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Kukula kwa tizilombo toyambitsa matenda m'mabala nthawi zambiri kumadziwonetsera ngati biofilms, zomwe zimasokoneza machiritso ndipo zimakhala zovuta kuzichotsa. Zovala zatsopano zasiliva zimati zimalimbana ndi matenda a mabala, koma mphamvu yawo yolimbana ndi biofilm komanso mphamvu yochiritsa popanda matenda sizidziwika kawirikawiri. Pogwiritsa ntchito zitsanzo za in vitro ndi in vivo za Staphylococcus aureus ndi Pseudomonas aeruginosa, tikunena za mphamvu ya zovala zopangira ma ion za Ag1+; Zovala za Ag1+ zokhala ndi ethylenediaminetetraacetic acid ndi benzethonium chloride (Ag1+/EDTA/BC), ndi zovala zokhala ndi silver nitrate (Ag oxysalts). , zomwe zimapanga ma ion a Ag1+, Ag2+ ndi Ag3+ kuti zithane ndi biofilm ya mabala komanso momwe imakhudzira machiritso. Zovala za Ag1+ sizinakhudze kwambiri biofilm ya mabala mu vitro ndi m'makoswe (C57BL/6j). Mosiyana ndi zimenezi, mchere wa Ag wokhala ndi mpweya ndi zodzoladzola za Ag1+/EDTA/BC zinachepetsa kwambiri kuchuluka kwa mabakiteriya opindulitsa mu biofilms mu vitro ndipo zinawonetsa kuchepa kwakukulu kwa mabakiteriya ndi zigawo za EPS mu biofilms ya mbewa. Zodzoladzola izi zinali ndi zotsatira zosiyana pa kuchiritsa mabala omwe ali ndi kachilombo ka biofilm komanso omwe alibe kachilombo ka biofilm, ndipo zodzoladzola za mchere wokhala ndi mpweya zimakhala ndi zotsatira zabwino kwambiri pa kubwezeretsa mabala, kukula kwa mabala, ndi kutupa poyerekeza ndi mankhwala oletsa ndi zodzoladzola zina zasiliva. Makhalidwe osiyanasiyana a physicochemical a zodzoladzola zasiliva akhoza kukhala ndi zotsatira zosiyana pa biofilm ya mabala ndi kuchiritsa, ndipo izi ziyenera kuganiziridwa posankha zodzoladzola zochizira mabala omwe ali ndi kachilombo ka biofilm.
Mabala osatha amatanthauzidwa kuti ndi "mabala omwe amalephera kupita patsogolo m'magawo abwinobwino a kuchira mwadongosolo komanso munthawi yake" 1. Mabala osatha amapangitsa kuti odwala ndi dongosolo lazachipatala azikhala ndi vuto la maganizo, chikhalidwe, komanso zachuma. Ndalama zomwe NHS imawononga pachaka pochiza mabala ndi matenda ena okhudzana ndi matendawa zikuyerekezeredwa kuti ndi £8.3 biliyoni mu 2017-182. Mabala osatha pakadali pano ndi vuto lalikulu ku United States, pomwe Medicare ikuyerekeza mtengo wapachaka wochizira odwala omwe ali ndi mabala pa $28.1–$96.8 biliyoni3.
Matenda ndi chinthu chachikulu chomwe chimaletsa kuchira kwa mabala. Matenda nthawi zambiri amaonekera ngati biofilms, zomwe zimapezeka mu 78% ya mabala osachiritsika omwe sachira. Biofilms zimapangidwa pamene tizilombo toyambitsa matenda timalumikizana mosasinthika pamalo, monga malo a mabala, ndipo timatha kusonkhana ndikupanga magulu opanga ma polymer akunja (EPS). Biofilm ya mabala imagwirizanitsidwa ndi kuwonjezeka kwa kutupa komwe kumabweretsa kuwonongeka kwa minofu, komwe kungachedwetse kapena kuletsa kuchira4. Kuwonjezeka kwa kuwonongeka kwa minofu kungakhale chifukwa cha kuchuluka kwa ntchito ya matrix metalloproteinases, collagenase, elastase ndi mitundu ya okosijeni yogwira ntchito5. Kuphatikiza apo, maselo otupa ndi biofilms okha ndi omwe amagwiritsa ntchito kwambiri mpweya ndipo motero angayambitse hypoxia ya minofu yakomweko, ndikuchepetsa maselo a mpweya wofunikira wofunikira kuti minofu ikonzedwe bwino6.
Ma biofilm okhwima amakhala olimba kwambiri ku mankhwala ophera majeremusi, zomwe zimafuna njira zamphamvu zowongolera matenda a biofilm, monga chithandizo chamakina chotsatiridwa ndi chithandizo chothandiza cha majeremusi. Chifukwa ma biofilm amatha kubwereranso mwachangu, mankhwala othandiza ophera majeremusi amatha kuchepetsa chiopsezo cha kupangidwanso pambuyo pochotsa majeremusi pa opaleshoni7.
Siliva imagwiritsidwa ntchito kwambiri mu ma dressing opha tizilombo toyambitsa matenda ndipo nthawi zambiri imagwiritsidwa ntchito ngati mankhwala oyamba a mabala omwe ali ndi kachilomboka. Pali ma dressing ambiri asiliva omwe amapezeka m'masitolo, iliyonse ili ndi kapangidwe kake ka siliva, kuchuluka kwake, ndi maziko ake. Kupita patsogolo kwa ma armband asiliva kwapangitsa kuti ma armband atsopano asiliva apangidwe. Mtundu wa siliva wachitsulo (Ag0) ndi wopanda mphamvu; Kuti ugwire ntchito yopha tizilombo toyambitsa matenda, uyenera kutaya elekitironi kuti upange siliva wa ionic (Ag1+). Ma dressing asiliva achikhalidwe amakhala ndi ma compounds asiliva kapena siliva wachitsulo omwe, akapezeka ndi madzi, amawola kuti apange ma Ag1+ ions. Ma Ag1+ ions awa amachitapo kanthu ndi selo la bakiteriya, kuchotsa ma elekitironi kuchokera kuzinthu zomangira kapena njira zofunika kwambiri kuti munthu akhale ndi moyo. Ukadaulo wokhala ndi patent wapangitsa kuti pakhale compound yatsopano yasiliva, Ag oxysalts (silver nitrate, Ag7NO11), yomwe imaphatikizidwa mu ma dressing a mabala. Mosiyana ndi siliva wachikhalidwe, kuwola kwa mchere wokhala ndi okosijeni kumapanga siliva wokhala ndi valence yapamwamba (Ag1+, Ag2+ ndi Ag3+). Kafukufuku wa mu vitro wasonyeza kuti kuchuluka kochepa kwa mchere wa siliva wokhala ndi mpweya ndi kothandiza kwambiri kuposa siliva wa ion imodzi (Ag1+) motsutsana ndi mabakiteriya opatsirana monga Pseudomonas aeruginosa, Staphylococcus aureus ndi Escherichia coli8,9. Mtundu wina watsopano wa siliva wopaka umaphatikizapo zosakaniza zina, zomwe ndi ethylenediaminetetraacetic acid (EDTA) ndi benzethonium chloride (BC), zomwe zimanenedwa kuti zimayang'ana biofilm EPS ndipo motero zimawonjezera kulowa kwa siliva mu biofilm. Ukadaulo watsopano wa siliva uwu umapereka njira zatsopano zowunikira biofilms ya mabala. Komabe, mphamvu ya maantibayotiki awa pa malo ovulala ndi kuchira kosadalira matenda ndikofunikira kuti zitsimikizire kuti sizipanga malo olakwika a mabala kapena kuchedwa kuchira. Nkhawa zokhudzana ndi poizoni wa siliva mu vitro zanenedwa ndi ma dressings angapo a siliva10,11. Komabe, poizoni wa mu vitro sunatanthauzidwebe kukhala poizoni wa mu vivo, ndipo ma dressings angapo a Ag1+ awonetsa mbiri yabwino yachitetezo12.
Apa, tinafufuza momwe ma dressing a carboxymethylcellulose okhala ndi ma formula atsopano a siliva amagwirira ntchito motsutsana ndi biofilm ya mabala mu vitro ndi mu vivo. Kuphatikiza apo, zotsatira za ma dressing awa pa chitetezo chamthupi ndi machiritso popanda matenda zinayesedwa.
Ma dressing onse omwe adagwiritsidwa ntchito anali ogulitsidwa. 3M Kerracel Gel Fiber Dressing (3M, Knutsford, UK) ndi dressing ya gel fiber ya 100% carboxymethylcellulose (CMC) yopanda ma antibiotic yomwe idagwiritsidwa ntchito ngati dressing yowongolera mu kafukufukuyu. Ma dressing atatu a CMC silver ophera ma antibiotic adayesedwa, omwe ndi 3M Kerracel Ag dressing (3M, Knutsford, UK), omwe ali ndi 1.7 wt%. oxygenated silver salt (Ag7NO11) mu ma ion apamwamba a valence silver (Ag1+, Ag2+ ndi Ag3+). Panthawi yowola ya Ag7NO11, ma ion a Ag1+, Ag2+ ndi Ag3+ amapangidwa mu chiŵerengero cha 1:2:4. Chovala chowonjezera cha Aquacel Ag chokhala ndi 1.2% siliva chloride (Ag1+) (ConvaTec, Deeside, UK) 13 ndi Chovala chowonjezera cha Aquacel Ag + chokhala ndi 1.2% siliva chloride (Ag1+), EDTA ndi benzethonium chloride (ConvaTec, Deeside, UK) 14.
Mitundu yomwe inagwiritsidwa ntchito mu kafukufukuyu inali Pseudomonas aeruginosa NCTC 10781 (Public Health England, Salisbury) ndi Staphylococcus aureus NCTC 6571 (Public Health England, Salisbury).
Mabakiteriya anakula usiku wonse mu msuzi wa Muller-Hinton (Oxoid, Altrincham, UK). Kenako, kumera kwa usiku wonse kunachepetsedwa ndi 1:100 mu msuzi wa Mueller-Hinton ndi 200 µl yokutidwa pa 0.2 µm Whatman cyclopore membranes (Whatman plc, Maidstone, UK) pa mbale za Mueller-Hinton agar (Sigma-Aldrich Company Ltd, Kent, Great Britain). Kupanga kwa biofilm yachikoloni pa 37°C kwa maola 24. Ma biofilm achikoloni awa adayesedwa kuti awone ngati ali ndi logarithmic shrinkage.
Dulani kuvala m'zidutswa za masikweya 3 cm2 ndikunyowetsa ndi madzi osadetsedwa. Ikani bandeji pamwamba pa biofilm ya koloni pa mbale ya agar. Mahekitala 24 aliwonse a biofilm adachotsedwa, ndipo mabakiteriya otheka mkati mwa biofilm (CFU/ml) adayesedwa poyezedwa (10−1 mpaka 10−7) mu msuzi wa Day-Angle neutralization (Merck-Millipore). Pambuyo pa maola 24 a incubation pa 37°C, kuwerengera kwa mbale zokhazikika kunachitika pa mbale za agar za Mueller-Hinton. Chithandizo chilichonse ndi nthawi yake zidachitika katatu, ndipo kuwerengera kwa mbale kudabwerezedwa pa kuchepetsedwa kulikonse.
Khungu la m'mimba mwa nkhumba limapezeka kuchokera ku nkhumba zazikazi za Large White mkati mwa mphindi 15 kuchokera pamene zinaphedwa motsatira miyezo ya European Union yotumiza kunja. Khungulo linametedwa ndi kutsukidwa ndi zopukutira mowa, kenako nkuzizira pa -80°C kwa maola 24 kuti khungu liume. Pambuyo posungunuka, zidutswa za khungu la 1 cm2 zinatsukidwa katatu ndi PBS, 0.6% sodium hypochlorite, ndi 70% ethanol kwa mphindi 20 nthawi iliyonse. Musanachotse khungu, chotsani ethanol yotsalayo poitsuka katatu mu PBS yoyera. Khungulo linakulitsidwa mu mbale ya zitsime 6 yokhala ndi nembanemba ya nayiloni ya 0.45-μm (Merck-Millipore) pamwamba ndi mapepala atatu onyowa (Merck-Millipore) okhala ndi 3 ml fetal bovine serum (Sigma) yowonjezera 10% Dulbecco's modified Eagle. Medium (Dulbecco's Modified Eagle Medium – Aldrich Ltd.).
Ma biofilm achikhalidwe adalimidwa monga momwe tafotokozera pa kafukufuku wokhudza kufalikira kwa biofilm. Pambuyo pokulitsa biofilm pa nembanemba kwa maola 72, biofilm idayikidwa pamwamba pa khungu pogwiritsa ntchito njira yothira mankhwala ndipo nembanembayo idachotsedwa. Kenako biofilm idayikidwa pa dermis ya nkhumba kwa maola ena 24 pa 37°C kuti biofilm ikule ndikumamatira pakhungu la nkhumba. Biofilm itatha kukula ndikulumikizidwa, dressing ya 1.5 cm2, yonyowetsedwa kale ndi madzi osungunuka, idayikidwa mwachindunji pamwamba pa khungu ndikuyikidwa pa 37°C kwa maola 24. Mabakiteriya okhazikika adawonedwa mwa kuyika utoto poyika mofanana PrestoBlue cell viability reagent (Invitrogen, Life Technologies, Paisley, UK) pamwamba pa apical ya explant iliyonse ndikuyiyika kwa mphindi 5. Gwiritsani ntchito kamera ya digito ya Leica DFC425 kuti mujambule zithunzi nthawi yomweyo pa maikulosikopu ya Leica MZ8. Madera amitundu ya pinki adayesedwa pogwiritsa ntchito pulogalamu ya Image Pro version 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (mediacy.com)). Kujambula maikulosikopu ya ma elekitironi kunachitika monga momwe tafotokozera pansipa.
Mabakiteriya omwe anakula usiku wonse anachepetsedwa ndi 1:100 mu msuzi wa Mueller-Hinton. 200 μl ya chikhalidwe inawonjezedwa ku 0.2 μm ya sterile Whatman cyclopore membrane (Whatman, Maidstone, UK) ndipo inayikidwa pa Mueller-Hinton agar. Mapepala a biofilm anaikidwa pa 37°C kwa maola 72 kuti alole kuti biofilm ikule bwino.
Pambuyo pa masiku atatu a biofilm yokhwima, bandeji ya 3 cm2 sikweya inayikidwa mwachindunji pa biofilm ndipo inayikidwa pa 37°C kwa maola 24. Bandejiyo atachotsedwa pamwamba pa biofilm, 1 ml ya PrestoBlue Cell Viability Reagent (Invitrogen, Waltham, MA) inawonjezedwa pamwamba pa biofilm iliyonse kwa masekondi 20. Malowo anaumitsidwa asanajambulidwe kusintha kwa mitundu pogwiritsa ntchito kamera ya digito ya Nikon D2300 (Nikon UK Ltd., Kingston, UK).
Konzani zokolola za usiku wonse pa Mueller-Hinton agar, sungani madera osiyanasiyana ku 10 ml ya msuzi wa Mueller-Hinton ndikuyika mu shaker pa 37°C (100 rpm). Pambuyo poika mu 100 rpm usiku wonse, zokololazo zinachepetsedwa ndi 1:100 mu msuzi wa Mueller-Hinton ndipo 300 µl zinapezeka pa 0.2 µm circular Whatman cyclopore membrane (Whatman International, Maidstone, UK) pa Mueller-Hinton agar ndikuyika mu 37°C mkati mwa maola 72. . Biofilm yokhwima inayikidwa pa bala monga tafotokozera pansipa.
Ntchito yonse ndi nyama idachitika ku University of Manchester pansi pa chilolezo cha polojekiti chovomerezedwa ndi Office of Animal Welfare and Ethical Review (P8721BD27) komanso motsatira malangizo ofalitsidwa ndi Home Office motsatira ASPA yosinthidwa ya 2012. Olemba onse adatsatira malangizo ofika. Mbewa za C57BL/6j za masabata asanu ndi atatu (Envigo, Oxon, UK) zidagwiritsidwa ntchito pa maphunziro onse a in vivo. Mbewa zidapatsidwa mankhwala oletsa ululu ndi isoflurane (Piramal Critical Care Ltd, West Drayton, UK) ndipo malo awo akumbuyo adametedwa ndikutsukidwa. Kenako mbewa iliyonse idapatsidwa bala la 2 × 6 mm pogwiritsa ntchito Stiefel biopsy punch (Schuco International, Hertfordshire, UK). Pa mabala omwe ali ndi kachilombo ka biofilm, ikani biofilm ya colonial ya maola 72 yomwe idakula pa nembanemba monga tafotokozera pamwambapa pa khungu la bala pogwiritsa ntchito njira yopatsirana nthawi yomweyo mutavulala ndikutaya nembanemba. Centimita imodzi ya kuvala imanyowetsedwa kale ndi madzi osabala kuti malo ovulala akhale onyowa. Ma dressing anaikidwa mwachindunji pa bala lililonse ndipo anaphimbidwa ndi 3M Tegaderm film (3M, Bracknell, UK) ndi Mastisol liquid glue (Eloquest Healthcare, Ferndale, MI) zomwe zinagwiritsidwa ntchito m'mbali kuti ziwonjezere kumatira. Buprenorphine (Animalcare, York, UK) inaperekedwa pamlingo wa 0.1 mg/kg ngati mankhwala ochepetsa ululu. Dulani mbewa patatha masiku atatu mutavulala pogwiritsa ntchito njira ya Schedule 1 ndikuchotsa, kudula pakati, ndikusunga malo a bala ngati pakufunika.
Kupaka utoto wa Hematoxylin (ThermoFisher Scientific) ndi eosin (ThermoFisher Scientific) kunachitika motsatira ndondomeko ya wopanga. Malo a bala ndi kubwezeretsanso kwa zilonda zinayesedwa pogwiritsa ntchito pulogalamu ya Image Pro version 10 (Media Cybernetics Inc, Rockville, MD).
Zigawo za minofu zinasungunuka mu xylene (ThermoFisher Scientific, Loughborough, UK), zinasinthidwa madzi ndi ethanol yokonzedwa ndi 100–50%, ndipo zinamizidwa pang'ono m'madzi oyeretsedwa (ThermoFisher Scientific). Immunohistochemistry inachitidwa pogwiritsa ntchito VectaStain Elite ABC PK-6104 kit (Vector Laboratories, Burlingame, CA) malinga ndi njira ya wopanga. Ma antibodies oyambira ku ma neutrophils NIMP-R14 (Thermofisher Scientific) ndi macrophages Ms CD107b Pure M3/84 (BD Biosciences, Wokingham, UK) anasungunuka 1:100 mu yankho lotsekereza ndikuwonjezeredwa pamwamba pake, kutsatiridwa ndi ma antibodies awiri Anti-, VectaStain ABC ndi Vector Nova Red Peroxidase (HRP) substrate kit (Vector Laboratories, Burlingame, CA) ndipo anaphatikizidwa ndi hematoxylin. Zithunzi zinapezedwa pogwiritsa ntchito maikulosikopu ya Olympus BX43 ndi kamera ya digito ya Olympus DP73 (Olympus, Southend-on-Sea, UK).
Zitsanzo za pakhungu zinaikidwa mu 2.5% glutaraldehyde ndi 4% formaldehyde mu 0.1 M HEPES (pH 7.4) kwa maola 24 pa 4°C. Zitsanzo zinachotsedwa madzi pogwiritsa ntchito ethanol yoyeretsedwa ndipo zinaumitsidwa mu CO2 pogwiritsa ntchito Quorum K850 critical point dryer (Quorum Technologies Ltd, Loughton, UK) ndipo zinapakidwa ndi sputter yokutidwa ndi gold-palladium alloy pogwiritsa ntchito Quorum SC7620 mini sputterer/glow discharge system. Zitsanzo zinajambulidwa pogwiritsa ntchito FEI Quanta 250 scanning electron microscope (ThermoFisher Scientific) kuti ziwonetse malo apakati pa bala.
Toto-1 iodide (2 μM) inagwiritsidwa ntchito pamwamba pa bala la mbewa ndipo inayikidwa kwa mphindi 5 pa kutentha kwa 37 °C (ThermoFisher Scientific) ndipo inachiritsidwa ndi Syto-60 (10 μM) pa kutentha kwa 37 °C (ThermoFisher Scientific). Zithunzi za Z-stack za mphindi 15 zinapangidwa pogwiritsa ntchito Leica TCS SP8.
Deta yobwerezabwereza ya zamoyo ndi yaukadaulo idalembedwa ndikusanthulidwa pogwiritsa ntchito pulogalamu ya Graphpad Prism V9 (GraphPad Software, La Jolla, CA). Kusanthula kwa njira imodzi kwa kusiyana ndi kufananiza kosiyanasiyana pogwiritsa ntchito mayeso a post hoc a Dunnett kunagwiritsidwa ntchito kuyesa kusiyana pakati pa chithandizo chilichonse ndi kuvala koletsa tizilombo toyambitsa matenda. P value <0.05 idawonedwa kuti ndi yofunika.
Kugwira ntchito kwa ma gel fibrous dressings a siliva kunayesedwa koyamba motsutsana ndi madera a biofilm a Staphylococcus aureus ndi Pseudomonas aeruginosa mu vitro. Ma dressings asiliva ali ndi mitundu yosiyanasiyana ya siliva: ma dressings achikhalidwe asiliva amapanga ma Ag1+ ions; ma dressings asiliva, omwe amatha kupanga ma Ag1+ ions atawonjezeredwa EDTA/BC, amatha kuwononga matrix a biofilm ndikuyika mabakiteriya ku siliva chifukwa cha mphamvu ya mabakiteriya a siliva. ma ions15 ndi ma dressings okhala ndi mchere wa Ag wokhala ndi okosijeni womwe umapanga ma Ag1+, Ag2+ ndi Ag3+. Kugwira ntchito kwake kunayerekezeredwa ndi dressing yosaletsa mabakiteriya yopangidwa kuchokera ku ulusi wopangidwa ndi gelled. Mabakiteriya otsala omwe ali mu biofilm adayesedwa maola 24 aliwonse kwa masiku 8 (Chithunzi 1). Pa tsiku la 5, biofilm idabwezeretsedwanso ndi 3.85 × 105S. Staphylococcus aureus kapena 1.22 × 105P. aeruginosa kuti ayese kuchira kwa biofilm. Poyerekeza ndi ma dressing osagwiritsa ntchito maantibayotiki, ma dressing a Ag1+ anali ndi mphamvu yochepa pa moyo wa mabakiteriya mu Staphylococcus aureus ndi Pseudomonas aeruginosa biofilms kwa masiku 5. Mosiyana ndi zimenezi, ma dressing okhala ndi mchere wa Ag ndi Ag1+ + EDTA/BC wokhala ndi mpweya anali othandiza kupha mabakiteriya mkati mwa biofilm mkati mwa masiku 5. Pambuyo pobaya mobwerezabwereza ndi mabakiteriya a planktonic pa tsiku la 5, palibe kubwezeretsa kwa biofilm komwe kunawonedwa (Chithunzi 1).
Kuyeza mabakiteriya okhazikika mu Staphylococcus aureus ndi Pseudomonas aeruginosa biofilms pambuyo polandira chithandizo ndi ma dressing asiliva. Ma colonies a biofilm a Staphylococcus aureus ndi Pseudomonas aeruginosa adachiritsidwa ndi ma dressing asiliva kapena ma dressing osaletsa mabakiteriya, ndipo chiwerengero cha mabakiteriya okhazikika omwe adatsala chidadziwika maola 24 aliwonse. Patatha masiku 5, biofilm idabwezeretsedwanso ndi 3.85×105S. Staphylococcus aureus kapena 1.22×105P. Ma colonies a bacterioplankton Pseudomonas aeruginosa adapangidwa payekhapayekha kuti awone momwe biofilm imakhalira. Ma graph akuwonetsa cholakwika cha +/- standard.
Kuti tiwone momwe ma dressing asiliva amakhudzira moyo wa biofilm, ma dressing anagwiritsidwa ntchito pa biofilms zokhwima zomwe zimamera pakhungu la nkhumba ex vivo. Pambuyo pa maola 24, dressing imachotsedwa ndipo biofilm imapakidwa utoto wabuluu wosinthika, womwe umasinthidwa ndi mabakiteriya amoyo kukhala pinki. Biofilms zomwe zimachiritsidwa ndi ma dressing olamulira zinali pinki, kusonyeza kukhalapo kwa mabakiteriya okhazikika mkati mwa biofilm (Chithunzi 2A). Mosiyana ndi zimenezi, biofilm yomwe imachiritsidwa ndi Ag oxysols dressing inali yabuluu makamaka, kusonyeza kuti mabakiteriya otsala pamwamba pa khungu la nkhumba anali mabakiteriya osakhazikika (Chithunzi 2B). Mitundu yosiyanasiyana ya buluu ndi pinki inawonedwa mu biofilms zomwe zimachiritsidwa ndi ma dressing okhala ndi Ag1+, kusonyeza kukhalapo kwa mabakiteriya okhazikika ndi osakhazikika mkati mwa biofilm (Chithunzi 2C), pomwe ma dressing a EDTA/BC omwe ali ndi Ag1+ anali abuluu kwambiri okhala ndi madontho ena a pinki, kusonyeza madera omwe sanakhudzidwe ndi dressing yasiliva (Chithunzi 2D). Kuyeza kwa madera ogwira ntchito (apinki) ndi osagwira ntchito (abuluu) kunasonyeza kuti chigamba chowongolera chinali 75% chogwira ntchito (Chithunzi 2E). Ma dressing a Ag1+ + EDTA/BC adachitika mofanana ndi ma dressing a mchere wa Ag omwe ali ndi mpweya, ndipo kuchuluka kwa moyo wa 13% ndi 14% kunachepa. Ma dressing a Ag1+ adachepetsanso kuthekera kwa mabakiteriya ndi 21%. Kenako ma biofilms awa adawonedwa pogwiritsa ntchito scanning electron microscopy (SEM). Pambuyo pa chithandizo ndi control dressing ndi Ag1+ dressing, wosanjikiza wa Pseudomonas aeruginosa unawonedwa ukuphimba khungu la nkhumba (Chithunzi 2F,H), pomwe pambuyo pa chithandizo ndi Ag1+ dressing, maselo ochepa a bakiteriya adapezeka ndipo maselo ochepa a bakiteriya adapezeka pansi pake. Ulusi wa Collagen ukhoza kuonedwa ngati kapangidwe ka minofu ya khungu la nkhumba (Chithunzi 2G). Pambuyo pa chithandizo ndi Ag1+ + EDTA/BC dressing, ma bacterial plaques ndi ma collagen fiber plaques omwe ali pansi pake adawoneka (Chithunzi 2I).
Kuwona mawonekedwe a Pseudomonas aeruginosa biofilm pambuyo pochiza siliva dressing. (A–D) Kutha kwa bakiteriya mu Pseudomonas aeruginosa biofilms yomwe imamera pakhungu la nkhumba kunawonetsedwa pogwiritsa ntchito utoto wa PrestoBlue viability maola 24 mutalandira chithandizo ndi siliva dressing kapena dressing yoletsa tizilombo toyambitsa matenda. Mabakiteriya amoyo ndi pinki, mabakiteriya osatha ndipo khungu la nkhumba ndi labuluu. (E) Kuyeza kwa Pseudomonas aeruginosa biofilms yomwe imamera pakhungu la nkhumba (pinki spot) pogwiritsa ntchito scanning electron microscopy Image Pro version 10 (FI) ndikuchiritsidwa ndi siliva dressing kapena siliva dressing yoletsa tizilombo toyambitsa matenda kwa maola 24. SEM scale bar = 5 µm. (J–M) Colonial biofilms inamera pa zosefera ndipo inapakidwa utoto wa PrestoBlue reactive patatha maola 24 atalandira siliva dressing.
Kuti adziwe ngati kukhudzana kwapafupi pakati pa ma dressing ndi biofilms kunakhudza momwe ma dressing amagwirira ntchito, ma biofilms achikhalidwe omwe adayikidwa pamalo athyathyathya adayikidwa ndi ma dressing kwa maola 24 kenako adapakidwa utoto wosinthika. Biofilm yosachiritsidwayo inali ya pinki wakuda (Chithunzi 2J). Mosiyana ndi ma biofilms omwe adapakidwa ndi ma dressing okhala ndi mchere wa Ag wopangidwa ndi okosijeni (Chithunzi 2K), ma biofilms omwe adapakidwa ndi ma dressing okhala ndi Ag1+ kapena Ag1+ + EDTA/BC adawonetsa mikwingwirima ya pinki (Chithunzi 2L,M). Mtundu wa pinki uwu umasonyeza kukhalapo kwa mabakiteriya otheka ndipo umagwirizanitsidwa ndi malo olumikizira mkati mwa dressing. Malo osokedwa awa amapanga malo akufa omwe amalola mabakiteriya mkati mwa biofilm kuti apulumuke.
Kuti aone momwe ma dressing asiliva amagwirira ntchito m'thupi, mabala otupa a mbewa omwe ali ndi matenda a S. aureus okhwima ndi P. aeruginosa biofilms adachiritsidwa ndi ma dressing oletsa tizilombo toyambitsa matenda kapena ma dressing asiliva. Pambuyo pa masiku atatu a chithandizo, kusanthula kwa zithunzi za macroscopic kunawonetsa kukula kwa mabala ang'onoang'ono akachiritsidwa ndi ma dressing amchere okometsedwa ndi mpweya poyerekeza ndi ma dressing oletsa tizilombo toyambitsa matenda ndi ma dressing ena asiliva (Chithunzi 3A-H). Kuti atsimikizire zomwe zapezekazi, mabala adasonkhanitsidwa ndipo malo a mabala ndi reepithelialization adayesedwa pa hematoxylin ndi zigawo za minofu yodetsedwa ndi eosin pogwiritsa ntchito image pro software version 10 (Chithunzi 3I-L).
Zotsatira za ma dressing asiliva pamwamba pa bala ndi kubwezeretsanso epithelialization ya mabala omwe ali ndi biofilms. (A–H) Maselo ang'onoang'ono omwe ali ndi biofilms ya Pseudomonas aeruginosa (A–D) ndi Staphylococcus aureus (E–H) patatha masiku atatu a chithandizo ndi dressing yosagwiritsa ntchito maantibayotiki, dressing ya Ag salt yokhala ndi mpweya, dressing ya Ag1+, ndi dressing ya Ag1+. Zithunzi zoyimira macroscopic. mabala a mbewa omwe ali ndi dressing ya Ag1+ + EDTA/BC. (IL) Woyimira matenda a Pseudomonas aeruginosa, magawo a histological okhala ndi hematoxylin ndi eosin, omwe amagwiritsidwa ntchito poyeza malo a bala ndi kukonzanso kwa epithelial. Kuyeza malo a bala (M, O) ndi kuchuluka kwa reepithelialization (N, P) ya mabala omwe ali ndi Pseudomonas aeruginosa (M, N) ndi Staphylococcus aureus (O, P) biofilms (pa gulu lililonse la chithandizo n = 12). Ma graph akuwonetsa cholakwika cha +/- standard. * amatanthauza p = < 0.05 ** amatanthauza p = < 0.01; Sikelo ya macroscopic = 2.5 mm, sikelo ya histological = 500 µm.
Kuyeza malo a bala m'mabala omwe ali ndi kachilombo ka Pseudomonas aeruginosa biofilm (Chithunzi 3M) kunawonetsa kuti mabala omwe amachiritsidwa ndi Ag oxysalts anali ndi kukula kwapakati kwa bala la 2.5 mm2, pomwe chovala chowongolera chosagwiritsa ntchito maantibayotiki chinali ndi kukula kwapakati kwa bala la 3.1 mm2, zomwe sizowona. zinafika pamlingo wofunikira (Chithunzi 3M). p = 0.423). Mabala omwe amachiritsidwa ndi Ag1+ kapena Ag1+ + EDTA/BC sanawonetse kuchepa kwa malo a bala (3.1 mm2 ndi 3.6 mm2, motsatana). Chithandizo ndi chovala chowongolera cha Ag chokhala ndi mpweya chinalimbikitsa kukonzanso epithelialization kwambiri kuposa chovala chowongolera chosagwiritsa ntchito maantibayotiki (34% ndi 15%, motsatana; p = 0.029) ndi Ag1+ kapena Ag1+ + EDTA/BC (10% ndi 11%) (Chithunzi 3N). . , motsatana).
Zochitika zofanana m'dera la bala ndi kukonzanso kwa epithelial zinawonedwa m'mabala omwe ali ndi S. aureus biofilms (Chithunzi 3O). Ma dressings okhala ndi mchere wa siliva wopangidwa ndi okosijeni adachepetsa dera la bala (2.0 mm2) ndi 23% poyerekeza ndi dressing yoletsa yoletsa mabakiteriya (2.6 mm2), ngakhale kuti kuchepa kumeneku sikunali kwakukulu (p = 0.304) (Chithunzi 3O). Kuphatikiza apo, dera la bala lomwe lili m'gulu la chithandizo cha Ag1+ linachepetsedwa pang'ono (2.4 mm2), pomwe bala lomwe linachiritsidwa ndi Ag1+ + EDTA/BC dressing silinachepetse dera la bala (2.9 mm2). Mchere wa okosijeni wa Ag unathandizanso kukonzanso epithelialization ya mabala omwe ali ndi S. aureus biofilm (31%) kwambiri kuposa omwe amachiritsidwa ndi ma dressings oletsa mabakiteriya (12%, p = 0.003) (Chithunzi 3P). Kupaka kwa Ag1+ (16%, p = 0.903) ndi Kupaka kwa Ag+1 + EDTA/BC (14%, p = 0.965) kunawonetsa kuchuluka kwa kukonzanso kwa epithelial mofanana ndi kulamulira.
Kuti tiwone momwe ma dressing asiliva amakhudzira biofilm matrix, Toto 1 ndi Syto 60 iodide staining idachitikira (Chithunzi 4). Toto 1 iodide ndi utoto wosalowa m'maselo womwe ungagwiritsidwe ntchito kuwonetsa molondola ma nucleic acids akunja kwa maselo, omwe ali ochulukirapo mu EPS ya biofilms. Syto 60 ndi utoto wolowa m'maselo womwe umagwiritsidwa ntchito ngati counterstain16. Kuwona kwa Toto 1 ndi Syto 60 iodide m'mabala omwe adalowetsedwa ndi biofilms za Pseudomonas aeruginosa (Chithunzi 4A-D) ndi Staphylococcus aureus (Chithunzi 4I-L) kunawonetsa kuti patatha masiku atatu chithandizo cha dressing, EPS mu biofilm idachepa kwambiri. Yokhala ndi mchere wopangidwa ndi okosijeni Ag ndi Ag1+ + EDTA/BC. Ma dressing a Ag1+ opanda zinthu zina zotsutsana ndi ma antibiotic anachepetsa kwambiri DNA yopanda maselo m'mabala omwe anabayidwa ndi Pseudomonas aeruginosa koma sanagwire bwino ntchito m'mabala omwe anabayidwa ndi Staphylococcus aureus.
Kujambula zithunzi za biofilm ya mabala mu vivo patatha masiku atatu a chithandizo pogwiritsa ntchito zodzoladzola zowongolera kapena zasiliva. Zithunzi zamkati za Pseudomonas aeruginosa (A–D) ndi Staphylococcus aureus (I–L) zopakidwa utoto ndi Toto 1 (wobiriwira) kuti muwone ma nucleic acids akunja kwa maselo, omwe ndi gawo la ma polima akunja kwa maselo. Kuti muike utoto mu nucleic acids, gwiritsani ntchito ma Syto 60 (ofiira). P. Kujambula ma microscopy a ma electron a mabala omwe ali ndi Pseudomonas aeruginosa (E–H) ndi Staphylococcus aureus (M–P) biofilms patatha masiku atatu a chithandizo pogwiritsa ntchito zodzoladzola zowongolera ndi zasiliva. SEM scale bar = 5 µm. Confocal imaging scale bar = 50 µm.
Kusanthula kwa maikulosikopu yamagetsi kunawonetsa kuti mbewa zomwe zinabayidwa ndi ma colonies a Pseudomonas aeruginosa (Chithunzi 4E-H) ndi Staphylococcus aureus (Chithunzi 4M-P) zinali ndi mabakiteriya ochepa kwambiri m'mabala awo atatha masiku atatu a chithandizo ndi zodzoladzola zonse zasiliva.
Kuti muwone momwe ma dressing asiliva amakhudzira kutupa kwa mabala m'makoswe omwe ali ndi kachilombo ka biofilm, magawo a mabala omwe ali ndi kachilombo ka biofilm omwe amachiritsidwa ndi ma dressing owongolera kapena asiliva kwa masiku atatu adapakidwa utoto wa immunohistochemical pogwiritsa ntchito ma antibodies apadera a ma neutrophils ndi macrophages. Kuzindikira kuchuluka kwa ma neutrophils ndi macrophages mkati. granulation tissue. Chithunzi 5). Ma dressing onse asiliva adachepetsa chiwerengero cha ma neutrophils ndi macrophages m'mabala omwe ali ndi kachilombo ka Pseudomonas aeruginosa poyerekeza ndi ma dressing owongolera osagwiritsa ntchito maantibayotiki patatha masiku atatu a chithandizo. Komabe, chithandizo ndi oxygenated silver salt dressing chinapangitsa kuti ma neutrophils (p = <0.0001) ndi ma macrophages (p = <0.0001) poyerekeza ndi ma dressing ena asiliva omwe adayesedwa (Chithunzi 5I,J). Ngakhale kuti Ag1+ + EDTA/BC inali ndi zotsatirapo zazikulu pa ma rolling, idachepetsa kuchuluka kwa ma neutrophil ndi macrophage pang'ono kuposa ma dressing a Ag1+. Mabala apakati omwe ali ndi kachilombo ka S. aureus biofilm adawonedwanso atapaka ndi Ag (p = <0.0001), Ag1+ (p = 0.0008) ndi Ag1++ EDTA/BC (p = 0.0043) oxisols poyerekeza ndi control . Zochitika zofananazi zikuwonetsedwa pa neutropenia. bandeji (Chithunzi 5K). Komabe, oxygenated Ag salt dressing yokha ndi yomwe idawonetsa kuchepa kwakukulu kwa macrophages mu minofu ya granulation poyerekeza ndi control m'mabala omwe ali ndi kachilombo ka S. aureus biofilms (p = 0.0339) (Chithunzi 5L).
Ma neutrophils ndi macrophages anayesedwa m'mabala omwe ali ndi kachilombo ka Pseudomonas aeruginosa ndi Staphylococcus aureus biofilms atatha masiku atatu a chithandizo ndi mankhwala oletsa tizilombo toyambitsa matenda kapena ma siliva. Ma Neutrophils (AD) ndi ma macrophages (EH) anayesedwa m'magawo a minofu okhala ndi ma antibodies apadera a ma neutrophils kapena ma macrophages. Kuyeza kwa ma neutrophils (I ndi K) ndi ma macrophages (J ndi L) m'mabala omwe ali ndi kachilombo ka Pseudomonas aeruginosa (I ndi J) ndi Staphylococcus aureus (K & L) biofilms. N = 12 pagulu lililonse. Ma graph akuwonetsa mean +/- standard error, significance values poyerekeza ndi mankhwala oletsa tizilombo toyambitsa matenda, * amatanthauza p = < 0.05, ** amatanthauza p = < 0.01; *** amatanthauza p = < 0.001; akusonyeza p = <0.0001).
Kenako tinayang'ana momwe ma dressing asiliva amakhudzira machiritso osakhudzana ndi matenda. Mabala osakhudzidwa ndi kachilomboka adachiritsidwa ndi dressing yoletsa mabakiteriya kapena dressing yasiliva kwa masiku atatu (Chithunzi 6). Pakati pa ma dressing asiliva omwe adayesedwa, mabala okha omwe adachiritsidwa ndi dressing yamchere yokhala ndi okosijeni adawoneka ochepa pazithunzi zazikulu kuposa mabala omwe adachiritsidwa ndi dressing (Chithunzi 6A-D). Kuyeza malo a bala pogwiritsa ntchito kusanthula kwa histological kunawonetsa kuti malo apakati a bala atalandira chithandizo ndi dressing ya Ag oxysols anali 2.35 mm2 poyerekeza ndi 2.96 mm2 ya mabala omwe adalandira chithandizo ndi gulu lowongolera, koma kusiyana kumeneku sikunafike pakufunika kwa ziwerengero (p = 0.488) (Chithunzi 6I). Mosiyana ndi izi, palibe kuchepa kwa malo a bala komwe kudawonedwa atalandira chithandizo ndi ma dressing a Ag1+ (3.38 mm2, p = 0.757) kapena Ag1+ + EDTA/BC (4.18 mm2, p = 0.054) poyerekeza ndi gulu lowongolera. Kuwonjezeka kwa kusinthika kwa epithelial kunawonedwa ndi Ag oxysol dressing poyerekeza ndi gulu lolamulira (30% vs. 22%, motsatana), ngakhale izi sizinafike pakufunika (p = 0.067), izi ndizofunikira kwambiri ndipo zimatsimikizira zotsatira zam'mbuyomu. Kuvala ndi oxysol kumalimbikitsa kubwezeretsanso epithelialization. -Kuchotsa epithelialization ya mabala osadwala17. Mosiyana ndi zimenezi, chithandizo ndi Ag1+ kapena Ag1+ + EDTA/BC dressings sichinagwire ntchito kapena chinawonetsa kuchepa kwa re-epithelialization poyerekeza ndi kulamulira.
Zotsatira za siliva wopaka mabala pa machiritso a mabala m'makoswe osadwala ndi kuchotsedwa kwathunthu. (AD) Zithunzi zosonyeza ma macroscopic a mabala atatha masiku atatu a chithandizo ndi chitsulo chosagwiritsa ntchito maantibayotiki ndi siliva wopaka mabala. (EH) Zigawo zoimira mabala zokhala ndi hematoxylin ndi eosin. Kuyeza malo a mabala (I) ndi kuchuluka kwa reepithelialization (J) zinawerengedwa kuchokera ku zigawo za histological pakati pa bala pogwiritsa ntchito pulogalamu yowunikira zithunzi (n = 11–12 pa gulu lililonse la chithandizo). Ma graph akuwonetsa cholakwika cha +/- standard. * zikutanthauza p = <0.05.
Silver wakhala akugwiritsidwa ntchito kwa nthawi yayitali ngati mankhwala opha tizilombo toyambitsa matenda pochiritsa mabala, koma njira zosiyanasiyana zoperekera mankhwala ndi njira zoperekera mankhwala zingayambitse kusiyana kwa mphamvu ya maantibayotiki 18. Komanso, mphamvu ya antibiofilm ya njira zina zoperekera mankhwala a siliva sizikumveka bwino. Ngakhale kuti chitetezo cha mthupi cha munthu chimakhala chothandiza kwambiri polimbana ndi mabakiteriya a planktonic, nthawi zambiri sichigwira ntchito bwino polimbana ndi biofilms19. Mabakiteriya a planktonic amachotsedwa mosavuta ndi macrophages, koma mkati mwa biofilms, maselo ophatikizidwa amabweretsa mavuto ena pochepetsa yankho la munthuyo mpaka kufika poti maselo a chitetezo cha mthupi amatha kudwala apoptosis ndikutulutsa zinthu zoyambitsa kutupa kuti awonjezere yankho la chitetezo cha mthupi20. Zawonedwa kuti ma leukocyte ena amatha kulowa mu biofilms21 koma sangathe kuphagocytose mabakiteriya chitetezochi chikachepa22. Njira yonse iyenera kugwiritsidwa ntchito kuthandizira chitetezo cha mthupi cha munthuyo polimbana ndi matenda a biofilm ya mabala. Kuchotsa mabala kungasokoneze biofilm ndikuchotsa katundu wambiri wa biofilm, koma chitetezo cha mthupi cha munthuyo sichingakhale chothandiza polimbana ndi biofilm yotsala, makamaka ngati chitetezo cha mthupi cha munthuyo chachepa. Motero, mankhwala ophera tizilombo toyambitsa matenda monga ma siliva odzola amatha kuthandizira chitetezo cha mthupi cha munthu ndikuchotsa matenda a biofilm. Kapangidwe kake, kuchuluka kwa mankhwala, kusungunuka, ndi gawo loperekera mankhwala zimatha kusintha mphamvu ya siliva yolimbana ndi mavairasi. M'zaka zaposachedwapa, kupita patsogolo kwa ukadaulo wokonza siliva kwapangitsa kuti ma siliva awa akhale ogwira mtima kwambiri9,23. Pamene ukadaulo wokongoletsa siliva ukupita patsogolo, ndikofunikira kumvetsetsa momwe ma siliva awa amagwirira ntchito poletsa matenda a bala, komanso chofunika kwambiri, momwe mitundu yamphamvu ya siliva imakhudzira malo ovulala ndi kuchira.
Mu kafukufukuyu, tinayerekeza mphamvu ya ma dressing awiri apamwamba a siliva ndi ma dressing a siliva wamba omwe amapanga ma Ag1+ ions motsutsana ndi biofilms pogwiritsa ntchito mitundu yosiyanasiyana ya in vitro ndi in vivo. Tinawonanso momwe ma dressing awa amakhudzira malo ovulala komanso machiritso osagwirizana ndi matenda. Kuti tichepetse mphamvu ya matrix yobereka, ma dressing onse a siliva omwe adayesedwa anali opangidwa ndi carboxymethylcellulose.
Kuwunika kwathu koyambirira kwa ma dressing asiliva awa motsutsana ndi ma biofilms achikhalidwe a Pseudomonas aeruginosa ndi Staphylococcus aureus kukuwonetsa kuti, mosiyana ndi ma dressing achikhalidwe a Ag1+, ma dressing awiri apamwamba asiliva, Ag1+ + EDTA/BC ndi mchere wa Ag wokhala ndi mpweya, amagwira ntchito pa 5. Amapha mabakiteriya a biofilm mkati mwa masiku ochepa. Kuphatikiza apo, ma dressing awa amaletsa kupangidwanso kwa biofilm akakumana ndi mabakiteriya a planktonic mobwerezabwereza. Ma dressing a Ag1+ anali ndi siliva chloride, siliva compound ndi base matrix yomweyo monga Ag1+ + EDTA/BC, ndipo anali ndi zotsatira zochepa pa moyo wa mabakiteriya mkati mwa biofilm panthawi yomweyi. Kuwona kuti ma dressing a Ag1+ + EDTA/BC anali othandiza kwambiri motsutsana ndi biofilm kuposa ma dressing a Ag1+ okhala ndi matrix yomweyo ndi siliva compound kumathandizira lingaliro lakuti zosakaniza zina zimafunika kuti ziwonjezere mphamvu ya siliva chloride motsutsana ndi biofilm, monga momwe zanenedwera kwina15. Zotsatirazi zikugwirizana ndi lingaliro lakuti BC ndi EDTA zimagwira ntchito yowonjezerapo pothandizira kuti ma dressing onse akhale ogwira ntchito komanso kuti kusakhalapo kwa gawoli mu ma dressing a Ag1+ mwina kunathandizira kulephera kuwonetsa mphamvu ya in vitro. Tapeza kuti ma dressing a Ag okhala ndi mpweya omwe amapanga ma Ag2+ ndi ma Ag3+ ion anali ndi mphamvu yolimbana ndi mabakiteriya kuposa Ag1+ komanso pamlingo wofanana ndi Ag1+ + EDTA/BC. Komabe, chifukwa cha mphamvu yayikulu ya redox, sizikudziwika kuti ma Ag3+ ion amakhalabe ogwira ntchito komanso ogwira ntchito motsutsana ndi ma biofilms a mabala ndipo motero akuyenera kufufuzidwa kwina. Kuphatikiza apo, pali ma dressing ambiri osiyanasiyana omwe amapanga ma Ag1+ ion omwe sanayesedwe mu kafukufukuyu. Ma dressing awa amapangidwa ndi ma compounds osiyanasiyana asiliva, kuchuluka, ndi ma base matrices, omwe angakhudze kuperekedwa kwa ma Ag1+ ion ndi mphamvu zawo motsutsana ndi ma biofilms. Ndikofunikiranso kudziwa kuti pali mitundu yosiyanasiyana ya in vitro ndi in vivo yomwe imagwiritsidwa ntchito poyesa mphamvu ya ma dressing a mabala motsutsana ndi ma biofilms. Mtundu wa chitsanzo chomwe chagwiritsidwa ntchito, komanso kuchuluka kwa mchere ndi mapuloteni omwe amagwiritsidwa ntchito mu ma models awa, zidzakhudza mphamvu ya ma dressing. Mu chitsanzo chathu cha in vivo, tinalola kuti biofilm ikule mu vitro kenako n’kuipititsa pamwamba pa bala. Chitetezo cha mbewa ya host chimagwira ntchito bwino polimbana ndi mabakiteriya a planktonic omwe amaikidwa pa bala, motero amapanga biofilm pamene bala likuchira. Kuwonjezera biofilm yokhwima pa bala kumachepetsa mphamvu ya chitetezo cha host ku kapangidwe ka biofilm mwa kulola biofilm yokhwima kudzikhazikitsa yokha mkati mwa bala lisanayambe kuchira. Chifukwa chake, chitsanzo chathu chimatithandiza kuwunika momwe ma dressing ophera maantibayotiki amagwirira ntchito pa biofilms yokhwima mabala asanayambe kuchira.
Tinapezanso kuti kuvala koyenera kunakhudza momwe ma dressing asiliva amagwirira ntchito pa biofilms zomwe zimamera m'thupi la nkhumba komanso pakhungu la nkhumba. Kukhudzana kwambiri ndi bala kumaonedwa kuti ndikofunikira kwambiri pakugwira ntchito bwino kwa dressing24,25. Ma dressing okhala ndi mchere wa Ag wokhala ndi mpweya anali pafupi ndi biofilms okhwima, zomwe zinapangitsa kuti chiwerengero cha mabakiteriya okhazikika mkati mwa biofilm chichepe kwambiri pambuyo pa maola 24. Mosiyana ndi zimenezi, akalandira ma dressing a Ag1+ ndi Ag1+ + EDTA/BC, mabakiteriya ambiri okhazikika anakhalabe. Ma dressing awa ali ndi ma stitches kutalika konse kwa dressing, zomwe zimapangitsa kuti malo akufa asagwirizane ndi biofilm. Mu maphunziro athu a in vitro, malo osakhudzana ndi mabakiteriya okhazikika awa adaletsa kuphedwa kwa mabakiteriya okhazikika mkati mwa biofilm. Tinayesa kuthekera kwa mabakiteriya kukhala ndi moyo patatha maola 24 okha chithandizo; Pakapita nthawi, pamene dressing imakhala yodzaza kwambiri, pakhoza kukhala malo ochepa akufa, zomwe zimachepetsa malo a mabakiteriya okhazikika awa. Komabe, izi zikuwonetsa kufunika kwa kapangidwe ka dressing, osati mtundu wa siliva wokha mu dressing.
Ngakhale maphunziro a in vitro ndi othandiza poyerekeza mphamvu ya ukadaulo wosiyanasiyana wa siliva, ndikofunikiranso kumvetsetsa zotsatira za ma dressing awa pa biofilms in vivo, komwe minofu ya wolandirayo ndi mayankho a chitetezo cha mthupi zimathandiza kuti ma dressing awa agwire bwino ntchito motsutsana ndi biofilms. Zotsatira za ma dressing awa pa biofilms zawonedwa pogwiritsa ntchito scanning electron microscopy ndi EPS staining ya biofilm pogwiritsa ntchito utoto wa intracellular ndi extracellular DNA. Tinapeza kuti patatha masiku atatu a chithandizo, ma dressing onse anali othandiza kuchepetsa DNA yopanda maselo m'mabala omwe ali ndi biofilm, koma Ag1+ dressing sinagwire bwino ntchito m'mabala omwe ali ndi Staphylococcus aureus. Scanning electron microscopy inawonetsanso kuti panali mabakiteriya ochepa kwambiri m'mabala omwe amachiritsidwa ndi ma dressing asiliva, ngakhale izi zinali zodziwika bwino ndi oxygenated Ag salt dressing ndi Ag1+ + EDTA/BC dressing poyerekeza ndi Ag1+. Deta iyi ikuwonetsa kuti ma dressing asiliva omwe adayesedwa anali ndi mphamvu zosiyanasiyana pa kapangidwe ka biofilm, koma palibe ma dressing asiliva omwe adakwanitsa kuthetsa biofilm, zomwe zikuthandizira kufunikira kwa njira yonse yochizira matenda a biofilm ya mabala; kugwiritsa ntchito mabande asiliva. Chithandizo chimayamba ndi kuchotsa zotsalira za thupi kuti zichotsedwe mu biofilm yambiri.
Mabala osatha nthawi zambiri amakhala ndi kutupa kwakukulu, ndipo maselo ochulukirapo otupa amakhalabe m'minofu ya bala kwa nthawi yayitali, zomwe zimapangitsa kuti minofu iwonongeke ndikuchepetsa mpweya wofunikira kuti maselo agwire bwino ntchito komanso kuti agwire bwino ntchito m'bala26. Ma biofilms amawonjezera malo olakwika a mabalawa mwa kusokoneza machiritso m'njira zosiyanasiyana, kuphatikizapo kuletsa kuchulukana kwa maselo ndi kusamuka kwawo komanso kuyambitsa ma cytokines oyambitsa kutupa27. Pamene ma dressing asiliva akuyamba kugwira ntchito bwino, ndikofunikira kumvetsetsa momwe amakhudzira malo ovulala komanso machiritso awo.
Chochititsa chidwi n'chakuti, ngakhale kuti ma dressing onse asiliva adakhudza kapangidwe ka biofilm, ma dressing a siliva okhala ndi mpweya wokwanira okha ndi omwe adawonjezera kubwezeretsanso kwa mabala omwe ali ndi kachilomboka. Deta iyi ikuthandizira zomwe tapeza kale17 ndi zomwe za Kalan et al. (2017)28, zomwe zidawonetsa chitetezo chabwino komanso poizoni wa mchere wa siliva wokhala ndi mpweya wokwanira, chifukwa kuchuluka kochepa kwa siliva kunali kothandiza motsutsana ndi biofilms.
Kafukufuku wathu wapano akuwonetsa kusiyana kwa ukadaulo wa siliva pakati pa ma dressing asiliva opha tizilombo toyambitsa matenda komanso momwe ukadaulowu umakhudzira malo ovulala komanso machiritso osakhudzana ndi matenda. Komabe, zotsatirazi zimasiyana ndi kafukufuku wakale wosonyeza kuti ma dressing a Ag1+ + EDTA/BC adathandizira machiritso a makutu a kalulu ovulala m'thupi. Komabe, izi zitha kukhala chifukwa cha kusiyana kwa mitundu ya nyama, nthawi yoyezera, ndi njira zogwiritsira ntchito mabakiteriya29. Pankhaniyi, kuyeza mabala kunatengedwa patatha masiku 12 kuchokera pamene kuvulala kunapangitsa kuti zosakaniza zogwira ntchito za dressing zigwire ntchito pa biofilm kwa nthawi yayitali. Izi zikuthandizidwa ndi kafukufuku yemwe adawonetsa kuti zilonda zam'miyendo zomwe zimayambitsidwa ndi Ag1+ + EDTA/BC poyamba zidakula kukula pambuyo pa sabata imodzi ya chithandizo, kenako pa masabata atatu otsatira a chithandizo ndi Ag1+ + EDTA/BC komanso mkati mwa masabata anayi akugwiritsa ntchito mankhwala osagwiritsa ntchito maantibayotiki. Ma dressing a CMC kuti achepetse kukula kwa zilonda30.
Mitundu ina ya siliva ndi kuchuluka kwake kwawonetsedwa kale kuti ndi poizoni mu vitro 11, koma zotsatira zake mu vitro sizitanthauza nthawi zonse kukhala zotsatirapo zoyipa mu vivo. Kuphatikiza apo, kupita patsogolo kwa ukadaulo wa siliva komanso kumvetsetsa bwino mankhwala asiliva ndi kuchuluka kwake mu zodzoladzola kwapangitsa kuti pakhale zodzoladzola zambiri zasiliva zotetezeka komanso zothandiza. Komabe, pamene ukadaulo wa zodzoladzola zasiliva ukupita patsogolo, ndikofunikira kumvetsetsa momwe zodzoladzolazi zimakhudzira malo ovulala31,32,33. Kale zidanenedwa kuti kuchuluka kwa re-epithelialization kukugwirizana ndi kuchuluka kwa ma macrophages a M2 oletsa kutupa poyerekeza ndi mtundu wa pro-inflammatory M1. Izi zidawonedwa mu chitsanzo cha mbewa cham'mbuyomu pomwe zodzoladzola za siliva za hydrogel zinkayerekezeredwa ndi siliva sulfadiazine ndi hydrogels zosakhala antimicrobial34.
Mabala osatha akhoza kusonyeza kutupa kwambiri ndipo zapezeka kuti kukhalapo kwa ma neutrophils ochulukirapo kungakhale kovulaza kuchira kwa mabala35. Mu kafukufuku wa mbewa zomwe zatha ma neutrophils, kukhalapo kwa ma neutrophils kunachedwetsa kubwezeretsedwanso kwa ma neutrophils. Kupezeka kwa ma neutrophils ochulukirapo kumabweretsa kuchuluka kwa ma protease ndi mitundu ya okosijeni yogwira ntchito, monga superoxide ndi hydrogen peroxide, zomwe zimagwirizanitsidwa ndi mabala osatha komanso ochira pang'onopang'ono37,38. Momwemonso, kuwonjezeka kwa chiwerengero cha ma macrophage, ngati sikulamuliridwa, kungayambitse kuchira kwa mabala mochedwa39. Kuwonjezeka kumeneku ndikofunikira kwambiri ngati ma macrophage sangathe kusintha kuchoka ku phenotype yoyambitsa kutupa kupita ku phenotype yoyambitsa kuchira, zomwe zimapangitsa kuti mabala alephere kutuluka mu gawo lotupa la kuchira40. Tinaona kuchepa kwa ma neutrophils ndi ma macrophages m'mabala omwe ali ndi kachilombo ka biofilm pambuyo pa masiku atatu a chithandizo ndi ma dressing onse asiliva, koma kuchepako kunali koonekera kwambiri ndi ma dressing amchere okongoletsedwa ndi mpweya. Kuchepa kumeneku kungakhale chifukwa cha momwe chitetezo cha mthupi chimayankhira ku siliva, momwe thupi limayankhira kuchepa kwa katundu, kapena kuti bala likuchira pambuyo pake ndipo motero maselo a chitetezo cha mthupi m'bala akuchepa. Kuchepetsa kuchuluka kwa maselo otupa m'bala kungasunge malo abwino ochiritsira mabala. Njira yomwe Ag oxysalts imagwirira ntchito pochiritsira popanda matenda sizikudziwika bwino, koma kuthekera kwa Ag oxysalts kupanga mpweya ndikuwononga hydrogen peroxide yoopsa, yomwe ndi mkhalapakati wa kutupa, kungafotokoze izi ndipo kumafuna kafukufuku wowonjezera17.
Mabala osachiritsika omwe ali ndi kachilombo osachiritsa nthawi zonse amakhala vuto kwa madokotala ndi odwala. Ngakhale kuti ma dressing ambiri amanena kuti ndi othandiza pochiza mabala, kafukufuku samayang'ana kwambiri zinthu zina zofunika zomwe zimakhudza chilengedwe cha mabala. Kafukufukuyu akuwonetsa kuti matekinoloje osiyanasiyana asiliva ali ndi mphamvu yosiyana yolimbana ndi mabakiteriya ndipo, chofunika kwambiri, zotsatira zosiyana pa malo ovulala ndi kuchira, mosasamala kanthu za matenda. Ngakhale kuti maphunziro awa a in vitro ndi in vivo akuwonetsa kuti ma dressing awa amagwira ntchito bwino pochiza matenda a mabala ndikulimbikitsa kuchira, mayeso olamulidwa mwachisawawa amafunika kuti awone momwe ma dressing awa amagwirira ntchito kuchipatala.
Ma data omwe agwiritsidwa ntchito ndi/kapena kufufuzidwa panthawi ya kafukufukuyu akupezeka kwa wolemba woyenerera ngati pakufunika kutero.
Nthawi yotumizira: Julayi-15-2024