-clot ebusweni bombhobho we-cotton fibers, i-polymer deposition phakathi kwe-Ct, i-erythrocyte morphology (i-shape), i-clot integrity, kunye ne-erythrocyte morphology phambi kwe-NFRCS.Iindawo ze-NFRCS ezingaphendulwanga kunye ne-Ch kunye ne-Cp ephathwayo kwiindawo ze-NFRCS ezifakwe ngegazi zihlolwe i-ion elemental (i-sodium, i-potassium, i-nitrogen, i-calcium, i-magnesium, i-zinc, ithusi kunye ne-selenium)33.Thelekisa iipesenti ze-ion phakathi kweesampulu eziphathwayo kunye nezingaphathwanga ukuze uqonde ukuqokelelwa kwe-ion elemental ngexesha lokwenziwa kwehlwili kunye ne-clot homogeneity.
Ubuninzi be-Cp HFFC yokwambathisa umphezulu kwi-Ct surface bunqunywe ngokusebenzisa i-FESEM.Amacandelo anqamlezayo e-Cp NFRCS aye anqunyulwa kwisakhelo kunye ne-sputter coated.Iisampulu ze-sputter ezibangelwayo zabonwa yi-FESEM kunye nobukhulu bengubo yomhlaba bulinganiswe 34 , 35 , 36 .
I-X-ray micro-CT inikeza i-high-resolution ye-3D ye-imaging engonakalisi kwaye ikuvumela ukuba ufunde ilungiselelo langaphakathi lesakhiwo se-NFRK.I-Micro-CT isebenzisa i-X-ray beam edlula isampuli ukurekhoda i-coefficient yendawo yokunciphisa umgca we-X-rays kwisampuli, enceda ukufumana ulwazi lwe-morphological.Indawo yangaphakathi ye-Ct kwi-Cp NFRCS kunye ne-Cp ye-NFRCS ephathwa ngegazi ihlolwe yi-micro-CT ukuze iqonde ukusebenza kakuhle kwe-absorption kunye ne-blood clotting phambi kwe-NFRCS37,38,39.Izakhiwo ze-3D zeesampuli ze-Cp NFRCS eziphathwe ngegazi kunye nezingaphathwanga zakhiwe ngokutsha kusetyenziswa i-micro-CT (V|tome|x S240, Phoenix, Germany).Ukusebenzisa i-VG STUDIO-MAX software version 2.2, imifanekiso emininzi ye-X-reyi ithathwe kwii-engile ezahlukeneyo (eyona nto i-360 ° coverage) ukuphuhlisa imifanekiso ye-3D ye-NFRCS.Idatha yentelekelelo eqokelelweyo yakhiwa kwakhona kwimifanekiso yevolumetric ye-3D kusetyenziswa isoftware ye-3D ScanIP Academic ehambelana nayo.
Ukongeza, ukuqonda ukuhanjiswa kwehlwili, i-20 µl yegazi elixutywe ngaphambili kunye ne-20 µl ye-0.2 M CaCl2 yongezwa kwi-NFRCS ukuqalisa ukuqiniswa kwegazi.Iisampulu ezilungisiweyo zishiywe ukuba lukhuni.Umphezulu we-NFRK uphathwe nge-0.5% glutaraldehyde kwaye yomiswe kwi-oven yomoya oshushu kwi-30-40 ° C kwi-30 min.Ihlwili legazi elenziwe kwi-NFRCS laskenwa, lakhiwa ngokutsha, kwaye umfanekiso we-3D wehlwili legazi wawubonwa.
Uvavanyo lwe-antibacterial lwenziwa kwi-Cp NFRCS (engcono xa kuthelekiswa ne-Ch NFRCS) usebenzisa indlela echazwe ngaphambili kunye nokuguqulwa okuncinci.Umsebenzi we-antibacterial we-Cp NFRCS kunye ne-Cp HFFC unqunywe ngokusebenzisa ii-microorganisms ezintathu ezahlukeneyo zokuvavanya [i-S.aureus (ibhaktheriya e-gram-positive), i-E.coli (ibhaktheriya e-gram-negative) kunye ne-white Candida (C.albicans)] ekhula kwi-agar kwi-Petri izitya kwi-incubator.I-inoculate ngokulinganayo i-50 ml yokunqunyanyiswa kwenkcubeko ye-bacterial diluted kwi-concentration ye-105-106 CFU ml-1 kwi-agar medium.Galela i-medium kwisitya sePetri kwaye uyivumele iqine.Amaqula ayenziwe kumphezulu we-agar plate ukuze azalise i-HFFC (i-3 imithombo ye-HFFC kunye ne-1 yokulawula okungalunganga).Yongeza i-200 µl HFFC kumaqula ama-3 kunye ne-200 µl pH 7.4 PBS ukuya kweyesine.Ngakolunye uhlangothi lwesitya se-petri, faka i-disk ye-12 mm Cp NFRCS kwi-agar eqinisiweyo kwaye ufumise nge-PBS (pH 7.4).I-Ciprofloxacin, i-ampicillin kunye neepilisi ze-fluconazole zithathwa njengemigangatho yereferensi ye-Staphylococcus aureus, i-Escherichia coli kunye ne-Candida albicans.Ukulinganisa indawo yokuthintela ngesandla kwaye uthathe umfanekiso wedijithali wendawo yokuthintela.
Emva kokuvunyelwa kweziko lokuziphatha, isifundo senziwe kwi-Kasturba Medical College yeMfundo kunye noPhando eManipal, eKarnataka, eningizimu yeIndiya.I-in vitro TEG yovavanyo lweprotocol iye yahlaziywa kwaye yavunywa yiKomiti yoBulungisa beZiko leKasturba Medical College, Manipal, Karnataka (IEC: 674/2020).Izifundo zafunyanwa kumatsha-ntliziyo anikela ngegazi (abaneminyaka eli-18 ukusa kwengama-55) kwindawo yokugcina igazi esibhedlele.Ukongezelela, ifom yemvume enolwazi yafunyanwa kumavolontiya ukuze kuqokelelwe iisampulu zegazi.I-TEG yoMthonyama (N-TEG) ​​yayisetyenziselwa ukufunda umphumo weCp HFFC ukwakheka kwigazi elipheleleyo elixutywe ne-sodium citrate.I-N-TEG iyaqatshelwa ngokubanzi ngendima yayo ekuvuseleleni i-point-care-care, eyenza iingxaki kwiiklinikhi ngenxa yokukwazi ukulibaziseka okubonakalayo kwiziphumo (uvavanyo lwe-coagulation yesiqhelo).Uhlalutyo lwe-N-TEG lwenziwa kusetyenziswa igazi elipheleleyo.Imvume enolwazi kunye nembali yonyango ecacileyo ifunyenwe kubo bonke abathathi-nxaxheba.Uphononongo aluzange lubandakanye abathathi-nxaxheba abane-hemostatic okanye iingxaki ze-thrombotic ezifana nokukhulelwa / emva kokubeleka okanye isifo sesibindi.Izifundo ezithatha amachiza achaphazela i-coagulation cascade nazo azifakwanga kuphononongo.Uvavanyo olusisiseko lwebhubhoratri (i-hemoglobin, ixesha le-prothrombin, i-thromboplastin esebenzayo kunye ne-platelet count) zenziwa kubo bonke abathathi-nxaxheba ngokweenkqubo eziqhelekileyo.I-N-TEG inquma i-blood clot viscoelasticity, isakhiwo sokuqala se-clot, ukusebenzisana kwamaqhekeza, ukuqiniswa kwe-clot, kunye ne-clot lysis.Uhlalutyo lwe-N-TEG lubonelela ngegraphical kunye nedatha yamanani kwiimpembelelo ezihlangeneyo zezinto ezininzi zeselula kunye neplasma.Uhlalutyo lwe-N-TEG lwenziwa kwimiqulu emibini eyahlukeneyo ye-Cp HFFC (10 µl kunye ne-50 µl).Ngenxa yoko, i-1 ml yegazi elipheleleyo kunye ne-citric acid yongezwa kwi-10 μl ye-Cp HFFC.Yongeza i-1 ml (Cp HFFC + igazi elicikiziweyo), 340 µl igazi elixutyiweyo ukuya kuma-20 µl 0.2 M CaCl2 equlethe isitya se-TEG.Emva koko, izitya ze-TEG zalayishwa kwi-TEG® 5000, e-US ukulinganisa i-R, K, i-alpha angle, i-MA, G, CI, TPI, EPL, LY 30% yeesampuli zegazi phambi kweCp HFFC41.
I-in vivo study protocol yahlaziywa kwaye yamkelwa yi-Institutional Animal Ethics Committee (IAEC), i-Kasturba School of Medicine, i-Manipal Institute of Higher Education, i-Manipal (IAEC/KMC/69/2020).Zonke iimvavanyo zezilwanyana zenziwa ngokuhambelana neengcebiso zeKomiti yoLawulo kunye nokuPhathwa kweMvavanyo yezilwanyana (CPCSEA).Zonke izifundo ze-NFRCS ze-vivo (2 × 2 cm2) zenziwa kwiigundane ze-Wistar zabasetyhini (ubunzima be-200 ukuya kwi-250 g).Zonke izilwanyana zazilungelelaniswa kwiqondo lokushisa kwe-24-26 ° C, izilwanyana zazinokufikelela ngokukhululekileyo kwi-standard standard kunye ne-ad libitum yamanzi.Zonke izilwanyana zahlulwe ngokungakhethiyo zangamaqela ahlukeneyo, iqela ngalinye lalinezilwanyana ezithathu.Zonke izifundo zenziwa ngokuhambelana neZifundo zezilwanyana: Ingxelo ye-In Vivo Experiments 43.Ngaphambi kokufunda, izilwanyana zatshatyalaliswa nge-intraperitoneal (ip) yokulawulwa komxube we-20-50 mg ye-ketamine (nge-1 kg yesisindo somzimba) kunye ne-2-10 mg ye-xylazine (nge-1 kg yesisindo somzimba).Emva kokuphonononga, umthamo wokuphuma kwegazi ubalwa ngokuvavanya umehluko phakathi kobunzima bokuqala kunye nokugqibela kweesampuli, ixabiso eliqhelekileyo elifunyenwe kwiimvavanyo ezintathu lithathwe njengomthamo wokuphuma kwegazi kwisampuli.
Imodeli yokunqunyulwa komsila wempuku yaphunyezwa ukuqonda amandla e-NFRCS ukulungelelanisa ukopha kwingozi, ukulwa, okanye ingozi yendlela (imodeli yokulimala).Sika i-50% yomsila nge-scalpel blade kwaye ubeke emoyeni i-15 s ukuqinisekisa ukopha okuqhelekileyo.Ukongezelela, iisampuli zokuvavanya zibekwe emsila wegundane ngokufaka uxinzelelo (Ct, Cs, Ch NFRCS kunye neCp NFRCS).Ukopha kunye ne-PCT zixelwe kwimizekelo yovavanyo (n = 3) 17,45.
Ukusebenza kwe-NFRCS yokulawula uxinzelelo ekulweni kwaphandwa kwimodeli ye-artery ye-femoral engaphezulu.I-artery femoral ibonakaliswe, ihlatywe nge-24G trocar, kwaye ikhuphe igazi ngaphakathi kwemizuzwana ye-15.Emva kokuba kubonwe ukopha okungalawulekiyo, isampuli yovavanyo ibekwe kwindawo yokugqobhoza ngoxinzelelo.Ngokukhawuleza emva kokusetyenziswa kwesampuli yovavanyo, ixesha lokuvala lirekhodwe kwaye ukusebenza kakuhle kwe-hemostatic kwabonwa kwimizuzu emi-5 elandelayo.Inkqubo efanayo yaphindwa ngeCs kunye ne-CT46.
UDowling et al.I-47 icebise imodeli yokulimala kwesibindi ukuvavanya amandla e-hemostatic kwi-hemostatic materials kumxholo wokuphuma kwegazi kwi-intraoperative.I-BCT irekhodwe kwiisampuli ze-Ct (ulawulo olubi), isakhelo seCs (ulawulo oluhle), iisampuli zeCh NFRCS, kunye neesampuli zeCp NFRCS.I-suprahepatic vena cava ye-rat yavezwa ngokwenza i-laparotomy ephakathi.Emva koko, inxalenye ekude ye-lobe ekhohlo inqunywe ngesikere.Yenza i-incision kwisibindi nge-scalpel blade kwaye uyivumele ukuba ikhuphe igazi imizuzwana embalwa.Ukulinganiswa ngokuchanekileyo kwe-Ch NFRCS kunye ne-Cp NFRCS iisampulu zovavanyo zibekwe kwindawo eyonakalisiweyo ngaphandle koxinzelelo oluhle kwaye i-BCT yarekhodwa.Iqela lokulawula (Ct) emva koko lifake uxinzelelo olulandelwa yiCs 30 s47 ngaphandle kokuphula ukulimala.
Kwi-vivo iimvavanyo zokupholisa amanxeba zenziwa kusetyenziswa imodeli yenxeba elikhethekileyo ukuvavanya iipropati zokupholisa inxeba kwii-NCFRCS eziphuhlisiwe ezisekelwe kwi-polymer.Iimodeli zamanxeba ahlanjululweyo zikhethwe kwaye zenziwa ngokweendlela ezipapashwe ngaphambili kunye nokuguqulwa okuncinci19,32,48.Zonke izilwanyana zaye zathotywa ii-anesthetism njengoko bekuchaziwe ngaphambili.Sebenzisa i-biopsy punch (12 mm) ukwenza isangqa esinzulu esisikekileyo eluswini lomqolo.Iziza ezilungisiweyo zezilonda zimbethe i-Cs (ulawulo oluhle), i-Ct (eqaphela ukuba i-cotton pads iphazamisa ukuphilisa), i-Ch NFRCS kunye ne-Cp NFRCS (iqela lovavanyo) kunye nolawulo olubi ngaphandle konyango.Ngosuku ngalunye lwesifundo, indawo yesilonda yayilinganiswa kuzo zonke iimpuku.Sebenzisa ikhamera yedijithali ukuthatha umfanekiso wendawo yenxeba kwaye ufake isambatho esitsha.Ipesenti yokuvalwa kwenxeba ilinganiswe ngale fomula ilandelayo:
Ngokusekelwe kwipesenteji yokuvalwa kwesilonda ngomhla we-12 wokufunda, ulusu lwegundane lweqela eligqwesileyo lachithwa ((Cp NFRCS) kunye neqela lokulawula) kwaye lifundwe nge-H & E staining kunye ne-Masson's trichrome staining. Ngokusekelwe kwipesenteji yokuvalwa kwesilonda ngomhla we-12 wokufunda, ulusu lwegundane lweqela eligqwesileyo lachithwa ((Cp NFRCS) kunye neqela lokulawula) kwaye lifundwe nge-H & E staining kunye ne-Masson's trichrome staining.Ngokusekelwe kwipesenti yokuvalwa kwesilonda ngomhla we-12 wokufunda, ulusu lwamagundane elona qela ligqwesileyo ((Cp NFRCS) kunye neqela lokulawula) lihlanjululwe kwaye lihlolwe ngokucoca nge-hematoxylin-eosin kunye 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)的大鼠皮肤和进海行H&EMas.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，和进行H&E扄)Iigundane kwiqela elingcono kakhulu ((i-Cp NFRCS) kunye namaqela okulawula) ahlanjululwe kwi-hematoxylin-eosin staining kunye ne-Masson's trichrome staining esekelwe kwipesenti yokuvalwa kwenxeba ngosuku lwe-12 yesifundo.Inkqubo yokucoca ephunyeziweyo yenziwa ngokweendlela ezichazwe ngaphambili49,50.Ngokufutshane, emva kokulungiswa kwi-10% ye-formalin, iisampulu zaphelelwa ngamanzi zisebenzisa uluhlu lwee-alcohols ezihleliweyo.Sebenzisa i-microtome ukufumana amacandelo abhityileyo (ama-5 µm ubukhulu) wethishu ekhutshiweyo.Amacandelo amancinci olawulo kunye ne-Cp NFRCS aphathwe nge-hematoxylin kunye ne-eosin ukufunda utshintsho lwe-histopathological.Ibala le-trichrome lika-Masson lasetyenziselwa ukufumanisa ukwakheka kwee-collagen fibrils.Iziphumo ezifunyenweyo zaphononongwa ngobumfama ziingcali zezifo.
Ukuzinza kweesampuli ze-Cp NFRCS zaphononongwa kwiqondo lokushisa legumbi (25 ° C ± 2 ° C / 60% RH ± 5%) kwiinyanga ze-1251.I-Cp NFRCS (ukuguqulwa kombala kunye nokukhula kwe-microbial) yahlolwa ngokubonakalayo kwaye yavavanyelwa ukunyanzeliswa kokugqoka kunye ne-BCT ngokweendlela ezingentla apha ezichazwe kwicandelo leMathiriyeli kunye neNdlela.
I-scalability kunye ne-reproducibility ye-Cp NFRCS ihlolwe ngokulungiselela i-Cp NFRCS kunye nobukhulu be-15 × 15 cm2.Ukongezelela, iisampulu ze-30 mg (n = 5) zikhutshwe kwiinqununu ezahlukeneyo ze-Cp NFRCS kunye ne-BCT yeesampuli ezifundwayo zavandlakanywa njengoko kuchazwe ngaphambili kwicandelo leNdlela.
Sizame ukuphuhlisa iimilo ezahlukeneyo kunye nolwakhiwo sisebenzisa imilo ye-Cp NFRCS kwizicelo ezahlukeneyo zebhayomedical.Iimilo ezinjalo okanye ulungelelwaniso lubandakanya i-conical swabs ye-nosebleeds, iinkqubo zamazinyo, kunye ne-cylindrical swabs yokopha kwelungu lobufazi.
Zonke iiseti zedatha zichazwa njengentsingiselo ± ukutenxa okusemgangathweni kwaye zahlalutywa yi-ANOVA kusetyenziswa i-Prism 5.03 (iGraphPad, iSan Diego, i-CA, i-USA) ilandelwa luvavanyo lothelekiso oluninzi lwe-Bonferroni (* p <0.05).
Zonke iinkqubo ezenziwa kwizifundo zabantu zihambelana nemigangatho yeZiko kunye neBhunga loPhando leSizwe, kunye neSibhengezo se-Helsinki 1964 kunye nezilungiso zayo ezilandelayo, okanye imigangatho efanayo yokuziphatha.Bonke abathathi-nxaxheba baxelelwa malunga neempawu zophononongo kunye nobume balo ngokuzithandela.Idatha yabathathi-nxaxheba ihlala iyimfihlo xa sele iqokelelwe.I-in vitro TEG yovavanyo lweprotocol iye yahlaziywa kwaye yavunywa yiKomiti yoBulungisa beZiko leKasturba Medical College, Manipal, Karnataka (IEC: 674/2020).Amavolontiya atyikitye imvume enolwazi yokuqokelela iisampuli zegazi.
Zonke iinkqubo ezenziwa kwizifundo zezilwanyana zenziwa ngokuhambelana neKastuba Faculty of Medicine, iManipal Institute of Higher Education, Manipal (IAEC/KMC/69/2020).Zonke iimvavanyo zezilwanyana eziyilwe zenziwe ngokuhambelana nezikhokelo zeKomiti yoLawulo kunye nokuPhathwa kweMvavanyo yezilwanyana (CPCSEA).Bonke ababhali balandela izikhokelo ze-ARIVE.
Imbonakalo ye-FTIR yazo zonke i-NFRCS yahlalutywa kwaye yathelekiswa ne-chitosan spectrum eboniswe kuMfanekiso 2A.Iimpawu eziphakamileyo ze-spectral ze-chitosan (ezirekhodiweyo) kwi-3437 cm-1 (i-OH kunye ne-NH yokwelula, i-overlap), i-2945 kunye ne-2897 cm-1 (i-CH stretching), i-1660 cm-1 (i-NH2 i-strain), i-1589 cm-1 (N-H bending), 1156 cm-i-bridge yesibini, i-1156 cm-ibhuloho yesibini i-hydroxyl), i-993 cm-1 (yolula i-CO, i-Bo-OH) 52.53.54.ITheyibhile eyoNgezelelweyo ye-S1 ibonisa amaxabiso okufunxa e-FTIR NFRCS e-chitosan (ingxelo), i-chitosan ecocekileyo, i-Cm, i-Ch, kunye ne-Cp.I-spectra ye-FTIR yazo zonke i-NFRCS (i-Cm, i-Ch kunye ne-Cp) ibonise iimpawu ezifanayo zokufunxa iibhendi njenge-chitosan ecocekileyo ngaphandle kotshintsho olubalulekileyo (Umfanekiso 2A).Iziphumo ze-FTIR ziqinisekisile ukungabikho kweekhemikhali okanye ukusebenzisana komzimba phakathi kweepolymers ezisetyenziselwa ukuphuhlisa i-NFRCS, ebonisa ukuba iipolymers ezisetyenzisiweyo zi-inert.
Iimpawu ze-in vitro ze-Cm NFRCS, i-Ch NFRCS, i-Cp NFRCS kunye ne-Cs.(A) imele i-spectra ye-FTIR edibeneyo yeengqungquthela ze-chitosan kunye ne-Cm NFRCS, i-Ch NFRCS kunye ne-Cp NFRCS phantsi koxinzelelo.(B) a) Umlinganiselo wegazi elipheleleyo le-NCFRCS Cm, Ch, Cp, kunye neCg (n = 3);Iisampulu ze-Ct zibonise i-BAR ephezulu kuba i-cotton swab inokusebenza okuphezulu kokufunxa;b) Igazi emva kokufunxa igazi Umzobo wesampulu efunliweyo.Ukubonakaliswa kwegraphical ye-BCT yesampula yovavanyo C (i-Cp NFRCS yayine-BCT engcono kakhulu (15 s, n = 3)). Idatha kwi-C, D, E, kunye ne-G yaboniswa njengentsingiselo ± SD, kwaye iibar zempazamo zimele i-SD, ***p <0.0001. Idatha kwi-C, D, E, kunye ne-G yaboniswa njengentsingiselo ± SD, kwaye iibar zempazamo zimele i-SD, ***p <0.0001. Данные в C, D, E и G представлены как среднее ± стандартное отклонение, а планки погрешностей представляют стандартное отклонение,0, ***p. Idatha ku-C, D, E, kunye no-G zinikwe njengentsingiselo ± ukutenxa okusemgangathweni, kwaye iibhari zeempazamo zimele ukutenxa okusemgangathweni, ***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 и G показаны как среднее значение ± стандартное отклонение, планки погрешностей представляют стандартное отклонение,00,00 Idatha kwi-C, D, E, kunye no-G iboniswa njengentsingiselo ± ukutenxa okusemgangathweni, iibar zempazamo zimele ukutenxa okusemgangathweni, ***p<0.0001.