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Ukukhula kweentsholongwane emanxebeni kudla ngokuzibonakalisa njengee-biofilms, eziphazamisa ukuphiliswa kwaye kunzima ukuzisusa. Ii-silver dressings ezintsha zithi zilwa nosulelo lwamanxeba, kodwa ukusebenza kwazo njenge-antibiofilm kunye nemiphumo yokuphiliswa engaxhomekekanga kusulelo ayaziwa ngokubanzi. Sisebenzisa iimodeli ze-in vitro kunye ne-in vivo ze-Staphylococcus aureus kunye ne-Pseudomonas aeruginosa, sixela ukusebenza kwee-Ag1+ ion-generating dressings; ii-Ag1+ dressings eziqulethe i-ethylenediaminetetraacetic acid kunye ne-benzethonium chloride (Ag1+/EDTA/BC), kunye nee-dressings eziqulethe i-silver nitrate (Ag oxysalts). , ezivelisa ii-Ag1+, Ag2+ kunye ne-Ag3+ ions ukulwa ne-biofilm yamanxeba kunye nempembelelo yayo ekuphilisweni. Ii-Ag1+ dressings zinempembelelo encinci kwi-biofilm yamanxeba kwi-vitro nakwiimpuku (C57BL/6j). Ngokwahlukileyo koko, iityuwa ze-Ag ezineoksijini kunye needressing ze-Ag1+/EDTA/BC zinciphise kakhulu inani leebhaktheriya ezisebenzayo kwi-biofilms kwi-vitro kwaye zabonisa ukwehla okukhulu kwizakhi zebhaktheriya kunye ne-EPS kwi-biofilms zamanxeba empuku. Ezi dressing zineziphumo ezahlukeneyo ekuphilisweni kwamanxeba ane-biofilm kunye nalawo angenayo i-biofilm, kunye needressing zetyuwa ezineoksijini ezineziphumo eziluncedo ngakumbi ekuvuseleleni kwakhona, ubungakanani benxeba, kunye nokudumba xa kuthelekiswa nonyango lolawulo kunye nezinye iidressing zesilivere. Iipropati ezahlukeneyo ze-physicochemical zeedressing zesilivere zinokuba neziphumo ezahlukeneyo kwi-biofilm yamanxeba kunye nokuphiliswa, kwaye oku kufuneka kuqwalaselwe xa kukhethwa idressing yokunyanga amanxeba aneoksijini.
Amanxeba angapheliyo achazwa “njengamanxeba angaphumeleliyo kumanqanaba aqhelekileyo okuphiliswa ngendlela ehlelekileyo nefanelekileyo” 1. Amanxeba angapheliyo adala umthwalo wengqondo, wentlalo nowezoqoqosho kwizigulana nakwinkqubo yezempilo. Inkcitho yonyaka ye-NHS ekunyangeni amanxeba kunye nezinye izifo ezinxulumene nawo iqikelelwa kwi-£8.3 yeebhiliyoni ngo-2017–182. Amanxeba angapheliyo nawo okwangoku ayingxaki enkulu eMelika, apho iMedicare iqikelela iindleko zonyaka zokunyanga izigulana ezinamanxeba kwi-$28.1–$96.8 yeebhiliyoni3.
Usulelo luyinto ephambili ethintela ukuphiliswa kwenxeba. Usulelo ludla ngokubonakala njengee-biofilms, ezikhoyo kwi-78% yamanxeba angapheliyo angapholiyo. Ii-biofilms zenzeka xa iintsholongwane zinamathela ngokungenakuguquguquka kwiindawo, ezifana nemiphezulu yamanxeba, kwaye zinokuhlangana zenze uluntu oluvelisa i-extracellular polymer (EPS). I-biofilm yamanxeba inxulunyaniswa nokwanda kwempendulo yokudumba ekhokelela kumonakalo wezicubu, okunokulibazisa okanye kuthintele ukuphiliswa4. Ukwanda komonakalo wezicubu kunokubangelwa ngokuyinxenye ngumsebenzi owandisiweyo wee-matrix metalloproteinases, i-collagenase, i-elastase kunye ne-reactive oxygen species5. Ngaphezu koko, iiseli ezivuvukalayo kunye nee-biofilms ngokwazo zidla kakhulu ioksijini kwaye ke ngoko zinokubangela i-hypoxia yezicubu zasekuhlaleni, zinciphise iiseli zeoksijini ebalulekileyo efunekayo ukuze kulungiswe izicubu ngempumelelo6.
Ii-biofilms ezivuthiweyo ziyakwazi ukumelana kakhulu nee-antimicrobial agents, zifuna amaqhinga aqatha okulawula usulelo lwe-biofilm, njengonyango loomatshini olulandelwa lunyango olusebenzayo lwe-antimicrobial. Ngenxa yokuba ii-biofilms zinokuphinda zikhule ngokukhawuleza, ii-antimicrobial ezisebenzayo zinokunciphisa umngcipheko wokwakheka kwakhona emva kokususwa kwe-antimicrobial ngotyando7.
Isilivere isetyenziswa kakhulu kwiimpahla zokugquma iintsholongwane kwaye isetyenziswa njengonyango lokuqala lwamanxeba anesifo esinganyangekiyo. Kukho iintlobo ezininzi zeempahla zesilivere ezithengiswayo, nganye inesakhiwo sesilivere esahlukileyo, uxinaniso, kunye nesiseko se-matrix. Ukuqhubela phambili kwiimpahla zesilivere kukhokelele ekuphuhlisweni kweempahla zesilivere ezintsha. Uhlobo lwesilivere oluyintsimbi (Ag0) alusebenzi; Ukuze kufezekiswe ukusebenza kwe-antimicrobial, kufuneka ilahlekelwe yi-electron ukuze yenze isilivere ye-ionic (Ag1+). Iimpahla zesilivere zemveli ziqulethe iikhompawundi zesilivere okanye isilivere yesinyithi ethi, xa ivezwe kulwelo, ibole ukuze yenze ii-ion ze-Ag1+. Ezi i-ion ze-Ag1+ zisabela kunye neseli yebhaktiriya, zisuse ii-electron kwizinto zokwakha okanye iinkqubo ezibalulekileyo ezifunekayo ukuze ziphile. Ubuchwepheshe obunelungelo lobunikazi bukhokelele ekuphuhlisweni kwekhompawundi entsha yesilivere, i-Ag oxysalts (i-nitrate yesilivere, i-Ag7NO11), equkwe kwiimpahla zokugquma iinxeba. Ngokungafaniyo nesilivere yendabuko, ukubola kweetyuwa ezineoksijini kuvelisa imo yesilivere ene-valence ephezulu (Ag1+, Ag2+ kunye ne-Ag3+). Izifundo ze-in vitro zibonise ukuba ubuninzi obuphantsi beetyuwa zesilivere ezineoksijini busebenza ngakumbi kune-single ion silver (Ag1+) ngokuchasene neebhaktheriya ezibangela izifo ezifana nePseudomonas aeruginosa, Staphylococcus aureus kunye ne-Escherichia coli8,9. Olunye uhlobo olutsha lwe-silver dressing luquka izithako ezongezelelweyo, ezizezi i-ethylenediaminetetraacetic acid (EDTA) kunye ne-benzethonium chloride (BC), ezixelwe ukuba zijolise kwi-biofilm EPS kwaye ngaloo ndlela zonyusa ukungena kwesilivere kwi-biofilm. Ezi teknoloji zintsha zesilivere zibonelela ngeendlela ezintsha zokujolisa kwi-biofilms zamanxeba. Nangona kunjalo, impembelelo yala mayeza okubulala iintsholongwane kwindawo yamanxeba kunye nokuphiliswa okungenalo usulelo kubalulekile ukuqinisekisa ukuba awenzi indawo yamanxeba engalunganga okanye ukulibaziseka kokuphiliswa. Iinkxalabo malunga ne-in vitro silver cytotoxicity ziye zaxelwa nge-silver dressings ezininzi10,11. Nangona kunjalo, i-in vitro cytotoxicity ayikaguqulelwa kwi-in vivo toxicity, kwaye ii-Ag1+ dressings ezininzi zibonakalise iprofayili elungileyo yokhuseleko12.
Apha, siphande ukusebenza kakuhle kwee-carboxymethylcellulose dressings eziqulethe iifomyula zesilivere ezintsha nxamnye ne-biofilm yamanxeba kwi-vitro nakwi-vivo. Ukongeza, imiphumo yezi dressings kwiimpendulo zomzimba kunye nokuphiliswa ngaphandle kosulelo ihlolwe.
Zonke ii-dressing ezisetyenzisiweyo bezifumaneka kurhwebo. I-3M Kerracel Gel Fiber Dressing (3M, Knutsford, UK) yi-non-antimicrobial 100% carboxymethylcellulose (CMC) gel fiber dressing esetyenziswe njenge-control dressing kolu phononongo. Ii-dressing zesilivere ze-CMC ezintathu ezilwa neentsholongwane zivavanyiwe, ezizezi i-3M Kerracel Ag dressing (3M, Knutsford, UK), equlethe i-1.7 wt%. ityuwa yesilivere eneoksijini (Ag7NO11) kwii-ion zesilivere ze-valence eziphezulu (Ag1+, Ag2+ kunye ne-Ag3+). Ngexesha lokubola kwe-Ag7NO11, ii-ion ze-Ag1+, Ag2+ kunye ne-Ag3+ zenziwa ngomlinganiselo we-1:2:4. I-Aquacel Ag i-extra dressing equlethe i-1.2% ye-silver chloride (Ag1+) (ConvaTec, Deeside, UK) 13 kunye ne-Aquacel Ag + i-extra dressing equlethe i-1.2% ye-silver chloride (Ag1+), i-EDTA kunye ne-benzethonium chloride (ConvaTec, Deeside, UK) 14.
Iintlobo ezisetyenzisiweyo kolu phononongo yayiyiPseudomonas aeruginosa NCTC 10781 (Public Health England, Salisbury) kunye neStaphylococcus aureus NCTC 6571 (Public Health England, Salisbury).
Iintsholongwane zakhuliswa ngobusuku bonke kumhluzi weMuller-Hinton (Oxoid, Altrincham, UK). Inkcubeko yobusuku bonke yaxutywa nge-1:100 kumhluzi weMueller-Hinton kunye ne-200 µl efakwe kwi-sterile 0.2 µm Whatman cyclopore membranes (Whatman plc, Maidstone, UK) kwiiplate zeMueller-Hinton agar (Sigma-Aldrich Company Ltd, Kent, Great Britain). ) Ukwakheka kwe-biofilm yamakoloniyali kuma-37°C kangangeeyure ezingama-24. Ezi biofilms zamakoloniyali zavavanywa ukuncipha kwe-logarithmic.
Sika i-dressing ibe ziingceba zesikwere eziyi-3 cm2 uze umanzise ngamanzi acocekileyo angena ntsholongwane. Beka i-bandage phezu kwe-biofilm yekoloni kwipleyiti ye-agar. Yonke i-24 ha ye-biofilm isusiwe, kwaye iintsholongwane eziphilayo ngaphakathi kwi-biofilm (CFU/ml) zilinganiswe ngokuxutywa ngokulandelelana (10−1 ukuya kwi-10−7) kwi-Day-Angle neutralization broth (Merck-Millipore). Emva kweeyure ezingama-24 zokuxutywa kwi-37°C, amanani eepleyiti aqhelekileyo enziwe kwiipleyiti ze-agar zeMueller-Hinton. Unyango ngalunye kunye nexesha lenziwe kathathu, kwaye amanani eepleyiti aphindaphindwa kumxube ngamnye.
Ulusu lwesisu sehagu lufunyanwa kwiihagu eziziimazi ezinkulu ezimhlophe kwimizuzu eli-15 emva kokuxhelwa ngokwemigangatho yokuthunyelwa kwamanye amazwe ye-European Union. Ulusu luchetyiwe lwaza lwacocwa ngee-alcohol wipes, lwaza lwafakwa efrijini kwi--80°C iiyure ezingama-24 ukuze luvuselele ulusu. Emva kokunyibilika, iziqwenga zesikhumba eziyi-1 cm2 zahlanjwa kathathu nge-PBS, i-0.6% sodium hypochlorite, kunye ne-70% ethanol imizuzu engama-20 ixesha ngalinye. Ngaphambi kokususa i-epidermis, susa nayiphi na i-ethanol eseleyo ngokuyihlamba kathathu kwi-PBS engenazintsholongwane. Ulusu lukhuliswe kwipleyiti eneziphelo ezi-6 ene-membrane yenylon enobukhulu obuyi-0.45-μm (Merck-Millipore) phezulu kunye nee-3 pads ezifunxayo (Merck-Millipore) ezine-3 ml ye-fetal bovine serum (Sigma) eyongezwe yi-10% ye-Dulbecco's modified Eagle. Medium (Dulbecco's Modified Eagle Medium – Aldrich Ltd.).
Ii-biofilms zasekoloniyali zakhuliswa njengoko kuchaziwe kwizifundo zokuvezwa kwe-biofilm. Emva kokukhulisa i-biofilm kwi-membrane iiyure ezingama-72, i-biofilm yafakwa kumphezulu wesikhumba kusetyenziswa i-sterile inoculation loop kwaye i-membrane yasuswa. I-biofilm yafakwa kwi-dermis yehagu iiyure ezingama-24 ezongezelelweyo kwi-37°C ukuvumela i-biofilm ukuba ikhule kwaye inamathele eluswini lwehagu. Emva kokuba i-biofilm ivuthiwe kwaye iqhotyoshelwe, i-dressing eyi-1.5 cm2, emanziswe ngamanzi acocekileyo acocekileyo, yafakwa ngqo kumphezulu wesikhumba yaza yafakwa kwi-37°C iiyure ezingama-24. Iibhaktheriya eziphilayo zabonwa ngokufaka i-reagent ye-PrestoBlue cell viability reagent (Invitrogen, Life Technologies, Paisley, UK) kumphezulu we-apical we-explant nganye kwaye uyifake imizuzu emi-5. Sebenzisa ikhamera yedijithali yeLeica DFC425 ukuze ubambe imifanekiso ngoko nangoko kwi-microscope yeLeica MZ8. Iindawo ezinombala opinki zalinganiswa kusetyenziswa isoftware ye-Image Pro version 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (mediacy.com)). Ukuskena nge-electron microscopy kwenziwe njengoko kuchaziwe ngezantsi.
Iintsholongwane ezikhuliswe ngobusuku bonke zaxutywa nge-1:100 kumhluzi weMueller-Hinton. I-200 μl yenkcubeko yongezwa kwi-0.2 μm Whatman cyclopore membrane ecocekileyo (Whatman, Maidstone, UK) yaza yabekwa kwi-agar yeMueller-Hinton. Iiplate zeBiofilm zafakwa kwi-37°C kangangeeyure ezingama-72 ukuvumela ukwakheka kwe-biofilm evuthiweyo.
Emva kweentsuku ezi-3 zokuvuthwa kwe-biofilm, ibhandeji eliyi-3 cm2 square lafakwa ngqo kwi-biofilm laza lafakwa kwi-37°C kangangeeyure ezingama-24. Emva kokususa ibhandeji kumphezulu we-biofilm, i-1 ml ye-PrestoBlue Cell Viability Reagent (Invitrogen, Waltham, MA) yongezwa kumphezulu we-biofilm nganye imizuzwana engama-20. Imiphezulu yomiswa ngaphambi kokuba kubhalwe utshintsho lombala kusetyenziswa ikhamera yedijithali ye-Nikon D2300 (Nikon UK Ltd., Kingston, UK).
Lungisa iinkcubeko zasebusuku kwi-agar yeMueller-Hinton, udlulisele iikholoni nganye kwi-10 ml yomhluzi weMueller-Hinton uze uzifake kwi-shaker kwi-37°C (100 rpm). Emva kokufunxa ubusuku bonke, inkcubeko yancitshiswa nge-1:100 kumhluzi weMueller-Hinton kwaye i-300 µl yabonwa kwi-0.2 µm circular Whatman cyclopore membrane (Whatman International, Maidstone, UK) kwi-agar yeMueller-Hinton yaza yafakwa kwi-37°C kwiiyure ezingama-72. . I-biofilm evuthiweyo yafakwa enxebeni njengoko kuchaziwe ngezantsi.
Lonke umsebenzi ngezilwanyana wenziwa kwiYunivesithi yaseManchester phantsi kwelayisenisi yeprojekthi evunyiweyo yiOfisi yeNtlalontle yeZilwanyana kunye noHlolo lweMithetho (P8721BD27) nangokuhambelana nezikhokelo ezipapashwe yiOfisi yaseKhaya phantsi kwe-ASPA ehlaziyiweyo ka-2012. Bonke ababhali balandele izikhokelo zokufika. Iimpuku ze-C57BL/6j ezineeveki ezisibhozo (Envigo, Oxon, UK) zasetyenziswa kuzo zonke izifundo ze-in vivo. Iimpuku zahlanjwa nge-isoflurane (Piramal Critical Care Ltd, West Drayton, UK) kwaye umphezulu wazo wangasemva wachetywa waza wacocwa. Impuku nganye emva koko yanikwa inxeba eliyi-2 × 6 mm lokususa inxeba kusetyenziswa i-Stiefel biopsy punch (Schuco International, Hertfordshire, UK). Kwizilonda ezine-biofilm, faka i-biofilm yekolonial yeeyure ezingama-72 ekhuliswe kwi-membrane njengoko kuchaziwe apha ngasentla kumaleko wesikhumba senxeba usebenzisa i-stainless injection loop kwangoko emva kokwenzakala kwaye ulahle i-membrane. Isentimitha enye yesikwere se-dressing imanziswa kwangaphambili ngamanzi acocekileyo ukugcina indawo yenxeba emanzi. Iibhandeji zifakwe ngqo kwinxeba ngalinye kwaye zigqunywe ngefilimu ye-3M Tegaderm (3M, Bracknell, UK) kunye ne-Mastisol liquid adhesive (Eloquest Healthcare, Ferndale, MI) ezifakwe emacaleni ukuze zinike ukunamathela okongeziweyo. I-Buprenorphine (Animalcare, York, UK) yanikwa uxinaniso lwe-0.1 mg/kg njengeyeza lokuthomalalisa iintlungu. Sika iimpuku emva kweentsuku ezintathu zenzakele usebenzisa indlela yeShedyuli 1 kwaye ususe, uhlukanise phakathi, kwaye ugcine indawo yenxeba njengoko kufuneka.
Ukupenda ngeHematoxylin (ThermoFisher Scientific) kunye ne-eosin (ThermoFisher Scientific) kwenziwe ngokwemigaqo yomenzi. Indawo yenxeba kunye nokuphinda kufakwe i-pithelialization kwalinganiswa kusetyenziswa isoftware ye-Image Pro version 10 (Media Cybernetics Inc, Rockville, MD).
Iinxalenye zezicubu zahluzwa nge-xylene (ThermoFisher Scientific, Loughborough, UK), zaphinda zamanziswa nge-ethanol eyi-100–50% elinganisiweyo, zaza zantywiliselwa okwethutyana emanzini anyibilikisiweyo (ThermoFisher Scientific). I-Immunohistochemistry yenziwe kusetyenziswa i-VectaStain Elite ABC PK-6104 kit (Vector Laboratories, Burlingame, CA) ngokwemigaqo yomenzi. Ii-antibodies eziphambili kwi-neutrophils NIMP-R14 (Thermofisher Scientific) kunye ne-macrophages Ms CD107b Pure M3/84 (BD Biosciences, Wokingham, UK) zaxutywa nge-1:100 kwisisombululo esivimbayo zaza zongezwa kumphezulu osikiweyo, zalandelwa zii-antibodies ezimbini ze-Anti-, VectaStain ABC kunye neVector Nova Red Peroxidase (HRP) substrate kit (Vector Laboratories, Burlingame, CA) zaza zachatshazelwa yi-hematoxylin. Imifanekiso ifunyenwe kusetyenziswa imakroskopu ye-Olympus BX43 kunye nekhamera yedijithali ye-Olympus DP73 (e-Olympus, eSouthend-on-Sea, e-UK).
Iisampulu zesikhumba zilungisiwe kwi-2.5% ye-glutaraldehyde kunye ne-4% ye-formaldehyde kwi-0.1 M HEPES (pH 7.4) iiyure ezingama-24 kwi-4°C. Iisampulu zasuswa amanzi kusetyenziswa i-ethanol elinganisiweyo zaza zomiswa kwi-CO2 kusetyenziswa i-Quorum K850 critical point dryer (Quorum Technologies Ltd, Loughton, UK) kwaye i-sputter igqunywe nge-gold-palladium alloy kusetyenziswa i-Quorum SC7620 mini sputterer/glow discharge system. Iisampulu zithathwe umfanekiso kusetyenziswa i-FEI Quanta 250 scanning electron microscope (ThermoFisher Scientific) ukuze kubonwe indawo ephakathi yenxeba.
I-Toto-1 iodide (2 μM) ifakwe kumphezulu wenxeba lempuku elisusiweyo yaza yafakwa imizuzu emi-5 kwi-37 °C (ThermoFisher Scientific) yaze yanyangwa ngeSyto-60 (10 μM) kwi-37 °C (ThermoFisher Scientific). Imifanekiso ye-Z-stack yemizuzu eli-15 yenziwe kusetyenziswa iLeica TCS SP8.
Idatha ephindaphindwayo yebhayoloji kunye neyobuchwephesha idweliswe kwaye yahlalutywa kusetyenziswa isoftware yeGraphpad Prism V9 (GraphPad Software, La Jolla, CA). Uhlalutyo lwendlela enye lokwahluka kunye nothelekiso oluninzi kusetyenziswa uvavanyo lwe-post hoc lukaDunnett lusetyenziselwe ukuvavanya umahluko phakathi konyango ngalunye kunye ne-non-antimicrobial control dressing. Ixabiso le-p <0.05 lithathwe njengelibalulekileyo.
Ukusebenza kakuhle kwee-silver gel fibrous dressings kwaqala kwavavanywa ngokuchasene nee-biofilm colonies ze-Staphylococcus aureus kunye ne-Pseudomonas aeruginosa in vitro. Ii-silver dressings ziqulathe iifomyula ezahlukeneyo zesilivere: ii-silver dressings zemveli zivelisa ii-Ag1+ ions; ii-silver dressings, ezinokuvelisa ii-Ag1+ ions emva kokongezwa kwe-EDTA/BC, zinokutshabalalisa i-biofilm matrix kwaye ziveze iibhaktheriya kwisilivere phantsi kwesiphumo sokulwa neentsholongwane sesilivere. ii-ions15 kunye nee-dressings eziqulethe iityuwa ze-Ag ezineoksijini ezivelisa ii-Ag1+, Ag2+ kunye ne-Ag3+. Ukusebenza kwayo kuthelekiswa ne-non-antimicrobial control dressing eyenziwe ngee-geled fibers. Iibhaktheriya eziseleyo ezisebenzayo kwi-biofilm zavavanywa rhoqo emva kweeyure ezingama-24 kangangeentsuku ezi-8 (Umfanekiso 1). Ngomhla wesi-5, i-biofilm yaphinda yafakwa kwi-3.85 × 105S. I-Staphylococcus aureus okanye i-1.22 × 105P. aeruginosa ukuvavanya ukubuyiselwa kwe-biofilm. Xa kuthelekiswa nee-dressings zokulawula ezingezizo ii-antimicrobial, ii-dressings ze-Ag1+ bezinefuthe elincinci ekusebenzeni kweebhaktheriya kwi-Staphylococcus aureus kunye ne-Pseudomonas aeruginosa biofilms kwiintsuku ezi-5. Ngokwahlukileyo koko, ii-dressings ezine-oxygenated Ag kunye ne-Ag1+ + EDTA/BC salts bezisebenza kakuhle ekubulaleni iibhaktheriya ngaphakathi kwi-biofilm kwiintsuku ezi-5. Emva kokugonyelwa ngokuphindaphindiweyo nge-planktonic bacteria ngomhla wesi-5, akukho kubuyiselwa kwe-biofilm okubonweyo (Umzobo 1).
Ukulinganiswa kweebhaktheriya ezisebenzayo kwi-Staphylococcus aureus kunye ne-Pseudomonas aeruginosa biofilms emva konyango ngee-silver dressings. Ii-biofilm colonies ze-Staphylococcus aureus kunye ne-Pseudomonas aeruginosa ziphathwe ngee-silver dressings okanye ii-non-antimicrobial control dressings, kwaye inani leebhaktheriya ezisebenzayo eziseleyo lichongiwe rhoqo emva kweeyure ezingama-24. Emva kweentsuku ezi-5, i-biofilm yaphinda yafakwa kwi-3.85×105S. I-Staphylococcus aureus okanye i-1.22×105P. Iikholoni ze-bacterioplankton Pseudomonas aeruginosa zenziwe ngokwahlukeneyo ukuze kuvavanywe ukubuyiselwa kwe-biofilm. Iigrafu zibonisa impazamo eqhelekileyo +/-.
Ukuze sibone impembelelo yeengubo zesilivere ekukhuleni kwe-biofilm, iingubo zifakwe kwi-biofilms ezivuthiweyo ezikhuliswe kwisikhumba sehagu ngaphandle kobomi. Emva kweeyure ezingama-24, i-dressing iyasuswa kwaye i-biofilm idaywe ngedayi eluhlaza okwesibhakabhaka, eguqulwa yibhaktiriya ephilayo ibe ngumbala opinki. Ii-Biofilms eziphathwe ngeengubo zokulawula zazipinki, nto leyo ebonisa ukuba kukho iibhaktiriya eziphilayo ngaphakathi kwi-biofilm (Umfanekiso 2A). Ngokwahlukileyo koko, i-biofilm ephathwe ngengubo ye-oxysols ye-Ag yayiluhlaza okwesibhakabhaka, nto leyo ebonisa ukuba iibhaktiriya eziseleyo kumphezulu wesikhumba sehagu yayiziibhaktiriya ezingaphiliyo (Umfanekiso 2B). Umbala oxutyiweyo oluhlaza okwesibhakabhaka nopinki wabonwa kwi-biofilms eziphathwe ngeengubo ze-Ag1+, nto leyo ebonisa ukuba kukho iibhaktiriya eziphilayo nezingaphiliyo ngaphakathi kwi-biofilm (Umfanekiso 2C), ngelixa iingubo ze-EDTA/BC eziqulethe i-Ag1+ zaziluhlaza okwesibhakabhaka kakhulu kunye namabala athile apinki. nto leyo ebonisa iindawo ezingachaphazelekanga yingubo yesilivere (Umfanekiso 2D). Ukulinganiswa kweendawo ezisebenzayo (ezipinki) nezingasebenziyo (eziluhlaza okwesibhakabhaka) kubonise ukuba i-control patch yayisebenza nge-75% (Umfanekiso 2E). Ii-Ag1+ + EDTA/BC dressings zenziwe ngendlela efanayo nee-Ag salt dressings ezine-oxygen, kunye namazinga okusinda angama-13% kunye nama-14%, ngokulandelelana. I-Ag1+ dressing ikwanciphise ukuphila kwebhaktheriya ngama-21%. Ezi biofilms zabonwa kusetyenziswa i-scanning electron microscopy (SEM). Emva konyango nge-control dressing kunye ne-Ag1+ dressing, kwabonwa umaleko wePseudomonas aeruginosa ugquma ulusu lwengulube (Umfanekiso 2F,H), ngelixa emva konyango nge-Ag1+ dressing, kwafunyanwa iiseli ezimbalwa zebhaktheriya kwaye kwafunyanwa iiseli ezimbalwa zebhaktheriya ngaphantsi. Iifayibha zeCollagen zinokuthathwa njengolwakhiwo lwezicubu zesikhumba sengulube (Umfanekiso 2G). Emva konyango nge-Ag1+ + EDTA/BC dressing, ii-bacterial plaques kunye ne-collagen fiber plaques ezingaphantsi kwazo zabonakala (Umfanekiso 2I).
Ukubona i-Pseudomonas aeruginosa biofilm emva konyango lwe-silver dressing. (A–D) Ukuphila kwebhaktiriya kwi-Pseudomonas aeruginosa biofilms ezikhuliswe kwisikhumba sengulube kuboniswe kusetyenziswa i-PrestoBlue viability dye kwiiyure ezingama-24 emva konyango ngee-silver dressings okanye ii-non-antimicrobial control dressings. Iibhaktiriya eziphilayo zipinki, iibhaktiriya ezingaphiliyo kwaye ulusu lwengulube luluhlaza okwesibhakabhaka. (E) Ukulinganiswa kwee-Pseudomonas aeruginosa biofilms ezikhuliswe kwisikhumba sengulube (ibala elipinki) kusetyenziswa i-scanning electron microscopy Image Pro version 10 (FI) kwaye ziphathwe nge-silver dressing okanye i-non-antimicrobial dressing iiyure ezingama-24. I-SEM scale bar = 5 µm. (J–M) Ii-colonial biofilms zakhula kwiifilitha kwaye zadaywa nge-PrestoBlue reactive dye emva kweeyure ezingama-24 zokufunxa ngee-silver dressings.
Ukuze kuchongwe ukuba ukudibana okusondeleyo phakathi kwee-dressings kunye ne-biofilms kuchaphazele ukusebenza kakuhle kwee-dressings, ii-biofilms zasekoloniyali ezibekwe kwindawo ethe tyaba zaphathwa ngee-dressings kangangeeyure ezingama-24 zaze zadaywa ngee-reactive dyes. I-biofilm enganyangwanga yayinombala opinki omnyama (Umfanekiso 2J). Ngokungafaniyo nee-biofilms eziphathwe ngee-dressings ezine-oxygenated Ag salts (Umfanekiso 2K), ii-biofilms eziphathwe ngee-dressings ezine-Ag1+ okanye i-Ag1+ + EDTA/BC zibonise imida ye-pink staining (Umfanekiso 2L,M). Lo mbala upinki ubonisa ubukho beebhaktheriya eziphilayo kwaye unxulunyaniswa nendawo ye-suture ngaphakathi kwe-dressing. Ezi ndawo zithungiweyo zenza izithuba ezifileyo ezivumela iibhaktheriya ngaphakathi kwe-biofilm ukuba ziphile.
Ukuvavanya ukusebenza kakuhle kwee-silver dressing in vivo, amanxeba akhutshwe ngokupheleleyo eempuku ezosulelwe yi-S. aureus evuthiweyo kunye ne-P. aeruginosa biofilms anyangwa ngee-non-antimicrobial control dressings okanye ii-silver dressings. Emva kweentsuku ezi-3 zonyango, uhlalutyo lomfanekiso we-macroscopic lubonise ubungakanani obuncinci bezilonda xa zinyangwa ngee-oxygenated salt dressings xa kuthelekiswa nee-non-antimicrobial control dressings kunye nezinye ii-silver dressings (Umfanekiso 3A-H). Ukuqinisekisa oku kuqatshelweyo, amanxeba aqokelelwe kwaye indawo yenxeba kunye nokuhlaziywa kwakhona kwalinganiswa kwiindawo zezicubu ezifakwe i-hematoxylin kunye ne-eosin kusetyenziswa isoftware ye-image pro version 10 (Umfanekiso 3I-L).
Impembelelo ye-silver dressings kumphezulu wenxeba kunye nokuphinda kufakwe i-epithelialization yamanxeba asuleleke yi-biofilms. (A–H) Iiseli ezincinci ezisuleleke yi-biofilms ze-Pseudomonas aeruginosa (A–D) kunye ne-Staphylococcus aureus (E–H) emva kweentsuku ezintathu zonyango nge-non-antimicrobial control dressing, i-oxygenated Ag salt dressing, i-Ag1+ dressing, kunye ne-Ag1+ dressing. Imifanekiso emele i-macroscopic. amanxeba eempuku ane-Ag1+ + EDTA/BC dressing. (IL) Imele i-Pseudomonas aeruginosa infection, iindawo ze-histological ezidaywe yi-hematoxylin kunye ne-eosin, ezisetyenziselwa ukulinganisa indawo yenxeba kunye nokuvuselelwa kwe-epithelial. Ukulinganiswa kwendawo yenxeba (M, O) kunye nepesenti ye-reepithelialization (N, P) yamanxeba asuleleke yi-Pseudomonas aeruginosa (M, N) kunye ne-Staphylococcus aureus (O, P) biofilms (ngeqela lonyango n = 12). Iigrafu zibonisa impazamo eqhelekileyo +/-. * kuthetha u-p = < 0.05 ** kuthetha u-p = < 0.01; Isikali se-macroscopic = 2.5 mm, isikali se-histological = 500 µm.
Ukulinganiswa kwendawo yenxeba kumanxeba asuleleke yi-Pseudomonas aeruginosa biofilm (Umfanekiso 3M) kubonise ukuba amanxeba aphathwe nge-Ag oxysalts ayenobukhulu obuphakathi benxeba obuyi-2.5 mm2, ngelixa i-non-antimicrobial control dressing yayinobukhulu obuphakathi benxeba obuyi-3.1 mm2, into engeyonyani. ifikelele kwinani elibalulekileyo (Umfanekiso 3M). p = 0.423). Amanxeba aphathwe nge-Ag1+ okanye i-Ag1+ + EDTA/BC awazange anciphise indawo yenxeba (3.1 mm2 kunye ne-3.6 mm2, ngokwahlukeneyo). Unyango nge-oxygenated Ag salt dressing lukhuthaze ukuphinda kusetyenziswe i-epithelialization ukuya kwinqanaba eliphezulu kune-non-antimicrobial control dressing (34% kunye ne-15%, ngokwahlukeneyo; p = 0.029) kunye ne-Ag1+ okanye i-Ag1+ + EDTA/BC (10% kunye ne-11%) (Umfanekiso 3N). . , ngokwahlukeneyo).
Iindlela ezifanayo kwindawo yenxeba kunye nokuvuselelwa kwe-epithelial zibonwe kumanxeba asuleleke yi-S. aureus biofilms (Umfanekiso 3O). Ii-dressings ezineetyuwa zesilivere ezineoksijini zinciphise indawo yenxeba (2.0 mm2) ngama-23% xa kuthelekiswa ne-control non-antimicrobial dressing (2.6 mm2), nangona oku kuncipha bekungabalulekanga (p = 0.304) (Umfanekiso 3O). Ukongeza, indawo yenxeba kwiqela lonyango lwe-Ag1+ incitshisiwe kancinci (2.4 mm2), ngelixa inxeba eliphathwe nge-Ag1+ + EDTA/BC dressing lingazange linciphise indawo yenxeba (2.9 mm2). Iityuwa zeoksijini ze-Ag nazo zikhuthaze ukuphinda kusetyenziswe ii-epithelialization zamanxeba asuleleke yi-S. aureus biofilm (31%) kakhulu kunalawo aphathwe ngee-non-antimicrobial control dressings (12%, p = 0.003) (Umfanekiso 3P). I-Ag1+ dressing (16%, p = 0.903) kunye ne-Ag+1 + EDTA/BC dressing (14%, p = 0.965) ibonise amanqanaba okuvuselelwa kwe-epithelial afana nolawulo.
Ukuze kubonwe impembelelo ye-silver dressings kwi-biofilm matrix, kwenziwe i-Toto 1 kunye ne-Syto 60 iodide staining (Umzobo 4). I-Toto 1 iodide yidayi engena-cell enokusetyenziswa ukubona ngokuchanekileyo ii-extracellular nucleic acids, ezifumaneka kakhulu kwi-EPS ye-biofilms. I-Syto 60 yidayi engena-cell esetyenziswa njenge-counterstain16. Ukuqwalaselwa kwe-Toto 1 kunye ne-Syto 60 iodide kumanxeba afakwe i-biofilms ye-Pseudomonas aeruginosa (Umfanekiso 4A-D) kunye ne-Staphylococcus aureus (Umfanekiso 4I-L) kubonise ukuba emva kweentsuku ezi-3 zonyango lwe-dressing, i-EPS kwi-biofilm yancitshiswa kakhulu. equlethe iityuwa ze-oxygenated Ag kunye ne-Ag1+ + EDTA/BC. Ii-Ag1+ dressings ezingenazo ezinye ii-antibiofilm ziye zanciphisa kakhulu i-DNA engenaseli kumanxeba afakwe i-Pseudomonas aeruginosa kodwa azizange zisebenze kakuhle kumanxeba afakwe i-Staphylococcus aureus.
Imifanekiso ye-biofilm yenxeba ekwi-vivo emva kweentsuku ezi-3 zonyango nge-control okanye i-silver dressings. Imifanekiso ye-Confocal ye-Pseudomonas aeruginosa (A–D) kunye ne-Staphylococcus aureus (I–L) epeyintwe nge-Toto 1 (eluhlaza) ukuze ubone i-extracellular nucleic acids, inxalenye ye-extracellular biofilm polymers. Ukuze upeyintwe i-intracellular nucleic acids, sebenzisa i-Syto 60 (ebomvu). ii-acids. P. Ukuskena i-electron microscopy yamanxeba asuleleke yi-Pseudomonas aeruginosa (E–H) kunye ne-Staphylococcus aureus (M–P) biofilms emva kweentsuku ezi-3 zonyango nge-control kunye ne-silver dressings. I-SEM scale bar = 5 µm. I-Confocal imaging scale bar = 50 µm.
I-scanning electron microscopy ibonise ukuba iimpuku ezifakwe ii-biofilm colonies zePseudomonas aeruginosa (Umfanekiso 4E-H) kunye neStaphylococcus aureus (Umfanekiso 4M-P) zazinebhaktheriya ezimbalwa kakhulu emanxebeni azo emva kweentsuku ezi-3 zonyango oluneempahla zesilivere zonke.
Ukuvavanya impembelelo ye-silver dressings ekuvuvukeni kwamanxeba kwiimpuku ezine-biofilm, amacandelo amanxeba ane-biofilm anyangwa nge-control okanye i-silver dressings kangangeentsuku ezi-3 adaywa nge-immunohistochemically kusetyenziswa ii-antibodies ezithile ze-neutrophils kunye ne-macrophages. Ukuchonga ubungakanani bee-neutrophils kunye ne-macrophages ngaphakathi. granulation tissue. Umfanekiso 5). Zonke ii-silver dressings zinciphise inani lee-neutrophils kunye ne-macrophages kumanxeba ane-Pseudomonas aeruginosa xa kuthelekiswa nee-non-antimicrobial control dressings emva kweentsuku ezintathu zonyango. Nangona kunjalo, unyango nge-oxygenated silver salt dressing lubangele ukwehla okukhulu kwee-neutrophils (p = <0.0001) kunye nee-macrophages (p = <0.0001) xa kuthelekiswa nezinye ii-silver dressings ezivavanyiweyo (Umfanekiso 5I,J). Nangona i-Ag1+ + EDTA/BC yayinempembelelo enkulu kwi-wound biofilm, yanciphisa amanqanaba e-neutrophil kunye ne-macrophage kancinci kune-Ag1+ dressing. Amanxeba aphakathi asuleleke yi-S. aureus biofilm nawo abonwe emva kokunxitywa nge-Ag (p = <0.0001), Ag1+ (p = 0.0008) kunye ne-Ag1++ EDTA/BC (p = 0.0043) oxisols xa kuthelekiswa ne-control . Iindlela ezifanayo ziyabonwa kwi-neutropenia. bandage (Umzobo 5K). Nangona kunjalo, kuphela i-oxygenated Ag salt dressing ebonise ukwehla okukhulu kwinani lee-macrophages kwizicubu ze-granulation xa kuthelekiswa nolawulo kumanxeba asuleleke yi-S. aureus biofilms (p = 0.0339) (Umfanekiso 5L).
IiNeutrophils kunye neemacrophages zilinganisiwe kwizilonda ezosulelwe yiPseudomonas aeruginosa kunye neStaphylococcus aureus biofilms emva kweentsuku ezi-3 zonyango nge-non-antimicrobial control okanye i-silver dressings. IiNeutrophils (AD) kunye neemacrophages (EH) zilinganisiwe kwiindawo zezicubu ezidaywe ngama-antibodies athile kwiineutrophils okanye iimacrophages. Ukulinganiswa kweeneutrophils (I kunye ne-K) kunye neemacrophages (J kunye ne-L) kwizilonda ezosulelwe yiPseudomonas aeruginosa (I kunye ne-J) kunye neStaphylococcus aureus (K & L) biofilms. N = 12 kwiqela ngalinye. Iigrafu zibonisa impazamo eqhelekileyo +/-, ixabiso lokubaluleka xa kuthelekiswa ne-non-antibacterial control dressing, * kuthetha p = < 0.05, ** kuthetha p = < 0.01; *** kuthetha p = < 0.001; kubonisa p = <0.0001).
Emva koko sivavanye impembelelo yee-silver dressings ekuphilisweni okuxhomekeke kusulelo. Amanxeba angosuleliyo aphathwe nge-non-antimicrobial control dressing okanye i-silver dressing kangangeentsuku ezi-3 (Umfanekiso 6). Phakathi kwee-silver dressings ezivavanyiweyo, kuphela amanxeba aphathwe nge-oxygenated salt dressing abonakala emancinci kwimifanekiso ye-macroscopic kunamanxeba aphathwe nge-control (Umfanekiso 6A-D). Ukulinganiswa kwendawo yenxeba kusetyenziswa uhlalutyo lwe-histological kubonise ukuba indawo yenxeba ephakathi emva konyango nge-Ag oxysols dressing yayiyi-2.35 mm2 xa kuthelekiswa ne-2.96 mm2 yamanxeba aphathwe ngeqela lolawulo, kodwa lo mahluko awuzange ufikelele kwinani lezibalo (p = 0.488) (Umfanekiso 6I). Ngokwahlukileyo koko, akukho kuncipha kwendawo yenxeba okubonwe emva konyango nge-Ag1+ (3.38 mm2, p = 0.757) okanye i-Ag1+ + EDTA/BC (4.18 mm2, p = 0.054) dressings xa kuthelekiswa neqela lolawulo. Ukwanda kokuvuselelwa kwe-epithelial kubonwe nge-Ag oxysol dressing xa kuthelekiswa neqela lolawulo (30% vs. 22%, ngokulandelanayo), nangona oku kungazange kufikelele kwixabiso elibalulekileyo (p = 0.067), oku kubaluleke kakhulu kwaye kuqinisekisa iziphumo zangaphambili. I-dressing ene-oxysols ikhuthaza ukuphinda kusetyenziswe i-epithelialization. -Ukufakelwa kwe-epithelialization yamanxeba angosuleleki17. Ngokwahlukileyo koko, unyango nge-Ag1+ okanye i-Ag1+ + EDTA/BC dressings aluzange lube nafuthe okanye lubonise ukwehla kokusetyenziswa kwe-epithelialization xa kuthelekiswa nolawulo.
Isiphumo sokubopha amanxeba esilivere ekupholisweni kwamanxeba kwiimpuku ezingasulelekiyo ngokususwa ngokupheleleyo. (AD) Imifanekiso emele i-macroscopic yamanxeba emva kweentsuku ezintathu zonyango nge-non-antimicrobial control dressing kunye ne-silver dressing. (EH) Amacandelo amele amanxeba afakwe i-hematoxylin kunye ne-eosin. Ubungakanani bendawo yenxeba (I) kunye nepesenti ye-reepithelialization (J) zibalwe kumacandelo e-histological embindini wenxeba kusetyenziswa isoftware yohlalutyo lomfanekiso (n = 11–12 kwiqela ngalinye lonyango). Iigrafu zibonisa impazamo ephakathi +/- eqhelekileyo. * kuthetha p = <0.05.
ISilver inembali ende yokusetyenziswa njengonyango lokulwa neentsholongwane ekuphilisweni kwamanxeba, kodwa iindlela ezahlukeneyo zokwenziwa kunye neendlela zokuhambisa zinokubangela umahluko ekusebenzeni kweentsholongwane 18. Ngaphezu koko, iipropati ze-antibiofilm zeenkqubo ezithile zokuhambisa isilivere aziqondwa ngokupheleleyo. Nangona impendulo yomzimba yomzi iyasebenza kakhulu kwi-planktonic bacteria, ngokubanzi ayisebenzi kangako kwi-biofilms19. Iibhaktheriya zePlanktonic zisuswa ngokulula yi-macrophages, kodwa ngaphakathi kwi-biofilms, iiseli ezihlanganisiweyo zibangela iingxaki ezongezelelweyo ngokunciphisa impendulo yomzi ukuya kwinqanaba apho iiseli zomzimba zinokudlula kwi-apoptosis kwaye zikhuphe izinto ezibangela ukudumba ukuze kuphuculwe impendulo yomzimba 20. Kuye kwabonwa ukuba ezinye ii-leukocytes zinokungena kwi-biofilms21 kodwa azikwazi ukuphagocytose iibhaktheriya xa olu khuselo lubuthathaka 22. Indlela epheleleyo kufuneka isetyenziswe ukuxhasa impendulo yomzimba yomzi ngokuchasene nosulelo lwe-biofilm yamanxeba. Ukususwa kwenxeba kunokuphazamisa ngokwasemzimbeni i-biofilm kwaye kususe uninzi lwe-bioburden, kodwa impendulo yomzimba yomzi isenokungasebenzi ngokuchasene ne-biofilm eseleyo, ngakumbi ukuba impendulo yomzimba yomzi ibuthathaka. Ngoko ke, unyango lwe-antimicrobial olufana ne-silver dressing lunokuxhasa impendulo yomzimba womzi kwaye luphelise usulelo lwe-biofilm. Ulwakhiwo, uxinaniso, ukunyibilika, kunye ne-substrate yokuhambisa zinokuchaphazela ukusebenza kwe-silver antimicrobial. Kwiminyaka yakutshanje, inkqubela phambili kubuchwepheshe bokucubungula isilivere yenze ezi dressing zisebenze ngakumbi9,23. Njengoko ubuchwepheshe be-silver dressing buhambela phambili, kubalulekile ukuqonda ukusebenza kwezi dressing ekulawuleni usulelo lwenxeba kwaye, okubaluleke ngakumbi, impembelelo yezi ntlobo zinamandla zesilivere kwindawo yenxeba kunye nokuphiliswa.
Kolu phononongo, sithelekise ukusebenza kakuhle kwee-silver dressings ezimbini eziphambili kunye nee-silver dressings zesiqhelo ezivelisa ii-Ag1+ ions ngokuchasene ne-biofilms zisebenzisa iimodeli ezahlukeneyo ze-in vitro kunye ne-in vivo. Sikwavavanyile nefuthe lale dressings kwindawo yenxeba kunye nokuphiliswa okuxhomekeke kusulelo. Ukunciphisa impembelelo ye-matrix yokuhanjiswa, zonke ii-silver dressings ezivavanyiweyo zenziwe nge-carboxymethylcellulose.
Uvavanyo lwethu lokuqala lwale mixobo yesilivere ngokuchasene ne-biofilms yekoloniyali yePseudomonas aeruginosa kunye neStaphylococcus aureus lubonisa ukuba, ngokungafaniyo nemixobo yendabuko ye-Ag1+, imixobo yesilivere emibini ephucukileyo, i-Ag1+ + EDTA/BC kunye neetyuwa ze-Ag ezineoksijini, zisebenza kwi-5. Zibulala ngempumelelo iibhaktheriya ze-biofilm kwiintsuku ezimbalwa. Ukongeza, ezi mixobo zithintela ukwakheka kwakhona kwe-biofilm xa zichaphazeleka ngokuphindaphindiweyo kwibhaktheriya ye-planktonic. I-Ag1+ dressing yayine-silver chloride, i-silver compound efanayo kunye ne-base matrix njenge-Ag1+ + EDTA/BC, kwaye yayinefuthe elincinci ekusebenzeni kwebhaktheriya ngaphakathi kwi-biofilm kwixesha elifanayo. Ukuqaphela ukuba i-Ag1+ + EDTA/BC dressing yayisebenza ngakumbi ngokuchasene ne-biofilm kune-Ag1+ dressing equlathe i-matrix efanayo kunye ne-silver compound ixhasa ingcamango yokuba izithako ezongezelelweyo ziyafuneka ukwandisa ukusebenza kwe-silver chloride ngokuchasene ne-biofilm, njengoko kuye kwaxelwa kwenye indawo15. Ezi ziphumo zixhasa ingcamango yokuba i-BC kunye ne-EDTA zidlala indima eyongezelelweyo enegalelo ekusebenzeni kakuhle kwe-dressing kwaye ukungabikho kwale nxalenye kwi-Ag1+ dressings kusenokuba negalelo ekungabonisini ukusebenza kakuhle kwe-in vitro. Sifumanise ukuba ii-Ag salt dressings ezivelisa i-Ag2+ kunye ne-Ag3+ ions zibonakalise ukusebenza okunamandla okulwa neentsholongwane kune-Ag1+ kwaye kumanqanaba afana ne-Ag1+ + EDTA/BC. Nangona kunjalo, ngenxa yamandla aphezulu e-redox, akucaci ukuba ii-Ag3+ ions zihlala zisebenza kwaye zisebenza ixesha elingakanani ngokuchasene ne-wound biofilms kwaye ngoko ke kufanelekile uphononongo olongezelelweyo. Ukongeza, kukho ii-dressings ezininzi ezahlukeneyo ezivelisa ii-Ag1+ ions ezingakhange zivavanywe kolu phononongo. Ezi dressings zenziwe ngee-compounds zesilivere ezahlukeneyo, ubuninzi, kunye ne-base matrices, ezinokuchaphazela ukuhanjiswa kwee-Ag1+ ions kunye nokusebenza kwazo ngokuchasene ne-biofilms. Kubalulekile ukuqaphela ukuba kukho iimodeli ezininzi ezahlukeneyo ze-in vitro kunye ne-in vivo ezisetyenziselwa ukuvavanya ukusebenza kakuhle kwe-wound dressings ngokuchasene ne-biofilms. Uhlobo lwemodeli esetyenzisiweyo, kunye nomxholo wetyuwa kunye neproteni yemidiya esetyenziswa kwezi modeli, ziya kuchaphazela ukusebenza kakuhle kwe-dressing. Kwimodeli yethu ye-in vivo, sivumele i-biofilm ukuba ivuthwe kwi-in vitro saza sayidlulisela kumphezulu wesikhumba senxeba. Impendulo yomzimba yempuku ephethwe yi-host iyasebenza kakhulu ngokuchasene neebhaktheriya ze-planktonic ezifakwe kwinxeba, ngaloo ndlela zenze i-biofilm njengoko inxeba liphola. Ukongezwa kwe-biofilm evuthiweyo kwinxeba kunciphisa ukusebenza kakuhle kwempendulo yomzimba womntu ekwenziweni kwe-biofilm ngokuvumela i-biofilm evuthiweyo ukuba izinze ngaphakathi kwenxeba ngaphambi kokuba kuqale ukuphiliswa. Ke ngoko, imodeli yethu isivumela ukuba sivavanye ukusebenza kakuhle kwe-antimicrobial dressings kwi-biofilms evuthiweyo ngaphambi kokuba amanxeba aqale ukuphola.
Sikwafumanise ukuba ukulingana kwempahla yokunxiba kuchaphazele ukusebenza kakuhle kwempahla yokunxiba yesilivere kwi-biofilms ezikhuliswe kwi-vitro kunye nolusu lwengulube. Ukudibana ngokusondeleyo nenxeba kuthathwa njengokubalulekileyo ekusebenzeni kakuhle kwempahla yokunxiba24,25. Iimpahla zokunxiba ezine-oxygen Ag salts zazisondelelene kakhulu ne-biofilms ezivuthiweyo, nto leyo eyabangela ukwehla okukhulu kwinani leebhaktheriya ezisebenzayo kwi-biofilm emva kweeyure ezingama-24. Ngokwahlukileyo koko, xa zinyangwa nge-Ag1+ kunye ne-Ag1+ + EDTA/BC dressings, kwasala inani elikhulu leebhaktheriya ezisebenzayo. Ezi mpahla zokunxiba ziqulathe imithungo kulo lonke ubude bempahla yokunxiba, nto leyo edala izithuba ezifileyo ezithintela ukudibana ngokusondeleyo ne-biofilm. Kwizifundo zethu ze-in vitro, ezi ndawo zingadibaniyo zithintele ukubulawa kweebhaktheriya ezisebenzayo kwi-biofilm. Sivavanye ukuphila kwebhaktheriya kuphela emva kweeyure ezingama-24 zonyango; Ngokuhamba kwexesha, njengoko impahla yokunxiba igcwala ngakumbi, kunokubakho indawo encinci efileyo, nto leyo eya kunciphisa indawo yezi bhaktheriya ezisebenzayo. Nangona kunjalo, oku kugxininisa ukubaluleka kokwakheka kwempahla yokunxiba, kungekuphela nje uhlobo lwesilivere kwimpahla yokunxiba.
Nangona izifundo ze-in vitro ziluncedo ekuthelekiseni ukusebenza kakuhle kweteknoloji ezahlukeneyo zesilivere, kubalulekile ukuqonda imiphumo yezi dressings kwi-biofilms in vivo, apho izicubu zomntu kunye nempendulo yomzimba zinegalelo ekusebenzeni kakuhle kwezi dressings nxamnye ne-biofilms. Impembelelo yezi dressings kwi-biofilms zamanxeba yabonwa kusetyenziswa i-scanning electron microscopy kunye ne-EPS staining ye-biofilm kusetyenziswa i-intracellular kunye ne-extracellular DNA dyes. Sifumanise ukuba emva kweentsuku ezi-3 zonyango, zonke ii-dressings zazisebenza kakuhle ekunciphiseni i-DNA engenaseli kumanxeba ane-biofilm, kodwa i-Ag1+ dressing yayingasebenzi kakuhle kumanxeba ane-Staphylococcus aureus. I-scanning electron microscopy ikwabonise ukuba iibhaktheriya ezimbalwa kakhulu zazikho kumanxeba aphathwe ngee-silver dressings, nangona oku kwakubonakala ngakumbi nge-oxygenated Ag salt dressing kunye ne-Ag1+ EDTA/BC dressing xa kuthelekiswa ne-Ag1+. Ezi datha zibonisa ukuba ii-silver dressings ezivavanyiweyo zazinefuthe elahlukeneyo kwisakhiwo se-biofilm, kodwa akukho nanye kwezi dressings zesilivere eyayikwazile ukuyiphelisa i-biofilm, ixhasa imfuneko yendlela epheleleyo yokunyanga usulelo lwe-biofilm yamanxeba; ukusetyenziswa kweebhanti zesilivere. Unyango luqalwa kukususwa kwesikhumba ukuze kususwe uninzi lwe-biofilm.
Amanxeba angapheliyo adla ngokuba kwimeko yokudumba okukhulu, kunye neeseli ezigqithisileyo zokudumba ezihlala kwizicubu zenxeba ixesha elide, nto leyo ebangela umonakalo kwizicubu kwaye inciphise ioksijini efunekayo ukuze i-metabolism yeseli isebenze kakuhle kwaye isebenze kwinxeba26. Iibhayofilm ziyayenza mandundu le ndawo inobungozi yamanxeba ngokuchaphazela kakubi ukuphiliswa ngeendlela ezahlukeneyo, kubandakanya ukuthintela ukwanda kweeseli kunye nokufuduka kunye nokusebenza kwee-cytokines ezibangela ukuvuvukala27. Njengoko ii-silver dressing zisebenza ngakumbi, kubalulekile ukuqonda impembelelo ezinayo kwindawo ebangela ukuvuvukala kunye nokuphiliswa.
Okunomdla kukuba, nangona zonke ii-silver dressings zichaphazele ukwakheka kwe-biofilm, ii-oxygenated silver salt dressings kuphela eziye zonyusa ukuphinda kufakwe la manxeba asulelekileyo. Ezi datha zixhasa iziphumo zethu zangaphambili17 kunye nezo zikaKalan et al. (2017)28, ezibonise ukhuseleko olufanelekileyo kunye neeprofayili zetyhefu yeetyuwa zesilivere ezine-oxygen, njengoko amazinga aphantsi esilivere ayesebenza ngokuchasene nee-biofilms.
Uphononongo lwethu lwangoku lubonisa umahluko kubuchwepheshe besilivere phakathi kwe-antimicrobial silver dressings kunye nempembelelo yale teknoloji kwindawo yenxeba kunye nokuphiliswa okungenalo usulelo. Nangona kunjalo, ezi ziphumo zahlukile kwizifundo zangaphambili ezibonisa ukuba i-Ag1+ + EDTA/BC dressing iphucule iiparameter zokuphiliswa kweendlebe zomvundla ezilimeleyo emzimbeni. Nangona kunjalo, oku kusenokubangelwa kukungafani kwiimodeli zezilwanyana, amaxesha okulinganisa, kunye neendlela zokusetyenziswa kwebhaktheriya29. Kule meko, ukulinganiswa kwenxeba kwathathwa kwiintsuku ezili-12 emva kokwenzakala ukuvumela izithako ezisebenzayo ze-dressing ukuba zisebenze kwi-biofilm ixesha elide. Oku kuxhaswa luphando olubonise ukuba izilonda zemilenze ezosulelekileyo zonyango ezinyangwe nge-Ag1+ + EDTA/BC ekuqaleni zanda ngobukhulu emva kweveki enye yonyango, kwaye emva koko kwiiveki ezi-3 ezilandelayo zonyango nge-Ag1+ + EDTA/BC kwaye kwiiveki ezi-4 zokusetyenziswa kwe-non-antimicrobial. ii-CMC dressings zokunciphisa ubungakanani bezilonda30.
Iintlobo ezithile kunye noxinzelelo lwesilivere ngaphambili zibonakaliswe njengezinobungozi kwi-vitro 11, kodwa ezi ziphumo kwi-vitro azisoloko ziguqulela kwimiphumo emibi kwi-vivo. Ukongeza, inkqubela phambili kwiteknoloji yesilivere kunye nokuqonda okungcono kweekhompawundi zesilivere kunye noxinzelelo kwii-dressings kukhokelele ekuphuhlisweni kwee-dressings ezininzi zesilivere ezikhuselekileyo nezisebenzayo. Nangona kunjalo, njengoko iteknoloji ye-dressing yesilivere iqhubela phambili, kubalulekile ukuqonda impembelelo yezi dressings kwindawo yamanxeba31,32,33. Kwaxelwa ngaphambili ukuba izinga elonyukayo lokuphinda kusetyenziswe i-epithelialization lihambelana nenani elonyukayo le-M2 macrophages ezichasene nokuvuvukala xa kuthelekiswa ne-phenotype ye-M1 ebangela ukuvuvukala. Oku kuphawulwe kwimodeli yangaphambili yegundane apho ii-dressings zesilivere ze-hydrogel zithelekiswa ne-silver sulfadiazine kunye ne-non-antimicrobial hydrogels34.
Amanxeba angapheliyo anokubonisa ukudumba okugqithisileyo kwaye kuye kwabonwa ukuba ubukho bee-neutrophils ezingaphezulu bunokuba yingozi ekuphilisweni kwamanxeba35. Kwisifundo esenziwe kwiimpuku eziphelelwe yi-neutrophils, ubukho bee-neutrophils buye balibazisa ukuphinda kubuyelwe epithelialization. Ubukho bee-neutrophils ezingaphezulu bukhokelela kumanqanaba aphezulu ee-proteases kunye neentlobo ze-oxygen ezisabelayo, ezifana ne-superoxide kunye ne-hydrogen peroxide, ezinxulunyaniswa namanxeba angapheliyo naphola kancinci37,38. Ngokufanayo, ukwanda kwamanani e-macrophage, ukuba akulawuleki, kunokukhokelela ekuphilisweni kwamanxeba okulibazisa39. Olu nyuko lubaluleke kakhulu ukuba ii-macrophages azikwazi ukutshintsha ukusuka kwi-phenotype ekhuthaza ukuvuvukala ukuya kwi-phenotype ekhuthaza ukupholisa, okubangela ukuba amanxeba angakwazi ukuphuma kwisigaba sokuvuvukala sokuphiliswa40. Sibone ukwehla kwee-neutrophils kunye nee-macrophages kumanxeba ane-biofilm emva kweentsuku ezi-3 zonyango ngazo zonke ii-silver dressings, kodwa ukwehla kwacaca ngakumbi ngee-oxygenated salt dressings. Oku kuncipha kusenokuba ngumphumo othe ngqo wempendulo yomzimba kwisilivere, impendulo ekunciphiseni umthwalo webhayoloji, okanye inxeba likwinqanaba lokugqibela lokuphiliswa kwaye ke ngoko iiseli zomzimba ezikwinxeba ziyancitshiswa. Ukunciphisa inani leeseli ezivuthayo enxebeni kunokugcina imeko-bume ilungele ukuphiliswa kwenxeba. Indlela esebenza ngayo indlela ii-oxysalts ze-Ag ezikhuthaza ngayo ukuphiliswa ngaphandle kosulelo ayicacanga, kodwa amandla e-oxysalts ze-Ag ukuvelisa ioksijini nokutshabalalisa amanqanaba ayingozi e-hydrogen peroxide, umlamli wokudumba, angachaza oku kwaye afune uphando olongezelelweyo17.
Amanxeba asulelekileyo angapholiyo ahlala ebangela ingxaki koogqirha nakwizigulane. Nangona ii-dressing ezininzi zithi ziyasebenza ekulweni neentsholongwane, uphando alusoloko lugxile kwezinye izinto eziphambili ezichaphazela imeko-bume yenxeba. Olu phononongo lubonisa ukuba ubuchwepheshe obahlukeneyo besilivere buneendlela ezahlukeneyo zokuthintela iintsholongwane kwaye, okubalulekileyo, iziphumo ezahlukeneyo kwindawo yenxeba kunye nokuphiliswa, ngaphandle kosulelo. Nangona ezi zifundo ze-in vitro kunye ne-in vivo zibonisa ukusebenza kakuhle kwezi dressing ekunyangeni usulelo lwenxeba kunye nokukhuthaza ukuphiliswa, iimvavanyo ezilawulwa ngokungacwangciswanga ziyafuneka ukuvavanya ukusebenza kakuhle kwezi dressing ekliniki.
Iiseti zedatha ezisetyenzisiweyo kunye/okanye ezihlalutyiweyo ngexesha lophando lwangoku ziyafumaneka kumbhali ohambelanayo xa kuceliwe ngokufanelekileyo.
Ixesha leposi: Julayi-15-2024