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Kutuluka magazi kosalamulirika ndi chimodzi mwa zifukwa zomwe zimayambitsa imfa.Kupeza hemostasis mwachangu kumatsimikizira kupulumuka kwa mutuwo ngati chithandizo choyamba pankhondo, ngozi zapamsewu, ndi ntchito zochepetsera imfa.Nanoporous fiber-reinforced composite scaffold (NFRCS) yochokera ku simple hemostatic film-forming composition (HFFC) monga gawo lopitirira likhoza kuyambitsa ndi kupititsa patsogolo hemostasis.Kukula kwa NFRCS kumatengera kapangidwe ka mapiko a tombolombo.Mapiko a dragonfly amapangidwa ndi mapiko opingasa ndi aatali, ndipo mapiko a mapiko amalumikizana wina ndi mnzake kuti asunge umphumphu wa microstructure.HFFC imavala mofanana pamwamba pa ulusi ndi filimu ya makulidwe a nanometer ndikugwirizanitsa makulidwe a thonje ogawidwa mwachisawawa (Ct) (gawo lobalalika) kuti apange mawonekedwe a nanoporous.Kuphatikizika kwa magawo opitilira ndi omwazika kumachepetsa mtengo wazinthu kakhumi poyerekeza ndi zinthu zomwe zimapezeka pamalonda.Zosinthidwa NFRCS (tampons kapena ma wristbands) zitha kugwiritsidwa ntchito m'njira zosiyanasiyana zamankhwala.Kafukufuku wa mu vivo watsimikiza kuti Cp NFRCS yopangidwa imayambitsa ndikuwonjezera njira yolumikizirana pamalo ogwiritsira ntchito.NFRCS imatha kusintha mawonekedwe a microenvironment ndikuchita pamlingo wa ma cell chifukwa cha kapangidwe kake ka nanoporous komwe kumapangitsa kuchira bwino kwa bala mu chitsanzo cha bala la excision.
Kutaya magazi kosalamulirika panthawi yankhondo, kupha anthu ovulala kapena pakachitika ngozi mwadzidzidzi kungayambitse moyo wa ovulala1.Izi zimabweretsanso kuwonjezereka kwapang'onopang'ono kwa mitsempha ya m'mitsempha, zomwe zimapangitsa kuti munthu azidwala matenda a hemorrhagic.Njira zoyenera zopewera kutaya magazi mkati ndi pambuyo pa opaleshoni zimaganiziridwa kuti zingakhale zoopsa2,3.Kuwonongeka kwa ziwiya zazikulu kumabweretsa kutaya magazi kwakukulu, zomwe zimapangitsa kuti chiwerengero cha imfa ≤ 50% pankhondo ndi 31% pa opaleshoni1.Kutaya magazi kwakukulu kumabweretsa kuchepa kwa thupi, zomwe zimachepetsa kutulutsa kwa mtima.Kuchuluka kwa zotumphukira zotumphukira kukana kwa mitsempha komanso kuwonongeka kwapang'onopang'ono kwa microcirculation kumabweretsa hypoxia mu ziwalo zothandizira moyo.Kuthamanga kwa magazi kungathe kuchitika ngati vutoli likupitirira popanda kuthandizira bwino1,4,5.Zovuta zina ndi monga kukwera kwa hypothermia ndi metabolic acidosis, komanso vuto la coagulation lomwe limalepheretsa kukomoka.Kugwedezeka kwakukulu kwa magazi kumalumikizidwa ndi chiopsezo chachikulu cha kufa6,7,8.Pakugwedezeka kwa kalasi ya III (kupita patsogolo), kuikidwa magazi n'kofunika kuti odwala apulumuke panthawi ya intraoperative ndi postoperative morbidity ndi imfa.Kuti tigonjetse zinthu zonse zomwe zili pamwambazi, tapanga nanoporous fiber-reinforced composite scaffold (NFRCS) yomwe imagwiritsa ntchito polima pang'ono (0.5%) pogwiritsa ntchito ma polima osungunuka amadzi omwe amasungunuka m'madzi.
Pogwiritsa ntchito fiber reinforcement, zinthu zotsika mtengo zimatha kupangidwa.Ulusi wopangidwa mwachisawawa umafanana ndi mmene mapiko a tombolombo amapangidwira, ndipo mapikowo amakhala opingasa komanso oimirira.Mitsempha yopingasa ndi yotalika ya phiko imalankhulana ndi nembanemba ya mapiko (mkuyu 1).NFRCS imakhala ndi Ct yolimbikitsidwa ngati scaffold system yokhala ndi mphamvu zabwino zakuthupi komanso zamakina (Chithunzi 1).Chifukwa cha kuthekera komanso luso laukadaulo, madokotala ochita opaleshoni amakonda kugwiritsa ntchito zida zoyezera ulusi wa thonje (Ct) panthawi ya maopaleshoni ndi mavalidwe. Choncho, poganizira ubwino wake wambiri, kuphatikizapo> 90% crystalline cellulose (imathandizira kupititsa patsogolo ntchito ya hemostatic), Ct inagwiritsidwa ntchito ngati chigoba cha NFRCS9,10. Choncho, poganizira ubwino wake wambiri, kuphatikizapo> 90% crystalline cellulose (imathandizira kupititsa patsogolo ntchito ya hemostatic), Ct inagwiritsidwa ntchito ngati chigoba cha NFRCS9,10. Следовательно, учитывая его многочисленные преимущества, в том числе > 90% кристаллической целлюлозы (участвущет в высположение ) пользовали в качестве скелетной системы NFRCS9,10. Choncho, chifukwa cha ubwino wake wambiri, kuphatikizapo> 90% crystalline cellulose (yomwe imakhudzidwa ndi ntchito yowonjezereka ya hemostatic), Ct inagwiritsidwa ntchito monga 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%Choncho, chifukwa cha ubwino wake wambiri, kuphatikizapo 90% crystalline cellulose (imathandizira kupititsa patsogolo ntchito ya hemostatic), Ct idagwiritsidwa ntchito ngati scaffold ya NFRCS9,10.Ct idakutidwa mwapang'onopang'ono (kupangidwa kwa filimu ya nano-thick) ndikulumikizana ndi hemostatic film-forming composition (HFFC).HFFC imachita ngati matrigel, yogwira Ct mosintha.Mapangidwe opangidwa amatumiza kupsinjika mkati mwa gawo lobalalika (kulimbitsa ulusi).Ndizovuta kupeza zida za nanoporous zokhala ndi mphamvu zamakina abwino pogwiritsa ntchito ma polima ochepa.Kuphatikiza apo, sikophweka kusinthira makonda osiyanasiyana amawumbidwe osiyanasiyana azachipatala.
Chithunzichi chikuwonetsa mawonekedwe a NFRCS potengera mawonekedwe a mapiko a tombolombo (A).Chithunzichi chikuwonetsa kufanana kwa mapiko a dragonfly (mitsempha yodutsana ndi yotalikirapo ya mapiko imalumikizana) ndi chithunzi chojambula cha Cp NFRCS (B).Kuyimira kwadongosolo kwa NFRCS.
Ma NFRC adapangidwa pogwiritsa ntchito HFFC ngati gawo lopitilira kuthana ndi malire omwe ali pamwambapa.HFFC imapangidwa ndi ma polima osiyanasiyana opangira filimu a hemostatic kuphatikiza chitosan (monga polima wamkulu wa hemostatic) wokhala ndi methylcellulose (MC), hydroxypropyl methylcellulose (HPMC 50 cp) ndi polyvinyl alcohol (PVA)) (125 kDa) monga polima wothandizira omwe amalimbikitsa mapangidwe a thrombus.mapangidwe.Kuphatikizika kwa polyvinylpyrrolidine K30 (PVP K30) kumathandizira kuyamwa kwa chinyezi cha NFRCS.Polyethylene glycol 400 (msomali 400) adawonjezedwa kuti apititse patsogolo kuphatikizika kwa polima polumikizana ndi polima.Mitundu itatu yosiyanasiyana ya HFFC ya hemostatic (Cm HFFC, Ch HFFC ndi Cp HFFC), yomwe ndi chitosan yokhala ndi MC (Cm), chitosan yokhala ndi HPMC (Ch), ndi chitosan yokhala ndi PVA (Cp), idagwiritsidwa ntchito ku Ct.Kafukufuku wosiyanasiyana wa in vitro ndi mu vivo watsimikizira ntchito ya machiritso a hemostatic ndi mabala a NFRCS.Zida zophatikizika zoperekedwa ndi NFRCS zitha kugwiritsidwa ntchito kusinthira makonda amitundu yosiyanasiyana kuti akwaniritse zosowa zenizeni.
Kuonjezera apo, NFRCS ikhoza kusinthidwa ngati bandeji kapena mpukutu kuti uphimbe malo onse ovulala a m'munsi ndi mbali zina za thupi.Makamaka pakuvulala kwa miyendo yakumenyana, kapangidwe kake ka NFRCS kangathe kusinthidwa kukhala theka la mkono kapena mwendo wathunthu (Supplementary Figure S11).NFRCS ikhoza kupangidwa kukhala chomangira chapamanja chokhala ndi minofu guluu, chomwe chingagwiritsidwe ntchito kuletsa magazi kuvulala koopsa m'manja.Cholinga chathu chachikulu ndikukhazikitsa NFRCS yokhala ndi polima pang'ono momwe tingathere yomwe ingathe kuperekedwa kwa anthu ambiri (pansi pa umphawi waumphawi) ndipo ikhoza kuikidwa mu chida choyamba chothandizira.Kapangidwe kosavuta, kothandiza, komanso kandalama, NFRCS imapindulitsa anthu amderalo ndipo imatha kukhudza dziko lonse lapansi.
Chitosan (molecular weight 80 kDa) ndi amaranth anagulidwa ku Merck, India.Hydroxypropyl methylcellulose 50 Cp, polyethylene glycol 400 ndi methylcellulose anagulidwa ku Loba Chemie Pvt.LLC, Mumbai.Polyvinyl mowa (mamolekyu olemera 125 kDa) (87-90% hydrolysed) anagulidwa ku National Chemicals, Gujarat.Polyvinylpyrrolidine K30 idagulidwa kuchokera ku Molychem, Mumbai, ma swabs osabala adagulidwa ku Ramaraju Surgery Cotton Mills Ltd., Tamil Nadu, ndi Milli Q water (Direct-Q3 water purification system, Merck, India) monga chonyamulira.
NFRCS idapangidwa pogwiritsa ntchito njira ya lyophilization11,12.Zolemba zonse za HFFC (Table 1) zidakonzedwa pogwiritsa ntchito makina oyambitsa makina.Konzani yankho la 0.5% la chitosan pogwiritsira ntchito 1% asidi acetic m'madzi mwa kusonkhezera mosalekeza pa 800 rpm pa choyambitsa makina.Kulemera kwenikweni kwa polima yodzaza yomwe yasonyezedwa mu Table 1 inawonjezeredwa ku yankho la chitosan ndikugwedezeka mpaka yankho lomveka bwino la polima litapezeka.PVP K30 ndi msomali 400 anawonjezedwa ku zotsatira kusakaniza mu ndalama zasonyezedwa Table 1, ndi kusonkhezera anapitilizidwa mpaka momveka viscous polima njira anapezedwa.The chifukwa kusamba wa polima njira anali sonicated kwa mphindi 60 kuchotsa atsekeredwa mpweya thovu ku polima osakaniza.Monga momwe tawonetsera mu Supplementary Figure S1 (b), Ct inagawidwa mofanana mu chitsime chilichonse cha mbale ya 6-chitsime (mold) yowonjezeredwa ndi 5 ml ya HFFC.
Mbale zisanu ndi chimodzi bwino sonicated kwa mphindi 60 kukwaniritsa yunifolomu wetting ndi kugawa HFFC mu Ct maukonde.Ndiye amaundana mbale zisanu ndi chimodzi bwino pa -20 ° C kwa maola 8-12.Ma mbale amaundana anali lyophilized kwa maola 48 kuti apeze mitundu yosiyanasiyana ya NFRCS.Njira yomweyi imagwiritsidwa ntchito popanga mawonekedwe ndi mapangidwe osiyanasiyana, monga ma tampons kapena ma cylindrical tampons, kapena mawonekedwe ena aliwonse ogwiritsira ntchito zosiyanasiyana.
Chitosan yoyezedwa bwino (80 kDa) (3%) imasungunuka mu 1% acetic acid pogwiritsa ntchito magnetic stirrer.Kuti chifukwa njira ya chitosan anawonjezera 1% msomali 400 ndi analimbikitsa kwa mphindi 30.Thirani yankholo mu chidebe cha lalikulu kapena lamakona anayi ndikuzizira pa -80 ° C kwa maola 12.Achisanu zitsanzo anali lyophilized kwa maola 48 kupeza porous Cs13.
NFRCS yopangidwa idayesedwa pogwiritsa ntchito Fourier transform infrared spectroscopy (FTIR) (Shimadzu 8400 s FTIR, Tokyo, Japan) kutsimikizira kuyanjana kwa mankhwala a chitosan ndi ma polima ena14,15.Mawonekedwe a FTIR (m'lifupi mwa mawonekedwe a spectral kuyambira 400 mpaka 4000 cm-1) a zitsanzo zonse zoyesedwa adapezedwa popanga masikelo 32.
Mlingo wa mayamwidwe amagazi (BAR) pamapangidwe onse adawunikidwa pogwiritsa ntchito njira yofotokozedwa ndi Chen et al.16 ndi zosintha pang'ono.Ma NFRK opangidwa mwazolemba zonse adawumitsidwa mu uvuni wa vacuum pa 105 ° C usiku wonse kuti achotse zosungunulira zotsalira.30 mg NFRCS (chitsanzo choyambirira cholemera - W0) ndi 30 mg Ct (kulamulira kwabwino) anayikidwa mu mbale zosiyana zomwe zili ndi premix ya 3.8% sodium citrate.Pazigawo zokonzedweratu za nthawi, mwachitsanzo 5, 10, 20, 30, 40 ndi 60 masekondi, NFRCS inachotsedwa ndipo malo awo amatsukidwa ndi magazi osatulutsidwa mwa kuika zitsanzo pa Ct kwa masekondi a 30.Kulemera komaliza kwa magazi otengedwa ndi NFRCS 16 kumaganiziridwa (W1) nthawi iliyonse.Kuwerengera kuchuluka kwa BAR pogwiritsa ntchito njira iyi:
Nthawi yotseka magazi (BCT) idatsimikiziridwa ndi Wang et al.17 .Nthawi yofunikira kwa magazi athunthu (magazi a makoswe osakanikirana ndi 3.8% sodium citrate) kuti atseke pamaso pa NFRCS adawerengedwa ngati BCT ya chitsanzo choyesera.Magawo osiyanasiyana a NFRCS (30 mg) adayikidwa mu 10 ml screw cap vials ndikumakulitsidwa pa 37°C.Magazi (0.5 ml) adawonjezedwa ku vial ndipo 0,3 ml ya 0.2 M CaCl2 adawonjezedwa kuti atsegule magazi.Pomaliza, tembenuzani vial masekondi 15 aliwonse (mpaka 180 °) mpaka kuundana kolimba.BCT yachitsanzoyo imayerekezedwa ndi kuchuluka kwa ma flips vails17,18.Kutengera ndi BCT, nyimbo ziwiri zabwino kwambiri zochokera ku NFRCS Cm, Ch ndi Cp zidasankhidwa kuti zipitilize maphunziro amunthu.
BCT ya Ch NFRCS ndi Cp NFRCS zolemba zidatsimikiziridwa ndikugwiritsa ntchito njira yomwe Li et al.19 .Ikani 15 x 15 mm2 Ch NFRCS, Cp NFRCS, ndi Cs (zowongolera zabwino) muzakudya zosiyana za Petri (37 °C).Magazi omwe ali ndi 3.8% sodium citrate adasakanizidwa ndi 0.2 M CaCl2 mu chiwerengero cha 10: 1 kuti ayambe kuyambitsa magazi.20 µl wa 0.2 M CaCl2 wosakaniza wa magazi a makoswe adayikidwa pachitsanzocho ndikuyikidwa m'mbale yopanda kanthu ya Petri.Kuwongolera kunali magazi otsanuliridwa m'mbale zopanda kanthu za Petri popanda Ct.Pazigawo zokhazikika za 0, 3, ndi 5 mphindi, siyani kutseka mwa kuwonjezera 10 ml ya madzi opangidwa ndi deionized (DI) ku chitsanzo chomwe chili ndi mbale popanda kusokoneza chivundikirocho.Ma erythrocyte (erythrocyte) osakhazikika amapita ku hemolysis pamaso pa madzi osungunuka ndikutulutsa hemoglobin.Hemoglobin pa nthawi zosiyanasiyana (HA(t)) inayesedwa pa 540 nm (λmax hemoglobin) pogwiritsa ntchito UV-Vis spectrophotometer.Mayamwidwe amtheradi a hemoglobini (AH(0)) mu 0 min ya 20 µl ya magazi mu 10 ml ya madzi a deionized adatengedwa ngati muyezo.Kukwezeka kwa hemoglobini (RHA) wamagazi olumikizana adawerengedwa kuchokera ku chiŵerengero cha HA (t)/HA(0) pogwiritsa ntchito mtanda womwewo wa magazi.
Pogwiritsa ntchito zojambula zojambula (Texture Pro CT V1.3 Build 15, Brookfield, USA), zomatira za NFRK ku minofu yowonongeka zinatsimikiziridwa.Kanikizani mbale ya cylindrical yotseguka pansi mkati mwa khungu la nkhumba (popanda mafuta osanjikiza).Zitsanzo (Ch NFRCS ndi Cp NFRCS) zinagwiritsidwa ntchito kudzera mu cannula mu nkhungu za cylindrical kuti apange kumatira pakhungu la nkhumba.Pambuyo pa kutsekemera kwa mphindi 3 pa kutentha kwa chipinda (RT) (25 ° C.), mphamvu zomatira za NFRCS zinalembedwa pamlingo wokhazikika wa 0.5 mm / sec.
Mbali yaikulu ya zosindikizira opaleshoni ndi kuonjezera magazi kuundana pamene kuchepetsa kutaya magazi.Coagulation yopanda kutaya mu NFRCS idawunikidwa pogwiritsa ntchito njira yomwe idasindikizidwa kale ndikusintha pang'ono 19.Pangani chubu cha microcentrifuge (2 ml) (m'mimba mwake 10 mm) ndi dzenje la 8 × 5 mm2 kumbali imodzi ya chubu la centrifuge (loyimira bala lotseguka).NFRCS imagwiritsidwa ntchito kutseka kutsegulira ndipo tepi imagwiritsidwa ntchito kusindikiza m'mphepete mwakunja.Onjezani 20 µl ya 0.2 M CaCl2 ku chubu cha microcentrifuge chomwe chili ndi 3.8% sodium citrate premix.Pambuyo pa mphindi 10, machubu a microcentrifuge adachotsedwa mu mbale ndipo kuwonjezeka kwa mbale kunatsimikiziridwa chifukwa cha kutuluka kwa magazi kuchokera ku NFRK (n = 3).Kutaya magazi Ch NFRCS ndi Cp NFRCS anafanizidwa ndi Cs.
Kukhulupirika konyowa kwa NFRCS kudatsimikiziridwa kutengera njira yofotokozedwa ndi Mishra ndi Chaudhary21 ndi zosintha zazing'ono.Ikani NFRCS mu botolo la 100 ml Erlenmeyer ndi madzi 50 ml ndi kuzungulira kwa 60 s popanda kupanga pamwamba.Kuyang'ana kowoneka ndi kuika patsogolo kwa zitsanzo za umphumphu wa thupi potengera kusonkhanitsa.
Mphamvu yomangirira ya HFFC ku Ct idaphunziridwa pogwiritsa ntchito njira zomwe zidasindikizidwa kale ndi zosintha zazing'ono.Kukhulupirika kwa zokutira pamwamba kunayesedwa powonetsa NFRK ku mafunde acoustic (zolimbikitsa zakunja) pamaso pa milliQ madzi (Ct).Zomwe zinapangidwa NFRCS Ch NFRCS ndi Cp NFRCS zinayikidwa mu beaker yodzazidwa ndi madzi ndi sonicated kwa 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, ndi 30 min, motero.Pambuyo poyanika, kusiyana kwa chiwerengero pakati pa kulemera koyambirira ndi komaliza kwa NFRCS kunagwiritsidwa ntchito kuwerengera peresenti ya kutaya kwa zinthu (HFFC).In vitro BCT idathandiziranso mphamvu zomangira kapena kutayika kwa zinthu zapamtunda.Kugwira ntchito bwino kwa HFFC kumangiriza ku Ct kumapereka magazi kukhazikika komanso zokutira zotanuka pamwamba pa Ct22.
Homogeneity ya NFRCS yopangidwa idatsimikiziridwa ndi BCT ya zitsanzo (30 mg) zotengedwa kuchokera kumalo osankhidwa mwachisawawa a NFRCS.Tsatirani ndondomeko ya BCT yomwe yatchulidwa kale kuti muwone ngati NFRCS ikutsatiridwa.Kuyandikira pakati pa zitsanzo zonse zisanu kumatsimikizira kuphimba kofanana ndi mawonekedwe a HFFC mu Ct mesh.
Malo okhudzana ndi magazi (NBCA) adatsimikiziridwa monga momwe adanenera kale ndi zosintha zina.Thirani magazi pomanga 20 µl ya magazi pakati pa malo awiri a Ct, Ch NFRCS, Cp NFRCS ndi Cs.Pambuyo pa ola la 1, magawo awiri a stent adalekanitsidwa ndikuyesa pamanja gawo la clot.Mtengo wapakati wa kubwereza katatu unkaganiziridwa kuti NBCA NFRCS19.
Kusanthula kwa Dynamic Vapor Sorption (DVS) kunagwiritsidwa ntchito poyesa mphamvu ya NFRCS kuti itenge madzi kuchokera ku chilengedwe chakunja kapena kuchokera kumalo ovulala omwe amachititsa kuti ayambe kusungunuka.DVS imayang'ana kapena kulemba kuchuluka kwa nthunzi ndi kutayika kwachitsanzo chokoka pogwiritsa ntchito ultra-sensitive balance yokhala ndi kusamvana kwakukulu kwa ±0.1 µg.Kuthamanga pang'ono kwa nthunzi (chinyezi chofanana) kumapangidwa ndi chowongolera zamagetsi zamagetsi kuzungulira chitsanzocho posakaniza mpweya wodzaza ndi wowuma. Malinga ndi malangizo a European Pharmacopeia, kutengera kuchuluka kwa chinyezi chomwe chatengedwa ndi zitsanzo, zitsanzo zidagawidwa m'magulu anayi (0-0.012% w/w-non-hygroscopic, 0.2-2% w/w pang'ono hygroscopic, 2-15% modekha hygroscopic, ndi> 35 hygroscopic) kwambiri. Malinga ndi malangizo a European Pharmacopeia, kutengera kuchuluka kwa chinyezi chomwe chimatengedwa ndi zitsanzo, zitsanzo zidagawidwa m'magulu 4 (0-0.012% w/w-non-hygroscopic, 0.2-2% w/w pang'ono hygroscopic, 2-15 % moderate hygroscopic, ndi> 315 hygroscopic) kwambiri.Malinga ndi malangizo a European Pharmacopoeia, malinga ndi kuchuluka kwa mayamwidwe a chinyezi ndi zitsanzo, zitsanzozo zidagawidwa m'magulu a 4 (0-0.012% w / w - non-hygroscopic, 0.2-2% w / w pang'ono hygroscopic, 2-15%).% умеренно гигроскопичен и > 15% очень гигроскопичен)23. % ya hygroscopic pang'ono ndi> 15% ya hygroscopic kwambiri)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/w-澎靿姽 靿姽 %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 分为 分为 类 %済/w-10 , , , 0.2-2% W/w 轻微, 2-15% 适度 吸湿 ，> 15 %非常吸湿）23.Malinga ndi malingaliro a European Pharmacopoeia, zitsanzo zimagawidwa m'makalasi 4 kutengera kuchuluka kwa chinyezi chomwe chimatengedwa ndi chitsanzo (0-0.012% ndi kulemera - non-hygroscopic, 0.2-2% ndi kulemera pang'ono hygroscopic, 2-15% kulemera).% умеренно гигроскопичен, > 15% очень гигроскопичен) 23. % ndi hygroscopic pang'ono, > 15% ya hygroscopic kwambiri) 23.Kuchita bwino kwa NFCS X NFCS ndi TsN NFCS kudatsimikiziridwa pa analyzer DVS TA TGA Q5000 SA.Panthawiyi, nthawi yothamanga, chinyezi chachibale (RH), ndi kulemera kwa nthawi yeniyeni pa 25 ° C24 zinapezedwa.Chinyezi chimawerengedwa ndi kusanthula kolondola kwa kuchuluka kwa NFRCS pogwiritsa ntchito equation iyi:
MC ndi chinyezi cha NFRCS.m1 - kulemera kouma kwa NSAIDs.m2 ndi nthawi yeniyeni ya NFRCS misa pa RH yoperekedwa.
Dera lonselo linayesedwa pogwiritsa ntchito kuyesa kwa nitrogen adsorption ndi nayitrogeni wamadzimadzi pambuyo pochotsa zitsanzo pa 25 °C kwa 10 h (<7 × 10-3 Torr). Dera lonselo linayesedwa pogwiritsa ntchito kuyesa kwa nitrogen adsorption ndi nayitrogeni wamadzimadzi pambuyo pochotsa zitsanzo pa 25 °C kwa 10 h (<7 × 10-3 Torr). Общая площадь поверхности оценивалась с помощью эксперимента по адсорбции азота жидким азотом после опорожнения образцов при 12 × 12 × 12 × 12 × 12 × 3 gawo). Malo onsewa adayesedwa pogwiritsa ntchito kuyesa kwa nitrogen adsorption ndi nayitrogeni wamadzimadzi pambuyo poti zitsanzo zidatsanulidwa pa 25 ° C kwa 10 h (<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。pa 25°C Werengani zambiri za momwe mungapangire zinthu zochititsa chidwi pa nkhani ya 20 z °C (< 7 × 10-3 тор). Malo onsewa adayesedwa pogwiritsa ntchito kuyesa kwa nitrogen adsorption ndi nayitrogeni wamadzimadzi pambuyo poti zitsanzo zidatsanulidwa kwa maola 10 pa 25°C (<7 x 10-3 torr).Malo onse, kuchuluka kwa pore ndi kukula kwa pore ya NFRCS kudatsimikiziridwa ndi Quantachrome kuchokera ku NOVA 1000e, Austria pogwiritsa ntchito pulogalamu ya RS 232.
Konzani 5% RBCs (saline ngati diluent) kuchokera m'magazi athunthu.Kenako tumizani aliquot ya HFFC (0.25 ml) ku mbale ya 96-chitsime ndi 5% RBC mass (0.1 ml).Yambani kusakaniza pa 37 ° C kwa mphindi 40.Kusakaniza kwa maselo ofiira a magazi ndi seramu kunkaonedwa kuti ndi njira yabwino yolamulira, komanso kusakaniza kwa saline ndi maselo ofiira a magazi monga kulamulira koipa.Hemagglutination inatsimikiziridwa molingana ndi mlingo wa Stajitzky.Mamba omwe akuperekedwawo ndi awa: + + + + zowunjikana zowunjikana;+ + + zosalala pansi zokhala ndi m’mphepete mwake;+ + mapadi osalala pansi okhala ndi m’mbali zong’ambika;+ mphete zofiira zopapatiza kuzungulira m’mphepete mwa mapepala osalala;- (zoipa) zowonekera zofiira batani 12 pakati pa chitsime chapansi.
The hemocompatibility wa NFRCS anaphunzira molingana ndi njira ya International Organisation for Standardization (ISO) (ISO10993-4, 1999)26,27.Njira ya gravimetric yofotokozedwa ndi Singh et al.Zosintha zazing'ono zidapangidwa kuti ziwone mapangidwe a thrombus pamaso pa kapena pamwamba pa NFRCS.500 mg ya Cs, Ch NFRCS ndi Cp NFRCS inayikidwa mu phosphate buffered saline (PBS) kwa maola 24 pa 37 ° C.Pambuyo pa maola 24, PBS inachotsedwa ndipo NFRCS inachiritsidwa ndi 2 ml ya magazi omwe ali ndi 3.8% sodium citrate.Pamwamba pa NFRCS, onjezerani 0.04 ml ya 0.1 M CaCl2 ku zitsanzo zokhala ndi incubated.Pambuyo pa mphindi 45, 5 ml ya madzi osungunuka anawonjezeredwa kuti asiye kusungunuka.Magazi osungunuka pamwamba pa NFRK adathandizidwa ndi 36-38% ya formaldehyde solution.Maunda opangidwa ndi formaldehyde amawumitsidwa ndikuyesedwa.Kuchuluka kwa thrombosis kunayesedwa powerengera kulemera kwa galasi popanda magazi ndi chitsanzo (kulamulira koipa) ndi galasi lokhala ndi magazi (kuwongolera kwabwino).
Monga chitsimikiziro choyambirira, zitsanzozo zinawonetsedwa pansi pa microscope ya kuwala kuti amvetse mphamvu ya HFFC yophimba pamwamba, Ct interconnected, ndi Ct network kupanga pores.Magawo owonda a Ch ndi Cp ochokera ku NFRCS adakonzedwa ndi tsamba la scalpel.Gawo lotsatila linayikidwa pa galasi la galasi, lophimbidwa ndi chophimba, ndipo m'mphepete mwake munakonzedwa ndi guluu.Zithunzi zokonzedwazo zidawonedwa pansi pa maikulosikopu ya kuwala ndipo zithunzi zidajambulidwa mokulira mosiyanasiyana.
Kuyika kwa polima mu Ct network kunawonetsedwa pogwiritsa ntchito ma microscopy a fluorescence kutengera njira yofotokozedwa ndi Rice et al.29. Mapangidwe a HFFC omwe amagwiritsidwa ntchito popangawo adasakanizidwa ndi utoto wa fulorosenti (amaranth), ndipo NFRCS (Ch & Cp) adakonzedwa motsatira njira yomwe tatchula kale. Mapangidwe a HFFC omwe amagwiritsidwa ntchito popangawo adasakanizidwa ndi utoto wa fulorosenti (amaranth), ndipo NFRCS (Ch & Cp) adakonzedwa motsatira njira yomwe tatchula kale.Mapangidwe a HFFC omwe amagwiritsidwa ntchito popanga adasakanizidwa ndi utoto wa fulorosenti (amaranth) ndi NFRCS (Ch ndi Cp) adapezedwa molingana ndi njira yomwe yatchulidwa kale.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））Mapangidwe a HFFC omwe adagwiritsidwa ntchito popanga adasakanizidwa ndi utoto wa fulorosenti (Amaranth) ndipo adalandira NFRCS (Ch ndi Cp), monga tafotokozera kale.Magawo owonda a NFRK adadulidwa kuchokera ku zitsanzo zomwe adapeza, kuyikidwa pazithunzi zamagalasi, ndikukutidwa ndi zokopa.Yang'anani zithunzi zomwe zakonzedwa pansi pa microscope ya fulorosenti pogwiritsa ntchito fyuluta yobiriwira (310-380 nm).Zithunzi zidatengedwa pakukula kwa 4x kuti amvetsetse maubwenzi a Ct komanso kuyika kowonjezera kwa polima mu network ya Ct.
Mawonekedwe a pamwamba a NFRCS Ch ndi Cp adatsimikiziridwa pogwiritsa ntchito microscope ya atomic force (AFM) yokhala ndi ultra-lap TESP cantilever mu tapping mode: 42 N / m, 320 kHz, ROC 2-5 nm, Bruker, Taiwan.Kukula kwapamtunda kudatsimikiziridwa ndi root mean square (RMS) pogwiritsa ntchito mapulogalamu (Scan Probe Image Processor).Malo osiyanasiyana a NFRCS anaperekedwa pazithunzi za 3D kuti ayang'ane kufanana kwapamwamba.Kupatuka koyenera kwa zigoli kudera lomwe mwapatsidwa kumatanthauzidwa ngati makulidwe a pamwamba.Equation ya RMS idagwiritsidwa ntchito kuwerengera kuuma kwapamwamba kwa NFRCS31.
Maphunziro opangidwa ndi FESEM adachitidwa pogwiritsa ntchito FESEM, SU8000, HI-0876-0003, Hitachi, Tokyo, kuti amvetsetse mawonekedwe a pamwamba a Ch NFRCS ndi Cp NFRCS, omwe adawonetsa bwino BCT kuposa Cm NFRCS.Phunziro la FESEM linachitidwa molingana ndi njira yofotokozedwa ndi Zhao et al.32 ndi zosintha zazing'ono.NFRCS 20 mpaka 30 mg Ch NFRCS ndi Cp NFRCS zinali zosakanikirana ndi 20 µl ya 3.8% sodium citrate yosakanikirana ndi magazi a makoswe.20 μl ya 0.2 M CaCl2 inawonjezedwa ku zitsanzo zowonongeka kwa magazi kuti ayambe kugwirizanitsa ndipo zitsanzozo zinakulungidwa kutentha kwa mphindi 10.Kuphatikiza apo, ma erythrocyte ochulukirapo adachotsedwa pamwamba pa NFRCS potsuka ndi saline.
Zitsanzo zotsatizanazi zinkachitidwa ndi 0.1% glutaraldehyde ndikuwumitsa mu uvuni wotentha wa 37 ° C kuti muchotse chinyezi.Zitsanzo zouma zidakutidwa ndikusanthula 32.Zithunzi zina zomwe zinapezedwa panthawi ya kusanthula zinali kupangidwa kwa magazi pamwamba pa ulusi wa thonje la munthu, polima polima pakati pa Ct, erythrocyte morphology (mawonekedwe), clot integrity, ndi erythrocyte morphology pamaso pa NFRCS.Madera a NFRCS osasamalidwa ndi Ch ndi Cp omwe amachitira madera a NFRCS opangidwa ndi magazi adafufuzidwa kuti apange ma ion elemental (sodium, potaziyamu, nayitrogeni, calcium, magnesium, zinki, mkuwa ndi selenium)33.Fananizani magawo oyambira a ma ion pakati pa zitsanzo zothandizidwa ndi zosagwiritsidwa ntchito kuti mumvetsetse kudziunjikira kwa ma ion panthawi yopanga magazi komanso kuphatikizika kwa clot.
Makulidwe a Cp HFFC opaka pamwamba pa Ct adatsimikiziridwa pogwiritsa ntchito FSEM.Magawo amtundu wa Cp NFRCS adadulidwa kuchokera pachimake ndikukutidwa ndi sputter.Zitsanzo zowonongeka za sputter zinawonedwa ndi FSEM ndipo makulidwe a pamwamba pake amayesedwa 34, 35, 36.
X-ray yaying'ono-CT imapereka chithunzithunzi chapamwamba cha 3D chosawononga ndikukulolani kuti muphunzire kakonzedwe kamkati ka NFRK.Micro-CT imagwiritsa ntchito mtengo wa X-ray womwe umadutsa pachitsanzo kuti ijambule gawo laling'ono la X-ray lachitsanzo, lomwe limathandiza kupeza chidziwitso cha morphological.Malo amkati a Ct mu Cp NFRCS ndi Cp NFRCS yopangidwa ndi magazi anayesedwa ndi micro-CT kuti amvetse bwino kuyamwa ndi kutsekeka kwa magazi pamaso pa NFRCS37,38,39.Mapangidwe a 3D a zitsanzo za Cp NFRCS zothandizidwa ndi magazi komanso zosagwiritsidwa ntchito zinamangidwanso pogwiritsa ntchito micro-CT (V|tome|x S240, Phoenix, Germany).Pogwiritsa ntchito pulogalamu ya VG STUDIO-MAX 2.2, zithunzi zingapo za X-ray zidatengedwa kuchokera kosiyanasiyana (koyenera kuphimba 360 °) kuti apange zithunzi za 3D za NFRCS.Zomwe zasonkhanitsidwa zidapangidwanso kukhala zithunzi za 3D volumetric pogwiritsa ntchito pulogalamu yosavuta yofananira ya 3D ScanIP Academic.
Kuonjezera apo, kuti amvetse kugawidwa kwa magazi, 20 µl ya premixed citrated magazi ndi 20 µl ya 0.2 M CaCl2 anawonjezedwa ku NFRCS kuti ayambe kutseka kwa magazi.Zitsanzo zokonzeka zimasiyidwa kuti ziwumitsidwe.Malo a NFRK adachitidwa ndi 0.5% glutaraldehyde ndikuwumitsa mu uvuni wotentha wa 30-40 ° C kwa 30 min.Chophimba cha magazi chomwe chinapangidwa pa NFRCS chinafufuzidwa, chinamangidwanso, ndipo chithunzi cha 3D cha magazi a magazi chinawonetsedwa.
Kuyeza kwa antibacterial kunachitika pa Cp NFRCS (yopambana kwambiri ndi Ch NFRCS) pogwiritsa ntchito njira yomwe tafotokozayi ndi zosintha zazing'ono.Ntchito ya antibacterial ya Cp NFRCS ndi Cp HFFC idatsimikiziridwa pogwiritsa ntchito tizilombo tating'ono tating'ono tating'ono tating'ono tating'ono toyesa [S.aureus (mabakiteriya a gram-positive), E.coli (mabakiteriya a gram-negative) ndi white Candida (C.albicans)] akukula pa agar mu Petri mbale mu chofungatira.Ikani 50 ml ya kuyimitsidwa kwa chikhalidwe cha bakiteriya mofananira pamlingo wa 105-106 CFU ml-1 pa sing'anga ya agar.Thirani sing'anga mu mbale ya Petri ndipo mulole kuti ikhale yolimba.Zitsime zidapangidwa pamwamba pa mbale ya agar kuti zidzaze ndi HFFC (zitsime zitatu za HFFC ndi 1 zowongolera zoyipa).Onjezani 200 µl HFFC ku zitsime zitatu ndi 200 µl pH 7.4 PBS pachitsime chachinayi.Kumbali ina ya mbale ya petri, ikani 12 mm Cp NFRCS disk pa agar yolimba ndikunyowetsa ndi PBS (pH 7.4).Mapiritsi a Ciprofloxacin, ampicillin ndi fluconazole amatengedwa ngati miyezo ya Staphylococcus aureus, Escherichia coli ndi Candida albicans.Yezerani madera oletsa pamanja ndi kutenga chithunzi cha digito cha zone yoletsa.
Pambuyo pa chivomerezo cha chikhalidwe cha chikhalidwe, phunziroli linachitidwa ku Kasturba Medical College of Education and Research ku Manipal, Karnataka, kum'mwera kwa India.Protocol yoyeserera ya in vitro TEG yawunikiridwa ndikuvomerezedwa ndi Institutional Ethics Committee ya Kasturba Medical College, Manipal, Karnataka (IEC: 674/2020).Nkhani zinatengedwa kuchokera kwa opereka mwazi odzifunira (azaka 18 mpaka 55) kuchokera ku nkhokwe yosungira mwazi m’chipatala.Kuonjezera apo, fomu yovomerezeka yodziwitsidwa inapezedwa kuchokera kwa odzipereka kuti atenge zitsanzo za magazi.Native TEG (N-TEG) ​​​​inagwiritsidwa ntchito pofufuza momwe Cp HFFC imapangidwira pamagazi athunthu osakanikirana ndi sodium citrate.N-TEG imadziwika kwambiri chifukwa cha ntchito yake yotsitsimutsa mfundo, zomwe zimabweretsa mavuto kwa asing'anga chifukwa cha kuthekera kwa kuchedwa kwachipatala pazotsatira (mayeso anthawi zonse a coagulation).Kusanthula kwa N-TEG kunkachitika pogwiritsa ntchito magazi athunthu.Chilolezo chodziwitsidwa komanso mbiri yakale yachipatala idapezedwa kwa onse omwe adatenga nawo gawo.Phunzirolo silinaphatikizepo omwe ali ndi vuto la hemostatic kapena thrombotic monga mimba / postpartum kapena matenda a chiwindi.Anthu omwe amamwa mankhwala omwe amakhudza coagulation cascade adachotsedwanso pa kafukufukuyu.Mayeso a labotale oyambira (hemoglobin, prothrombin time, activated thromboplastin ndi platelet count) adachitidwa kwa onse omwe adatenga nawo gawo molingana ndi njira zokhazikika.N-TEG imatsimikizira kuti magazi a clot viscoelasticity, mapangidwe oyambirira a magazi, kuyanjana kwa tinthu, kulimbitsa magazi, ndi clot lysis.Kusanthula kwa N-TEG kumapereka chidziwitso chazithunzi komanso manambala pazotsatira zamagulu angapo a ma cell ndi plasma.Kusanthula kwa N-TEG kunachitika pamitundu iwiri yosiyana ya Cp HFFC (10 µl ndi 50 µl).Zotsatira zake, 1 ml ya magazi athunthu okhala ndi citric acid adawonjezeredwa ku 10 μl ya Cp HFFC.Onjezani 1 ml (Cp HFFC + citrated blood), 340 µl magazi osakaniza mpaka 20 µl 0.2 M CaCl2 okhala ndi TEG mbale.Pambuyo pake, mbale za TEG zidakwezedwa mu TEG® 5000, US kuyeza R, K, alpha angle, MA, G, CI, TPI, EPL, LY 30% ya zitsanzo za magazi pamaso pa Cp HFFC41.
Njira yophunzirira mu vivo idawunikiridwa ndikuvomerezedwa ndi Institutional Animal Ethics Committee (IAEC), Kasturba School of Medicine, Manipal Institute of Higher Education, Manipal (IAEC/KMC/69/2020).Zoyeserera zonse za nyama zidachitika motsatira malingaliro a Komiti Yoyang'anira ndi Kuyang'anira Kuyesa Kwa Zinyama (CPCSEA).Maphunziro onse a vivo NFRCS (2 × 2 cm2) adachitidwa pa makoswe aakazi a Wistar (olemera 200 mpaka 250 g).Zinyama zonse zidazolowera kutentha kwa 24-26 ° C, nyamazo zinali ndi mwayi wopeza chakudya chokwanira komanso madzi ad libitum.Nyama zonse zidagawidwa mwachisawawa m'magulu osiyanasiyana, gulu lililonse limakhala ndi nyama zitatu.Maphunziro onse anachitidwa molingana ndi Maphunziro a Zinyama: Lipoti la In Vivo Experiments 43 .Phunziroli lisanachitike, nyamazo zinagwidwa ndi anesthetized ndi intraperitoneal (ip) kayendetsedwe ka chisakanizo cha 20-50 mg ya ketamine (pa 1 kg ya kulemera kwa thupi) ndi 2-10 mg wa xylazine (pa 1 kg ya kulemera kwa thupi).Pambuyo pa phunzirolo, kuchuluka kwa magazi kunawerengedwa poyesa kusiyana pakati pa kulemera koyambirira ndi komaliza kwa zitsanzo, mtengo wapakati womwe umapezeka kuchokera ku mayesero atatuwo unatengedwa ngati kuchuluka kwa magazi kwa chitsanzo.
Njira yodulira mchira wa makoswe idakhazikitsidwa kuti imvetsetse kuthekera kwa NFRCS kuwongolera magazi pamavuto, kumenya, kapena ngozi yapamsewu (chitsanzo chovulaza).Dulani 50% ya mchira ndi tsamba la scalpel ndikuyika mumlengalenga kwa masekondi 15 kuti muwonetsetse kuti magazi abwinobwino.Kuonjezera apo, zitsanzo zoyesera zinayikidwa pa mchira wa makoswe pogwiritsa ntchito kukakamiza (Ct, Cs, Ch NFRCS ndi Cp NFRCS).Kukhetsa magazi ndi PCT zidanenedwa pazoyeserera (n = 3) 17,45.
Kuchita bwino kwa kuwongolera kuthamanga kwa NFRCS pankhondo kudafufuzidwa pachitsanzo cha mtsempha wachikazi wapamwamba kwambiri.Mitsempha yachikazi imawululidwa, imakhomedwa ndi 24G trocar, ndikutuluka magazi mkati mwa masekondi a 15.Pambuyo pakutuluka kwa magazi kosalamulirika, chitsanzo choyesera chimayikidwa pamalo okhomerera ndikukakamiza.Atangogwiritsa ntchito chitsanzo choyesera, nthawi yotseka inalembedwa ndipo mphamvu ya hemostatic inawonedwa kwa mphindi 5 zotsatira.Njira yomweyo idabwerezedwanso ndi Cs ndi CT46.
Dowling et al.47 anakonza chitsanzo chovulaza chiwindi kuti awone mphamvu ya hemostatic ya zipangizo za hemostatic ponena za kutuluka kwa magazi mkati mwa opaleshoni.BCT inalembedwa pa zitsanzo za Ct (zoletsa zoipa), Cs framework (positive control), zitsanzo za Ch NFRCS, ndi zitsanzo za Cp NFRCS.The suprahepatic vena cava wa khoswe adawonekera pochita laparotomy yapakati.Pambuyo pake, mbali yakutali ya lobe yakumanzere idadulidwa ndi lumo.Pangani chocheka m'chiwindi ndi tsamba la scalpel ndikusiya magazi kwa masekondi angapo.Kuyeza molondola kwa Ch NFRCS ndi Cp NFRCS zitsanzo zoyesedwa zinayikidwa pamtunda wowonongeka popanda kukakamizidwa kwabwino ndipo BCT inalembedwa.Gulu lolamulira (Ct) kenako linagwiritsa ntchito kukakamiza kutsatiridwa ndi Cs 30 s47 popanda kuphwanya chovulalacho.
Mu vivo machiritso a mabala adayesedwa pogwiritsa ntchito chitsanzo cha bala kuti ayese machiritso a mabala a NFRCSs opangidwa ndi polima.Zitsanzo za zilonda zowonongeka zinasankhidwa ndikuchitidwa molingana ndi njira zofalitsidwa kale ndi zosinthidwa zazing'ono19,32,48.Zinyama zonse zidagonekedwa monga momwe tafotokozera kale.Gwiritsani ntchito nkhonya ya biopsy (12 mm) kuti mupange mozungulira mozungulira pakhungu lakumbuyo.Malo okonzekera mabala adavekedwa ndi Cs (kuwongolera zabwino), Ct (pozindikira kuti mapepala a thonje amasokoneza machiritso), Ch NFRCS ndi Cp NFRCS (gulu loyesera) ndi kulamulira kolakwika popanda chithandizo chilichonse.Pa tsiku lililonse la phunzirolo, dera la balalo linkayezedwa ndi makoswe onse.Gwiritsani ntchito kamera ya digito kuti mujambule malo a bala ndikuvala chovala chatsopano.Chiwerengero cha kutsekedwa kwa chilonda chinayesedwa ndi njira iyi:
Kutengera kuchuluka kwa kutsekedwa kwa bala pa tsiku la 12 la phunzirolo, khungu la makoswe a gulu labwino kwambiri linadulidwa ((Cp NFRCS) ndi gulu lolamulira) ndikuphunziridwa ndi madontho a H & E ndi Masson's trichrome staining. Kutengera kuchuluka kwa kutsekedwa kwa bala pa tsiku la 12 la phunzirolo, khungu la makoswe a gulu labwino kwambiri linadulidwa ((Cp NFRCS) ndi gulu lolamulira) ndikuphunziridwa ndi madontho a H & E ndi Masson's trichrome staining.Malingana ndi kuchuluka kwa chilonda chotsekedwa pa tsiku la 12 la phunzirolo, khungu la makoswe a gulu labwino kwambiri ((Cp NFRCS) ndi gulu lolamulira) linadulidwa ndikuyesedwa ndi kudetsa ndi hematoxylin-eosin ndi Masson's trichrome.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，进海行H&EMas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)Makoswe omwe ali m'gulu labwino kwambiri ((Cp NFRCS) ndi magulu olamulira) adadulidwa chifukwa cha madontho a hematoxylin-eosin ndi madontho a Masson's trichrome potengera kutsekedwa kwa bala pa tsiku la 12 la kafukufukuyu.Njira yodetsa yomwe idakhazikitsidwa idachitika motsatira njira zomwe zafotokozedwa kale49,50.Mwachidule, atatha kukonza mu 10% formalin, zitsanzozo zidachotsedwa madzi pogwiritsa ntchito ma alcohols angapo.Gwiritsani ntchito microtome kuti mupeze magawo owonda (5 µm wokhuthala) a minofu yodulidwayo.Magawo ocheperako owongolera ndi Cp NFRCS adathandizidwa ndi hematoxylin ndi eosin kuti aphunzire kusintha kwa histopathological.Madontho atatu a Masson adagwiritsidwa ntchito kuti azindikire mapangidwe a collagen fibrils.Zotsatira zomwe zidapezedwa zidaphunziridwa mwachimbulimbuli ndi akatswiri azachipatala.
Kukhazikika kwa zitsanzo za Cp NFRCS kunaphunziridwa kutentha (25 ° C ± 2 ° C / 60% RH ± 5%) kwa miyezi 1251.Cp NFRCS (kusinthika kwapamtunda ndi kukula kwa tizilombo tating'onoting'ono) idawunikidwa ndi kuyesedwa kuti ikhale yolimbana ndi kuvala ndi BCT malinga ndi njira zomwe zili pamwambazi zomwe zafotokozedwa mugawo la Zida ndi Njira.
Kuchulukana ndi kuberekanso kwa Cp NFRCS kunayesedwa pokonzekera Cp NFRCS ndi kukula kwa 15 × 15 cm2.Kuonjezera apo, zitsanzo za 30 mg (n = 5) zinatulutsidwa kuchokera kumagulu osiyanasiyana a Cp NFRCS ndipo BCT ya zitsanzo zomwe zaphunziridwa zinayesedwa monga momwe tafotokozera kale mu gawo la Njira.
Tayesa kupanga mawonekedwe ndi mapangidwe osiyanasiyana pogwiritsa ntchito zolemba za Cp NFRCS pazogwiritsa ntchito zosiyanasiyana zamankhwala.Maonekedwe kapena masinthidwe oterowo amaphatikiza ma conical swabs a mphuno, njira zamano, ndi ma cylindrical swabs pakutulutsa magazi kumaliseche.
Ma seti onse a data amawonetsedwa ngati ± kupatuka kokhazikika ndipo adawunikidwa ndi ANOVA pogwiritsa ntchito Prism 5.03 (GraphPad, San Diego, CA, USA) kutsatiridwa ndi mayeso ofananitsa angapo a Bonferroni (* p <0.05).
Njira zonse zomwe zinkachitika m'maphunziro a anthu zinali zogwirizana ndi mfundo za Institute ndi National Research Council, komanso Declaration of Helsinki 1964 ndi zosintha zake, kapena miyezo yofanana ya makhalidwe.Onse omwe adatenga nawo mbali adadziwitsidwa za zomwe zachitika pa kafukufukuyu komanso momwe adadzifunira.Deta ya omwe atenga nawo mbali imakhalabe yachinsinsi ikasonkhanitsidwa.Protocol yoyeserera ya in vitro TEG yawunikiridwa ndikuvomerezedwa ndi Institutional Ethics Committee ya Kasturba Medical College, Manipal, Karnataka (IEC: 674/2020).Odzipereka adasaina chilolezo chodziwitsidwa kuti atenge zitsanzo za magazi.
Njira zonse zomwe zidachitika pamaphunziro a nyama zidachitika motsatira Kastuba Faculty of Medicine, Manipal Institute of Higher Education, Manipal (IAEC/KMC/69/2020).Zoyeserera zonse za nyama zomwe zidapangidwa zidachitika motsatira malangizo a Komiti Yoyang'anira ndi Kuyang'anira Kuyesa Kwa Zinyama (CPCSEA).Olemba onse amatsatira malangizo a ARRIVE.
Mawonekedwe a FTIR a NFRCS onse adawunikidwa ndikufaniziridwa ndi mawonekedwe a chitosan omwe akuwonetsedwa pachithunzi 2A.Mawonekedwe apamwamba a chitosan (olembedwa) pa 3437 cm-1 (OH ndi NH kutambasula, kuphatikizika), 2945 ndi 2897 cm-1 (CH kutambasula), 1660 cm-1 (NH2 kupsyinjika), 1589 cm-1 (N-H kupinda), 1O 106 cm - mlatho wachiwiri, 1156 cm-mlatho hydroxyl), 993 cm-1 (kutambasula CO, Bo-OH) 52.53.54.Supplementary Table S1 ikuwonetsa mayamwidwe a FTIR NFRCS a chitosan (reporter), chitosan choyera, Cm, Ch, ndi Cp.Mawonekedwe a FTIR a NFRCS onse (Cm, Ch ndi Cp) adawonetsa magulu omwewo amayamwa ngati chitosan choyera popanda kusintha kwakukulu (mkuyu 2A).Zotsatira za FTIR zidatsimikizira kusakhalapo kwa kuyanjana kwamankhwala kapena thupi pakati pa ma polima omwe amagwiritsidwa ntchito popanga NFRCS, kuwonetsa kuti ma polima omwe amagwiritsidwa ntchito ndi opanda.
Mu vitro mawonekedwe a Cm NFRCS, Ch NFRCS, Cp NFRCS ndi Cs.(A) imayimira mawonekedwe a FTIR ophatikizidwa a chitosan ndi Cm NFRCS, Ch NFRCS ndi Cp NFRCS pansi pa kukanidwa.(B) a) Kuchuluka kwa magazi a NFRCS Cm, Ch, Cp, ndi Cg (n = 3);Zitsanzo za Ct zimasonyeza BAR yapamwamba chifukwa swab ya thonje ili ndi mphamvu yoyamwa kwambiri;b) Magazi pambuyo poyamwa magazi Chithunzi cha chitsanzo chomwe chatengedwa.Kuwonetsera kwazithunzi za BCT ya chitsanzo choyesa C (Cp NFRCS inali ndi BCT yabwino kwambiri (15 s, n = 3)). Deta mu C, D, E, ndi G adawonetsedwa ngati akutanthauza ± SD, ndipo zolakwitsa zikuyimira SD, ***p <0.0001. Deta mu C, D, E, ndi G adawonetsedwa ngati akutanthauza ± SD, ndipo zolakwitsa zikuyimira SD, ***p <0.0001. Данные в C, D, E ndi G представлены как среднее ± стандартное отклонение, а планки погрешностей представляют стандартное отклонение,0,0. Deta mu C, D, E, ndi G imawonetsedwa ngati ± kupatuka kokhazikika, ndipo zolakwitsa zikuyimira kupatuka kokhazikika, ***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 & G показаны как среднее значение значение ± стандартное отклонение, планки погрешностей представляют стандартное отклонение,0,00,000 Deta mu C, D, E, ndi G ikuwonetsedwa ngati ± kupatuka kokhazikika, zolakwitsa zikuyimira kupatuka kokhazikika, ***p<0.0001.