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Ukopha okungalawulekiyo yenye yezona zinto ziphambili ezibangela ukufa. Ukufikelela kwi-hemostasis ekhawulezileyo kuqinisekisa ukusinda komntu njengoncedo lokuqala ngexesha lokulwa, iingozi zendlela, kunye nemisebenzi yokunciphisa ukufa. I-scaffold ye-nanoporous fiber-reinforced composite (NFRCS) ephuma kwi-composition elula ye-hemostatic film-forming (HFFC) njengesigaba esiqhubekayo inokubangela kwaye iphucule i-hemostasis. Uphuhliso lwe-NFRCS lusekelwe kuyilo lwephiko le-dragonfly. Ulwakhiwo lwephiko le-dragonfly lunamaphiko anqamlezileyo nawade, kwaye ii-membrane zephiko zidibene ukuze kugcinwe ukuthembeka kwesakhiwo se-microstructure. I-HFC igubungela ngokulinganayo umphezulu we-fiber ngefilimu yobukhulu be-nanometer kwaye iqhagamshela ubukhulu be-cotton esasazwe ngokungacwangciswanga (Ct) (isigaba esisasazekileyo) ukwenza isakhiwo se-nanoporous. Ukudibanisa izigaba eziqhubekayo nezisasazekileyo kunciphisa ixabiso lemveliso ngokuphindwe kalishumi xa kuthelekiswa neemveliso ezithengiswayo. I-NFRCS eguquliweyo (ii-tampons okanye ii-wristbands) zingasetyenziswa kwiindlela ezahlukeneyo ze-biomedical. Izifundo ze-in vivo zigqibe kwelokuba i-Cp NFRCS ephuhlisiweyo ivuselela kwaye iphucule inkqubo yokujiya kwindawo yokufaka isicelo. I-NFRCS inokuguqula imeko-bume yeseli kwaye isebenze kwinqanaba leeseli ngenxa yesakhiwo sayo esineembobo ezi-nano okubangela ukuphiliswa ngcono kwenxeba kwimodeli yenxeba lokususwa.
Ukopha okungalawulwayo ngexesha lokulwa, ngexesha lotyando kunye neemeko ezingxamisekileyo kunokubangela ingozi enkulu kubomi babantu abenzakeleyo1. Ezi meko zikhokelela ekwandeni ngokubanzi kokuxhathisa kwemithambo yegazi ejikeleze umzimba, okukhokelela ekothukeni kwegazi. Amanyathelo afanelekileyo okulawula ukopha ngexesha nasemva kotyando athathwa njengobungozi kubomi2,3. Ukonakala kwemithambo emikhulu kukhokelela ekulahlekelweni kwegazi okukhulu, okubangela izinga lokufa eliyi-≤ 50% ekulweni kunye ne-31% ngexesha lotyando1. Ukulahleka kwegazi okukhulu kukhokelela ekwehleni komthamo womzimba, okunciphisa ukuphuma kwentliziyo. Ukwanda kokuxhathisa kwemithambo yegazi ejikeleze umzimba kunye nokuphazamiseka okuqhubekayo kwe-microcirculation kukhokelela kwi-hypoxia kwizitho ezixhasa ubomi. Ukopha kwegazi kunokwenzeka ukuba imeko iyaqhubeka ngaphandle kokungenelela okusebenzayo1,4,5. Ezinye iingxaki ziquka ukuqhubela phambili kwe-hypothermia kunye ne-metabolic acidosis, kunye nesifo sokugabha esithintela inkqubo yokugabha. Ukopha kwegazi okukhulu kunxulunyaniswa nomngcipheko ophezulu wokufa6,7,8. Kwi-grade III (progressive) shock, ukufakelwa igazi kubalulekile ukuze isigulana sisinde ngexesha lokugula nokufa ngaphakathi kotyando nasemva kotyando. Ukuze soyise zonke ezi meko zingentla ezisongela ubomi, siye sayila i-nanoporous fiber-reinforced composite scaffold (NFRCS) esebenzisa i-polymer concentration encinci (0.5%) sisebenzisa indibaniselwano ye-water-soluble hemostatic polymers.
Ngokusebenzisa ifayibha yokuqinisa, iimveliso ezingabizi kakhulu zinokuphuhliswa. Iifayibha ezicwangciswe ngokungacwangciswanga zifana nolwakhiwo lwephiko ledragonfly, zilungelelaniswe yimicu ethe tye nethe nkqo emaphikweni. Imithambo enqamlezileyo nemide yephiko inxibelelana ne-membrane yephiko (Umzobo 1). I-NFRCS yenziwe yi-Ct eqinisiweyo njengenkqubo ye-scaffold enamandla angcono omzimba nawoomatshini (Umfanekiso 1). Ngenxa yokufikeleleka kunye nobugcisa, oogqirha bakhetha ukusebenzisa ii-cotton thread gauges (Ct) ngexesha lotyando kunye nee-dressing. Ngenxa yoko, xa sicinga ngeenzuzo zayo ezininzi, kuquka i-crystalline cellulose engaphezulu kwe-90% (enika amandla okuphucula umsebenzi we-hemostatic), i-Ct yasetyenziswa njengenkqubo yamathambo ye-NFRCS9,10. Ngenxa yoko, xa sicinga ngeenzuzo zayo ezininzi, kuquka i-crystalline cellulose engaphezulu kwe-90% (enika amandla okuphucula umsebenzi we-hemostatic), i-Ct yasetyenziswa njengenkqubo yamathambo ye-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. Ngoko ke, ngenxa yeenzuzo zayo ezininzi, kuquka i-cellulose ekristale engaphezulu kwama-90% (ebandakanyeka kumsebenzi owandisiweyo we-hemostatic), i-Ct yasetyenziswa njengenkqubo yamathambo ye-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,用的骨架系统。因此，考虑到它的多重益处，包括> 90%Ngoko ke, ngenxa yeenzuzo zayo ezininzi, kuquka i-crystalline cellulose engaphezulu kwe-90% (inceda ukuphucula umsebenzi we-hemostatic), i-Ct yasetyenziswa njenge-scaffold ye-NFRCS9,10.I-Ct yayigqunywe ngaphezulu (kwabonwa ukwakheka kwefilimu enobukhulu obufana ne-nano) kwaye idityaniswe nolwakhiwo lwefilimu oluyi-hemostatic (HFFC). I-HFC isebenza njenge-matrigel, ibambe i-Ct ebekwe ngokungacwangciswanga kunye. Uyilo oluphuhlisiweyo ludlulisela uxinzelelo ngaphakathi kwesigaba esisasazekileyo (iifayibha zokuqinisa). Kunzima ukufumana izakhiwo ezinamanzi aqinileyo kusetyenziswa ubuninzi bepolymer obuncinci. Ukongeza, akulula ukwenza ngokwezifiso iimold ezahlukeneyo kwizicelo ezahlukeneyo ze-biomedical.
Lo mfanekiso ubonisa umzobo woyilo lwe-NFRCS olusekwe kwisakhiwo sephiko le-dragonfly (A). Lo mfanekiso ubonisa uthelekiso oluthelekisayo lwesakhiwo sephiko le-dragonfly (imithambo edibanisayo nemide yephiko idibene) kunye ne-photomicrograph ye-Cp NFRCS (B). Umzobo we-Schematic we-NFRCS.
Ii-NFRC zaphuhliswa kusetyenziswa i-HFC njengesigaba esiqhubekayo ukujongana nemida engentla. I-HFC yenziwe ngee-polymers ezahlukeneyo ze-hemostatic ezenza ifilimu kuquka i-chitosan (njenge-polymer ephambili ye-hemostatic) ene-methylcellulose (MC), i-hydroxypropyl methylcellulose (HPMC 50 cp) kunye ne-polyvinyl alcohol (PVA)) (125 kDa) njenge-polymer exhasayo ekhuthaza ukwakheka kwe-thrombus. Ukongezwa kwe-polyvinylpyrrolidine K30 (PVP K30) kuphucule amandla okufunxa umswakama we-NFRCS. I-Polyethylene glycol 400 (PEG 400) yongezwa ukuphucula ukudibana kwe-polymer kwii-polymer blends ezidibeneyo. Ii-compositions ezintathu ezahlukeneyo ze-HFCC hemostatic (Cm HFCC, Ch HFCC kunye ne-Cp HFCC), ezizezi i-chitosan ene-MC (Cm), i-chitosan ene-HPMC (Ch), kunye ne-chitosan ene-PVA (Cp), zasetyenziswa kwi-Ct. Izifundo ezahlukeneyo zokuchaza i-in vitro kunye ne-in vivo ziqinisekisile umsebenzi wokuphilisa amanxeba kunye nokuphola kwe-NFRCS. Izixhobo ezidityanisiweyo ezinikezelwa yi-NFRCS zingasetyenziselwa ukwenza ngokwezifiso iintlobo ezahlukeneyo ze-scaffolding ukuhlangabezana neemfuno ezithile.
Ukongeza, i-NFRCS ingatshintshwa njengebhandishi okanye i-roll ukuze igubungele yonke indawo yokwenzakala kwimilenze esezantsi kunye nezinye iindawo zomzimba. Ngokukodwa kwizilonda zemilenze yokulwa, uyilo lwe-NFRCS oluyilwe lunokutshintshwa lube yingalo encinci okanye umlenze opheleleyo (Umfanekiso Ongezelelweyo S11). I-NFRCS ingenziwa ibe yibhanti yesandla eneglue yethishu, enokusetyenziselwa ukunqanda ukopha ngenxa yokwenzakala okukhulu kwesandla. Injongo yethu ephambili kukuphuhlisa i-NFRCS ene-polymer encinci kangangoko inokuhanjiswa kuluntu oluninzi (ngaphantsi komgca wobuhlwempu) kwaye enokufakwa kwikhithi yoncedo lokuqala. I-NFRCS ilula, isebenza kakuhle, kwaye inoqoqosho kuyilo, inceda uluntu lwasekuhlaleni kwaye inokuba nefuthe kwihlabathi liphela.
I-Chitosan (ubunzima bemolekyuli obuyi-80 kDa) kunye ne-amaranth zithengwe eMerck, eIndiya. I-Hydroxypropyl methylcellulose 50 Cp, i-polyethylene glycol 400 kunye ne-methylcellulose zithengwe kwiLoba Chemie Pvt. LLC, eMumbai. I-Polyvinyl alcohol (ubunzima bemolekyuli obuyi-125 kDa) (87-90% hydrolysed) ithengwe kwiNational Chemicals, eGujarat. I-Polyvinylpyrrolidine K30 ithengwe kwiMolychem, eMumbai, ii-swabs ezingenamafutha zithengwe kwiRamaraju Surgery Cotton Mills Ltd., eTamil Nadu, ngamanzi eMilli Q (inkqubo yokucoca amanzi eDirect-Q3, eMerck, eIndiya) njengomthwali.
I-NFRCS yaphuhliswa kusetyenziswa indlela ye-lyophilization11,12. Zonke izinto ze-HFCC (Itheyibhile 1) zalungiswa kusetyenziswa i-mechanical stirrer. Lungisa isisombululo se-0.5% se-chitosan usebenzisa i-1% ye-acetic acid emanzini ngokuxubha rhoqo kwi-800 rpm kwi-mechanical stirrer. Ubunzima obuchanekileyo be-polymer elayishiweyo obuboniswe kwiTheyibhile 1 bongezwe kwisisombululo se-chitosan kwaye baxubha de kwafunyanwa isisombululo se-polymer esicacileyo. I-PVP K30 kunye ne-PEG 400 zongezwa kumxube ophumayo kwizixa eziboniswe kwiTheyibhile 1, kwaye ukuxubha kwaqhubeka de kwafunyanwa isisombululo se-polymer esicacileyo. Ukuhlamba okuphumayo kwesisombululo se-polymer kwafakwa i-sonic imizuzu engama-60 ukususa amaqamza omoya avalelekileyo kumxube we-polymer. Njengoko kuboniswe kwi-Supplementary Figure S1(b), i-Ct yasasazwa ngokulinganayo emthonjeni ngamnye wepleyiti ye-6-well (mold) eyongezwe yi-5 ml ye-HFCC.
Ipleyiti yemithombo emithandathu ifakelwe i-sonic imizuzu engama-60 ukuze ifumane ukumanzisa kunye nokusasazwa okufanayo kwe-HFCC kwinethiwekhi ye-Ct. Emva koko ifake ipleyiti yemithombo emithandathu kwi--20°C iiyure ezi-8-12. Iipleyiti zokuqandisa zifakwe i-lyophilized iiyure ezingama-48 ukuze kufunyanwe iindlela ezahlukeneyo ze-NFRCS. Inkqubo efanayo isetyenziselwa ukuvelisa iimilo ezahlukeneyo kunye nezakhiwo, ezifana neetampons okanye ii-cylindrical tampons, okanye nayiphi na enye imo yezicelo ezahlukeneyo.
I-chitosan elinganiswe ngokuchanekileyo (80 kDa) (3%) inyibilikiswa kwi-1% ye-acetic acid kusetyenziswa i-magnetic stirrer. Kwisisombululo esiphumayo se-chitosan kongezwe i-1% PEG 400 yaza yaxutywa imizuzu engama-30. Galela isisombululo esiphumayo kwisitya esisikwere okanye esixande uze usenze sibe shushu kwi--80°C iiyure ezili-12. Iisampulu eziqandisiweyo zixutywe ne-lyophilized iiyure ezingama-48 ukuze kufunyanwe i-Cs13 eneembobo.
I-NFRCS ephuhlisiweyo yenziwe uvavanyo kusetyenziswa i-Fourier transform infrared spectroscopy (FTIR) (Shimadzu 8400 s FTIR, Tokyo, Japan) ukuqinisekisa ukuhambelana kweekhemikhali kwe-chitosan nezinye ii-polymers14,15. Ii-spectra ze-FTIR (ububanzi boluhlu lwee-spectral ukusuka kwi-400 ukuya kwi-4000 cm-1) zazo zonke iisampulu ezivavanyiweyo zifunyenwe ngokwenza iiskeni ezingama-32.
Izinga lokufunxwa kwegazi (i-BAR) kuzo zonke iifomyula livavanywe kusetyenziswa indlela echazwe nguChen et al. 16 kunye notshintsho oluncinci. Ii-NFRKs eziphuhlisiweyo zazo zonke iifomyula zomiswa kwi-oven ye-vacuum kwi-105°C ubusuku bonke ukuze kususwe i-solvent eseleyo. I-30 mg ye-NFRCS (ubunzima besampulu yokuqala - i-W0) kunye ne-30 mg ye-Ct (ulawulo oluhle) zibekwe kwizitya ezahlukeneyo eziqulethe i-premix ye-3.8% ye-sodium citrate. Ngamaxesha amiselweyo, oko kukuthi, imizuzwana emi-5, 10, 20, 30, 40 kunye ne-60, ii-NFRCS zisusiwe kwaye ubuso bazo bucocwe kwigazi elingafunxwanga ngokubeka iisampuli kwi-Ct imizuzwana engama-30. Ubunzima bokugqibela begazi elifunxwa yi-NFRCS 16 buqwalaselwe (i-W1) kwindawo nganye yexesha. Bala ipesenti ye-BAR usebenzisa le fomyula ilandelayo:
Ixesha lokuqunjelwa kwegazi (BCT) limiselwe njengoko kubikwe nguWang et al. 17. Ixesha elifunekayo ukuze igazi elipheleleyo (igazi lempuku elixutywe kwangaphambili ne-3.8% sodium citrate) liqumbe phambi kwe-NFRCS libalwe njenge-BCT yesampulu yovavanyo. Iinxalenye ezahlukeneyo ze-NFRCS (30 mg) zibekwe kwiibhotile ze-screw cap eziyi-10 ml zaza zafakwa kwi-37°C. Igazi (0.5 ml) longezwa kwi-vial kwaye i-0.3 ml ye-0.2 M CaCl2 yongezwa ukuze kusebenze ukuqunjelwa kwegazi. Okokugqibela, guqula i-vial rhoqo emva kwemizuzwana eli-15 (ukuya kuthi ga kwi-180°) de kube yi-clot eqinileyo. I-BCT yesampulu iqikelelwa ngenani le-flips vails17,18. Ngokusekelwe kwi-BCT, kukhethwe iindlela ezimbini ezifanelekileyo ezivela kwi-NFRCS Cm, Ch kunye ne-Cp ukuze kufundwe ngakumbi.
I-BCT ye-Ch NFRCS kunye ne-Cp NFRCS compositions imiselwe ngokusebenzisa indlela echazwe nguLi et al. 19. Beka i-15 x 15 mm2 Ch NFRCS, i-Cp NFRCS, kunye ne-Cs (ulawulo oluhle) kwizitya zePetri ezahlukeneyo (37 °C). Igazi eliqulethe i-3.8% sodium citrate lixutywe ne-0.2 M CaCl2 kwi-10:1 volume ratio ukuqala inkqubo yokuqina kwegazi. I-20 µl yomxube wegazi lempuku we-0.2 M CaCl2 yafakwa kumphezulu wesampuli yaza yafakwa kwisitya sePetri esingenanto. Ulawulo lwagalelwa igazi kwizitya zePetri ezingenanto ngaphandle kwe-Ct. Ngamaxesha athile emizuzu eyi-0, 3, kunye ne-5, yeka ukuqina ngokongeza i-10 ml yamanzi e-deionized (DI) kwisampuli equlethe isitya ngaphandle kokuphazamisa i-clot. Ii-erythrocytes ezingafakwanga (ii-erythrocytes) ziyafa xa kukho amanzi a-deionized kwaye zikhuphe i-hemoglobin. I-Hemoglobin ngamaxesha ahlukeneyo (HA(t)) ilinganiswe kwi-540 nm (λmax hemoglobin) kusetyenziswa i-UV-Vis spectrophotometer. Ukufunxwa ngokupheleleyo kwe-hemoglobin (AH(0)) kwi-0 min ye-20 µl yegazi kwi-10 ml yamanzi acocekileyo kuthathwe njengomgangatho omiselweyo. I-relative hemoglobin uptake (RHA) yegazi elijiyileyo ibalwe ukusuka kumlinganiselo we-HA(t)/HA(0) kusetyenziswa iqela elifanayo legazi.
Kusetyenziswa i-texture analyzer (Texture Pro CT V1.3 Build 15, Brookfield, USA), iimpawu zokuncamathelisa ze-NFRK kwizicubu ezonakeleyo zafunyanwa. Cinezela isitya esisilinda esivulekileyo esingaphantsi kwesikhumba sengulube (ngaphandle komaleko wamafutha). Iisampuli (Ch NFRCS kunye ne-Cp NFRCS) zafakwa nge-cannula kwiimolds zesilinda ukuze zenze ukunamathela eluswini lwengulube. Emva kokufunxwa kwemizuzu emi-3 kubushushu begumbi (RT) (25° C.), amandla okuncamathelisa e-NFRCS arekhodwa ngesantya esingaguqukiyo se-0.5 mm/sec.
Uphawu oluphambili lwezitywini zotyando kukwandisa ukujiya kwegazi ngelixa kunciphisa ukulahleka kwegazi. Ukujiya okungenalahleko kwi-NFRCS kuhlolwe kusetyenziswa indlela epapashwe ngaphambili enotshintsho oluncinci 19. Yenza ityhubhu ye-microcentrifuge (2 ml) (ububanzi bangaphakathi obuyi-10 mm) enombobo oyi-8 × 5 mm2 kwelinye icala letyhubhu ye-centrifuge (emele inxeba elivulekileyo). I-NFRCS isetyenziselwa ukuvala ukuvulwa kwaye iteyiphu isetyenziselwa ukuvala imiphetho engaphandle. Yongeza i-20 µl ye-0.2 M CaCl2 kwityhubhu ye-microcentrifuge equlethe i-3.8% sodium citrate premix. Emva kwemizuzu eli-10, iityhubhu ze-microcentrifuge zisusiwe kwizitya kwaye ukwanda kobunzima bezitya kwamiselwa ngenxa yokuphuma kwegazi kwi-NFRK (n = 3). Ukulahleka kwegazi i-Ch NFRCS kunye ne-Cp NFRCS zithelekiswa ne-Cs.
Ukunyaniseka okumanzi kwe-NFRCS kumiselwe ngokusekelwe kwindlela echazwe nguMishra noChaudhary21 ngotshintsho oluncinci. Beka i-NFRCS kwiflaski ye-Erlenmeyer eyi-100 ml kunye namanzi angama-50 ml uze uyijikelezise imizuzwana engama-60 ngaphandle kokwenza umphezulu. Ukuhlolwa ngokubonakalayo kunye nokubekwa phambili kweesampuli ukuze kubekho ukuthembeka ngokomzimba ngokusekelwe ekuqokelelweni.
Amandla okubopha e-HFCC kwi-Ct afundwe kusetyenziswa iindlela ezipapashwe ngaphambili kunye notshintsho oluncinci. Ukunyaniseka kwengubo yomphezulu kuhlolwe ngokutyhila i-NFRK kumaza e-acoustic (isikhuthazo sangaphandle) xa kukho amanzi e-milliQ (Ct). I-NFRCS Ch NFRCS kunye ne-Cp NFRCS eziphuhlisiweyo zibekwe kwibheki egcwele amanzi kwaye zafakwa i-sonicated imizuzu eyi-1, 2, 3, 4, 5, 6, 7, 8, 9, 10, kunye ne-30, ngokulandelanayo. Emva kokomiswa, umahluko wepesenti phakathi kobunzima bokuqala kunye nobokugqibela be-NFRCS wasetyenziselwa ukubala ipesenti yokulahleka kwezinto (HFC). I-In vitro BCT ixhase ngakumbi amandla okubopha okanye ukulahleka kwezinto eziphezulu. Ukusebenza kakuhle kwe-HFCC yokubopha kwi-Ct kubonelela ngokujiya kwegazi kunye nokwaleka okuthambileyo kumphezulu we-Ct22.
Ukulingana kwe-NFRCS ephuhlisiweyo kumiselwe yi-BCT yeesampuli (30 mg) ezithathwe kwiindawo eziqhelekileyo ezikhethwe ngokungacwangciswanga ze-NFRCS. Landela inkqubo ye-BCT ekhankanyiweyo ngaphambili ukuze umisele ukuthotyelwa kwe-NFRCS. Ukusondela phakathi kwazo zonke iisampuli ezintlanu kuqinisekisa ukugqunywa komphezulu okufanayo kunye nokufakwa kwe-HFC kwi-Ct mesh.
Indawo yoqhagamshelwano lwegazi (NBCA) ifunyenwe njengoko bekuxeliwe ngaphambili ngotshintsho oluthile. Qhoboshela igazi ngokucinezela i-20 µl yegazi phakathi kwemiphezulu emibini ye-Ct, i-Ch NFRCS, i-Cp NFRCS kunye ne-Cs. Emva kweyure e-1, iindawo ezimbini ze-stent zahlulwa kwaye zalinganiswa ngesandla indawo yehlwili. Ixabiso eliqhelekileyo lokuphindaphinda kathathu lithathwe njenge-NBCA NFRCS19.
Uhlalutyo lweDynamic Vapor Sorption (DVS) lusetyenziselwe ukuvavanya ukusebenza kakuhle kwe-NFRCS ekufunxeni amanzi kwindawo engaphandle okanye kwindawo yokwenzakala enoxanduva lokuqalisa ukujiya. I-DVS ivavanya okanye irekhoda ukufunxwa komphunga kunye nokulahleka kwisampulu ngokwegravimetric isebenzisa ibhalansi enobuthathaka kakhulu enesisombululo sobunzima se-±0.1 µg. Uxinzelelo lomphunga olungaphelelanga (umswakama oqhelekileyo) lwenziwa ngumlawuli wokuhamba kobunzima be-elektroniki ojikeleze isampulu ngokuxuba iigesi ezigcweleyo nezomileyo. Ngokwemigaqo ye-European Pharmacopeia, ngokusekelwe kwipesenti yokufunxwa komswakama ziisampuli, iisampuli zahlulwe zahlulwe zangamahlelo ama-4 (0–0.012% w/w− non-hygroscopic, 0.2–2% w/w hygroscopic kancinci, 2–15% hygroscopic ephakathi, kunye >15% hygroscopic kakhulu)23. Ngokwemigaqo ye-European Pharmacopeia, ngokusekelwe kwipesenti yokufunxwa komswakama ziisampuli, iisampuli zahlulwe zangamacandelo ama-4 (0–0.012% w/w− non-hygroscopic, 0.2–2% w/w hygroscopic kancinci, 2–15% hygroscopic ephakathi, kunye >15% hygroscopic kakhulu)23.Ngokuhambelana neengcebiso ze-European Pharmacopoeia, kuxhomekeke kwipesenti yokufunxwa komswakama yiisampulu, iisampulu zahlulwe zangamacandelo ama-4 (0–0.012% w/w – ezingezizo ezi-hygroscopic, 0.2–2% w/w ezi-hygroscopic kancinci, 2–15%).% умеренно гигроскопичен и > 15% очень гигроскопичен)23. % i-hygroscopic ephakathi kwaye > 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2 类（0-0.012% w/w-水比、靿家 2.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 樣品 分为分为分為类% （0（w-1010 ivenkile yokuthengisela i-%-0.性 , , , , 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 zahlulwe zibe ziiklasi ezi-4 ngokuxhomekeke kwipesenti yokufuma okufunxwa yisampuli (0-0.012% ngobunzima – engekho hygroscopic, 0.2-2% ngobunzima obuyi-hygroscopic kancinci, 2-15% ngobunzima).% умеренно гигроскопичен, > 15 % очень гигроскопичен) 23. % ethambileyo kancinci, > 15% ethambileyo kakhulu) 23.Ukusebenza kakuhle kwe-NFCS X NFCS kunye ne-TsN NFCS kumiselwe kwi-analyzer DVS TA TGA Q5000 SA. Ngexesha lale nkqubo, ixesha lokusebenza, ukufuma okuqhelekileyo (RH), kunye nobunzima besampuli yexesha langempela kwi-25°C24 kufunyenwe. Umxholo womswakama ubalwa ngohlalutyo oluchanekileyo lwe-NFRCS mass usebenzisa i-equation elandelayo:
I-MC yi-NFRCS ukufuma. m1 - ubunzima obomileyo be-NSAIDs. m2 bubunzima be-NFRCS bexesha langempela kwi-RH ethile.
Ummandla wonke womphezulu uqikelelwe kusetyenziswa uvavanyo lokufunxwa kwe-nitrogen nge-nitrogen engamanzi emva kokukhupha iisampulu kwi-25 °C kangangeeyure ezili-10 (< 7 × 10–3 Torr). Ummandla wonke womphezulu uqikelelwe kusetyenziswa uvavanyo lokufunxwa kwe-nitrogen nge-nitrogen engamanzi emva kokukhupha iisampulu kwi-25 °C kangangeeyure ezili-10 (< 7 × 10–3 Torr). Общая площадь поверхности оценивалась с помощью эксперимента по адсорбции азота жидким азотом после опорожнения образцов ° при120 × 10–3 Торр). Ummandla wonke womphezulu uqikelelwe kusetyenziswa uvavanyo lokufunxwa kwe-nitrogen nge-nitrogen engamanzi emva kokuba iisampulu zithululwe kwi-25°C kangangeeyure ezili-10 (< 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 образ0 при 25 ° C (< 7 × 10-3 тор). Ummandla wonke womphezulu uqikelelwe kusetyenziswa uvavanyo lokufunxwa kwe-nitrogen nge-nitrogen engamanzi emva kokuba iisampulu zikhutshwe iiyure ezili-10 kuma-25°C (< 7 x 10-3 torr).Ubungakanani bomphezulu opheleleyo, ubungakanani bembobo kunye nobukhulu bembobo ye-NFRCS buchongiwe ngeQuantachrome evela kwi-NOVA 1000e, e-Austria kusetyenziswa isoftware ye-RS 232.
Lungisa ii-RBC ezi-5% (ityuwa njenge-diluent) ezivela egazini liphela. Emva koko dlulisela i-aliquot ye-HFCC (0.25 ml) kwipleyiti ye-96-well kunye ne-5% RBC mass (0.1 ml). Faka umxube kwi-37°C imizuzu engama-40. Umxube weeseli ezibomvu zegazi kunye ne-serum uthathwe njengolawulo oluhle, kunye nomxube we-saline kunye neeseli ezibomvu zegazi njengolawulo olubi. I-Hemagglutination imiselwe ngokwesikali se-Stajitzky. Izikali ezicetywayo zezi zilandelayo: + + + + ii-aggregates ezixineneyo ze-granular; + + + ii-bottom pads ezigudileyo ezinee-curved edges; + + ii-bottom pads ezigudileyo ezinee-edges ezikrazukileyo; + ii-red rings ezimxinwa ezijikeleze imiphetho yee-smooth pads; – (negative) iqhosha elibomvu eli-discrete 12 embindini womthombo ongezantsi.
Ukuhambelana kwegazi ne-NFRCS kufundwe ngokwendlela ye-International Organisation for Standardization (ISO) (ISO10993-4, 1999)26,27. Indlela ye-gravimetric echazwe nguSingh et al. Kwenziwe utshintsho oluncinci ukuvavanya ukwakheka kwe-thrombus phambi okanye phezu komphezulu we-NFRCS. I-500 mg ye-Cs, i-Ch NFRCS kunye ne-Cp NFRCS zafakwa kwi-phosphate buffered saline (PBS) iiyure ezingama-24 kwi-37°C. Emva kweeyure ezingama-24, i-PBS yasuswa kwaye i-NFRCS yanyangwa nge-2 ml yegazi equlethe i-3.8% sodium citrate. Kumphezulu we-NFRCS, yongeza i-0.04 ml ye-0.1 M CaCl2 kwiisampuli ezifakwe kwi-incubated. Emva kwemizuzu engama-45, kongezwa i-5 ml yamanzi acociweyo ukuze kumiswe ukujiya. Igazi elijiweyo kumphezulu we-NFRK lanyangwa ngesisombululo se-formaldehyde esingu-36-38%. Amahlwili afakwe i-formaldehyde omisiwe aze alinganiswe. Ipesenti ye-thrombosis iqikelelwe ngokubala ubunzima beglasi ngaphandle kwegazi kunye nesampuli (ulawulo olungalunganga) kunye neglasi enegazi (ulawulo oluchanekileyo).
Njengobungqina bokuqala, iisampulu zabonwa nge-microscope ebonakalayo ukuze kuqondwe amandla okugqunywa komphezulu we-HFCC, i-Ct edibeneyo, kunye nenethiwekhi ye-Ct yokwenza ii-pores. Amacandelo amancinci e-Ch kunye ne-Cp avela kwi-NFRCS anqunyulwa nge-scalpel blade. Icandelo eliphumayo labekwa kwi-slide yeglasi, ligqunywe nge-coverlip, kwaye imiphetho yaqiniswa ngeglue. Ii-slides ezilungisiweyo zajongwa nge-microscope ebonakalayo kwaye iifoto zathathwa ngokwahluka kobukhulu.
Ukufakwa kwepolymer kwiinethiwekhi zeCt kubonwe kusetyenziswa i-fluorescence microscopy ngokusekelwe kwindlela echazwe nguRice et al.29. Umxube we-HFCC osetyenziselwe le fomyula uxutywe nedayi ye-fluorescent (amaranth), kwaye i-NFRCS (Ch & Cp) yalungiswa ngokwendlela ekhankanyiweyo ngaphambili. Umxube we-HFCC osetyenziselwe le fomyula uxutywe nedayi ye-fluorescent (amaranth), kwaye i-NFRCS (Ch & Cp) yalungiswa ngokwendlela ekhankanyiweyo ngaphambili.Umxube we-HFCC osetyenziselwe ukwenza umxube uxutywe nedayi ye-fluorescent (amaranth) kwaye i-NFRCS (Ch kunye ne-Cp) yafunyanwa ngokwendlela ekhankanyiweyo 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）Umxube we-HFCC osetyenzisiweyo kule fomyula uxutywe nedayi ye-fluorescent (i-Amaranth) kwaye wafumana i-NFRCS (i-Ch kunye ne-Cp), njengoko bekutshiwo ngaphambili.Iinxalenye ezincinci ze-NFRK zisikwe kwiisampuli ezifunyenweyo, zabekwa kwiislayidi zeglasi, zaza zagqunywa ngezigqubuthelo. Jonga izilayidi ezilungisiweyo phantsi kwe-microscope ye-fluorescent usebenzisa isihluzo esiluhlaza (310-380 nm). Imifanekiso ithathwe kwi-4x magnification ukuqonda ubudlelwane be-Ct kunye nokufakwa kwe-polymer engaphezulu kwinethiwekhi ye-Ct.
I-surface topography ye-NFRCS Ch kunye ne-Cp imiselwe kusetyenziswa i-atomic force microscope (AFM) ene-ultra-sharp TESP cantilever kwimo yokuthepha: 42 N/m, 320 kHz, ROC 2-5 nm, eBruker, eTaiwan. Uburhabaxa bomphezulu bumiselwe yi-root mean square (RMS) kusetyenziswa isoftware (Scanning Probe Image Processor). Iindawo ezahlukeneyo ze-NFRCS zenziwe kwimifanekiso ye-3D ukujonga ubufana bomphezulu. Ukuphambuka okuqhelekileyo kwenqaku kwindawo ethile kuchazwa njengoburhabaxa bomphezulu. I-RMS equation isetyenziselwe ukulinganisa uburhabaxa bomphezulu be-NFRCS31.
Izifundo ezisekelwe kwi-FESEM zenziwe kusetyenziswa i-FESEM, i-SU8000, i-HI-0876-0003, eHitachi, eTokyo, ukuze kuqondwe imo yomphezulu we-Ch NFRCS kunye ne-Cp NFRCS, ebonise i-BCT engcono kune-Cm NFRCS. Uphononongo lwe-FESEM lwenziwe ngokwendlela echazwe nguZhao et al. 32 kunye notshintsho oluncinci. I-NFRCS 20 ukuya kwi-30 mg ye-Ch NFRCS kunye ne-Cp NFRCS zaxutywa kwangaphambili nge-20 µl ye-3.8% ye-sodium citrate exutywe kwangaphambili negazi leempuku. I-20 μl ye-0.2 M CaCl2 yongezwa kwiisampuli eziphathwe ngegazi ukuze kuqaliswe ukujiya kwaye iisampuli zafakwa kwiqondo lobushushu begumbi imizuzu eli-10. Ukongeza, ii-erythrocytes ezingaphezulu zasuswa kumphezulu we-NFRCS ngokuhlanjwa nge-saline.
Iisampulu ezilandelayo zanyangwa nge-0.1% glutaraldehyde zaza zomiswa kwi-oven yomoya oshushu kwi-37°C ukususa umswakama. Iisampulu ezomileyo zagqunywa zaza zahlalutywa 32. Eminye imifanekiso efunyenwe ngexesha lohlalutyo yayikukwakheka kwe-clot kumphezulu weefayibha zomqhaphu ngamnye, ukufakwa kwepolymer phakathi kwe-Ct, imo ye-erythrocyte (imilo), ukuthembeka kwe-clot, kunye nemo ye-erythrocyte phambi kwe-NFRCS. Iindawo ze-NFRCS ezinganyangwanga kunye neendawo ze-NFRCS eziphathwe nge-Ch kunye ne-Cp ezifakwe igazi zaskenwa ukuze kufunyanwe ii-ions ze-elemental (i-sodium, i-potassium, i-nitrogen, i-calcium, i-magnesium, i-zinc, i-copper kunye ne-selenium)33. Thelekisa iipesenti ze-ion ze-elemental phakathi kweesampulu ezinyangiweyo kunye nezinganyangwanga ukuze uqonde ukuqokelelwa kwe-ion ze-elemental ngexesha lokwakheka kwe-clot kunye ne-homogeneity ye-clot.
Ubukhulu bomphezulu we-Cp HFCC kumphezulu we-Ct buchongiwe kusetyenziswa i-FESEM. Amacandelo anqamlezileyo e-Cp NFRCS asikwe kwisakhelo kwaye agqunywe nge-sputter. Iisampulu zomphezulu we-sputter ezivelileyo zibonwe yi-FESEM kwaye ubukhulu bomphezulu we-sputter bulinganiswe ngama-34, 35, 36.
I-X-ray micro-CT inika umfanekiso we-3D ongengonakalisiyo onesisombululo esiphezulu kwaye ikuvumela ukuba ufunde ulungelelwaniso lwangaphakathi lwe-NFRK. I-Micro-CT isebenzisa umqadi we-X-ray odlula kwisampulu ukurekhoda i-local linear attenuation coefficient ye-X-rays kwisampulu, enceda ukufumana ulwazi lwemo. Indawo yangaphakathi ye-Ct kwi-Cp NFRCS kunye ne-Cp NFRCS ephathwe ngegazi ihlolwe yi-micro-CT ukuqonda ukusebenza kakuhle kokufunxa kunye nokuxinana kwegazi phambi kwe-NFRCS37,38,39. Izakhiwo ze-3D zeesampulu ze-Cp NFRCS eziphathwe ngegazi nezinganyangwanga zakhiwe ngokutsha kusetyenziswa i-micro-CT (V|tome|x S240, ePhoenix, eJamani). Kusetyenziswa isoftware ye-VG STUDIO-MAX version 2.2, imifanekiso emininzi ye-X-ray ithathwe kwii-engile ezahlukeneyo (ngokufanelekileyo ukugubungela i-360°) ukuphuhlisa imifanekiso ye-3D ye-NFRCS. Idatha eqokelelweyo yoqikelelo yakhiwe ngokutsha kwimifanekiso ye-3D volumetric kusetyenziswa isoftware elula ye-3D ScanIP Academic.
Ukongeza, ukuze kuqondwe ukusasazwa kwehlwili, i-20 µl yegazi elixutywe kwangaphambili kunye ne-20 µl ye-0.2 M CaCl2 zongezwa kwi-NFRCS ukuqalisa ukujiya kwegazi. Iisampuli ezilungisiweyo zishiywa ziqinile. Umphezulu we-NFRK uphathwe nge-0.5% glutaraldehyde kwaye womiswa kwi-oven yomoya oshushu kwi-30–40°C imizuzu engama-30. Ihlwili legazi elenziwe kwi-NFRCS laskenwa, lakhiwa ngokutsha, kwaye umfanekiso we-3D wehlwili legazi wabonwa.
Uvavanyo lwe-antibacterial lwenziwe kwi-Cp NFRCS (ngcono xa kuthelekiswa ne-Ch NFRCS) kusetyenziswa indlela echazwe ngaphambili kunye notshintsho oluncinci. Umsebenzi we-antibacterial we-Cp NFRCS kunye ne-Cp HFCC uchongiwe kusetyenziswa ii-microorganisms ezintathu ezahlukeneyo zovavanyo [i-S.aureus (i-gram-positive bacteria), i-E.coli (i-gram-negative bacteria) kunye ne-Candida emhlophe (i-C.albicans)] ekhula kwi-agar kwizitya zePetri kwi-incubator. Faka i-50 ml ye-diluted bacterial culture suspension kwi-concentration ye-105-106 CFU ml-1 kwi-agar medium. Galela i-medium kwisitya sePetri uze uyivumele iqine. Ii-wells zenziwe phezu kwepleyiti ye-agar ukuze zizaliswe yi-HFCC (ii-wells ezi-3 ze-HFCC kunye ne-1 yokulawula okungalunganga). Yongeza i-200 µl HFCC kwi-wells ezi-3 kunye ne-200 µl pH 7.4 PBS kwi-well yesi-4. Kwelinye icala lesitya sepetri, beka idiski ye-12 mm Cp NFRCS kwi-agar eqinileyo uze umanzise nge-PBS (pH 7.4). Iipilisi zeCiprofloxacin, i-ampicillin kunye ne-fluconazole zithathwa njengemigangatho yokubonisa iStaphylococcus aureus, i-Escherichia coli kunye neCandida albicans. Linganisa indawo yokuthintela ngesandla kwaye uthathe umfanekiso wedijithali wendawo yokuthintela.
Emva kokuvunywa kwemigaqo yokuziphatha kwesikhungo, olu phononongo lwenziwe kwiKasturba Medical College of Education and Research eManipal, eKarnataka, kumazantsi eIndiya. Inkqubo yovavanyo lwe-in vitro TEG ihlolwe kwaye yavunywa yiKomiti yeZimilo zeZiko yeKasturba Medical College, eManipal, eKarnataka (IEC: 674/2020). Izifundo zifunyenwe kubanikeli begazi abazithandelayo (abaneminyaka eli-18 ukuya kwengama-55) kwibhanki yegazi yesibhedlele. Ukongeza, ifomu yemvume enolwazi ifunyenwe kubanikeli begazi abazithandelayo ukuze kuqokelelwe iisampuli zegazi. I-Native TEG (N-TEG) ​​​​yasetyenziswa ukufunda isiphumo se-Cp HFCC formula egazini elipheleleyo elixutywe ne-sodium citrate. I-N-TEG yaziwa ngokubanzi ngendima yayo ekuvuseleleni indawo yokhathalelo, edala iingxaki koogqirha ngenxa yokuba kunokwenzeka ukuba iziphumo zingabi buhlungu kakhulu (uvavanyo lwe-coagulation oluqhelekileyo). Uhlalutyo lwe-N-TEG lwenziwe kusetyenziswa igazi elipheleleyo. Imvume enolwazi kunye nembali yezonyango eneenkcukacha zifunyenwe kubo bonke abathathi-nxaxheba. Olu phononongo aluzange lubandakanye abathathi-nxaxheba abaneengxaki ze-hemostatic okanye ze-thrombotic ezifana nokukhulelwa/ukukhulelwa okanye isifo sesibindi. Abantu abasebenzisa amayeza achaphazela i-coagulation cascade nabo abafakwanga kuphando. Uvavanyo olusisiseko lwelabhoratri (i-hemoglobin, ixesha le-prothrombin, i-activated thromboplastin kunye ne-platelet count) lwenziwe kubo bonke abathathi-nxaxheba ngokweenkqubo eziqhelekileyo. I-N-TEG imisela i-blood clot viscoelasticity, ulwakhiwo lwe-clot yokuqala, ukusebenzisana kwamasuntswana, ukuqinisa i-clot, kunye ne-clot lysis. Uhlalutyo lwe-N-TEG lubonelela ngedatha yemizobo kunye neyamanani kwimiphumo edibeneyo yezinto ezininzi zeseli kunye ne-plasma. Uhlalutyo lwe-N-TEG lwenziwe kwiivolumu ezimbini ezahlukeneyo ze-Cp HFCC (10 µl kunye ne-50 µl). Ngenxa yoko, i-1 ml yegazi elipheleleyo eline-citric acid yongezwa kwi-10 μl ye-Cp HFCC. Yongeza i-1 ml (Cp HFCC + igazi elicitrated), i-340 µl yegazi elixutyiweyo kwi-20 µl 0.2 M CaCl2 equlethe isitya se-TEG. Emva koko, izitya ze-TEG zafakwa kwi-TEG® 5000, e-US ukulinganisa i-R, i-K, i-alpha angle, i-MA, i-G, i-CI, i-TPI, i-EPL, i-LY i-30% yeesampuli zegazi xa kukho i-Cp HFFC41.
Iprotokholi yokufunda ngaphakathi emzimbeni ihlolwe yaza yavunywa yiKomiti yeZimilo zeZilwanyana zeZiko (IAEC), iKasturba School of Medicine, iManipal Institute of Higher Education, eManipal (IAEC/KMC/69/2020). Zonke iimvavanyo zezilwanyana zenziwe ngokweengcebiso zeKomiti yoLawulo nokuLawulwa koVavanyo lweZilwanyana (CPCSEA). Zonke izifundo ze-NFRCS ngaphakathi emzimbeni (2 × 2 cm2) zenziwe kwiigundane zeWistar eziziimazi (ezinobunzima obuyi-200 ukuya kwi-250 g). Zonke izilwanyana zaqhelana nobushushu obuyi-24-26°C, izilwanyana zazinokufikelela simahla ekutyeni okuqhelekileyo namanzi ngokukhawuleza. Zonke izilwanyana zahlulwe ngokungacwangciswanga zangamaqela ahlukeneyo, iqela ngalinye lalinazo izilwanyana ezintathu. Zonke izifundo zenziwe ngokweZifundo zeZilwanyana: Ingxelo yoVavanyo lwe-In Vivo 43. Ngaphambi kophando, izilwanyana zanikwa i-anesthesia ngokufakwa kwe-intraperitoneal (ip) komxube we-20-50 mg ye-ketamine (nge-1 kg yobunzima bomzimba) kunye ne-2-10 mg ye-xylazine (nge-1 kg yobunzima bomzimba). Emva kophando, umthamo wokopha wabalwa ngokuvavanya umahluko phakathi kobunzima bokuqala nobokugqibela beesampuli, ixabiso eliqhelekileyo elifunyenwe kwiimvavanyo ezintathu lathathwa njengomthamo wokopha wesampuli.
Imodeli yokunqunyulwa komsila wempuku yasetyenziswa ukuze kuqondwe amandla e-NFRCS okulawula ukopha xa kusenzeka ingozi, ukulwa, okanye ingozi yendlela (imodeli yokwenzakala). Sika i-50% yomsila nge-scalpel blade uze uyibeke emoyeni imizuzwana eli-15 ukuqinisekisa ukopha okuqhelekileyo. Ukongeza, iisampulu zovavanyo zabekwa emsileni wempuku ngokusebenzisa uxinzelelo (Ct, Cs, Ch NFRCS kunye neCp NFRCS). Ukopha kunye ne-PCT zaxelwa kwiisampulu zovavanyo (n ​​= 3)17,45.
Ukusebenza kakuhle kolawulo loxinzelelo lwe-NFRCS ekulweni kuphandwe kwimodeli ye-femoral artery engaphezulu. I-femoral artery iyavezwa, ibhobozwe nge-24G trocar, ize yophe kwimizuzwana eli-15. Emva kokuba kubonwe ukopha okungalawulekiyo, isampuli yovavanyo ibekwa kwindawo yokubhoboza ngoxinzelelo olusetyenzisiweyo. Emva nje kokusetyenziswa kwesampuli yovavanyo, ixesha lokujiya kwegazi larekhodwa kwaye ukusebenza kakuhle kwe-hemostatic kwabonwa kwimizuzu emi-5 elandelayo. Inkqubo efanayo yaphindwa nge-Cs kunye ne-Ct46.
UDowling nabanye 47 bacebise imodeli yokwenzakala kwesibindi ukuvavanya amandla okuhambisa igazi kwizinto ezihambisa igazi kwimeko yokopha ngaphakathi kotyando. I-BCT yarekhodwa kwiisampuli ze-Ct (ulawulo olungalunganga), ulwakhiwo lwe-Cs (ulawulo oluhle), iisampuli ze-Ch NFRCS, kunye neesampuli ze-Cp NFRCS. I-suprahepatic vena cava yempuku yavezwa ngokwenza i-median laparotomy. Emva koko, inxalenye ekude yelobe yasekhohlo yasikwa ngesikere. Yenza isikere kwisibindi nge-scalpel blade kwaye uyivumele yophe imizuzwana embalwa. Iisampuli zovavanyo ze-Ch NFRCS kunye ne-Cp NFRCS ezilinganiswe ngokuchanekileyo zabekwa kumphezulu owonakeleyo ngaphandle koxinzelelo oluhle kwaye i-BCT yarekhodwa. Iqela lolawulo (Ct) emva koko lasebenzisa uxinzelelo olulandelwa yi-Cs 30 s47 ngaphandle kokwaphula ukwenzakala.
Uvavanyo lokuphiliswa kwamanxeba emzimbeni we-in vivo lwenziwe kusetyenziswa imodeli yenxeba elisusayo ukuvavanya iimpawu zokuphiliswa kwamanxeba kwi-NFRCSs eziphuhlisiweyo ezisekelwe kwi-polymer. Iimodeli zamanxeba asusayo zikhethwe kwaye zenziwa ngokweendlela ezipapashwe ngaphambili kunye notshintsho oluncinci19,32,48. Zonke izilwanyana zathotywa i-anesthesia njengoko kuchaziwe ngaphambili. Sebenzisa i-biopsy punch (12 mm) ukwenza ukusika okujikelezayo okunzulu eluswini lomqolo. Iindawo zamanxeba ezilungisiweyo zanxitywa nge-Cs (ulawulo oluhle), i-Ct (ukuqonda ukuba ii-cotton pads ziphazamisa ukuphiliswa), i-Ch NFRCS kunye ne-Cp NFRCS (iqela lovavanyo) kunye nolawulo olungalunganga ngaphandle konyango. Kusuku ngalunye lophando, indawo yenxeba yayilinganiswa kuzo zonke iimpuku. Sebenzisa ikhamera yedijithali ukuthatha umfanekiso wendawo yenxeba kwaye ufake i-dressing entsha. Ipesenti yokuvalwa kwenxeba yalinganiswa ngale fomyula ilandelayo:
Ngokusekelwe kwipesenti yokuvalwa kwenxeba ngomhla we-12 wophando, ulusu lwempuku lweqela elingcono lwasuswa ((Cp NFRCS) kunye neqela lolawulo) lwaze lwafundwa yi-H&E staining kunye ne-Masson's trichrome staining. Ngokusekelwe kwipesenti yokuvalwa kwenxeba ngomhla we-12 wophando, ulusu lwempuku lweqela elingcono lwasuswa ((Cp NFRCS) kunye neqela lolawulo) lwaze lwafundwa yi-H&E staining kunye ne-Masson's trichrome staining.Ngokusekelwe kwipesenti yokuvalwa kwenxeba ngomhla we-12 wophando, ulusu lweempuku zeqela elingcono ((Cp NFRCS) kunye neqela lolawulo) lwasuswa lwaza lwahlolwa ngokufaka i-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，进行H&E染色和Masson三色染色研究。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)Iimpuku ezikwiqela elingcono ((Cp NFRCS) kunye namaqela olawulo) zasuswa ukuze kufakwe i-hematoxylin-eosin staining kunye ne-Masson's trichrome staining ngokusekelwe kwipesenti yokuvalwa kwenxeba ngosuku lwe-12 lophando.Inkqubo yokudaya esetyenzisiweyo yenziwe ngokweendlela ezichazwe ngaphambili49,50. Okwethutyana, emva kokufakwa kwi-10% formalin, iisampulu zasuswa amanzi kusetyenziswa uthotho lwee-alcohols ezilinganisiweyo. Sebenzisa i-microtome ukufumana amacandelo amancinci (5 µm ubukhulu) kwizicubu ezikhutshiweyo. Amacandelo amancinci okulandelelana kunye ne-Cp NFRCS anyangwa nge-hematoxylin kunye ne-eosin ukuze kufundwe utshintsho lwe-histopathological. I-Masson's trichrome stain yasetyenziswa ukufumanisa ukwakheka kwe-collagen fibrils. Iziphumo ezifunyenweyo zafundwa ngokungaboniyo ziingcali ze-pathologists.
Uzinzo lweesampuli ze-Cp NFRCS lufundwe kubushushu begumbi (25°C ± 2°C/60% RH ± 5%) kangangeenyanga ezili-1251. I-Cp NFRCS (ukutshintsha kombala womphezulu kunye nokukhula kweentsholongwane) ihlolwe ngokubonakalayo kwaye yavavanywa ukuba ayigugi kwaye ayigugi ngokweendlela ezichazwe ngasentla kwicandelo leZixhobo kunye neeNdlela.
Ukwanda kunye nokuphindaphinda kwe-Cp NFRCS kuhlolwe ngokulungiselela i-Cp NFRCS enobukhulu obuyi-15×15 cm2. Ukongeza, iisampulu ezingama-30 mg (n = 5) zasuswa kwiinxalenye ezahlukeneyo ze-Cp NFRCS kwaye i-BCT yeesampulu ezifundweyo yavavanywa njengoko kuchaziwe ngaphambili kwicandelo leeNdlela.
Sizamile ukuphuhlisa iimilo ezahlukeneyo kunye nezakhiwo sisebenzisa ukwakheka kwe-Cp NFRCS kwiindlela ezahlukeneyo zonyango. Ezi zakhiwo okanye ulwakhiwo ziquka ii-conical swabs zokopha empumlweni, iinkqubo zamazinyo, kunye nee-cylindrical swabs zokopha eluswini.
Zonke iiseti zedatha zichazwa njenge-mean ± standard deviation kwaye zahlalutywa yi-ANOVA kusetyenziswa iPrism 5.03 (GraphPad, San Diego, CA, USA) zilandelwa luvavanyo lweBonferroni lokuthelekisa okuninzi (*p<0.05).
Zonke iinkqubo ezenziwa kwizifundo zabantu bezihambelana nemigangatho yeZiko kunye neBhunga loPhando leSizwe, kunye neSibhengezo saseHelsinki sika-1964 kunye nezilungiso zaso ezalandelayo, okanye imigangatho yokuziphatha efanayo. Bonke abathathi-nxaxheba baxelelwe ngeempawu zolu phando kunye nohlobo lwalo lokuzithandela. Idatha yabathathi-nxaxheba ihlala iyimfihlo xa sele iqokelelwe. Iprotokholi yovavanyo lwe-TEG ye-in vitro ihlolwe kwaye yavunywa yiKomiti yeZimilo zeZiko yeKholeji yezoNyango yaseKasturba, eManipal, eKarnataka (IEC: 674/2020). Amavolontiya atyikitye imvume enolwazi yokuqokelelwa kweesampuli zegazi.
Zonke iinkqubo ezenziwe kwizifundo zezilwanyana zenziwe ngokuhambelana neKastuba Faculty of Medicine, Manipal Institute of Higher Education, Manipal (IAEC/KMC/69/2020). Zonke iimvavanyo zezilwanyana ezenziweyo zenziwe ngokuhambelana nezikhokelo zeKomiti yoLawulo kunye nokuLawulwa koVavanyo lweZilwanyana (CPCSEA). Bonke ababhali balandela izikhokelo ze-ARRIVE.
Iispectra ze-FTIR zazo zonke ii-NFRCS zihlalutywe zaza zathelekiswa ne-chitosan spectrum eboniswe kuMfanekiso 2A. Iincopho ze-spectral eziphawulekayo ze-chitosan (ezirekhodiweyo) kwi-3437 cm-1 (OH kunye ne-NH stretching, overlap), 2945 kunye ne-2897 cm-1 (CH stretching), 1660 cm-1 (NH2 strain), 1589 cm-1 (N–H bending ), 1157 cm-1 (bridge stretch O-), 1067 cm-1 (stretch C–O, secondary hydroxyl), 993 cm-1 (stretch CO, Bo-OH) 52.53.54. Itheyibhile eyongeziweyo S1 ibonisa amaxabiso e-FTIR NFRCS absorption spectrum ye-chitosan (reporter), i-pure chitosan, i-Cm, i-Ch, kunye ne-Cp. Iispectra ze-FTIR zazo zonke ii-NFRCS (Cm, Ch kunye ne-Cp) zibonise iibhendi zokufunxa ezifanayo njenge-chitosan ecocekileyo ngaphandle kotshintsho olubalulekileyo (Umzobo 2A). Iziphumo ze-FTIR ziqinisekisile ukungabikho konxibelelwano lweekhemikhali okanye lwendalo phakathi kwee-polymers ezisetyenziselwa ukuphuhlisa i-NFRCS, nto leyo ebonisa ukuba ii-polymers ezisetyenzisiweyo azinamsebenzi.
Ukuchazwa kwe-in vitro kwe-Cm NFRCS, i-Ch NFRCS, i-Cp NFRCS kunye ne-Cs. (A) imele ii-spectra ze-FTIR ezidibeneyo zezinto ezidityanisiweyo ze-chitosan kunye ne-Cm NFRCS, i-Ch NFRCS kunye ne-Cp NFRCS phantsi koxinzelelo. (B) a) Izinga lokufunxwa kwegazi elipheleleyo le-NFRCS Cm, Ch, Cp, kunye ne-Cg (n = 3); Iisampulu ze-Ct zibonise i-BAR ephezulu kuba i-cotton swab inokusebenza okuphezulu kokufunxwa; b) Igazi emva kokufunxwa kwegazi Umfanekiso wesampulu efunxwayo. Ukubonakaliswa kwemizobo ye-BCT yesampulu yovavanyo C (i-Cp NFRCS yayine-BCT engcono kakhulu (15 s, n = 3)). Idatha kwi-C, D, E, kunye ne-G iboniswe njenge-± SD ephakathi, kwaye ii-error bars zimele i-SD, ***p < 0.0001. Idatha kwi-C, D, E, kunye ne-G iboniswe njenge-± SD ephakathi, kwaye ii-error bars 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 ekwi-C, D, E, kunye no-G iboniswa njenge-avareji ± ukuphambuka okuqhelekileyo, kwaye ii-error bars zimele ukuphambuka okuqhelekileyo, ***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 откло0ние,00 Idatha ekwi-C, D, E, kunye no-G iboniswa njenge-avareji ± ukuphambuka okuqhelekileyo, ii-error bars zimele ukuphambuka okuqhelekileyo, ***p<0.0001.