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Ukukhula kwamagciwane emanxebeni kuvame ukuzibonakalisa njenge-biofilms, ephazamisa ukuphulukiswa futhi kunzima ukuyiqeda. Ama-silver dressing amasha athi alwa nokutheleleka kwamanxeba, kodwa ukusebenza kwawo kwe-antibiofilm kanye nemiphumela yokuphulukisa engaxhomekile ekuthelelekeni ngokuvamile akwaziwa. Sisebenzisa amamodeli e-in vitro kanye ne-in vivo biofilm e-Staphylococcus aureus kanye ne-Pseudomonas aeruginosa, sibika ukusebenza kahle kwama-Ag1+ ion-generating dressings; ama-Ag1+ dressings aqukethe i-ethylenediaminetetraacetic acid kanye ne-benzethonium chloride (Ag1+/EDTA/BC), kanye nama-dressing aqukethe i-silver nitrate (Ag oxysalts). , akhiqiza ama-Ag1+, Ag2+ kanye nama-Ag3+ ion ukulwa ne-wound biofilm kanye nomphumela wayo ekuphulukisweni. Ama-Ag1+ dressing abe nemiphumela emincane ku-wound biofilm in vitro kanye namagundane (C57BL/6j). Ngokuphambene nalokho, usawoti we-Ag one-oxygen kanye nokugqoka kwe-Ag1+/EDTA/BC kunciphise kakhulu inani lamabhaktheriya asebenzayo kuma-biofilms in vitro futhi kwabonisa ukwehla okukhulu kwezingxenye zamabhaktheriya kanye ne-EPS kuma-biofilms amanxeba egundane. Lezi zingubo zinemiphumela ehlukene ekwelapheni amanxeba atheleleke nge-biofilm kanye nalawo angenayo i-biofilm, kanti ukugqoka kukasawoti one-oxygen kunemiphumela enenzuzo kakhulu ekuvuseleleni kabusha, usayizi wamanxeba, kanye nokuvuvukala uma kuqhathaniswa nokwelashwa kokulawula kanye nezinye izingubo zesiliva. Izakhiwo ezahlukene ze-physicochemical zezingubo zesiliva zingaba nemiphumela ehlukene kuma-biofilm amanxeba kanye nokuphulukiswa, futhi lokhu kufanele kucatshangelwe lapho kukhethwa ingubo yokwelapha amanxeba atheleleke nge-biofilm.
Amanxeba angapheli achazwa ngokuthi “amanxeba ahluleka ukuqhubekela phambili ezigabeni ezijwayelekile zokuphulukiswa ngendlela ehlelekile nefike ngesikhathi” 1. Amanxeba angapheli adala umthwalo wengqondo, wezenhlalo kanye nowezomnotho ezigulini kanye nohlelo lwezempilo. Imali esetshenziswa yi-NHS minyaka yonke ekwelapheni amanxeba kanye nezinye izifo ezihlobene nawo ilinganiselwa ku-£8.3 billion ngo-2017-182. Amanxeba angapheli nawo njengamanje ayinkinga enkulu e-United States, lapho i-Medicare ilinganisela izindleko zonyaka zokwelapha iziguli ezinamanxeba ku-$28.1–$96.8 billion3.
Ukutheleleka kuyisici esiyinhloko esivimbela ukuphulukiswa kwenxeba. Ukutheleleka kuvame ukubonakala njenge-biofilms, ekhona kuma-78% amanxeba angapholi angalapheki. Ama-biofilms akheka lapho ama-microorganisms enamathela ngokungenakulungiseka ezindaweni, njengezindawo zamanxeba, futhi angahlangana ukuze akhe imiphakathi ekhiqiza i-extracellular polymer (EPS). I-biofilm yamanxeba ihlotshaniswa nokusabela okwandayo kokuvuvukala okuholela ekulimaleni kwezicubu, okungabambezela noma kuvimbele ukuphulukiswa4. Ukwanda komonakalo wezicubu kungase kubangelwe ngokwengxenye umsebenzi owandisiwe wama-matrix metalloproteinases, i-collagenase, i-elastase kanye nezinhlobo ze-oxygen ezisabelayo5. Ngaphezu kwalokho, amaseli okuvuvukala kanye nama-biofilms ngokwawo angabathengi abaphezulu be-oxygen futhi ngenxa yalokho angabangela i-hypoxia yezicubu zendawo, aqede amaseli omoya-mpilo obalulekile odingekayo ukuze kulungiswe izicubu ngempumelelo6.
Ama-biofilm avuthiwe amelana kakhulu nama-antimicrobial agents, adinga amasu anamandla okulawula ukutheleleka kwe-biofilm, njengokwelashwa ngomshini okulandelwa ukwelashwa okusebenzayo kwe-antimicrobial. Ngenxa yokuthi ama-biofilm angaphinde avuseleleke ngokushesha, ama-antimicrobial asebenzayo anganciphisa ingozi yokwakheka kabusha ngemva kokususwa kwezicubu ngokuhlinzwa7.
Isiliva lisetshenziswa kakhulu ekufakweni kwama-antimicrobial futhi livame ukusetshenziswa njengokwelashwa kokuqala kwamanxeba athelelekile angapheli. Kunezinhlobo eziningi zesiliva ezithengiswayo, ngayinye equkethe ukwakheka kwesiliva okuhlukile, ukuhlushwa, kanye nesisekelo. Intuthuko kuma-armbands esiliva iholele ekuthuthukisweni kwama-armbands esiliva amasha. Uhlobo lwesiliva oluyinsimbi (Ag0) alusebenzi; Ukuze kufezwe ukusebenza kahle kwama-antimicrobial, kumele ilahlekelwe yi-electron ukuze yakhe isiliva le-ionic (Ag1+). Ama-armbands esiliva endabuko aqukethe ama-compounds esiliva noma isiliva lensimbi, lapho evezwa oketshezini, aqhekeke ukuze akhe ama-ion e-Ag1+. Lawa ma-ion e-Ag1+ asabela neseli lebhaktheriya, asuse ama-electron ezingxenyeni zesakhiwo noma ezinqubweni ezibalulekile ezidingekayo ukuze kusindiswe. Ubuchwepheshe obunelungelo lobunikazi buholele ekuthuthukisweni kwe-compound entsha yesiliva, i-Ag oxysalts (i-nitrate yesiliva, i-Ag7NO11), efakiwe ekufakweni kwama-armbands. Ngokungafani nesiliva lendabuko, ukubola kosawoti oqukethe umoya-mpilo kukhiqiza izimo zesiliva ezine-valence ephezulu (Ag1+, Ag2+ kanye ne-Ag3+). Izifundo ze-in vitro zikhombisile ukuthi amazinga aphansi kasawoti wesiliva one-oxygen asebenza kangcono kunesiliva elilodwa (Ag1+) ngokumelene namagciwane abangela izifo njenge-Pseudomonas aeruginosa, i-Staphylococcus aureus kanye ne-Escherichia coli8,9. Olunye uhlobo olusha lwe-silver dressing luhlanganisa izithako ezengeziwe, okuyi-ethylenediaminetetraacetic acid (EDTA) kanye ne-benzethonium chloride (BC), okubikwa ukuthi zihlasela i-biofilm EPS futhi ngaleyo ndlela zandise ukungena kwesiliva ku-biofilm. Lezi zobuchwepheshe ezintsha zesiliva zinikeza izindlela ezintsha zokuhlasela ama-biofilm amanxeba. Kodwa-ke, umthelela walezi zidakamizwa zokulwa namagciwane endaweni yamanxeba kanye nokuphulukiswa okungaxhomekeki ekuthelelekeni kubalulekile ukuqinisekisa ukuthi azidali indawo yamanxeba engathandeki noma zilibazise ukuphulukiswa. Ukukhathazeka mayelana ne-cytotoxicity yesiliva ye-in vitro kubikwe nge-silver dressing eziningana10,11. Kodwa-ke, i-cytotoxicity ye-in vitro ayikahunyushwa ibe ubuthi be-in vivo, futhi ama-Ag1+ dressing amaningana abonise iphrofayili enhle yokuphepha12.
Lapha, sihlole ukusebenza kahle kwezindwangu ze-carboxymethylcellulose eziqukethe izinhlobo ezintsha zesiliva ngokumelene ne-biofilm yesilonda ngaphakathi kwe-vitro kanye ne-in vivo. Ngaphezu kwalokho, imiphumela yalezi zindwangu ezisabela ekuzivikeleni komzimba nasekuphulukisweni ngaphandle kokutheleleka yahlolwa.
Zonke izindwangu ezisetshenzisiwe zazitholakala kwezentengiselwano. I-3M Kerracel Gel Fiber Dressing (3M, Knutsford, UK) iyindwangu ye-gel fiber engeyona elwa namagciwane engu-100% carboxymethylcellulose (CMC) esetshenziswe njengendwangu yokulawula kulolu cwaningo. Kuhlolwe izindwangu ezintathu zesiliva ze-CMC ezilwa namagciwane, okuyi-3M Kerracel Ag dressing (3M, Knutsford, UK), equkethe usawoti wesiliva one-oxygen ongu-1.7 wt%. (Ag7NO11) kuma-ion esiliva e-valence aphezulu (Ag1+, Ag2+ kanye ne-Ag3+). Ngesikhathi sokubola kwe-Ag7NO11, ama-ion e-Ag1+, Ag2+ kanye ne-Ag3+ akhiwa ngesilinganiso esingu-1:2:4. I-Aquacel Ag Extra dressing equkethe i-1.2% silver chloride (Ag1+) (ConvaTec, Deeside, UK) 13 kanye ne-Aquacel Ag + Extra dressing equkethe i-1.2% silver chloride (Ag1+), i-EDTA kanye ne-benzethonium chloride (ConvaTec, Deeside, UK) 14.
Izinhlobo ezisetshenziswe kulolu cwaningo kwakuyi-Pseudomonas aeruginosa NCTC 10781 (Public Health England, Salisbury) kanye ne-Staphylococcus aureus NCTC 6571 (Public Health England, Salisbury).
Amabhaktheriya akhuliswa ngobusuku obubodwa emhluzini we-Muller-Hinton (Oxoid, Altrincham, UK). Isiko lobusuku obubodwa sabe sesincishiswa ngo-1:100 emhluzini we-Mueller-Hinton kanye no-200 µl ofakwe kuma-membrane e-Whatman cyclopore ahlanzekile angu-0.2 µm (Whatman plc, Maidstone, UK) kuma-plate e-agar e-Mueller-Hinton (Sigma-Aldrich Company Ltd, Kent, Great Britain). ) Ukwakheka kwe-biofilm yamakoloni ku-37°C amahora angama-24. Lawa ma-biofilm amakoloni ahlolwe ukuncipha kwe-logarithmic.
Sika i-dressing ibe yizicucu zesikwele ezingama-3 cm2 bese umanzisa ngamanzi ahlanzekile ahlanzekile. Beka ibhandishi phezu kwe-biofilm yekoloni epuletini le-agar. Wonke amahektha angu-24 e-biofilm asusiwe, futhi amabhaktheriya asebenzayo ngaphakathi kwe-biofilm (CFU/ml) alinganiswa ngokuncibilikisa okulandelanayo (10−1 kuya ku-10−7) ku-Day-Angle neutralization broth (Merck-Millipore). Ngemva kwamahora angu-24 okufukamela ku-37°C, ukubalwa kwamapuleti okujwayelekile kwenziwa kumapuleti e-agar e-Mueller-Hinton. Ukwelashwa ngakunye kanye nesikhathi kwenziwa kathathu, futhi ukubalwa kwamapuleti kwaphindwa ngokuncibilikisa ngakunye.
Isikhumba sesisu sengulube sitholakala ezingulubeni zensikazi ezinkulu ezimhlophe zingakapheli imizuzu eyi-15 zihlatshiwe ngokuvumelana nezindinganiso zokuthumela kwamanye amazwe ze-European Union. Isikhumba sagundwa futhi sahlanzwa ngeziphuphu zotshwala, sabe sesiqandisiwe ku--80°C amahora angama-24 ukuze siqede amandla esikhumbeni. Ngemva kokuncibilika, izingcezu zesikhumba eziyi-1 cm2 zagezwa kathathu nge-PBS, i-0.6% sodium hypochlorite, kanye ne-70% ethanol imizuzu engama-20 isikhathi ngasinye. Ngaphambi kokususa i-epidermis, susa noma iyiphi i-ethanol esele ngokugeza izikhathi ezi-3 ku-PBS engcolile. Isikhumba sakhuliswa epuletini elinamanzi ayi-6 eline-membrane ye-nylon engu-0.45-μm ubukhulu (Merck-Millipore) phezulu kanye nama-pad amathathu okumunca (Merck-Millipore) aqukethe i-3 ml ye-fetal bovine serum (Sigma) engezwe nge-10% Dulbecco's modified Eagle. Medium (Dulbecco's Modified Eagle Medium – Aldrich Ltd.).
Ama-biofilm ekoloni akhuliswa njengoba kuchaziwe ezifundweni zokuchayeka kwe-biofilm. Ngemva kokukhulisa i-biofilm ku-membrane amahora angu-72, i-biofilm yafakwa ebusweni besikhumba kusetshenziswa i-sterile inoculation loop kwase kususwa i-membrane. I-biofilm yabe isifakwa ku-dermis yengulube amahora angu-24 engeziwe ku-37°C ukuze ivumele i-biofilm ukuthi ikhule futhi inamathele esikhumbeni sengulube. Ngemva kokuba i-biofilm isivuthiwe futhi inamathiselwe, i-dressing engu-1.5 cm2, efakwe amanzi ahlanzekile ahlanzekile, yafakwa ngqo ebusweni besikhumba futhi yafakwa ku-37°C amahora angu-24. Amabhaktheriya aphilayo abonakala ngokufaka umbala ngokufaka ngokufanayo i-PrestoBlue cell viability reagent (Invitrogen, Life Technologies, Paisley, UK) ebusweni obuphezulu besitshalo ngasinye bese uyibeka ku-incubation imizuzu emi-5. Sebenzisa ikhamera yedijithali yeLeica DFC425 ukuze uthwebule izithombe ngokushesha ku-microscope yeLeica MZ8. Izindawo ezinombala opinki zalinganiswa kusetshenziswa isofthiwe ye-Image Pro version 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (mediacy.com)). Ukuskena nge-electron microscopy kwenziwa njengoba kuchaziwe ngezansi.
Amabhaktheriya akhuliswe ubusuku bonke ancibilikiswa nge-1:100 kumhluzi we-Mueller-Hinton. Ama-200 μl esiko afakwe ku-0.2 μm Whatman cyclopore membrane engcolile (Whatman, Maidstone, UK) futhi abekwa ku-agar ye-Mueller-Hinton. Amapuleti e-biofilm afakwa ku-37°C amahora angu-72 ukuze kuvunyelwe ukwakheka kwe-biofilm evuthiwe.
Ngemva kwezinsuku ezi-3 zokuvuthwa kwe-biofilm, ibhandishi lesikwele esingu-3 cm2 lafakwa ngqo kwi-biofilm futhi lafakwa ekushiseni okungu-37°C amahora angama-24. Ngemva kokususa ibhandishi ebusweni be-biofilm, i-1 ml ye-PrestoBlue Cell Viability Reagent (Invitrogen, Waltham, MA) yanezelwa ebusweni be-biofilm ngayinye imizuzwana engama-20. Izindawo zomile ngaphambi kokuba kuqoshwe izinguquko zombala kusetshenziswa ikhamera yedijithali ye-Nikon D2300 (Nikon UK Ltd., Kingston, UK).
Lungisa ukukhuliswa kwasebusuku ku-agar kaMueller-Hinton, udlulisele amakoloni ngamanye ku-10 ml yomhluzi kaMueller-Hinton bese ufukamela ku-shaker ku-37°C (100 rpm). Ngemva kokufukamela ebusuku, ukukhuliswa kwancishiswa ngo-1:100 kumhluzi kaMueller-Hinton kwathi u-300 µl wabonakala ku-0.2 µm circular Whatman cyclopore membrane (Whatman International, Maidstone, UK) ku-agar kaMueller-Hinton futhi kwafukamela ku-37°C zingakapheli amahora angu-72. . I-biofilm evuthiwe yafakwa esilondeni njengoba kuchaziwe ngezansi.
Wonke umsebenzi nezilwane wenziwa eNyuvesi yaseManchester ngaphansi kwelayisensi yephrojekthi evunyelwe yiHhovisi Lezenhlalakahle Yezilwane kanye Nokubuyekezwa Kokuziphatha (P8721BD27) nangokuhambisana neziqondiso ezishicilelwe yiHhovisi Lasekhaya ngaphansi kwe-ASPA ebuyekeziwe ka-2012. Bonke ababhali balandele iziqondiso zokufika. Amagundane e-C57BL/6j anamasonto ayisishiyagalombili (Envigo, Oxon, UK) asetshenziswa kuzo zonke izifundo ze-in vivo. Amagundane ahlinzwa nge-isoflurane (Piramal Critical Care Ltd, West Drayton, UK) futhi ubuso bawo bangemuva baphucwa futhi bahlanzwa. Igundane ngalinye labe selinikwa inxeba lokususa elingu-2 × 6 mm kusetshenziswa i-Stiefel biopsy punch (Schuco International, Hertfordshire, UK). Kumanxeba atheleleke nge-biofilm, faka i-biofilm yamahora angu-72 ekhuliswe kulwelwesi njengoba kuchaziwe ngenhla kungqimba lwesikhumba senxeba usebenzisa i-loop yokugoma engcolile ngokushesha ngemva kokulimala bese ulahla ulwelwesi. Isentimitha elilodwa lesikwele lokugqoka limanziswa kusengaphambili ngamanzi angcolile ukuze kugcinwe indawo yenxeba emanzi. Ama-dressing afakwa ngqo enxebeni ngalinye futhi ambozwe ngefilimu ye-3M Tegaderm (3M, Bracknell, UK) kanye ne-Mastisol liquid adhesive (Eloquest Healthcare, Ferndale, MI) efakwe emaphethelweni ukuze kuhlinzekwe ukunamathela okwengeziwe. I-Buprenorphine (Animalcare, York, UK) yanikezwa ngesilinganiso esingu-0.1 mg/kg njengesiqeda ubuhlungu. Sika amagundane ezinsukwini ezintathu ngemva kokulimala usebenzisa indlela yeShejuli 1 bese ususa, uhlukanise phakathi, bese ugcina indawo yenxeba njengoba kudingeka.
Ukupenda nge-Hematoxylin (ThermoFisher Scientific) kanye ne-eosin (ThermoFisher Scientific) kwenziwa ngokwenqubo yomenzi. Indawo yenxeba kanye nokulungiswa kabusha kwalinganiswa kusetshenziswa isofthiwe ye-Image Pro version 10 (Media Cybernetics Inc, Rockville, MD).
Izingxenye zezicubu zahlanzwa nge-xylene (ThermoFisher Scientific, Loughborough, UK), zaphinda zamanziswa nge-ethanol engu-100–50% esezingeni, futhi zacwiliswa isikhashana emanzini acwengekile (ThermoFisher Scientific). I-Immunohistochemistry yenziwe kusetshenziswa i-VectaStain Elite ABC PK-6104 kit (Vector Laboratories, Burlingame, CA) ngokwephrothokholi yomenzi. Ama-antibodies ayinhloko kuma-neutrophils i-NIMP-R14 (Thermofisher Scientific) kanye nama-macrophages i-Ms CD107b Pure M3/84 (BD Biosciences, Wokingham, UK) ahlanjululwe nge-1:100 esixazululweni esivimbayo futhi afakwa endaweni eqoshiwe, kwalandela ama-antibodies amabili i-Anti-, VectaStain ABC kanye ne-Vector Nova Red Peroxidase (HRP) substrate kit (Vector Laboratories, Burlingame, CA) futhi ahlanjululwe nge-hematoxylin. Izithombe zatholakala kusetshenziswa imakroskopu ye-Olympus BX43 kanye nekhamera yedijithali ye-Olympus DP73 (e-Olympus, eSouthend-on-Sea, e-UK).
Amasampula esikhumba aqiniswe ku-2.5% glutaraldehyde kanye no-4% formaldehyde ku-0.1 M HEPES (pH 7.4) amahora angama-24 ku-4°C. Amasampula ancishiswa amanzi kusetshenziswa i-ethanol esezingeni eliphezulu futhi omiswa ku-CO2 kusetshenziswa i-Quorum K850 critical point dryer (Quorum Technologies Ltd, Loughton, UK) kanye ne-sputter embozwe nge-gold-palladium alloy kusetshenziswa i-Quorum SC7620 mini sputterer/glow discharge system. Amasampula athathwe izithombe kusetshenziswa i-FEI Quanta 250 scanning electron microscope (ThermoFisher Scientific) ukuze kubonwe iphuzu eliphakathi lenxeba.
I-Toto-1 iodide (2 μM) yafakwa endaweni yenxeba legundane elikhishwe futhi yafakwa esikhumbeni imizuzu emi-5 ku-37 °C (ThermoFisher Scientific) futhi yaphathwa nge-Syto-60 (10 μM) ku-37 °C (ThermoFisher Scientific). Izithombe ze-Z-stack zemizuzu eyi-15 zadalwa kusetshenziswa i-Leica TCS SP8.
Idatha ephindaphindwayo yezinto eziphilayo kanye nezobuchwepheshe yahlelwa futhi yahlaziywa kusetshenziswa isofthiwe ye-Graphpad Prism V9 (GraphPad Software, La Jolla, CA). Ukuhlaziywa kwendlela eyodwa kokuhlukahluka ngokuqhathanisa okuningi kusetshenziswa ukuhlolwa kwe-post hoc kukaDunnett kwasetshenziswa ukuhlola umehluko phakathi kokwelashwa ngakunye kanye ne-non-antimicrobial control dressing. Inani le-p <0.05 labhekwa njengelibalulekile.
Ukusebenza kahle kwezingubo zesiliva ezine-fibrous gel kwahlolwa okokuqala ngokumelene namakholoni e-biofilm e-Staphylococcus aureus kanye ne-Pseudomonas aeruginosa in vitro. Izingubo zesiliva ziqukethe amafomula ahlukene esiliva: izingubo zesiliva zendabuko zikhiqiza ama-ion e-Ag1+; izingubo zesiliva, ezingakhiqiza ama-ion e-Ag1+ ngemva kokufakwa kwe-EDTA/BC, zingabhubhisa i-matrix ye-biofilm futhi ziveze amabhaktheriya esiliva ngaphansi komphumela wokulwa namagciwane we-ions yesiliva15 kanye nezingubo eziqukethe usawoti we-Ag one-oxygen okhiqiza ama-ion e-Ag1+, Ag2+ kanye ne-Ag3+. Ukusebenza kwayo kuqhathaniswa nengubo yokulawula engeyona i-antimicrobial eyenziwe ngemicu ye-gelled. Amabhaktheriya asele asebenzayo ngaphakathi kwe-biofilm ahlolwa njalo emahoreni angama-24 izinsuku eziyi-8 (Isithombe 1). Ngosuku lwesi-5, i-biofilm yaphinde yafakwa ku-3.85 × 105S. I-Staphylococcus aureus noma i-1.22 × 105P. aeruginosa ukuhlola ukululama kwe-biofilm. Uma kuqhathaniswa nezingubo zokulawula ezingezona ezokulwa namagciwane, izambatho ze-Ag1+ zibe nomthelela omncane ekusebenzeni kwamagciwane ku-Staphylococcus aureus kanye ne-Pseudomonas aeruginosa biofilms ezinsukwini ezi-5. Ngokuphambene nalokho, izambatho eziqukethe usawoti we-Ag kanye ne-Ag1+ + EDTA/BC one-oxygen zazisebenza kahle ekubulaleni amagciwane ngaphakathi kwe-biofilm zingakapheli izinsuku ezi-5. Ngemva kokugonywa okuphindaphindiwe ngamagciwane e-planktonic ngosuku lwesi-5, akukho ukubuyiselwa kwe-biofilm okwabonwa (Isithombe 1).
Ukulinganiswa kwamagciwane aphilayo ku-Staphylococcus aureus kanye ne-Pseudomonas aeruginosa biofilms ngemva kokwelashwa nge-silver dressings. Amakholoni e-Biofilm e-Staphylococcus aureus kanye ne-Pseudomonas aeruginosa aphathwe nge-silver dressings noma nge-non-antimicrobial control dressings, kanti inani lamagciwane aphilayo asele lanqunywa njalo emahoreni angama-24. Ngemva kwezinsuku ezi-5, i-biofilm yaphinde yafakwa ku-3.85×105S. I-Staphylococcus aureus noma i-1.22×105P. Amakholoni e-bacterioplankton Pseudomonas aeruginosa akhiwa ngawodwana ukuze kuhlolwe ukululama kwe-biofilm. Amagrafu abonisa iphutha elijwayelekile +/- elijwayelekile.
Ukuze sibone umphumela wezingubo zesiliva ekuphileni kwe-biofilm, izingubo zokugqoka zifakwe kuma-biofilm avuthiwe akhuliswe esikhumbeni sengulube ngaphandle kokuphila. Ngemva kwamahora angu-24, ingubo yokugqoka iyasuswa bese i-biofilm igcotshwa ngodayi oluhlaza okwesibhakabhaka osabelayo, oguqulwa amabhaktheriya aphilayo abe umbala opinki. Ama-biofilm aphathwe ngezingubo zokulawula ayepinki, okubonisa ukuthi kukhona amabhaktheriya aphilayo ngaphakathi kwe-biofilm (Isithombe 2A). Ngokuphambene nalokho, i-biofilm ephathwe ngengubo ye-oxysols ye-Ag yayiluhlaza okwesibhakabhaka ngokuyinhloko, okubonisa ukuthi amabhaktheriya asele ebusweni besikhumba sengulube ayengamabhaktheriya angaphili (Isithombe 2B). Umbala oxubile oluhlaza okwesibhakabhaka nopinki wabonwa kuma-biofilm aphathwe ngezingubo ze-Ag1+, okubonisa ukuthi kukhona amabhaktheriya aphilayo nangenali ngaphakathi kwe-biofilm (Isithombe 2C), kanti izingubo ze-EDTA/BC eziqukethe i-Ag1+ zaziluhlaza okwesibhakabhaka kakhulu zinamabala athile apinki. okubonisa izindawo ezingathintekile yingubo yesiliva (Isithombe 2D). Ukulinganiswa kwezindawo ezisebenzayo (ezipinki) nezingasebenzi (eziluhlaza okwesibhakabhaka) kubonise ukuthi i-control patch yayisebenza ngo-75% (Isithombe 2E). Ukufakwa kwe-Ag1+ + EDTA/BC kwenziwe ngendlela efanayo nokufakwa kwe-Ag salt okunomoya-mpilo, ngamazinga okusinda angu-13% no-14%, ngokulandelana. Ukufakwa kwe-Ag1+ nakho kunciphise ukusebenza kwamagciwane ngo-21%. Lawa ma-biofilm abe esebonwa kusetshenziswa i-scanning electron microscopy (SEM). Ngemva kokwelashwa nge-control dressing kanye ne-Ag1+ dressing, kwabonakala ungqimba lwe-Pseudomonas aeruginosa lumboza isikhumba sengulube (Isithombe 2F,H), kanti ngemva kokwelashwa nge-Ag1+ dressing, kwatholakala amangqamuzana ambalwa ebhaktheriya futhi kwatholakala amangqamuzana ambalwa ebhaktheriya ngaphansi. Imicu ye-Collagen ingabhekwa njengesakhiwo sezicubu zesikhumba sengulube (Isithombe 2G). Ngemva kokwelashwa nge-Ag1+ + EDTA/BC dressing, kwabonakala ama-bacterial plaque kanye nama-collagen fiber plaque angaphansi (Isithombe 2I).
Ukubona ngeso lengqondo i-Pseudomonas aeruginosa biofilm ngemva kokwelashwa nge-silver dressing. (A–D) Ukuphila kwamagciwane ku-Pseudomonas aeruginosa biofilms ezikhule esikhumbeni sengulube kwabonakala ngeso lengqondo kusetshenziswa i-PrestoBlue viability dye emahoreni angu-24 ngemva kokwelashwa nge-silver dressing noma i-non-antimicrobial control dressings. Amagciwane aphilayo apinki, amagciwane angaphili kanti isikhumba sengulube siluhlaza okwesibhakabhaka. (E) Ukulinganiswa kwe-Pseudomonas aeruginosa biofilms ezikhule esikhumbeni sengulube (ibala elipinki) kusetshenziswa i-scanning electron microscopy Image Pro version 10 (FI) futhi zelashwa nge-silver dressing noma i-non-antimicrobial control dressing amahora angu-24. Ibha yesikali se-SEM = 5 µm. (J–M) Ama-biofilms asekoloni akhule ezihlungini futhi agcotshwa nge-PrestoBlue reactive dye ngemva kwamahora angu-24 okufukamela nge-silver dressing.
Ukuze kutholakale ukuthi ukuxhumana okuseduze phakathi kwezindwangu kanye nama-biofilm kuthinte ukusebenza kahle kwezindwangu, ama-biofilm amakoloni abekwe endaweni eyisicaba aphathwa ngezindwangu amahora angama-24 bese egcotshwa ngodayi abasabelayo. I-biofilm engaphathwanga yayinombala opinki omnyama (Isithombe 2J). Ngokungafani nama-biofilm aphathwe ngezindwangu eziqukethe usawoti we-Ag one-oxygen (Isithombe 2K), ama-biofilm aphathwe ngezindwangu eziqukethe i-Ag1+ noma i-Ag1+ + EDTA/BC abonise amabhande okudaya okupinki (Isithombe 2L, M). Lo mbala opinki ukhombisa ukuba khona kwamagciwane aphilayo futhi uhlotshaniswa nendawo yokuthungwa ngaphakathi kwesindwangu. Lezi zindawo ezithungwe ngaphakathi zidala izikhala ezifile ezivumela amagciwane ngaphakathi kwe-biofilm ukuthi asinde.
Ukuze kuhlolwe ukusebenza kahle kwezingubo zesiliva emzimbeni, izilonda ezikhishwe ngokuphelele zamagundane atheleleke nge-S. aureus evuthiwe kanye ne-P. aeruginosa biofilms zaphathwa ngezingubo zokulawula ezingezona ezilwa namagciwane noma izambatho zesiliva. Ngemva kwezinsuku ezi-3 zokwelashwa, ukuhlaziywa kwesithombe se-macroscopic kwabonisa ubukhulu obuncane bezilonda lapho kuphathwa ngezingubo zosawoti ezine-oxygen uma kuqhathaniswa nezingubo zokulawula ezingezona ezilwa namagciwane kanye nezinye izambatho zesiliva (Isithombe 3A-H). Ukuqinisekisa lokhu okubonwe, izilonda zavunwa futhi indawo yenxeba kanye nokuvuselelwa kabusha kwalinganiswa ezingxenyeni zezicubu ezinemibala ye-hematoxylin kanye ne-eosin kusetshenziswa isofthiwe yesithombe pro version 10 (Isithombe 3I-L).
Umphumela wokufakwa kwesiliva ebusweni benxeba kanye nokufakwa kabusha kwe-epithelialization yamanxeba atheleleke nge-biofilms. (A–H) Amaseli amancane atheleleke nge-biofilms ye-Pseudomonas aeruginosa (A–D) kanye ne-Staphylococcus aureus (E–H) ngemva kwezinsuku ezintathu zokwelashwa nge-non-antimicrobial control dressing, i-oxygenated Ag salt dressing, i-Ag1+ dressing, kanye ne-Ag1+. Izithombe ezimele i-macroscopic. amanxeba amagundane ane-Ag1+ + EDTA/BC dressing. (IL) Ukutheleleka kwe-Pseudomonas aeruginosa okumele, izingxenye ze-histological ezifakwe i-hematoxylin kanye ne-eosin, ezisetshenziselwa ukulinganisa indawo yenxeba kanye nokuvuselelwa kwe-epithelial. Ukulinganiswa kwendawo yenxeba (M, O) kanye nephesenti lokufakwa kabusha kwe-reepithelialization (N, P) yamanxeba atheleleke nge-Pseudomonas aeruginosa (M, N) kanye ne-Staphylococcus aureus (O, P) biofilms (ngeqembu lokwelashwa n = 12). Amagrafu abonisa iphutha elijwayelekile +/-. * kusho u-p = < 0.05 ** kusho u-p = < 0.01; Isikali se-macroscopic = 2.5 mm, isikali se-histological = 500 µm.
Ukulinganiswa kwendawo yenxeba emanxebeni atheleleke nge-Pseudomonas aeruginosa biofilm (Isithombe 3M) kubonise ukuthi amanxeba aphathwe nge-Ag oxysalts ayenosayizi wenxeba omaphakathi ongu-2.5 mm2, kuyilapho i-non-antimicrobial control dressing yayinosayizi wenxeba omaphakathi ongu-3.1 mm2, okungelona iqiniso. kufinyelele ukubaluleka kwezibalo (Isithombe 3M). p = 0.423). Amanxeba aphathwe nge-Ag1+ noma i-Ag1+ + EDTA/BC awazange abonise ukwehla endaweni yenxeba (3.1 mm2 kanye no-3.6 mm2, ngokulandelana). Ukwelashwa nge-oxygenated Ag salt dressing kukhuthaze ukuvuselelwa kabusha kwe-epithelialization ngezinga elikhulu kune-non-antimicrobial control dressing (34% kanye no-15%, ngokulandelana; p = 0.029) kanye ne-Ag1+ noma i-Ag1+ + EDTA/BC (10% kanye no-11%) (Isithombe 3N). . , ngokulandelana).
Ukuthambekela okufanayo endaweni yenxeba kanye nokuvuselelwa kwe-epithelial kwabonwa emanxebeni atheleleke nge-S. aureus biofilms (Isithombe 3O). Ukufakwa kosawoti wesiliva one-oxygen kunciphisa indawo yenxeba (2.0 mm2) ngo-23% uma kuqhathaniswa nokufakwa kosawoti okungeyona i-antimicrobial (2.6 mm2), yize lokhu kuncipha kwakungabonakali (p = 0.304) (Isithombe 3O). Ngaphezu kwalokho, indawo yenxeba eqenjini lokwelashwa kwe-Ag1+ yancishiswa kancane (2.4 mm2), kuyilapho inxeba elashwe nge-Ag1+ + EDTA/BC dressing lingayinciphisi indawo yenxeba (2.9 mm2). Usawoti we-oxygen we-Ag nawo wakhuthaza ukuvuselelwa kabusha kwamanxeba atheleleke nge-S. aureus biofilm (31%) ngezinga elikhulu kunalawo aphathwe nge-non-antimicrobial control dressings (12%, p = 0.003) (Isithombe 3P). Ukugqoka kwe-Ag1+ (16%, p = 0.903) kanye nokugqoka kwe-Ag+1 + EDTA/BC (14%, p = 0.965) kubonise amazinga okuvuselelwa kwe-epithelial afana nokulawula.
Ukuze kubonwe umphumela wokufakwa kwesiliva ku-matrix ye-biofilm, kwenziwa ukudaya kwe-Toto 1 kanye ne-Syto 60 iodide (Isithombe 4). I-Toto 1 iodide iyidayi engangenwa yiseli engasetshenziswa ukubona ngokunembile ama-nucleic acid angaphandle kweseli, atholakala kakhulu ku-EPS yama-biofilms. I-Syto 60 iyidayi engangenwa yiseli esetshenziswa njenge-counterstain16. Ukubonwa kwe-Toto 1 kanye ne-Syto 60 iodide emanxebeni afakwe ama-biofilms e-Pseudomonas aeruginosa (Isithombe 4A-D) kanye ne-Staphylococcus aureus (Isithombe 4I-L) kubonise ukuthi ngemva kwezinsuku ezi-3 zokwelashwa kokufakwa, i-EPS ku-biofilm yehliswe kakhulu. equkethe usawoti one-oxygen Ag kanye ne-Ag1+ + EDTA/BC. Ukufakwa kwe-Ag1+ ngaphandle kwezingxenye ezengeziwe ze-antibiofilm kunciphisa kakhulu i-DNA engenamaseli emanxebeni afakwe i-Pseudomonas aeruginosa kodwa akusebenzanga kahle emanxebeni afakwe i-Staphylococcus aureus.
Izithombe ze-biofilm yesilonda ezithathwe emzimbeni ngemuva kwezinsuku ezi-3 zokwelashwa nge-control noma i-silver dressing. Izithombe ze-confocal ze-Pseudomonas aeruginosa (A–D) kanye ne-Staphylococcus aureus (I–L) ezifakwe i-Toto 1 (eluhlaza) ukuze ubone ngeso lengqondo ama-nucleic acid angaphandle, ingxenye yama-polymer e-extracellular biofilm. Ukuze ulahle ama-nucleic acid angaphakathi kweseli, sebenzisa ama-acid e-Syto 60 (abomvu). P. Ukuskena i-electron microscopy yamanxeba atheleleke nge-Pseudomonas aeruginosa (E–H) kanye ne-Staphylococcus aureus (M–P) biofilms ngemuva kwezinsuku ezi-3 zokwelashwa nge-control kanye ne-silver dressing. Ibha yesikali se-SEM = 5 µm. Ibha yesikali se-Confocal imaging = 50 µm.
Ukuskena nge-electron microscopy kubonise ukuthi amagundane afakwe ama-biofilm colonies e-Pseudomonas aeruginosa (Isithombe 4E-H) kanye ne-Staphylococcus aureus (Isithombe 4M-P) ayenamabhaktheriya ambalwa kakhulu ezilonda zawo ngemva kwezinsuku ezi-3 zokwelashwa ngazo zonke izithasiselo zesiliva.
Ukuze kuhlolwe umphumela wezingubo zesiliva ekuvuvukeni kwamanxeba emagundwini atheleleke nge-biofilm, izingxenye zamanxeba atheleleke nge-biofilm aphathwe ngezingubo zokulawula noma zesiliva izinsuku ezi-3 zafakwa umbala we-immunohistochemically kusetshenziswa ama-antibodies akhethekile kuma-neutrophil nama-macrophage. Ukunqunywa kobuningi bama-neutrophil nama-macrophage ngaphakathi. izicubu ze-granulation. Isithombe 5). Zonke izingubo zesiliva zinciphise inani lama-neutrophil nama-macrophage emanxebeni atheleleke nge-Pseudomonas aeruginosa uma kuqhathaniswa nezingubo zokulawula ezingezona ezilwa namagciwane ngemva kwezinsuku ezintathu zokwelashwa. Kodwa-ke, ukwelashwa nge-oxygenated silver salt dressing kwaholela ekunciphiseni okukhulu kwama-neutrophil (p = <0.0001) nama-macrophage (p = <0.0001) uma kuqhathaniswa nezinye izingubo zesiliva ezihlolwe (Isithombe 5I,J). Nakuba i-Ag1+ + EDTA/BC yayinethonya elikhulu ku-wound biofilm, yanciphisa amazinga e-neutrophil nama-macrophage ngezinga elincane kune-Ag1+ dressing. Amanxeba aphakathi atheleleke nge-S. aureus biofilm nawo abonwe ngemuva kokufakwa ama-oxisols e-Ag (p = <0.0001), Ag1+ (p = 0.0008) kanye ne-Ag1++ EDTA/BC (p = 0.0043) uma kuqhathaniswa ne-control . Kuye kwabonakala izitayela ezifanayo ze-neutropenia. bandage (Isithombe 5K). Kodwa-ke, ukufakwa kwe-oxygenated Ag salt dressing kuphela okubonise ukwehla okukhulu kwenani lama-macrophage ezicutshini ze-granulation uma kuqhathaniswa nokulawulwa kwamanxeba atheleleke nge-S. aureus biofilms (p = 0.0339) (Isithombe 5L).
Ama-neutrophils nama-macrophage alinganiswa emanxebeni atheleleke nge-Pseudomonas aeruginosa kanye ne-Staphylococcus aureus biofilms ngemva kwezinsuku ezi-3 zokwelashwa nge-non-antimicrobial control noma i-silver dressing. Ama-neutrophils (AD) kanye nama-macrophage (EH) alinganiswa ngezigaba zezicubu ezifakwe ama-antibodies akhethekile kuma-neutrophils noma ama-macrophage. Ukulinganiswa kwama-neutrophils (I kanye no-K) kanye nama-macrophage (J kanye no-L) emanxebeni atheleleke nge-Pseudomonas aeruginosa (I kanye no-J) kanye ne-Staphylococcus aureus (K & L) biofilms. N = 12 ngeqembu ngalinye. Amagrafu abonisa iphutha elijwayelekile +/-, amanani okubaluleka uma kuqhathaniswa ne-non-antibacterial control dressing, * kusho p = < 0.05, ** kusho p = < 0.01; *** kusho p = < 0.001; kubonisa p = <0.0001).
Sabe sesihlola umphumela wezinsimbi zesiliva ekwelapheni okungaxhomekeki ekuthelelekeni. Amanxeba okukhipha angethelelekile aphathwe ngensimbi yokulawula engeyona elwa namagciwane noma ngensimbi yesiliva izinsuku ezi-3 (Isithombe 6). Phakathi kwezinsimbi zesiliva ezihlolwe, amanxeba kuphela aphathwe ngensimbi yosawoti enomoya-mpilo abonakala emancane ezithombeni ezinkulu kunamanxeba aphathwe ngensimbi yokulawula (Isithombe 6A-D). Ukulinganiswa kwendawo yenxeba kusetshenziswa ukuhlaziywa kwe-histological kubonise ukuthi indawo yenxeba emaphakathi ngemva kokwelashwa ngensimbi yokubopha i-Ag oxysols yayingu-2.35 mm2 uma kuqhathaniswa no-2.96 mm2 wamanxeba aphathwe ngeqembu lokulawula, kodwa lo mehluko awuzange ufinyelele ukubaluleka kwezibalo (p = 0.488) (Isithombe 6I). Ngokuphambene nalokho, akukho ukwehla kwendawo yenxeba okwabonwa ngemuva kokwelashwa ngezinsimbi zokubopha i-Ag1+ (3.38 mm2, p = 0.757) noma i-Ag1+ + EDTA/BC (4.18 mm2, p = 0.054) uma kuqhathaniswa neqembu lokulawula. Ukwanda kokuvuselelwa kwe-epithelial kwabonwa nge-Ag oxysol dressing uma kuqhathaniswa neqembu lokulawula (30% vs. 22%, ngokulandelana), yize lokhu kungafinyelelanga ukubaluleka (p = 0.067), lokhu kubaluleke kakhulu futhi kuqinisekisa imiphumela yangaphambilini. I-dressing ene-oxysol ikhuthaza ukuvuselelwa kabusha kwe-epithelialization. -Ukuvuselelwa kwezilonda ezingathelelekile17. Ngokuphambene nalokho, ukwelashwa nge-Ag1+ noma i-Ag1+ + EDTA/BC dressings akuzange kube nomthelela noma kubonise ukwehla kokuvuselelwa kabusha kwe-epithelialization uma kuqhathaniswa nokulawulwa.
Umphumela wokufakwa kwenxeba lesiliva ekwelapheni inxeba emagundwini angathelelekile ngokususwa okuphelele. (AD) Izithombe ezimele ama-macroscopic zamanxeba ngemva kwezinsuku ezintathu zokwelashwa nge-dressing yokulawula engeyona i-antimicrobial kanye ne-dressing yesiliva. (EH) Izingxenye ezimele zamanxeba ezifakwe i-hematoxylin ne-eosin. Ukulinganiswa kwendawo yenxeba (I) kanye nephesenti lokubuyiselwa kwezicubu (J) kubalwe kusukela ezingxenyeni ze-histological maphakathi nenxeba kusetshenziswa isofthiwe yokuhlaziya isithombe (n = 11–12 ngeqembu lokwelashwa). Amagrafu abonisa iphutha elijwayelekile +/-. * kusho p = <0.05.
I-Silver inomlando omude wokusetshenziswa njengokwelashwa ngama-antimicrobial ekwelapheni amanxeba, kodwa izindlela eziningi ezahlukene zokwenziwa kanye nezindlela zokulethwa zingabangela umehluko ekusebenzeni kwama-antimicrobial 18. Ngaphezu kwalokho, izakhiwo zama-antibiofilm zezinhlelo ezithile zokulethwa kwesiliva aziqondakali ngokugcwele. Nakuba impendulo yomzimba yomnikazi iphumelela kakhulu ngokumelene namabhaktheriya e-planktonic, ngokuvamile ayisebenzi kahle ngokumelene nama-biofilms19. Amabhaktheriya e-planktonic asuswa kalula yi-macrophages, kodwa ngaphakathi kwama-biofilms, amaseli ahlanganisiwe adala izinkinga ezengeziwe ngokunciphisa impendulo yomnikazi kuze kube yilapho amaseli omzimba engabhekana ne-apoptosis futhi akhulule izici zokuvuvukala ukuze kuthuthukiswe impendulo yomzimba yokuzivikela 20. Kuye kwaphawulwa ukuthi amanye ama-leukocyte angangena kuma-biofilms21 kodwa awakwazi ukubhubhisa amabhaktheriya uma lokhu kuzivikela sekuphazamisekile 22. Indlela ephelele kufanele isetshenziswe ukusekela impendulo yomzimba yomnikazi ngokumelene nokutheleleka kwe-biofilm yamanxeba. Ukususwa kwamanxeba kungaphazamisa i-biofilm ngokomzimba futhi kususe iningi le-bioburden, kodwa impendulo yomzimba yomnikazi ingase ingasebenzi ngokumelene ne-biofilm esele, ikakhulukazi uma impendulo yomzimba yomnikazi iphazamisekile. Ngakho-ke, ukwelashwa okulwa namagciwane njenge-silver dressing kungasekela impendulo yomzimba womnikazi futhi kuqede ukutheleleka kwe-biofilm. Ukwakheka, ukuhlushwa, ukuncibilika, kanye ne-substrate yokulethwa kungathonya ukusebenza kahle kwesiliva ekulweni namagciwane. Eminyakeni yamuva nje, intuthuko kwezobuchwepheshe bokucubungula isiliva yenze lezi dressing zisebenza kangcono9,23. Njengoba ubuchwepheshe be-silver dressing buthuthuka, kubalulekile ukuqonda ukusebenza kahle kwalezi dressing ekulawuleni ukutheleleka kwenxeba futhi, okubaluleke kakhulu, umthelela walezi zinhlobo ezinamandla zesiliva endaweni yenxeba kanye nokuphulukiswa.
Kulolu cwaningo, siqhathanise ukusebenza kahle kwezindwangu ezimbili zesiliva ezithuthukisiwe nezindwangu zesiliva ezivamile ezikhiqiza ama-ion e-Ag1+ ngokumelene nama-biofilms sisebenzisa amamodeli ahlukene e-in vitro kanye ne-in vivo. Siphinde sahlola umphumela walezi zindwangu endaweni yenxeba kanye nokuphulukiswa okungaxhomekeki ekuthelelekeni. Ukuze kuncishiswe ithonya le-matrix yokulethwa, zonke izindwangu zesiliva ezivivinyiwe zazakhiwe nge-carboxymethylcellulose.
Ukuhlolwa kwethu kokuqala kwalezi zingubo zesiliva ngokumelene ne-biofilms yamakoloni ye-Pseudomonas aeruginosa kanye ne-Staphylococcus aureus kukhombisa ukuthi, ngokungafani nezingubo zendabuko ze-Ag1+, izingubo zesiliva ezimbili ezithuthukisiwe, i-Ag1+ + EDTA/BC kanye nosawoti we-Ag one-oxygen, zisebenza kahle ku-5. Zibulala ngempumelelo amabhaktheriya e-biofilm ezinsukwini ezimbalwa. Ngaphezu kwalokho, lezi zingubo zivimbela ukwakheka kabusha kwe-biofilm lapho kuvezwa ngokuphindaphindiwe kuma-bacteria e-planktonic. I-Ag1+ dressing yayiqukethe i-silver chloride, i-silver compound efanayo kanye ne-base matrix njenge-Ag1+ + EDTA/BC, futhi yayinomthelela olinganiselwe ekusebenzeni kwama-bacteria ngaphakathi kwe-biofilm esikhathini esifanayo. Ukuqaphela ukuthi i-Ag1+ + EDTA/BC dressing yayisebenza kangcono ngokumelene ne-biofilm kune-Ag1+ dressing equkethe i-matrix efanayo kanye ne-silver compound kusekela umbono wokuthi izithako ezengeziwe ziyadingeka ukwandisa ukusebenza kwe-silver chloride ngokumelene ne-biofilm, njengoba kubikwe kwenye indawo15. Le miphumela isekela umqondo wokuthi i-BC ne-EDTA zidlala indima eyengeziwe ekufakeni isandla ekusebenzeni kahle kokugqoka kanye nokuthi ukungabikho kwalesi sakhi kuma-Ag1+ dressings kungenzeka kube nomthelela ekwehlulekeni kokubonisa ukusebenza kahle kwe-in vitro. Sithole ukuthi ama-Ag salt dressing aqukethe umoya-mpilo akhiqiza ama-Ag2+ nama-Ag3+ ions abonise ukusebenza kahle okunamandla okulwa namagciwane kune-Ag1+ futhi emazingeni afana ne-Ag1+ + EDTA/BC. Kodwa-ke, ngenxa yamandla aphezulu e-redox, akucaci ukuthi ama-Ag3+ ions ahlala isikhathi esingakanani esebenza futhi esebenza kahle ngokumelene nama-wound biofilms ngakho-ke kufanele kuqhutshekwe nocwaningo. Ngaphezu kwalokho, kunezinhlobo eziningi ezahlukene zama-Ag3+ ions ezingazange zihlolwe kulolu cwaningo. Lezi zinhlobo zama-dressing zakhiwe ngama-compound esiliva ahlukene, amazinga okugxila, kanye nama-base matrices, angathonya ukulethwa kwama-Ag1+ ions kanye nokusebenza kwawo ngokumelene nama-biofilms. Kubalulekile futhi ukuqaphela ukuthi kunezinhlobo eziningi ezahlukene zama-in vitro nama-in vivo model asetshenziswa ukuhlola ukusebenza kahle kwama-wound dressing ngokumelene nama-biofilms. Uhlobo lwemodeli esetshenzisiwe, kanye nokuqukethwe kosawoti kanye namaprotheni kwemidiya esetshenziswa kula mamodeli, kuzothinta ukusebenza kahle kwe-dressing. Kumodeli yethu ye-in vivo, sivumele i-biofilm ukuthi ivuthwe ngaphakathi kwe-vitro bese siyidlulisela ebusweni besikhumba sesilonda. Impendulo yokuzivikela yegundane eliyibambayo isebenza kahle ngokumelene namagciwane e-planktonic asetshenziswa enxebeni, ngaleyo ndlela yakha i-biofilm njengoba isilonda siphola. Ukwengezwa kwe-biofilm evuthiwe enxebeni kunciphisa ukusebenza kahle kwempendulo yokuzivikela yomnikazi ekwakhekeni kwe-biofilm ngokuvumela i-biofilm evuthiwe ukuthi izinze ngaphakathi kwesilonda ngaphambi kokuba ukwelashwa kuqale. Ngakho-ke, imodeli yethu isivumela ukuthi sihlole ukusebenza kahle kwezingubo zokulwa namagciwane kuma-biofilm avuthiwe ngaphambi kokuba amanxeba aqale ukuphola.
Sithole nokuthi ukulingana kwezingubo kuthonye ukusebenza kahle kwezingubo zesiliva kuma-biofilms akhuliswe ngaphakathi kwe-vitro kanye nesikhumba sengulube. Ukuxhumana eduze nenxeba kubhekwa njengokubalulekile ekusebenzeni kahle kwezingubo zokulwa namagciwane24,25. Izingubo zokumboza eziqukethe usawoti we-Ag one-oxygen zazisondelene kakhulu nama-biofilms avuthiwe, okwaholela ekunciphiseni okukhulu kwenani lamabhaktheriya asebenzayo ngaphakathi kwe-biofilm ngemva kwamahora angama-24. Ngokuphambene nalokho, lapho kwelashwa ngezingubo zokumboza ze-Ag1+ kanye ne-Ag1+ + EDTA/BC, kwasala inani elikhulu lamabhaktheriya asebenzayo. Lezi zingubo zokumboza ziqukethe imithungo kulo lonke ubude bengubo, okudala izikhala ezifile ezivimbela ukuxhumana eduze ne-biofilm. Ezifundweni zethu ze-in vitro, lezi zindawo ezingathintani zivimbele ukubulawa kwamabhaktheriya asebenzayo ngaphakathi kwe-biofilm. Sihlole ukuphila kwamabhaktheriya kuphela ngemva kwamahora angama-24 okwelashwa; Ngokuhamba kwesikhathi, njengoba ingubo igcwala kakhulu, kungase kube nendawo encane efile, kunciphisa indawo yala mabhaktheriya asebenzayo. Kodwa-ke, lokhu kugcizelela ukubaluleka kokwakheka kwengubo, hhayi nje uhlobo lwesiliva ebhandeni.
Nakuba izifundo ze-in vitro ziwusizo ekuqhathaniseni ukusebenza kahle kobuchwepheshe obuhlukene besiliva, kubalulekile futhi ukuqonda imiphumela yalezi zingubo zokugqoka kuma-biofilms in vivo, lapho izicubu zomsingathi kanye nezimpendulo zomzimba zifaka isandla ekusebenzeni kahle kwezingubo zokugqoka ngokumelene nama-biofilms. Umphumela walezi zingubo zokugqoka kuma-biofilms amanxeba wabonwa kusetshenziswa i-scanning electron microscopy kanye ne-EPS staining ye-biofilm kusetshenziswa amadayi e-DNA angaphakathi kweseli kanye nangaphandle. Sithole ukuthi ngemva kwezinsuku ezi-3 zokwelashwa, zonke izingubo zokugqoka zazisebenza kahle ekunciphiseni i-DNA engenamaseli emanxebeni atheleleke nge-biofilm, kodwa ingubo ye-Ag1+ yayingasebenzi kahle emanxebeni atheleleke nge-Staphylococcus aureus. I-scanning electron microscopy iphinde yabonisa ukuthi amabhaktheriya ambalwa kakhulu ayekhona emanxebeni aphathwe ngezingubo zesiliva, yize lokhu kwakugqame kakhulu ngengubo ye-Ag salt ene-oxygenated kanye nengubo ye-Ag1+ + EDTA/BC uma kuqhathaniswa nengubo ye-Ag1+. Le datha ikhombisa ukuthi izingubo zesiliva ezivivinyiwe zazinezinga elihlukahlukene lomthelela esakhiweni se-biofilm, kodwa azikho izingubo zesiliva ezakwazi ukuqeda i-biofilm, zisekela isidingo sendlela ephelele yokwelapha ukutheleleka kwe-biofilm yamanxeba; ukusetshenziswa kwamabhande esiliva. Ukwelashwa kwandulelwa ukususwa kwengxenye enkulu ye-biofilm.
Amanxeba angapheli avame ukuba sesimweni sokuvuvukala okukhulu, lapho amangqamuzana okuvuvukala amaningi ehlala ezicutshini zenxeba isikhathi eside, okubangela ukulimala kwezicubu futhi kuqede umoya-mpilo odingekayo ukuze kusebenze kahle i-metabolism yamaseli kanye nokusebenza kahle enxebeni26. Ama-biofilms ayenza ibe yingozi le ndawo yenxeba ngokuthinta kabi ukuphulukiswa ngezindlela ezahlukahlukene, okuhlanganisa ukuvimbela ukwanda kwamaseli kanye nokufuduka kanye nokusebenza kwama-cytokines abangela ukuvuvukala27. Njengoba izinsimbi zesiliva zisebenza kahle kakhulu, kubalulekile ukuqonda umthelela ezinawo endaweni yenxeba kanye nokuphulukiswa.
Ngokuthakazelisayo, nakuba zonke izithasiselo zesiliva zithinte ukwakheka kwe-biofilm, izithasiselo zesiliva ezinosawoti wesiliva kuphela ezandisa ukuvuselelwa kabusha kwalezi zilonda ezinegciwane. Le datha isekela okutholakele kwethu kwangaphambilini17 kanye noKalan et al. (2017)28, okubonise ukuphepha okuhle kanye namaphrofayili obuthi kasawoti wesiliva one-oxygen, njengoba amazinga aphansi esiliva ayesebenza kahle ngokumelene nama-biofilm.
Ucwaningo lwethu lwamanje luqokomisa umehluko kubuchwepheshe besiliva phakathi kwezingubo zesiliva ezilwa namagciwane kanye nomthelela walobu buchwepheshe endaweni yamanxeba kanye nokuphulukiswa okungaxhomekeki ekuthelelekeni. Kodwa-ke, le miphumela ihlukile ezifundweni zangaphambilini ezibonisa ukuthi ingubo ye-Ag1+ + EDTA/BC ithuthukise imingcele yokuphulukisa yezindlebe zonogwaja ezilimele emzimbeni. Kodwa-ke, lokhu kungase kube ngenxa yomehluko kumamodeli ezilwane, izikhathi zokulinganisa, kanye nezindlela zokusebenzisa amagciwane29. Kulesi simo, izilinganiso zamanxeba zathathwa ezinsukwini eziyi-12 ngemuva kokulimala ukuze kuvunyelwe izithako ezisebenzayo zengubo ukuthi zisebenze ku-biofilm isikhathi eside. Lokhu kusekelwa ucwaningo olukhombisile ukuthi izilonda zemilenze ezinegciwane ezelashwe nge-Ag1+ + EDTA/BC ekuqaleni zanda ngobukhulu ngemva kwesonto elilodwa lokwelashwa, kwabe sekuba phakathi kwamasonto amathathu alandelayo okwelashwa nge-Ag1+ + EDTA/BC kanye naphakathi kwamasonto ama-4 okusetshenziswa kwezidakamizwa ezingezona ezilwa namagciwane. Izingubo ze-CMC zokunciphisa usayizi wezilonda30.
Izinhlobo ezithile kanye nokugcwala kwesiliva kuye kwabonakala ngaphambilini ukuthi kuyingozi kakhulu ku-vitro 11, kodwa le miphumela ye-in vitro ayisho njalo imiphumela emibi ku-vivo. Ngaphezu kwalokho, intuthuko kwezobuchwepheshe besiliva kanye nokuqonda kangcono amakhemikhali esiliva kanye nokugcwala ekugqokeni kuye kwaholela ekuthuthukisweni kwezingubo eziningi zesiliva eziphephile nezisebenzayo. Kodwa-ke, njengoba ubuchwepheshe bezingubo zesiliva buthuthuka, kubalulekile ukuqonda umthelela walezi zingubo endaweni yamanxeba31,32,33. Kwabikwa ngaphambilini ukuthi izinga elikhuphukile lokuvuselelwa kabusha kwe-epithelialization lihambisana nengxenye ekhuphukile yama-macrophage e-M2 alwa nokuvuvukala uma kuqhathaniswa ne-phenotype ye-M1 ebangela ukuvuvukala. Lokhu kwaphawulwa kumodeli yegundane yangaphambilini lapho izingubo zesiliva ze-hydrogel ziqhathaniswa ne-silver sulfadiazine kanye nama-hydrogel angewona ama-antimicrobial34.
Amanxeba angapheli angase abonise ukuvuvukala okweqile futhi kuye kwaphawulwa ukuthi ukuba khona kwama-neutrophil amaningi kungaba yingozi ekuphilisweni kwamanxeba35. Ocwaningweni olwenziwe kumagundane aphelelwe yi-neutrophil, ukuba khona kwama-neutrophil kulibazise ukuvuselelwa kabusha kwama-neutrophil. Ukuba khona kwama-neutrophil amaningi kuholela emazingeni aphezulu ama-protease kanye nezinhlobo ze-oxygen ezisabelayo, njenge-superoxide ne-hydrogen peroxide, ezihlotshaniswa namanxeba angapheli futhi aphulukisayo kancane37,38. Ngokufanayo, ukwanda kwenani lama-macrophage, uma kungalawulwa, kungaholela ekuphilisweni kwamanxeba okulibazisekile39. Lokhu kwanda kubaluleke kakhulu uma ama-macrophage engakwazi ukushintsha kusuka ku-phenotype ekhuthaza ukuvuvukala aye ku-phenotype ekhuthaza ukupholisa, okuholela ekutheni amanxeba ahluleke ukuphuma esigabeni sokuvuvukala sokuphulukiswa40. Sibone ukwehla kwama-neutrophil nama-macrophage ezilonda ezitheleleke nge-biofilm ngemuva kwezinsuku ezi-3 zokwelashwa ngazo zonke izinsimbi zesiliva, kodwa ukwehla kwabonakala kakhulu ngezinsimbi zikasawoti ezine-oxygen. Lokhu kwehla kungaba umphumela oqondile wempendulo yomzimba esiliva, impendulo ekunciphiseni umthwalo we-bio, noma inxeba lisesigabeni sokugcina sokuphulukiswa ngakho-ke amangqamuzana omzimba enxebani ancishisiwe. Ukunciphisa inani lamangqamuzana okuvuvukala enxebani kungagcina indawo evumela ukuphulukiswa kwenxeba. Indlela yokusebenza kwe-Ag oxysalts ekhuthaza ngayo ukuphulukiswa okungaxhomekeki ekuthelelekeni ayicaci, kodwa ikhono lama-Ag oxysalts lokukhiqiza umoya-mpilo nokubhubhisa amazinga ayingozi e-hydrogen peroxide, umlamuleli wokuvuvukala, lingachaza lokhu futhi lidinga ucwaningo olwengeziwe17.
Amanxeba angenwe yileli gciwane angapholi abangela inkinga kodokotela kanye neziguli. Nakuba ama-dressing amaningi ethi asebenza kahle ekulweni namagciwane, ucwaningo alugxili kwezinye izici ezibalulekile ezithonya indawo encane yamanxeba. Lolu cwaningo lubonisa ukuthi ubuchwepheshe obuhlukene besiliva busebenza kahle ngokulwa namagciwane futhi, okubaluleke kakhulu, imiphumela ehlukene endaweni yamanxeba kanye nokuphulukiswa kwawo, ngaphandle kokutheleleka. Nakuba lezi zifundo ze-in vitro kanye ne-in vivo zibonisa ukusebenza kahle kwala ma-dressing ekwelapheni ukutheleleka kwamanxeba kanye nokukhuthaza ukuphulukiswa, kudingeka izivivinyo ezilawulwa ngokungahleliwe ukuze kuhlolwe ukusebenza kahle kwala ma-dressing emtholampilo.
Amasethi edatha asetshenzisiwe kanye/noma ahlaziywe ngesikhathi socwaningo lwamanje ayatholakala kumbhali ohambisanayo uma ecelwa ngokufanele.
Isikhathi sokuthunyelwe: Julayi-15-2024