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Cov kab mob me me uas loj hlob hauv cov qhov txhab feem ntau pom nws tus kheej ua cov biofilms, uas cuam tshuam rau kev kho thiab nyuaj rau tshem tawm. Cov khaub ncaws nyiaj tshiab thov kom tawm tsam cov kab mob qhov txhab, tab sis lawv cov tshuaj tua kab mob thiab cov teebmeem kho mob uas tsis cuam tshuam rau kev kis kab mob feem ntau tsis paub. Siv cov qauv biofilm hauv vitro thiab hauv vivo ntawm Staphylococcus aureus thiab Pseudomonas aeruginosa, peb tshaj tawm qhov ua tau zoo ntawm Ag1+ ion-generating dressings; Ag1+ dressings uas muaj ethylenediaminetetraacetic acid thiab benzethonium chloride (Ag1+/EDTA/BC), thiab cov khaub ncaws uas muaj silver nitrate (Ag oxysalts). , uas tsim cov Ag1+, Ag2+ thiab Ag3+ ions los tawm tsam cov biofilm qhov txhab thiab nws cov teebmeem rau kev kho. Ag1+ dressings muaj cov teebmeem tsawg kawg nkaus rau cov biofilm qhov txhab hauv vitro thiab hauv nas (C57BL/6j). Qhov sib txawv, cov ntsev Ag uas muaj oxygenated thiab Ag1+/EDTA/BC dressings tau txo cov kab mob uas muaj sia nyob hauv biofilms hauv vitro thiab tau qhia txog kev txo qis ntawm cov kab mob thiab EPS hauv cov biofilms qhov txhab nas. Cov dressings no muaj cov teebmeem sib txawv ntawm kev kho cov qhov txhab uas muaj kab mob biofilm thiab cov qhov txhab uas tsis muaj kab mob biofilm, nrog rau cov dressings ntsev uas muaj oxygenated muaj cov teebmeem zoo dua rau kev rov ua dua tshiab, qhov loj ntawm qhov txhab, thiab kev o piv rau kev kho mob tswj thiab lwm yam kev hnav khaub ncaws nyiaj. Cov khoom sib txawv ntawm cov khaub ncaws nyiaj yuav muaj cov teebmeem sib txawv ntawm cov biofilm qhov txhab thiab kev kho, thiab qhov no yuav tsum tau xav txog thaum xaiv cov khaub ncaws rau kev kho cov qhov txhab uas muaj kab mob biofilm.
Cov qhov txhab uas mob ntev ntev yog txhais tias yog "cov qhov txhab uas tsis kho tau zoo li qub raws li kev cai thiab raws sijhawm." 1. Cov qhov txhab uas mob ntev ntev ua rau muaj kev nyuaj siab rau lub hlwb, kev sib raug zoo, thiab kev lag luam rau cov neeg mob thiab lub cev kho mob. Kev siv nyiaj txhua xyoo ntawm NHS rau kev kho cov qhov txhab thiab cov kab mob sib txuas yog kwv yees li ntawm £ 8.3 billion hauv xyoo 2017–182. Cov qhov txhab uas mob ntev ntev kuj yog qhov teeb meem tseem ceeb hauv Tebchaws Meskas, nrog rau Medicare kwv yees tus nqi txhua xyoo ntawm kev kho cov neeg mob uas muaj qhov txhab ntawm $ 28.1–$ 96.8 billion3.
Kev kis kab mob yog ib qho tseem ceeb uas ua rau cov qhov txhab tsis zoo. Feem ntau cov kab mob tshwm sim ua cov biofilms, uas muaj nyob rau hauv 78% ntawm cov qhov txhab uas tsis kho tau ntev. Cov biofilms tsim thaum cov kab mob me me txuas nrog cov nto, xws li cov nto qhov txhab, thiab tuaj yeem sib sau ua ke los tsim cov zej zog tsim cov polymer extracellular (EPS). Cov biofilm qhov txhab muaj feem cuam tshuam nrog kev ua kom mob ntxiv ua rau cov ntaub so ntswg puas tsuaj, uas tuaj yeem ncua lossis tiv thaiv kev kho mob4. Kev nce ntxiv ntawm kev puas tsuaj ntawm cov ntaub so ntswg tej zaum yuav yog vim muaj kev ua haujlwm ntau ntxiv ntawm matrix metalloproteinases, collagenase, elastase thiab reactive oxygen species5. Ntxiv mus, cov hlwb inflammatory thiab biofilms lawv tus kheej yog cov neeg siv oxygen ntau thiab yog li ntawd tuaj yeem ua rau cov ntaub so ntswg hauv zos hypoxia, ua rau cov hlwb tsis muaj cov pa oxygen tseem ceeb uas xav tau rau kev kho cov ntaub so ntswg zoo6.
Cov biofilms uas laus lawm tiv taus cov tshuaj tua kab mob heev, xav tau cov tswv yim nruj heev los tswj cov kab mob biofilm, xws li kev kho mob siv tshuab ua raws li kev kho mob tua kab mob zoo. Vim tias cov biofilms tuaj yeem rov tsim dua sai sai, cov tshuaj tua kab mob zoo tuaj yeem txo qhov kev pheej hmoo ntawm kev rov tsim dua tom qab phais mob debridement7.
Nyiaj siv ntau zuj zus rau hauv cov ntaub qhwv tiv thaiv kab mob thiab feem ntau siv ua thawj kab kev kho mob rau cov qhov txhab uas kis tau kab mob ntev. Muaj ntau cov ntaub qhwv nyiaj muaj nyob hauv khw, txhua daim muaj cov khoom sib txawv ntawm cov nyiaj, qhov concentration, thiab cov matrix hauv paus. Kev nce qib hauv cov ntaub qhwv caj npab nyiaj tau ua rau muaj kev tsim cov ntaub qhwv caj npab nyiaj tshiab. Cov hlau ntawm nyiaj (Ag0) yog inert; Yuav kom ua tiav cov tshuaj tua kab mob zoo, nws yuav tsum poob ib lub electron los tsim cov nyiaj ionic (Ag1+). Cov ntaub qhwv nyiaj ib txwm muaj cov khoom sib xyaw nyiaj lossis cov nyiaj hlau uas, thaum raug kua, lwj mus ua cov ions Ag1+. Cov ions Ag1+ no ua rau cov kab mob cell, tshem tawm cov electrons ntawm cov khoom siv lossis cov txheej txheem tseem ceeb uas tsim nyog rau kev ciaj sia. Cov thev naus laus zis patented tau ua rau muaj kev tsim cov khoom sib xyaw nyiaj tshiab, Ag oxysalts (silver nitrate, Ag7NO11), uas suav nrog hauv cov ntaub qhwv qhov txhab. Tsis zoo li cov nyiaj ib txwm muaj, kev lwj ntawm cov ntsev uas muaj oxygen ua rau cov xwm txheej ntawm nyiaj nrog valence siab dua (Ag1+, Ag2+ thiab Ag3+). Cov kev tshawb fawb hauv vitro tau qhia tias cov ntsev nyiaj uas muaj oxygenated tsawg tsawg muaj txiaj ntsig zoo dua li cov nyiaj ib leeg (Ag1+) tiv thaiv cov kab mob pathogenic xws li Pseudomonas aeruginosa, Staphylococcus aureus thiab Escherichia coli8,9. Lwm hom kev hnav khaub ncaws nyiaj tshiab suav nrog cov khoom xyaw ntxiv, uas yog ethylenediaminetetraacetic acid (EDTA) thiab benzethonium chloride (BC), uas tau tshaj tawm tias tsom mus rau biofilm EPS thiab yog li ua rau cov nyiaj nkag mus rau hauv biofilm ntau dua. Cov thev naus laus zis nyiaj tshiab no muab cov txoj hauv kev tshiab los tsom mus rau cov biofilms qhov txhab. Txawm li cas los xij, qhov cuam tshuam ntawm cov tshuaj tua kab mob no rau qhov chaw mob thiab kev kho mob tsis muaj kev kis kab mob yog qhov tseem ceeb kom ntseeg tau tias lawv tsis tsim ib puag ncig qhov txhab tsis zoo lossis qeeb kho. Kev txhawj xeeb txog kev lom nyiaj hauv vitro tau tshaj tawm nrog ntau cov khaub ncaws nyiaj10,11. Txawm li cas los xij, kev lom nyiaj hauv vitro tseem tsis tau txhais ua kev lom hauv vivo, thiab ntau cov khaub ncaws Ag1+ tau ua pov thawj tias muaj kev nyab xeeb zoo12.
Ntawm no, peb tau tshawb nrhiav qhov ua tau zoo ntawm cov tshuaj pleev carboxymethylcellulose uas muaj cov qauv nyiaj tshiab tiv thaiv cov biofilm qhov txhab hauv vitro thiab hauv vivo. Tsis tas li ntawd, cov teebmeem ntawm cov tshuaj pleev no rau kev tiv thaiv kab mob thiab kev kho mob yam tsis muaj kev kis kab mob tau raug soj ntsuam.
Txhua daim ntaub qhwv uas siv yog muaj muag hauv khw. 3M Kerracel Gel Fiber Dressing (3M, Knutsford, UK) yog ib daim ntaub qhwv gel fiber 100% uas tsis yog tshuaj tua kab mob uas tau siv ua daim ntaub qhwv tswj hauv txoj kev tshawb fawb no. Peb daim ntaub qhwv nyiaj CMC uas tua kab mob tau raug soj ntsuam, uas yog 3M Kerracel Ag dressing (3M, Knutsford, UK), uas muaj 1.7 wt%. oxygenated silver salt (Ag7NO11) hauv cov valence silver ions siab dua (Ag1+, Ag2+ thiab Ag3+). Thaum lub sijhawm lwj ntawm Ag7NO11, Ag1+, Ag2+ thiab Ag3+ ions tau tsim nyob rau hauv qhov sib piv ntawm 1:2:4. Aquacel Ag Extra dressing muaj 1.2% silver chloride (Ag1+) (ConvaTec, Deeside, UK) 13 thiab Aquacel Ag + Extra dressing muaj 1.2% silver chloride (Ag1+), EDTA thiab benzethonium chloride (ConvaTec, Deeside, UK) 14.
Cov kab mob siv hauv kev tshawb fawb no yog Pseudomonas aeruginosa NCTC 10781 (Public Health England, Salisbury) thiab Staphylococcus aureus NCTC 6571 (Public Health England, Salisbury).
Cov kab mob tau loj hlob ib hmos hauv Muller-Hinton kua zaub (Oxoid, Altrincham, UK). Tom qab ntawd, cov kab lis kev cai ib hmos tau diluted 1:100 hauv Mueller-Hinton kua zaub thiab 200 µl plated rau ntawm sterile 0.2 µm Whatman cyclopore membranes (Whatman plc, Maidstone, UK) rau ntawm Mueller-Hinton agar plates (Sigma-Aldrich Company Ltd, Kent, Great Britain). ) Kev tsim cov biofilm colonial ntawm 37°C rau 24 teev. Cov biofilms colonial no tau sim rau logarithmic shrinkage.
Txiav daim ntaub qhwv ua tej daim plaub fab 3 cm2 thiab muab dej deionized uas tsis muaj kab mob ntxuav ua ntej. Muab daim ntaub qhwv tso rau saum daim biofilm ntawm lub colony ntawm lub phaj agar. Txhua 24 ha ntawm biofilm raug tshem tawm, thiab cov kab mob uas muaj sia nyob hauv biofilm (CFU/ml) raug ntsuas los ntawm kev sib xyaw ua ke (10−1 txog 10−7) hauv Day-Angle neutralization broth (Merck-Millipore). Tom qab 24 teev ntawm kev sib xyaw ua ke ntawm 37°C, cov phaj txheem suav tau ua rau ntawm Mueller-Hinton agar phaj. Txhua qhov kev kho mob thiab lub sijhawm tau ua tiav hauv peb daim, thiab cov phaj suav tau rov ua dua rau txhua qhov kev sib xyaw ua ke.
Cov tawv nqaij npua plab tau txais los ntawm cov npua poj niam Dawb Loj hauv 15 feeb tom qab tua raws li cov qauv xa tawm ntawm European Union. Cov tawv nqaij raug txiav thiab ntxuav nrog cov ntaub so cawv, tom qab ntawd khov ntawm -80 ° C rau 24 teev kom ua rau cov tawv nqaij tsis muaj zog. Tom qab yaj, 1 cm2 daim tawv nqaij tau ntxuav peb zaug nrog PBS, 0.6% sodium hypochlorite, thiab 70% ethanol rau 20 feeb txhua zaus. Ua ntej tshem cov epidermis, tshem tawm cov ethanol seem los ntawm kev ntxuav 3 zaug hauv PBS sterile. Cov tawv nqaij tau cog rau hauv lub phaj 6-qhov nrog daim nyias nyias nylon 0.45-μm-tuab (Merck-Millipore) rau saum thiab 3 daim ntaub nqus dej (Merck-Millipore) uas muaj 3 ml fetal bovine serum (Sigma) ntxiv nrog 10% Dulbecco's modified Eagle. Medium (Dulbecco's Modified Eagle Medium - Aldrich Ltd.).
Cov biofilms colonial tau loj hlob raws li tau piav qhia rau kev tshawb fawb txog kev raug biofilm. Tom qab cog cov biofilm rau ntawm daim nyias nyias rau 72 teev, cov biofilm tau siv rau ntawm daim tawv nqaij siv lub voj voog sterile inoculation thiab daim nyias nyias tau raug tshem tawm. Tom qab ntawd cov biofilm tau incubated rau ntawm tus npua lub dermis rau ntxiv 24 teev ntawm 37 ° C kom cov biofilm loj hlob thiab lo rau ntawm daim tawv nqaij npua. Tom qab cov biofilm tau loj hlob thiab txuas, ib daim ntaub qhwv 1.5 cm2, uas tau ua ntej ntub nrog dej distilled sterile, tau siv ncaj qha rau ntawm daim tawv nqaij thiab incubated ntawm 37 ° C rau 24 teev. Cov kab mob muaj sia nyob tau pom los ntawm kev staining los ntawm kev siv PrestoBlue cell viability reagent (Invitrogen, Life Technologies, Paisley, UK) rau ntawm qhov chaw apical ntawm txhua qhov explant thiab incubating nws rau 5 feeb. Siv lub koob yees duab digital Leica DFC425 los ntes cov duab tam sim ntawd ntawm Leica MZ8 microscope. Cov cheeb tsam xim liab tau ntsuas siv Image Pro software version 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (mediacy.com)). Kev ntsuas electron microscopy tau ua raws li tau piav qhia hauv qab no.
Cov kab mob uas loj hlob thaum hmo ntuj tau raug diluted 1:100 hauv Mueller-Hinton kua zaub. 200 μl ntawm cov kab lis kev cai tau ntxiv rau hauv sterile 0.2 μm Whatman cyclopore membrane (Whatman, Maidstone, UK) thiab plated rau ntawm Mueller-Hinton agar. Biofilm plates tau incubated ntawm 37°C rau 72 teev kom tso cai rau biofilm tsim.
Tom qab 3 hnub ntawm biofilm maturation, ib daim ntaub qhwv 3 cm2 square tau muab tso ncaj qha rau ntawm biofilm thiab incubated ntawm 37°C rau 24 teev. Tom qab tshem daim ntaub qhwv tawm ntawm qhov chaw biofilm, 1 ml ntawm PrestoBlue Cell Viability Reagent (Invitrogen, Waltham, MA) tau ntxiv rau ntawm qhov chaw ntawm txhua biofilm rau 20 vib nas this. Cov chaw tau ziab ua ntej cov xim hloov pauv tau kaw siv Nikon D2300 digital koob yees duab (Nikon UK Ltd., Kingston, UK).
Npaj cov kab mob thaum hmo ntuj ntawm Mueller-Hinton agar, hloov cov kab mob ib pawg mus rau 10 ml Mueller-Hinton broth thiab incubate rau ntawm lub shaker ntawm 37°C (100 rpm). Tom qab incubation thaum hmo ntuj, cov kab mob tau diluted 1:100 hauv Mueller-Hinton broth thiab 300 µl tau pom rau ntawm 0.2 µm circular Whatman cyclopore membrane (Whatman International, Maidstone, UK) ntawm Mueller-Hinton agar thiab incubated ntawm 37°C hauv 72 teev. . Cov biofilm laus tau siv rau ntawm qhov txhab raws li tau piav qhia hauv qab no.
Txhua txoj haujlwm nrog tsiaj txhu tau ua tiav ntawm University of Manchester raws li daim ntawv tso cai project uas tau pom zoo los ntawm Office of Animal Welfare and Ethical Review (P8721BD27) thiab raws li cov lus qhia uas tau tshaj tawm los ntawm Home Office raws li 2012 kho dua tshiab ASPA. Txhua tus kws sau ntawv tau ua raws li cov lus qhia tuaj txog. Cov nas C57BL/6j uas muaj yim lub lis piam (Envigo, Oxon, UK) tau siv rau txhua qhov kev tshawb fawb hauv vivo. Cov nas tau muab tshuaj loog nrog isoflurane (Piramal Critical Care Ltd, West Drayton, UK) thiab lawv cov qhov chaw dorsal tau raug txiav thiab ntxuav. Txhua tus nas tau muab qhov txhab 2 × 6 hli excisional siv Stiefel biopsy punch (Schuco International, Hertfordshire, UK). Rau cov qhov txhab uas muaj kab mob biofilm, thov siv 72-teev colonial biofilm uas loj hlob ntawm daim nyias nyias raws li tau piav qhia saum toj no rau txheej dermal ntawm qhov txhab siv lub voj voog sterile inoculation tam sim ntawd tom qab raug mob thiab pov tseg daim nyias nyias. Ib square centimeter ntawm kev hnav khaub ncaws tau ua ntej ntub nrog dej sterile kom tswj tau qhov chaw ntub dej. Cov ntaub qhwv tau muab tso ncaj qha rau txhua qhov txhab thiab npog nrog 3M Tegaderm zaj duab xis (3M, Bracknell, UK) thiab Mastisol kua nplaum (Eloquest Healthcare, Ferndale, MI) siv nyob ib ncig ntawm cov npoo kom muab kev nplaum ntxiv. Buprenorphine (Animalcare, York, UK) tau muab rau ntawm qhov concentration ntawm 0.1 mg / kg ua tshuaj loog. Tua nas peb hnub tom qab raug mob siv txoj kev Schedule 1 thiab tshem tawm, txiav ib nrab, thiab khaws cia thaj chaw txhab raws li qhov xav tau.
Kev pleev xim rau Hematoxylin (ThermoFisher Scientific) thiab eosin (ThermoFisher Scientific) tau ua raws li cov txheej txheem ntawm cov chaw tsim khoom. Thaj chaw raug mob thiab kev rov ua dua tshiab tau ntsuas siv Image Pro software version 10 (Media Cybernetics Inc, Rockville, MD).
Cov ntu ntawm cov ntaub so ntswg tau dewaxed hauv xylene (ThermoFisher Scientific, Loughborough, UK), rov ua kom huv nrog 100–50% ethanol, thiab muab tso rau hauv dej deionized ib ntus (ThermoFisher Scientific). Kev tshuaj ntsuam immunohistochemistry tau ua tiav siv VectaStain Elite ABC PK-6104 kit (Vector Laboratories, Burlingame, CA) raws li cov txheej txheem ntawm cov chaw tsim khoom. Cov tshuaj tiv thaiv thawj zaug rau neutrophils NIMP-R14 (Thermofisher Scientific) thiab macrophages Ms CD107b Pure M3/84 (BD Biosciences, Wokingham, UK) tau diluted 1:100 hauv cov tshuaj thaiv thiab ntxiv rau qhov chaw txiav, ua raws li 2 cov tshuaj tiv thaiv Anti-, VectaStain ABC thiab Vector Nova Red Peroxidase (HRP) substrate kit (Vector Laboratories, Burlingame, CA) thiab counterstained nrog hematoxylin. Cov duab tau txais los ntawm kev siv lub tshuab tsom iav Olympus BX43 thiab lub koob yees duab digital Olympus DP73 (Olympus, Southend-on-Sea, UK).
Cov qauv tawv nqaij tau kho rau hauv 2.5% glutaraldehyde thiab 4% formaldehyde hauv 0.1 M HEPES (pH 7.4) rau 24 teev ntawm 4 ° C. Cov qauv tau qhuav dej siv cov ethanol qib thiab ziab hauv CO2 siv lub tshuab ziab Quorum K850 (Quorum Technologies Ltd, Loughton, UK) thiab sputter coated nrog kub-palladium alloy siv lub tshuab Quorum SC7620 mini sputterer/glow discharge system. Cov qauv tau thaij duab siv lub tshuab FEI Quanta 250 scanning electron microscope (ThermoFisher Scientific) kom pom qhov chaw nruab nrab ntawm qhov txhab.
Toto-1 iodide (2 μM) tau siv rau ntawm qhov txhab nas uas raug txiav tawm thiab muab tso rau 5 feeb ntawm 37 ° C (ThermoFisher Scientific) thiab kho nrog Syto-60 (10 μM) ntawm 37 ° C (ThermoFisher Scientific). Cov duab Z-stack 15-feeb tau tsim los ntawm kev siv Leica TCS SP8.
Cov ntaub ntawv rov ua dua ntawm cov kab mob thiab kev siv tshuab tau muab tso ua pawg thiab tshuaj xyuas siv Graphpad Prism V9 software (GraphPad Software, La Jolla, CA). Kev tshuaj xyuas ib txoj kev ntawm qhov sib txawv nrog ntau qhov kev sib piv siv Dunnett qhov kev xeem tom qab hoc tau siv los sim qhov sib txawv ntawm txhua txoj kev kho mob thiab kev hnav khaub ncaws tswj tsis yog tshuaj tua kab mob. Tus nqi p <0.05 tau suav tias yog qhov tseem ceeb.
Qhov ua tau zoo ntawm cov tshuaj pleev xim nyiaj gel fibrous tau raug soj ntsuam thawj zaug tawm tsam cov kab mob biofilm ntawm Staphylococcus aureus thiab Pseudomonas aeruginosa hauv vitro. Cov tshuaj pleev xim nyiaj muaj cov mis sib txawv ntawm cov nyiaj: cov tshuaj pleev xim nyiaj ib txwm tsim cov ions Ag1+; cov tshuaj pleev xim nyiaj, uas tuaj yeem tsim cov ions Ag1+ tom qab ntxiv EDTA/BC, tuaj yeem rhuav tshem cov biofilm matrix thiab ua rau cov kab mob raug nyiaj nyob rau hauv cov nyhuv antibacterial ntawm cov nyiaj. ions15 thiab cov tshuaj pleev xim uas muaj cov ntsev Ag oxygenated uas tsim cov ions Ag1+, Ag2+ thiab Ag3+. Nws qhov ua tau zoo tau piv nrog cov tshuaj pleev xim tsis yog antimicrobial tswj uas ua los ntawm cov fibers gelled. Cov kab mob uas tseem muaj sia nyob hauv biofilm tau raug soj ntsuam txhua 24 teev rau 8 hnub (Daim Duab 1). Nyob rau hnub 5, cov biofilm tau rov ua dua nrog 3.85 × 105S. Staphylococcus aureus lossis 1.22 × 105P. aeruginosa los soj ntsuam kev rov qab biofilm. Piv rau cov tshuaj uas tsis yog tshuaj tua kab mob, cov tshuaj Ag1+ muaj qhov cuam tshuam tsawg kawg nkaus rau kev muaj sia nyob ntawm cov kab mob hauv Staphylococcus aureus thiab Pseudomonas aeruginosa biofilms dhau 5 hnub. Qhov sib txawv, cov tshuaj uas muaj cov ntsev Ag thiab Ag1+ + EDTA/BC uas muaj oxygenated tau ua haujlwm zoo hauv kev tua cov kab mob hauv biofilm hauv 5 hnub. Tom qab rov ua dua cov kab mob planktonic rau hnub 5, tsis pom muaj kev kho dua tshiab ntawm biofilm (Daim Duab 1).
Kev ntsuas tus nqi ntawm cov kab mob uas muaj sia nyob hauv Staphylococcus aureus thiab Pseudomonas aeruginosa biofilms tom qab kho nrog cov nyiaj dressings. Biofilm pawg ntawm Staphylococcus aureus thiab Pseudomonas aeruginosa tau kho nrog cov nyiaj dressings lossis cov tshuaj tiv thaiv tsis yog antimicrobial, thiab tus naj npawb ntawm cov kab mob uas muaj sia nyob tau txiav txim siab txhua 24 teev. Tom qab 5 hnub, cov biofilm tau rov ua dua nrog 3.85 × 105S. Staphylococcus aureus lossis 1.22 × 105P. Cov pawg ntawm bacterioplankton Pseudomonas aeruginosa tau tsim ib tus zuj zus los ntsuas kev rov qab biofilm. Cov duab qhia qhia qhov nruab nrab +/- tus qauv yuam kev.
Yuav kom pom tau qhov cuam tshuam ntawm cov nyiaj hnav khaub ncaws rau biofilm viability, cov khaub ncaws tau siv rau cov biofilms laus uas cog rau ntawm daim tawv nqaij npua ex vivo. Tom qab 24 teev, cov khaub ncaws raug tshem tawm thiab cov biofilm raug pleev xim nrog cov xim xiav reactive, uas cov kab mob uas muaj sia nyob metabolized mus rau xim liab. Cov biofilms kho nrog cov khaub ncaws tswj yog xim liab, qhia tias muaj cov kab mob nyob hauv biofilm (Daim Duab 2A). Qhov sib txawv, cov biofilm kho nrog Ag oxysols dressing feem ntau yog xiav, qhia tias cov kab mob seem ntawm qhov chaw ntawm daim tawv nqaij npua yog cov kab mob tsis muaj sia nyob (Daim Duab 2B). Cov xim xiav thiab liab sib xyaw tau pom hauv cov biofilms kho nrog Ag1 +-muaj cov khaub ncaws, qhia tias muaj cov kab mob muaj sia nyob thiab tsis muaj sia nyob hauv biofilm (Daim Duab 2C), thaum EDTA/BC hnav khaub ncaws uas muaj Ag1 + feem ntau yog xiav nrog qee qhov chaw liab. qhia tias thaj chaw tsis cuam tshuam los ntawm cov nyiaj hnav khaub ncaws (Daim Duab 2D). Kev ntsuas ntawm cov chaw nquag (xim liab) thiab tsis nquag (xiav) qhia tau hais tias thaj chaw tswj yog 75% nquag (Daim Duab 2E). Cov ntaub qhwv Ag1+ + EDTA/BC ua haujlwm zoo ib yam li cov ntaub qhwv ntsev Ag uas muaj oxygenated, nrog rau cov feem pua ntawm kev ciaj sia ntawm 13% thiab 14%, raws li. Cov ntaub qhwv Ag1+ kuj txo qhov muaj sia nyob ntawm cov kab mob los ntawm 21%. Cov biofilms no tau pom los ntawm kev siv scanning electron microscopy (SEM). Tom qab kho nrog cov ntaub qhwv tswj thiab cov ntaub qhwv Ag1+, ib txheej ntawm Pseudomonas aeruginosa tau pom npog daim tawv nqaij npua (Daim Duab 2F, H), thaum tom qab kho nrog cov ntaub qhwv Ag1+, ob peb lub hlwb kab mob tau pom thiab ob peb lub hlwb kab mob tau pom hauv qab. Cov fibers collagen tuaj yeem suav hais tias yog cov qauv ntaub so ntswg ntawm daim tawv nqaij npua (Daim Duab 2G). Tom qab kho nrog cov ntaub qhwv Ag1+ + EDTA/BC, cov kab mob plaques thiab cov plaques fiber hauv qab tau pom (Daim Duab 2I).
Kev Pom Tau Pseudomonas aeruginosa biofilm tom qab kev kho mob hnav nyiaj. (A-D) Kev muaj sia nyob ntawm cov kab mob hauv Pseudomonas aeruginosa biofilms uas loj hlob ntawm daim tawv nqaij npua tau pom tau siv PrestoBlue viability dye 24 teev tom qab kev kho mob nrog cov khaub ncaws nyiaj lossis cov khaub ncaws tsis tswj kab mob. Cov kab mob uas muaj sia nyob yog xim liab, cov kab mob uas tsis muaj sia nyob thiab daim tawv nqaij npua yog xim xiav. (E) Kev ntsuas ntawm Pseudomonas aeruginosa biofilms uas loj hlob ntawm daim tawv nqaij npua (qhov chaw liab) siv scanning electron microscopy Image Pro version 10 (FI) thiab kho nrog cov khaub ncaws nyiaj lossis cov khaub ncaws tsis tswj kab mob rau 24 teev. SEM scale bar = 5 µm. (J-M) Colonial biofilms loj hlob ntawm cov lim dej thiab tau pleev xim nrog PrestoBlue reactive dye tom qab 24 teev ntawm kev sib xyaw nrog cov khaub ncaws nyiaj.
Txhawm rau txiav txim siab seb qhov kev sib cuag ze ntawm cov ntaub qhwv thiab cov biofilms puas cuam tshuam rau qhov ua tau zoo ntawm cov ntaub qhwv, cov biofilms colonial uas muab tso rau ntawm qhov chaw tiaj tiaj tau kho nrog cov ntaub qhwv rau 24 teev thiab tom qab ntawd pleev xim nrog cov xim reactive. Cov biofilm tsis kho yog xim liab tsaus (Daim Duab 2J). Sib piv rau cov biofilms kho nrog cov ntaub qhwv uas muaj cov ntsev Ag oxygenated (Daim Duab 2K), cov biofilms kho nrog cov ntaub qhwv uas muaj Ag1+ lossis Ag1+ + EDTA/BC qhia txog cov xim liab (Daim Duab 2L, M). Cov xim liab no qhia txog qhov muaj cov kab mob uas muaj sia nyob thiab cuam tshuam nrog thaj chaw suture hauv cov ntaub qhwv. Cov chaw xaws no tsim cov chaw tuag uas tso cai rau cov kab mob hauv biofilm kom muaj sia nyob.
Txhawm rau ntsuam xyuas qhov ua tau zoo ntawm cov ntaub qhwv nyiaj hauv vivo, cov qhov txhab uas raug txiav tawm tag nrho ntawm cov nas uas muaj tus kab mob S. aureus thiab P. aeruginosa biofilms uas laus lawm tau kho nrog cov ntaub qhwv uas tsis yog tshuaj tua kab mob lossis cov ntaub qhwv nyiaj. Tom qab 3 hnub ntawm kev kho mob, kev tshuaj xyuas duab macroscopic qhia tau tias qhov txhab me dua thaum kho nrog cov ntaub qhwv ntsev oxygenated piv rau cov ntaub qhwv uas tsis yog tshuaj tua kab mob thiab lwm cov ntaub qhwv nyiaj (Daim Duab 3A-H). Txhawm rau kom paub meej cov kev soj ntsuam no, cov qhov txhab tau sau thiab thaj chaw qhov txhab thiab reepithelialization tau ntsuas ntawm cov ntaub so ntswg hematoxylin thiab eosin siv image pro software version 10 (Daim Duab 3I-L).
Cov nyhuv ntawm cov nyiaj hnav khaub ncaws rau ntawm qhov txhab thiab rov ua dua cov qhov txhab uas muaj cov biofilms. (A-H) Cov hlwb me me uas muaj cov biofilms ntawm Pseudomonas aeruginosa (A-D) thiab Staphylococcus aureus (E-H) tom qab peb hnub ntawm kev kho mob nrog cov khaub ncaws tswj tsis yog antimicrobial, cov khaub ncaws oxygenated Ag ntsev, cov khaub ncaws Ag1+, thiab cov khaub ncaws Ag1+. Cov duab macroscopic sawv cev. cov qhov txhab ntawm nas nrog Ag1+ + EDTA/BC hnav khaub ncaws. (IL) Tus sawv cev Pseudomonas aeruginosa tus kab mob, cov ntu histological stained nrog hematoxylin thiab eosin, siv los ntsuas qhov chaw txhab thiab epithelial regeneration. Kev ntsuas qhov chaw txhab (M, O) thiab feem pua ntawm reepithelialization (N, P) ntawm cov qhov txhab uas muaj cov Pseudomonas aeruginosa (M, N) thiab Staphylococcus aureus (O, P) biofilms (ib pawg kho mob n = 12). Cov duab qhia qhov nruab nrab +/- qhov yuam kev txheem. * txhais tau tias p = < 0.05 ** txhais tau tias p = < 0.01; Macroscopic scale = 2.5 mm, histological scale = 500 µm.
Kev ntsuas qhov chaw ntawm qhov txhab hauv cov qhov txhab uas muaj kab mob Pseudomonas aeruginosa biofilm (Daim Duab 3M) qhia tau tias cov qhov txhab kho nrog Ag oxysalts muaj qhov nruab nrab qhov txhab loj ntawm 2.5 mm2, thaum cov khaub ncaws tswj tsis yog tshuaj tua kab mob muaj qhov nruab nrab qhov txhab loj ntawm 3.1 mm2, uas tsis yog qhov tseeb. tau mus txog qhov tseem ceeb ntawm kev suav lej (Daim Duab 3M). p = 0.423). Cov qhov txhab kho nrog Ag1+ lossis Ag1+ + EDTA/BC tsis qhia txog kev txo qis hauv thaj chaw qhov txhab (3.1 mm2 thiab 3.6 mm2, feem). Kev kho nrog oxygenated Ag ntsev dressing txhawb kev rov ua epithelialization ntau dua li cov khaub ncaws tswj tsis yog tshuaj tua kab mob (34% thiab 15%, feem; p = 0.029) thiab Ag1+ lossis Ag1+ + EDTA/BC (10% thiab 11%) (Daim Duab 3N). . , feem).
Cov qauv zoo sib xws hauv thaj chaw qhov txhab thiab kev rov ua dua tshiab ntawm cov epithelial tau pom hauv cov qhov txhab uas muaj kab mob S. aureus biofilms (Daim Duab 3O). Cov ntaub qhwv uas muaj cov ntsev nyiaj oxygenated txo thaj chaw qhov txhab (2.0 mm2) los ntawm 23% piv rau cov ntaub qhwv uas tsis yog tshuaj tua kab mob (2.6 mm2), txawm hais tias qhov kev txo qis no tsis tseem ceeb (p = 0.304) (Daim Duab 3O). Tsis tas li ntawd, thaj chaw qhov txhab hauv pawg kho Ag1+ tau txo me ntsis (2.4 mm2), thaum qhov txhab kho nrog Ag1+ + EDTA/BC ntaub qhwv tsis txo thaj chaw qhov txhab (2.9 mm2). Cov ntsev oxygen ntawm Ag kuj txhawb kev rov ua dua tshiab ntawm cov qhov txhab uas muaj kab mob S. aureus biofilm (31%) ntau dua li cov uas kho nrog cov ntaub qhwv uas tsis yog tshuaj tua kab mob (12%, p = 0.003) (Daim Duab 3P). Kev hnav khaub ncaws Ag1+ (16%, p = 0.903) thiab kev hnav khaub ncaws Ag+1 + EDTA/BC (14%, p = 0.965) qhia tau tias cov epithelial regeneration zoo ib yam li kev tswj hwm.
Yuav kom pom tau qhov cuam tshuam ntawm cov nyiaj hnav khaub ncaws rau ntawm biofilm matrix, Toto 1 thiab Syto 60 iodide staining tau ua tiav (Daim Duab 4). Toto 1 iodide yog cov xim zas uas tsis nkag tau rau hauv lub cell uas siv tau los pom qhov tseeb ntawm cov nucleic acids sab nraud, uas muaj ntau nyob rau hauv EPS ntawm biofilms. Syto 60 yog cov xim zas uas nkag tau rau hauv lub cell siv ua cov xim tiv thaiv16. Kev soj ntsuam ntawm Toto 1 thiab Syto 60 iodide hauv cov qhov txhab uas muaj biofilms ntawm Pseudomonas aeruginosa (Daim Duab 4A-D) thiab Staphylococcus aureus (Daim Duab 4I-L) qhia tau tias tom qab 3 hnub ntawm kev kho mob hnav khaub ncaws, EPS hauv biofilm tau txo qis heev. Muaj cov ntsev oxygenated Ag thiab Ag1+ + EDTA/BC. Ag1+ hnav khaub ncaws uas tsis muaj cov khoom siv antibiofilm ntxiv tau txo qis cov DNA tsis muaj cell hauv cov qhov txhab uas muaj Pseudomonas aeruginosa tab sis tsis muaj txiaj ntsig zoo hauv cov qhov txhab uas muaj Staphylococcus aureus.
Kev thaij duab hauv vivo ntawm cov qhov txhab biofilm tom qab 3 hnub ntawm kev kho mob nrog kev tswj hwm lossis cov khaub ncaws nyiaj. Cov duab Confocal ntawm Pseudomonas aeruginosa (A-D) thiab Staphylococcus aureus (I-L) pleev xim nrog Toto 1 (ntsuab) kom pom cov extracellular nucleic acids, ib feem ntawm extracellular biofilm polymers. Txhawm rau pleev xim rau intracellular nucleic acids, siv Syto 60 (liab). acids. P. Scanning electron microscopy ntawm cov qhov txhab uas muaj kab mob Pseudomonas aeruginosa (E-H) thiab Staphylococcus aureus (M-P) biofilms tom qab 3 hnub ntawm kev kho mob nrog kev tswj hwm thiab cov khaub ncaws nyiaj. SEM scale bar = 5 µm. Confocal imaging scale bar = 50 µm.
Kev siv tshuab hluav taws xob scanning qhia tau hais tias cov nas uas tau txhaj tshuaj nrog cov kab mob Pseudomonas aeruginosa (Daim Duab 4E-H) thiab Staphylococcus aureus (Daim Duab 4M-P) muaj cov kab mob tsawg dua hauv lawv cov qhov txhab tom qab 3 hnub ntawm kev kho mob nrog txhua yam nyiaj dressings.
Txhawm rau soj ntsuam qhov cuam tshuam ntawm cov nyiaj hnav khaub ncaws rau qhov txhab o hauv cov nas uas muaj kab mob biofilm, cov ntu ntawm cov qhov txhab uas muaj kab mob biofilm kho nrog kev tswj hwm lossis cov nyiaj hnav khaub ncaws rau 3 hnub tau raug immunohistochemically stained siv cov tshuaj tiv thaiv tshwj xeeb rau neutrophils thiab macrophages. Kev txiav txim siab ntau ntawm neutrophils thiab macrophages sab hauv. cov ntaub so ntswg granulation. Daim duab 5). Txhua cov nyiaj hnav khaub ncaws txo cov lej ntawm neutrophils thiab macrophages hauv cov qhov txhab uas muaj kab mob Pseudomonas aeruginosa piv rau cov tsis yog tshuaj tua kab mob tswj tom qab peb hnub ntawm kev kho mob. Txawm li cas los xij, kev kho mob nrog cov nyiaj hnav khaub ncaws oxygenated ua rau muaj kev txo qis ntau dua hauv neutrophils (p = <0.0001) thiab macrophages (p = <0.0001) piv rau lwm cov nyiaj hnav khaub ncaws sim (Daim duab 5I, J). Txawm hais tias Ag1+ + EDTA/BC muaj cov nyhuv ntau dua rau qhov txhab biofilm, nws txo cov neutrophil thiab macrophage theem kom tsawg dua li Ag1+ hnav khaub ncaws. Cov qhov txhab nruab nrab uas muaj kab mob S. aureus biofilm kuj tau pom tom qab hnav khaub ncaws nrog Ag (p = <0.0001), Ag1+ (p = 0.0008) thiab Ag1++ EDTA/BC (p = 0.0043) oxisols piv rau kev tswj hwm. Cov qauv zoo sib xws tau pom rau neutropenia. bandage (Daim Duab 5K). Txawm li cas los xij, tsuas yog cov tshuaj oxygenated Ag ntsev hnav khaub ncaws qhia txog kev txo qis ntawm cov macrophages hauv cov ntaub so ntswg granulation piv rau kev tswj hwm hauv cov qhov txhab uas muaj kab mob S. aureus biofilms (p = 0.0339) (Daim Duab 5L).
Cov neutrophils thiab macrophages tau ntsuas hauv cov qhov txhab uas muaj kab mob Pseudomonas aeruginosa thiab Staphylococcus aureus biofilms tom qab 3 hnub ntawm kev kho mob nrog kev tswj tsis yog tshuaj tua kab mob lossis cov khaub ncaws nyiaj. Neutrophils (AD) thiab macrophages (EH) tau ntsuas hauv cov nqaij ntu uas muaj cov tshuaj tiv thaiv tshwj xeeb rau neutrophils lossis macrophages. Kev ntsuas ntawm neutrophils (I thiab K) thiab macrophages (J thiab L) hauv cov qhov txhab uas muaj kab mob Pseudomonas aeruginosa (I thiab J) thiab Staphylococcus aureus (K & L) biofilms. N = 12 ib pawg. Cov duab qhia qhov nruab nrab +/- qhov yuam kev tus qauv, qhov tseem ceeb piv rau kev hnav khaub ncaws tsis tswj kab mob, * txhais tau tias p = <0.05, ** txhais tau tias p = <0.01; *** txhais tau tias p = <0.001; qhia p = <0.0001).
Tom qab ntawd peb tau soj ntsuam qhov cuam tshuam ntawm cov ntaub qhwv nyiaj rau kev kho mob uas tsis muaj kev kis kab mob. Cov qhov txhab uas tsis muaj kev kis kab mob tau kho nrog cov ntaub qhwv uas tsis yog tshuaj tua kab mob lossis cov ntaub qhwv nyiaj rau 3 hnub (Daim Duab 6). Ntawm cov ntaub qhwv nyiaj uas tau sim, tsuas yog cov qhov txhab uas kho nrog cov ntaub qhwv ntsev oxygenated zoo li me dua ntawm cov duab macroscopic dua li cov qhov txhab uas kho nrog cov tswj (Daim Duab 6A-D). Kev ntsuas qhov chaw ntawm qhov txhab siv kev tshuaj xyuas histological qhia tau hais tias qhov nruab nrab ntawm qhov txhab tom qab kho nrog Ag oxysols dressing yog 2.35 mm2 piv rau 2.96 mm2 rau cov qhov txhab uas kho nrog pawg tswj, tab sis qhov sib txawv no tsis tau mus txog qhov tseem ceeb ntawm kev suav lej (p = 0.488) (Daim Duab 6I). Qhov sib piv, tsis muaj kev txo qis ntawm thaj chaw qhov txhab tau pom tom qab kho nrog Ag1+ (3.38 mm2, p = 0.757) lossis Ag1+ + EDTA/BC (4.18 mm2, p = 0.054) cov ntaub qhwv piv rau pawg tswj. Kev tsim kho dua tshiab ntawm epithelial tau pom nrog Ag oxysol dressing piv rau pawg tswj (30% vs. 22%, feem), txawm hais tias qhov no tsis tau mus txog qhov tseem ceeb (p = 0.067), qhov no yog qhov tseem ceeb heev thiab lees paub cov txiaj ntsig yav dhau los. Kev hnav khaub ncaws nrog oxysols txhawb nqa kev rov ua dua epithelialization. -Epithelization ntawm cov qhov txhab tsis muaj kab mob17. Hauv kev sib piv, kev kho mob nrog Ag1 + lossis Ag1 + EDTA / BC dressings tsis muaj txiaj ntsig lossis qhia tias txo qis re-epithelialization piv rau pawg tswj.
Cov nyhuv ntawm cov nyiaj qhwv qhov txhab rau kev kho qhov txhab hauv cov nas tsis muaj kab mob nrog kev txiav tawm tag nrho. (AD) Cov duab macroscopic sawv cev ntawm cov qhov txhab tom qab peb hnub ntawm kev kho mob nrog cov tshuaj tiv thaiv tsis yog tshuaj tua kab mob thiab cov tshuaj nyiaj qhwv. (EH) Cov seem ntawm qhov txhab uas muaj xim nrog hematoxylin thiab eosin. Kev ntsuas qhov chaw ntawm qhov txhab (I) thiab feem pua ntawm reepithelialization (J) tau suav los ntawm cov seem histological ntawm qhov nruab nrab ntawm qhov txhab siv cov software tshuaj xyuas duab (n = 11–12 ib pawg kho mob). Cov duab qhia qhov nruab nrab +/- qhov yuam kev txheem. * txhais tau tias p = <0.05.
Nyiaj muaj keeb kwm ntev ntawm kev siv ua tshuaj tua kab mob hauv kev kho qhov txhab, tab sis ntau cov qauv sib txawv thiab cov txheej txheem xa khoom yuav ua rau muaj qhov sib txawv ntawm kev ua haujlwm tiv thaiv kab mob 18. Ntxiv mus, cov khoom siv antibiofilm ntawm cov txheej txheem xa khoom nyiaj tshwj xeeb tsis tau nkag siab tag nrho. Txawm hais tias tus tswv tsev lub cev tiv thaiv kab mob zoo tiv thaiv cov kab mob planktonic, nws feem ntau tsis zoo tiv thaiv biofilms19. Cov kab mob planktonic yooj yim phagocytosed los ntawm macrophages, tab sis hauv biofilms, cov hlwb sib sau ua ke ua rau muaj teeb meem ntxiv los ntawm kev txwv tus tswv tsev teb rau qhov uas cov hlwb tiv thaiv kab mob tuaj yeem dhau los ua apoptosis thiab tso tawm cov yam ntxwv proinflammatory los txhim kho kev tiv thaiv kab mob20. Nws tau pom tias qee cov leukocytes tuaj yeem nkag mus rau biofilms21 tab sis tsis tuaj yeem phagocytose cov kab mob thaum qhov kev tiv thaiv no raug cuam tshuam22. Yuav tsum siv txoj hauv kev holistic los txhawb tus tswv tsev lub cev tiv thaiv kab mob tiv thaiv kab mob biofilm qhov txhab. Kev tshem tawm qhov txhab tuaj yeem cuam tshuam lub cev biofilm thiab tshem tawm feem ntau ntawm bioburden, tab sis tus tswv tsev lub cev tiv thaiv kab mob yuav tsis ua haujlwm tiv thaiv biofilm seem, tshwj xeeb tshaj yog tias tus tswv tsev lub cev tiv thaiv kab mob raug cuam tshuam. Yog li ntawd, cov tshuaj tua kab mob xws li cov ntaub qhwv nyiaj tuaj yeem txhawb nqa lub cev tiv thaiv kab mob thiab tshem tawm cov kab mob biofilm. Kev sib xyaw, kev sib sau ua ke, kev yaj, thiab cov khoom xa tuaj yeem cuam tshuam rau qhov ua tau zoo ntawm cov nyiaj tua kab mob. Nyob rau hauv xyoo tas los no, kev nce qib hauv kev siv tshuab ua cov nyiaj tau ua rau cov ntaub qhwv no muaj txiaj ntsig zoo dua9,23. Raws li cov thev naus laus zis hnav nyiaj tau nce qib, nws yog ib qho tseem ceeb kom nkag siab txog qhov ua tau zoo ntawm cov ntaub qhwv no hauv kev tswj cov kab mob qhov txhab thiab, qhov tseem ceeb dua, qhov cuam tshuam ntawm cov hom nyiaj muaj zog no rau qhov chaw mob thiab kev kho mob.
Hauv kev tshawb fawb no, peb tau piv qhov ua tau zoo ntawm ob daim ntaub qhwv nyiaj siab heev nrog cov ntaub qhwv nyiaj ib txwm muaj uas tsim cov ions Ag1+ tawm tsam cov biofilms siv cov qauv sib txawv hauv vitro thiab hauv vivo. Peb kuj tau soj ntsuam qhov cuam tshuam ntawm cov ntaub qhwv no rau qhov chaw mob thiab kev kho mob tsis muaj kev kis kab mob. Txhawm rau kom txo qhov cuam tshuam ntawm cov khoom xa tuaj, txhua daim ntaub qhwv nyiaj uas tau sim yog tsim los ntawm carboxymethylcellulose.
Peb qhov kev ntsuam xyuas ua ntej ntawm cov khaub ncaws nyiaj no tawm tsam cov biofilms colonial ntawm Pseudomonas aeruginosa thiab Staphylococcus aureus qhia tau hais tias, tsis zoo li cov khaub ncaws Ag1+ ib txwm muaj, ob qho khaub ncaws nyiaj siab heev, Ag1+ + EDTA/BC thiab cov ntsev Ag oxygenated, muaj txiaj ntsig ntawm 5. Tua cov kab mob biofilm tau zoo hauv ob peb hnub. Tsis tas li ntawd, cov khaub ncaws no tiv thaiv kev rov tsim dua ntawm biofilm thaum raug cov kab mob planktonic ntau zaus. Cov khaub ncaws Ag1+ muaj cov tshuaj silver chloride, tib cov tshuaj silver thiab cov khoom siv matrix li Ag1+ + EDTA/BC, thiab muaj qhov cuam tshuam tsawg rau kev muaj sia nyob ntawm cov kab mob hauv biofilm dhau lub sijhawm tib yam. Kev soj ntsuam tias cov khaub ncaws Ag1+ + EDTA/BC muaj txiaj ntsig zoo dua tawm tsam biofilm dua li cov khaub ncaws Ag1+ uas muaj tib lub matrix thiab cov tshuaj silver txhawb nqa lub tswv yim tias cov khoom xyaw ntxiv yuav tsum tau ua kom muaj txiaj ntsig ntawm cov tshuaj silver chloride tawm tsam biofilm, raws li tau tshaj tawm lwm qhov15. Cov txiaj ntsig no txhawb nqa lub tswv yim tias BC thiab EDTA ua lub luag haujlwm ntxiv uas pab txhawb rau kev hnav khaub ncaws zoo thiab tias qhov tsis muaj cov khoom no hauv Ag1+ cov khaub ncaws tej zaum yuav ua rau tsis pom kev ua haujlwm hauv vitro. Peb pom tias cov khaub ncaws oxygenated Ag ntsev tsim cov Ag2+ thiab Ag3+ ions tau pom tias muaj zog tiv thaiv kab mob ntau dua li Ag1+ thiab ntawm cov qib zoo ib yam li Ag1+ + EDTA/BC. Txawm li cas los xij, vim muaj peev xwm redox siab, nws tsis meej tias Ag3+ ions tseem ua haujlwm ntev npaum li cas thiab ua haujlwm tau zoo tiv thaiv cov biofilms qhov txhab thiab yog li ntawd tsim nyog kawm ntxiv. Tsis tas li ntawd, muaj ntau yam khaub ncaws sib txawv uas tsim cov Ag1+ ions uas tsis tau sim hauv kev tshawb fawb no. Cov khaub ncaws no yog tsim los ntawm cov khoom sib xyaw nyiaj sib txawv, cov concentration, thiab cov matrices hauv paus, uas yuav cuam tshuam rau kev xa cov Ag1+ ions thiab lawv cov haujlwm tiv thaiv biofilms. Nws kuj tsim nyog sau tseg tias muaj ntau yam qauv sib txawv hauv vitro thiab hauv vivo siv los ntsuas qhov ua tau zoo ntawm cov khaub ncaws qhov txhab tiv thaiv biofilms. Hom qauv siv, nrog rau cov ntsev thiab cov protein ntawm cov xov xwm siv hauv cov qauv no, yuav cuam tshuam rau qhov ua tau zoo ntawm cov khaub ncaws. Hauv peb tus qauv in vivo, peb tau tso cai rau cov biofilm kom loj hlob hauv vitro thiab tom qab ntawd hloov mus rau qhov chaw ntawm qhov txhab. Cov tshuaj tiv thaiv kab mob ntawm tus nas tswj hwm muaj txiaj ntsig zoo rau cov kab mob planktonic uas siv rau qhov txhab, yog li tsim cov biofilm thaum qhov txhab kho. Kev ntxiv cov biofilm laus rau qhov txhab txwv tsis pub cov tshuaj tiv thaiv kab mob ntawm tus tswv tsev rau kev tsim cov biofilm los ntawm kev tso cai rau cov biofilm laus kom tsim nws tus kheej hauv qhov txhab ua ntej kho tau. Yog li, peb tus qauv tso cai rau peb los soj ntsuam qhov ua tau zoo ntawm cov tshuaj tua kab mob ntawm cov biofilm laus ua ntej qhov txhab pib kho.
Peb kuj pom tias qhov hnav khaub ncaws haum rau qhov ua tau zoo ntawm cov nyiaj hnav khaub ncaws rau ntawm cov biofilms uas cog hauv vitro thiab daim tawv nqaij npua. Kev sib cuag ze nrog qhov txhab yog qhov tseem ceeb rau qhov ua tau zoo ntawm cov tshuaj tua kab mob ntawm cov khaub ncaws24,25. Cov khaub ncaws uas muaj cov ntsev Ag oxygenated nyob ze nrog cov biofilms laus, ua rau muaj kev txo qis ntawm cov kab mob uas muaj sia nyob hauv biofilm tom qab 24 teev. Qhov sib txawv, thaum kho nrog Ag1 + thiab Ag1 + + EDTA / BC hnav khaub ncaws, muaj ntau cov kab mob uas muaj sia nyob tseem tshuav. Cov khaub ncaws no muaj cov xov txuas raws tag nrho qhov ntev ntawm cov khaub ncaws, uas tsim cov chaw tuag uas tiv thaiv kev sib cuag ze nrog biofilm. Hauv peb cov kev tshawb fawb hauv vitro, cov chaw tsis sib cuag no tiv thaiv kev tua cov kab mob uas muaj sia nyob hauv biofilm. Peb tau soj ntsuam cov kab mob uas muaj sia nyob tsuas yog tom qab 24 teev ntawm kev kho mob; Raws li lub sijhawm, thaum cov khaub ncaws ua rau ntau dua, tej zaum yuav muaj qhov chaw tuag tsawg dua, txo qhov chaw rau cov kab mob uas muaj sia nyob no. Txawm li cas los xij, qhov no qhia txog qhov tseem ceeb ntawm cov khoom sib xyaw ntawm cov khaub ncaws, tsis yog tsuas yog hom nyiaj hauv cov khaub ncaws.
Txawm hais tias kev tshawb fawb hauv vitro muaj txiaj ntsig zoo rau kev sib piv cov txiaj ntsig ntawm cov thev naus laus zis nyiaj sib txawv, nws tseem ceeb heev kom nkag siab txog cov teebmeem ntawm cov ntaub qhwv no rau ntawm biofilms hauv vivo, qhov twg cov ntaub so ntswg thiab cov lus teb ntawm lub cev tiv thaiv kab mob pab txhawb rau qhov ua tau zoo ntawm cov ntaub qhwv tiv thaiv biofilms. Cov teebmeem ntawm cov ntaub qhwv no rau ntawm cov biofilms qhov txhab tau pom los ntawm kev siv scanning electron microscopy thiab EPS staining ntawm biofilm siv cov xim DNA hauv thiab sab nraud. Peb pom tias tom qab 3 hnub ntawm kev kho mob, txhua qhov kev hnav khaub ncaws tau ua haujlwm zoo hauv kev txo cov DNA tsis muaj cell hauv cov qhov txhab uas muaj kab mob biofilm, tab sis cov ntaub qhwv Ag1+ tsis muaj txiaj ntsig zoo hauv cov qhov txhab uas muaj kab mob Staphylococcus aureus. Scanning electron microscopy kuj qhia tau tias muaj cov kab mob tsawg dua nyob rau hauv cov qhov txhab kho nrog cov ntaub qhwv nyiaj, txawm hais tias qhov no tau pom tseeb dua nrog cov ntaub qhwv oxygenated Ag thiab cov ntaub qhwv Ag1+ + EDTA/BC piv rau cov ntaub qhwv Ag1+. Cov ntaub ntawv no qhia tau tias cov ntaub qhwv nyiaj uas tau sim muaj ntau yam cuam tshuam rau cov qauv ntawm cov biofilm, tab sis tsis muaj ib qho ntaub qhwv nyiaj twg uas tuaj yeem tshem tawm cov biofilm, txhawb nqa qhov xav tau kev kho mob tag nrho rau kev kis kab mob ntawm cov qhov txhab biofilm; siv cov hlua khi tes nyiaj. Kev kho mob yog ua ntej los ntawm kev tshem tawm cov biofilm kom tshem tawm feem ntau ntawm cov biofilm.
Cov qhov txhab uas mob ntev feem ntau nyob rau hauv qhov xwm txheej ntawm kev o hnyav, nrog rau cov hlwb o ntau dhau uas tseem nyob hauv cov nqaij ntawm qhov txhab rau lub sijhawm ntev, ua rau cov nqaij puas tsuaj thiab ua rau cov pa oxygen uas xav tau rau kev ua haujlwm ntawm cov hlwb thiab kev ua haujlwm hauv qhov txhab26. Cov biofilms ua rau qhov chaw mob no hnyav dua los ntawm kev cuam tshuam tsis zoo rau kev kho mob ntau yam, suav nrog kev txwv tsis pub cov hlwb loj hlob thiab kev tsiv teb tsaws chaw thiab kev ua haujlwm ntawm cov cytokines proinflammatory27. Thaum cov ntaub qhwv nyiaj ua haujlwm tau zoo dua, nws yog ib qho tseem ceeb kom nkag siab txog qhov cuam tshuam uas lawv muaj rau qhov chaw mob thiab kev kho mob.
Txawm hais tias txhua yam kev hnav khaub ncaws nyiaj cuam tshuam rau cov biofilm muaj pes tsawg leeg, tsuas yog cov kev hnav khaub ncaws ntsev oxygenated xwb thiaj ua rau cov qhov txhab uas muaj kab mob no rov qab zoo li qub. Cov ntaub ntawv no txhawb nqa peb cov kev tshawb pom yav dhau los17 thiab cov ntawm Kalan et al. (2017)28, uas tau qhia txog kev nyab xeeb thiab kev lom ntawm cov ntsev oxygenated, vim tias cov nyiaj tsawg dua tau zoo rau cov biofilms.
Peb txoj kev tshawb fawb tam sim no qhia txog qhov sib txawv ntawm cov thev naus laus zis nyiaj ntawm cov tshuaj pleev xim nyiaj tua kab mob thiab qhov cuam tshuam ntawm cov thev naus laus zis no rau qhov chaw mob thiab kev kho mob tsis muaj kev kis kab mob. Txawm li cas los xij, cov txiaj ntsig no txawv ntawm cov kev tshawb fawb yav dhau los uas qhia tias Ag1+ + EDTA/BC hnav khaub ncaws txhim kho cov kev kho mob ntawm cov pob ntseg luav raug mob hauv vivo. Txawm li cas los xij, qhov no yuav yog vim muaj qhov sib txawv ntawm cov qauv tsiaj, lub sijhawm ntsuas, thiab cov txheej txheem siv kab mob 29. Hauv qhov no, kev ntsuas qhov txhab tau coj 12 hnub tom qab raug mob kom tso cai rau cov khoom xyaw nquag ntawm cov khaub ncaws ua haujlwm rau ntawm biofilm dhau lub sijhawm ntev dua. Qhov no tau txhawb nqa los ntawm kev tshawb fawb uas qhia tau tias cov kab mob ceg uas tau kho nrog Ag1+ + EDTA/BC pib loj hlob tom qab ib lub lim tiam ntawm kev kho mob, thiab tom qab ntawd dhau 3 lub lim tiam tom ntej ntawm kev kho mob nrog Ag1+ + EDTA/BC thiab hauv 4 lub lim tiam ntawm kev siv cov tshuaj tsis yog tshuaj tua kab mob. CMC hnav khaub ncaws kom txo qhov loj ntawm cov kab mob 30.
Qee hom thiab cov concentration ntawm cov nyiaj tau pom tias yog cytotoxic hauv vitro 11, tab sis cov txiaj ntsig hauv vitro no tsis tas li txhais ua cov teebmeem tsis zoo hauv vivo. Tsis tas li ntawd, kev nce qib hauv kev siv tshuab nyiaj thiab kev nkag siab zoo dua ntawm cov khoom sib xyaw nyiaj thiab cov concentration hauv cov ntaub qhwv tau ua rau muaj kev tsim ntau cov ntaub qhwv nyiaj muaj kev nyab xeeb thiab zoo. Txawm li cas los xij, thaum cov thev naus laus zis hnav nyiaj nce qib, nws yog ib qho tseem ceeb kom nkag siab txog qhov cuam tshuam ntawm cov ntaub qhwv no rau qhov chaw mob31,32,33. Nws tau tshaj tawm ua ntej tias qhov nce ntxiv ntawm kev rov ua dua tshiab sib raug rau qhov nce ntxiv ntawm cov macrophages tiv thaiv kev o M2 piv rau cov phenotype pro-inflammatory M1. Qhov no tau sau tseg hauv tus qauv nas yav dhau los qhov twg cov ntaub qhwv nyiaj hydrogel raug piv nrog cov nyiaj sulfadiazine thiab cov hydrogels tsis-antimicrobial34.
Cov qhov txhab ntev ntev yuav ua rau muaj kev o thiab nws tau pom tias qhov muaj cov neutrophils ntau dhau yuav ua rau qhov txhab kho tsis zoo35. Hauv kev tshawb fawb hauv cov nas uas tsis muaj neutrophil, qhov muaj neutrophils qeeb reepithelialization. Qhov muaj cov neutrophils ntau dhau ua rau muaj cov proteases thiab cov tshuaj oxygen reactive, xws li superoxide thiab hydrogen peroxide, uas cuam tshuam nrog cov qhov txhab ntev ntev thiab kho qeeb37,38. Ib yam li ntawd, kev nce ntxiv ntawm cov lej macrophage, yog tias tsis tswj hwm, tuaj yeem ua rau qhov txhab kho qeeb39. Qhov kev nce no tseem ceeb heev yog tias macrophages tsis tuaj yeem hloov pauv los ntawm tus qauv pro-inflammatory mus rau tus qauv pro-healing, ua rau cov qhov txhab tsis tawm ntawm theem inflammatory ntawm kev kho40. Peb tau pom qhov txo qis ntawm neutrophils thiab macrophages hauv cov qhov txhab uas muaj kab mob biofilm tom qab 3 hnub ntawm kev kho mob nrog txhua cov nyiaj hnav khaub ncaws, tab sis qhov txo qis tau pom tseeb dua nrog cov ntsev oxygenated. Qhov kev txo qis no tej zaum yuav yog qhov tshwm sim ncaj qha ntawm kev tiv thaiv kab mob rau nyiaj, kev teb rau kev txo qis bioburden, lossis qhov txhab nyob rau theem tom qab ntawm kev kho thiab yog li ntawd cov hlwb tiv thaiv kab mob hauv qhov txhab raug txo qis. Kev txo cov lej ntawm cov hlwb o hauv qhov txhab yuav tswj tau ib puag ncig uas zoo rau kev kho qhov txhab. Lub tshuab ua haujlwm ntawm Ag oxysalt txhawb kev kho mob tsis muaj kev kis kab mob tsis meej, tab sis lub peev xwm ntawm Ag oxysalts los tsim cov pa oxygen thiab rhuav tshem cov qib hydrogen peroxide uas ua rau mob, tus neeg sawv cev ntawm kev o, yuav piav qhia qhov no thiab xav tau kev kawm ntxiv17.
Cov qhov txhab uas kis kab mob mus ntev thiab tsis kho tau zoo ua rau muaj teeb meem rau cov kws kho mob thiab cov neeg mob. Txawm hais tias muaj ntau daim ntaub qhwv hais tias muaj txiaj ntsig zoo rau kev tua kab mob, kev tshawb fawb tsis tshua tsom mus rau lwm yam tseem ceeb uas cuam tshuam rau qhov chaw ib puag ncig ntawm qhov txhab. Kev tshawb fawb no qhia tau hais tias cov thev naus laus zis nyiaj sib txawv muaj txiaj ntsig zoo rau kev tua kab mob thiab, qhov tseem ceeb, muaj cov teebmeem sib txawv rau qhov chaw ib puag ncig thiab kev kho mob, tsis hais txog kev kis kab mob. Txawm hais tias cov kev tshawb fawb hauv vitro thiab hauv vivo no qhia tau tias cov ntaub qhwv no zoo rau kev kho cov kab mob qhov txhab thiab txhawb kev kho mob, kev sim tshuaj ntsuam xyuas yog qhov xav tau los ntsuas qhov ua tau zoo ntawm cov ntaub qhwv no hauv chaw kho mob.
Cov ntaub ntawv siv thiab/lossis tshuaj xyuas thaum lub sijhawm kawm tam sim no muaj los ntawm tus sau ntawv sib xws thaum thov.
Lub sijhawm tshaj tawm: Lub Xya Hli-15-2024