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Ukukhula kwama-microbial emanxebeni kuvame ukuzibonakalisa njenge-biofilms, okuphazamisa ukuphulukiswa futhi kunzima ukuqeda. Izingubo ezintsha zesiliva zithi zilwa nokulwa nezifo zezilonda, kepha imiphumela yazo ye-antibiofilm kanye nokutheleleka okuzimele ngokuvamile akwaziwa. Usebenzisa amamodeli e-vitro naseVivo biofilm amamodeli we-staphylococcus aureus kanye ne-pseudomonas aeruginosa, sibika ukusebenza ngempumelelo kokugqoka kwe-AG1 + ion-adalula ukugqoka; I-AG1 + Izingubo eziqukethe i-ethyleniaminettetteteTefy acid ne-benzethonium chloride (AG1 + EDTA / BC), nokugqoka okuqukethe i-nitrate yesiliva (ag oxysalts). , ekhiqiza i-AG1 +, i-AG2 + ne-AG3 + ama-Ion ukulwa ne-biofilm nomphumela wayo ekuphulukisweni. Izingubo zokugqoka ze-AG1 + zazinemiphumela emincane kwi-Biofilm eVitro naseMice (C57BL / 6J). Ngokuphambene nalokho, usawoti we-ag oxygenated kanye ne-ag1 + / edta / BC ukugqoka kunciphise inani lamagciwane asebenzayo kuma-biofilms e-vitro futhi kukhombise ukuncishiswa okuthe xaxa ezingxenyeni ze-bacterial kanye ne-APS ku-biofilms yegundane. Lezi zingubo zazinemiphumela ehlukile ekuphulukisweni kwamanxeba okutheleleka athelelekile futhi angewona ama-biofilm, ngokugqoka usawoti oxygen, nokuvuvukala okuqhathaniswa nokuvuselelwa kwezindlela zokwelapha nezinye izigqoko zesiliva. Izakhiwo ezahlukahlukene ze-physicochemical zezingubo zesiliva zingaba nemiphumela ehlukile kwi-biofilm nokuphulukisa, futhi lokhu kufanele kubhekwe lapho kukhethwa ukugqoka amanxeba atheleleke ngegciwane le-biofilm.
Amanxeba angapheli achazwa ngokuthi "amanxeba ahlulekile ukuthuthuka ngezigaba ezijwayelekile zokuphulukiswa ngendlela ehlelekile nangesikhathi esifanele" 1. Amanxeba angapheli adala umthwalo wezengqondo, wezenhlalo nakwezomnotho ezigulini kanye nohlelo lwezempilo. Ukuchitha imali yonyaka ye-NHS ekwelapheni amanxeba kanye nama-comorbidies ahambisana nakho kulinganiselwa ku- £ 8.3 billion ngo-2017-182. Amanxeba angapheli nawo njengamanje uyinkinga ecindezelayo e-United States, ne-Medicare eveza izindleko zonyaka zokwelapha iziguli ezinenxeba ngo- $ 28.1- $ 96.8 billion3.
Ukutheleleka kuyinto enkulu evimbela ukuphulukiswa kwezilonda. Ukutheleleka kuvame ukubonakala njenge-biofilms, okukhona kumanxeba angapheli angapholi. Ifomu le-biofilms lapho ama-microorganisms ehlangana ngokungenakuphikiswa ukuze afakwe endaweni enezinsilo Isilonda Biofilm sihlotshaniswa nokuphendula okuvuvukala kokuvuvukala okuholela ekulimaleni kwezicubu, okungabambezeleka noma ukuvimba ukuphulukiswa4. Ukwanda komonakalo wezicubu kungahle kube khona ngokwengxenye ukuya ekwandeni kwemisebenzi ye-matrix metalpoproteases, i-collagenase, elastase kanye ne-oxygen perfol5. Ngaphezu kwalokho, amaseli wokuvuvukala kanye nama-biofilms ngokwawo abathengi abaphezulu be-oxygen ngakho-ke kungadala ama-hypoxia asendaweni, anciphise amaseli we-oksijini ebalulekile adingekayo ukuze kulungiswe ama-app alungisa izicubu ezisebenzayo6.
Ama-biofilms avuthiwe amelana kakhulu nama-antimicrobial agents, adinga amasu anolaka ukulawula ukutheleleka kwe-biofilm, njengokuphathwa kwemishini kulandelwa ukwelashwa okusebenzayo kwe-antimicrobial. Ngoba ama-biofilms angavuselela ngokushesha, ama-antimicrobials asebenzayo anganciphisa ubungozi bokwakheka kabusha ngemuva kokungcola okuhlinzayo7.
Isiliva liya ngokuya lisetshenziswa ezingutsheni ze-antimicrobial futhi livame ukusetshenziswa njengokwelashwa komugqa wokuqala wamanxeba angenagciwane athelelekile. Kunezingubo zokugqoka zesiliva eziningi ezitholakalayo, ngayinye equkethe ukwakheka kwesiliva okuhlukile, okuhlushwa, kanye ne-matrix yasekuqaleni. Intuthuko emaphethelweni esiliva aholele ekuthuthukisweni kwezingubo ezintsha zesiliva. Uhlobo lwensimbi lwesiliva (AG0) luvukile; Ukufinyelela ukusebenza kwe-antimicrobial, kufanele kulahlekelwe i-elektroni ukwakha isiliva ye-ionic (AG1 +). Izingubo zesiliva zesiliva ziqukethe amakhompiyutha wesiliva noma isiliva lensimbi, lapho kuvezwa uketshezi, ukubola kukha ukwakha i-AG1 + i-Ion. Lawa a-AG1 + I-Ion asabela ngeseli le-bacterial, asuse ama-elektroni avela ezingxenyeni ezihlelekile noma izinqubo ezibucayi ezidingekayo ukuze zisinde. Ubuchwepheshe obunelungelo lobunikazi buholele ekuthuthukisweni kwengxenye entsha yesiliva, ag oxysalts (isiliva i-nitrate, i-AG7NO11), efakiwe ekugqokeni amanxeba. Ngokungafani nesiliva yendabuko, ukuwohloka kosawoti okunama-oksijini aveza izizwe zesiliva ngobuqili obuphakeme (AG1 +, AG2 + ne-AG3 +). Ezifundweni ze-vitro zikhombisile ukuthi ukugxila okuphansi kosawoti wesiliva oxygedated kusebenza kakhulu kunesiliva elilodwa (AG1 +) ngokumelene namagciwane e-pathogenic afana ne-pseudomonas aeruginosa, staphylococcus aureus ne-escherhia coli8,9. Olunye uhlobo olusha lokugqoka lwesiliva luhlanganisa izithako ezengeziwe, okungukuthi i-ethyleniaminetetraetetracetic acid (Edta) kanye ne-benzethonium chlorium (BC), okubikwa ukuthi kubhekiswe ku-biofilm eps bese kukhulisa ukungena kwesiliva ku-biofilm. Lezi ziliva ezintsha zinikeza izindlela ezintsha zokuqondisa izilonda ze-biofilms. Kodwa-ke, umthelela walezi zinti zokulwa nezilonda emgqonyeni wenxeba nasekuthelelekeni kwezifo ezizimele kubalulekile ukuqinisekisa ukuthi azidali imfashini engathandeki noma ukubambezeleka. Ukukhathazeka mayelana ne-vitro yesiliva cytotoxicity kubikwe ngezingubo eziningana zesiliva10,11. Kodwa-ke, eVitro Cytotoxicity ayisahunyushelwe ku-Vivo Forexity, futhi izingubo eziningana ze-AG1 + zikhombise iphrofayili enhle yokuphepha12.
Lapha, siphenya ukusebenza ngempumelelo kwezingubo zokugqoka ze-carboxymethylceseose eziqukethe ukwakheka kwesiliva kwesiliva ngokulwa ne-biofilm eVitro naseVivo. Ngaphezu kwalokho, imiphumela yalezi zindlela zokugqoka eziphendula izimpendulo zokuzivikela nokuphulukisa ngokutheleleka kwahlolwa.
Zonke izingubo ezisetshenzisiwe zazitholakala ngokuthengisa. 3M Kerracel Gel Fibred Creshing (3M, Knutsford, UK) yi-cargene gelbobial 100% Carl gel fiber egqoke ukugqoka njengokugqoka okulawulwa kulolu cwaningo. Izingubo ezintathu zesiliva ze-Antimicrobial CMC zahlolwa, okungukuthi okungukuthi okungu-3M Kerracel AG Ukugqoka (3M, Knutsford, UK), okuqukethe 1.7 wt%. Usawoti wesiliva ophefumulelwe ophefumulelwe (AG7NO11) ngobuqili obuphezulu besiliva ion (AG1 +, AG2 + ne-AG3 +). Ngesikhathi sokubola kwe-AG7NO11, i-AG1 +, i-AG2 + ne-AG3 + ama-Ion yakheka ngesilinganiso esingu-1: 2: 4. I-Aquel Ag Ukugqoka okungeziwe okuqukethe i-1,2% isiliva (AG1 +) (i-convatec, deeside, i-UK) i-chloride engu-1.2%
Izingqinamba ezisetshenziswe kulolu cwaningo kwakuyi-pseudomonas aeruginosa NCTC 10781 (I-Public Health England, iSalisbury) noStaphbury aureus NCTC 6571 (I-Public Health England, Salisbury).
Amagciwane akhuliswe ubusuku bonke kumhluzi we-muller-hinton (OXOID, Altrincham, UK). Isiko ubusuku bonke labe selihlanjululwe umhluzi ongu-1: 100 kuMnu. ). ) Ukwakheka kwamakholoni biofilm ku-37 ° C amahora angama-24. Lawa ma-biofilms amakoloni ahlolwe ngokuqothuka kwe-logarithmic.
Sika ukugqoka kube yizicucu ezi-3 cm2 square futhi u-pre-moinken namanzi ayinyumba. Beka ibhandeshi ngaphezulu kwe-biofilm yekoloni epuletini le-agar. Njalo kwasuswa ama-biofilm njalo, futhi amagciwane asebenzayo ngaphakathi kwe-biofilm (CFU / ML) ahlukaniswe yi-dilution ye-serial (10-1 kuya ku-10-7) kumhluzi wokungathathi hlangothi kosuku (MENCK-millipore). Ngemuva kwamahora angama-24 wokufunga ku-37 ° C, kwenziwa izibalo zepuleti ezijwayelekile kumapuleti e-Mueller-Hinton Agar. Ukwelashwa ngakunye kanye nephuzu lesikhathi kwenziwa ngamathathu, kanti amabala epuleti aphindwayo nge-dilution ngayinye.
Isikhumba sengulube sesisu sitholakala ezingulubeni ezinkulu ezimhlophe zabesifazane kungakapheli imizuzu eyi-15 yokuhlatshwa ngokuya ngamazinga okuthumela ezinyunyana european. Isikhumba saphucwa futhi sahlanzwa ngotshwala, sabe sesifrikelwa ku--80 ° C amahora angama-24 ukugwema isikhumba. Ngemuva kokuncipha, izingcezu zesikhumba ezi-1 cm2 zagezwa amahlandla amathathu nge-PBS, 0.6% Sodium Hypochlorite, ne-70% ethanol imizuzu engama-20 isikhathi ngasinye. Ngaphambi kokususa i-epidermis, susa noma iyiphi i-ethanol esele ngokugeza izikhathi ezi-3 kuma-pbs oyinyumba. Isikhumba sasiklanyelwe epuletini elinama-6 nge-0,45-μm-nye nolwelwesi lwe-nylon Ukhozi. I-Medium (Dulbecco's Modified Eagle Medium - Aldrich Ltd.).
Ama-Biofilms angamaKoloni akhule njengoba kuchazwe izifundo zokuvezwa kwe-biofilm. Ngemuva kokwenza isiko le-biofilm ku-membrane amahora angama-72, i-biofilm yasetshenziswa endaweni yesikhumba isebenzisa i-locunt ye-inoculation futhi ulwelwesi lwalususwa. I-biofilm yabe isifakwa e-dermis yengulube yamahora angama-24 angezekile ku-37 ° C ukuvumela i-biofilm ukuthi ivuthwe futhi ilandele isikhumba sengulube. Ngemuva kokuthi i-biofilm yasuswa futhi yanamathiselwa, ukugqoka okungu-1.5 cm2, kwafakwa ngaphambili ngamanzi afakwe emanzini, kwasetshenziswa ngqo endaweni yesikhumba futhi kwafakwa ku-37 ° C amahora angama-37 amahora angama-24 amahora angama-24 amahora angama-24 amahora angama-24. Amagciwane asebenzayo abonakale sengathi aboniswe ngokufaka izicelo ngokulingana nge-pestoblue cell realent realent (i-invitrogen, impilo, ubuchwepheshe, i-paisley, uk) endaweni eyodwa yokuhlonza futhi ayivuselele imizuzu emi-5. Sebenzisa ikhamera ye-Leica Dfc425 yedijithali ukuze uthwebule izithombe ngokushesha kwi-Leica MZCOPE Microscope. Izindawo ezinemibala enombala zisetshenzisiwe kusetshenziswa i-Image Pro Software Version 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (Mediacy.com). Ukuskena i-elektroni microscopy yenziwa njengoba kuchazwe ngezansi.
Amagciwane akhuliswe ubusuku bonke ahlanjululwe 1: 100 eMueller-Hinton Umhluzi. I-200 μL yesiko lengezwe ku-velile 0.2 μm Whatch cyclopore membrane (What Monman, Mainstone, UK) kanye nePurted ku-Mueller-Hinton Agar. Amapuleti e-biofilm afakwe ku-37 ° C amahora angama-72 ukuvumela ukwakheka kwe-biofilm evuthiwe.
Ngemuva kwezinsuku ezi-3 zokuvuthwa kwe-biofilm, i-bandage engu-3 CM2 yabekwa ngqo kwi-biofilm futhi yafakwa ku-37 ° C amahora angama-24. Ngemuva kokususa i-bandage kusuka ebusweni be-biofilm, 1 ml we-prestoblue cell realent realent (i-invitrogen, e-waltham, i-MA) yengezwe ebusweni be-biofilm ngayinye imizuzwana engama-20. Izindawo zomisiwe ngaphambi kokuqoshwa kwezinguquko zemibala kubhalwe kusetshenziswa ikhamera ye-nikon D2300 Digital Camera (Nikon UK Ltd., Kingston, UK).
Lungiselela amasiko ubusuku bonke ku-Mueller-Hinton Agar, udlulise amakoloni ngamanye kuya ku-10 ml Mueller-Hinton Umhluzi futhi ufaka ku-Shaker ku-37 ° C (100 RPM). Ngemuva kokufakwa ebusuku, isiko lahlanjululwe 1: I-100 eMueller-Hinton Umhluzi futhi u-300 μl wayindilinga ku-stclopopore memblopore (Whandy International Agar e-Mueller-Hinton Agar futhi afakwe ku-37 ° C kungakapheli amahora angama-37 . . I-biofilm evuthiwe yasetshenziswa esilondeni njengoba kuchazwe ngezansi.
Wonke umsebenzi nezilwane wenziwa e-University of Manchester ngaphansi kwelayisensi yephrojekthi evunyelwe yiHhovisi Lezenhlalakahle Yezilwane kanye nokubukelwa kokuziphatha (P871bd27) nangokuhambisana nemihlahlandlela eshicilelwe yihhovisi lasekhaya ngaphansi kwe-2012 aspa esebukeziwe. Bonke ababhali banamathela kwimihlahlandlela yokufika. I-C57BL / 6J Mice Elevel-6 / 6J Mice (Enving, OxON, UK) yayisetshenziselwa bonke abaseVIVO Study. Amagundane abulawa nge-isoflurane (Piramal Crivere Care Ltd, West Drayton, ek) kanye nezindawo zazo ze-dorsal zaphuzwa futhi zahlanzwa. Igundane ngalinye labe selunikezwe isilonda esijabulisayo se-2 × 6 mm sisebenzisa isilonda se-stiefy biopsy (Schuco International, Hertfordshire, UK). Kwamanxeba athelelekile e-biofilm, faka amanxeba e-biofilm angama-72 akhule ngolwelwesi njengoba kuchaziwe ngenhla kungqimba le-dermal yesilonda ngokushesha ngemuva kokulimala nokulahla ulwelwesi. Isentimitha elilodwa lesikwele sokugqoka kwangaphambi kokuswakama ngamanzi ayinyumba ukuze alondoloze indawo enomswakama. Ukugqoka kwakusetshenziswa ngqo esilondeni ngasinye futhi kwambozwa nge-3M tegaderm Film (3M, Bracknell, UK) kanye ne-Mastisol Health Adhesive (i-Eloquest Health. I-Buprenorphine (Animalcare, York, UK) waphathwe ekuhlolweni kwe-0.1 mg / kg njenge-analgesic. Cindezela amagundane ezinsukwini ezintathu ngemuva kokulimala usebenzisa indlela eyi-1 bese ususa, uhlukanise, bese ugcina indawo yokulonda njengoba kudingeka.
IHemaToxylin (Thermofoisher Science) ne-Eosin (Thermofoisher Science) yenziwe ngokwenziwe ngokweprotokol yomenzi. Indawo ye-WLANDE ne-regeepikelialization yahlukaniswa kusetshenziswa i-Sithombe Pro Software Version 10 (Media Cybernetics Inc, Rockville, MD).
Izingxenye zezicubu zazenziwa ngamakhilomitha e-Xylene (i-thermofoisher, e-UK), zivuselele i-ethanol engu-100-50%, futhi zibhange ngamanzi kafushane (i-thermofoisher yesayensi). I-ImmunohistocheMemistry yenziwa kusetshenziswa iVectastain Elite ABC PK-6104 KIT (i-Vector Laboratories, i-Burlilingilingling, CA) ngokuya nge-protocol yomenzi. Ama-antibodies aphambili kuma-neutrophils NIMP-R14 (Thermofoisher Science) kanye neMacrophage MS CD107B amsulwa m3 / 84 (BD Bioscience, i-UK) ahlanjululwe, afakwe ku-2 antibodies anti-, iVectastain I-ABC neVector Nova Red Peroxidase (HRP) substrate kit (i-vector laworatories, i-burlilinglingling, CA) futhi ifakwe kabusha nge-hematoxylin. Izithombe zatholakala zisebenzisa ama-microscope ama-Olympus BX43 kanye nekhamera ye-Olympus DP73 Digital Camera (Olympus, Southend-ON-Sea, UK).
Amasampula wesikhumba ahlelwe ngo-2,5% ama-glutaraldehyde kanye ne-4% ye-formaldehyde ku-0,1 m Hepes (PH 7.4) amahora angama-24 ku-4 ° C. Amasampula aphenya ngamanzi aphenyisiwe esebenzisa i-ethanol ehleliwe futhi omiswe kwi-CO2 esebenzisa i-quorum k850 esibucayi i-Tmer Creater (i-quedon, ek) kanye ne-sputter esebenzisa i-quorum sc7620 uhlelo lokukhipha. Izinhlobo ze-specimenters zizimisele kusetshenziswa i-FEI Quanta 250 Ukuskena ama-scroncop sconcope angama-250 e-elektroni (Thermofeisher Science) ukubona ngeso lengqondo iphuzu eliphakathi nendawo.
I-Toto-1 iodide (2 μm) yasetshenziswa ebusweni obukhulu bemfashini yegundane futhi ifakwe amaminithi ama-5 ku-37 ° C (Thermofoisher Science) futhi iphathwe nge-syto-60 (10 μm) ku-37 ° C (Thermofoisher Science). Izithombe zemizuzu engu-15 ze-Z-Stack zadalwa kusetshenziswa i-Leica TCS SP8.
Idatha ye-biological and technical futhi yahlaziywa futhi yahlaziywa kusetshenziswa isoftware ye-graphpad Prism V9 (i-Chrazpad Software, uLa Jolla, CA). Ukuhlaziywa kwendlela eyodwa yokuhlukahluka nokuqhathaniswa okuningi kusetshenziswa ukuhlolwa kwe-Dunnett Post Host HOC kusetshenziselwa ukuhlola umehluko phakathi kokwelashwa ngakunye kanye nokugqoka okungasho lutho. Inani le-P <0.05 libhekwa njengelibalulekile.
Ukusebenza kahle kwezingubo zokugqoka ze-gel fibrous kuqala kuhlolwe kuqala ngokumelene nama-biofilm colonies we-staphylococcus aureus kanye ne-pseudomonas aeruginosa eVitro. Ukugqoka kwesiliva kuqukethe amafomula ahlukile wesiliva: Izingubo zesiliva zendabuko zikhiqiza i-AG1 + i-Ion; Izingubo zesiliva, ezingakhiqiza i-AG1 + i-Ion ngemuva kokungezwa kwe-EDTA / BC, zingabhubhisa i-matrix ye-biofilm futhi ziveze amagciwane esiliva ngaphansi komphumela we-antibacterial wesiliva. I-Ions15 nokugqoka okuqukethe usawoti wakwa-AG okhiqiza i-AG1 +, i-AG2 + ne-AG3 + i-Ion. Ukusebenza kwayo kwaqhathaniswa nokugqoka okungekho okuqukethwe okungekho emthethweni okwenziwe ngemicu ye-golled. Kusele ama-bacterium asebenzayo ngaphakathi kwe-biofilm njalo emahoreni angama-24 izinsuku eziyi-8 (Umdwebo 1). Ngosuku 5, i-biofilm yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde itfolwe nge-3.85 × 105s. I-Staphylococcus aureus noma ngo-1.22 × 105p. I-Aeruginosa ukuhlola ukululama kwe-biofilm. Uma kuqhathaniswa nokugqoka okungekho oku-antimicrobial control, ukugqoka kwa-AG1 + kwakunomphumela omncane ngokusebenza kwamagciwane eStaphylococcus aureus nama-pseudomonas aerugilm kanye nama-aerudomonas aeruginms angaphezu kwezinsuku ezi-5. Ngokuphambene, ukugqoka okuqukethe i-AG ne-AG1 + + e-Edta / i-BC usawoti kwakusebenza ngokubulala amagciwane ngaphakathi kwe-biofilm kungakapheli izinsuku ezi-5. Ngemuva kokuphindaphindeka kwe-inoculation nama-bacteria ama-planktonic ngosuku 5, akukho ukubuyiselwa kwe-biofilm okubonwe (umdwebo 1).
Ukuqwashiswa kwamagciwane asebenzayo eStaphylococcus aureus kanye ne-pseudomonas aeruginle BIOFILLMS ngemuva kokwelashwa ngezingubo zesiliva. Amakoloni we-Biofilm weStaphylococcus aureus kanye ne-pseudomonas aeruginosa aphathwe ngezingubo zesiliva noma ukugqoka okungekho emthethweni, kanti inani lamagciwane asebenzayo asele anqunywe njalo emahoreni angama-24. Ngemuva kwezinsuku ezi-5, i-biofilm yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde yaphinde itfolwe nge-3.85 × 105s. I-Staphylococcus aureus noma ngo-1.22 × 105p. Amakoloni amaBacterioPlankton Pseudomonas aeruginosa akhiwa ngawodwana ukuhlola ukululama kwe-biofilm. Amagrafu akhombisa kusho ukuthi:
Ukubona ngeso lengqondo umphumela wokugqoka kwesiliva kuma-biofilm asebenzayo, ukugqoka kwasetshenziswa kuma-biofilms avuthiwe akhule ku-porcine Skin Ex Vivo. Ngemuva kwamahora angama-24, ukugqoka kuyasuswa futhi i-biofilm igcwele udayi osebenzayo osheshayo, ohlelwa ngamagciwane aphilayo ngombala opinki. Ama-Biofilms aphathwe ngezingubo zokulawula ayepinki, ekhombisa ukuba khona kwamagciwane asebenzayo ngaphakathi kwe-biofilm (Umdwebo 2a). Ngokuphambene, i-biofilm ephathwe nge-AG Oxysols yokugqoka yayihlaza okwesibhakabhaka, okukhombisa ukuthi ama-bacteria asele ebusweni besikhumba sengulube ayengamagciwane angenakubonwa (umfanekiso 2b). Umbala oxubile ohlaza okwesibhakabhaka no-pink waqashelwa nge-biofilms ephethwe nge-AG1 + -Containg dress yokugqoka, ekhombisa ukuba khona kwamagciwane asebenzayo nawokungasebenzi ngaphakathi kwe-biofilm (Umdwebo 2c), kanti ukugqoka okuqukethe i-AG1. okukhombisa izindawo ezingathinteki ngokugqoka kwesiliva (Umdwebo 2D). Izindawo ezisebenzayo ze-Active (Pink) nezindawo ezingasebenzi (eziluhlaza okwesibhakabhaka) zibonisa ukuthi isichibi sokulawula sasisebenza ngo-75% (Umdwebo 2e). Ukugqoka kwe-AG1 + + Edta / BC okwenziwe ngendlela efanayo nokugqoka kukasawoti we-oxygedged, ngamanani okusinda angu-13% no-14%, ngokulandelana. Ukugqoka kwe-AG1 + kunciphise ukusebenza kwama-bacterial nge-21%. Lezi biofilms zabe sezigcinwa zisebenzisa ukuskena i-elektroni microscopy (SEM). Ngemuva kokwelashwa ngokugqoka okulawulwa kanye nokugqoka kwe-AG1 +, ungqimba we-pseudomonas aeruginosa waqashelwa ukumboza isikhumba se-porcine (isithombe 2f, ambalwa amaseli angamagciwane atholwe futhi ambalwa amaseli angamagciwane atholwe ngaphansi. I-Collagen Fibers ingabhekwa njengesakhiwo sezicubu zesikhumba se-porcine (Umdwebo 2g). Ngemuva kokwelashwa nge-AG1 + + Edta / BC ukugqoka, ama-plaque ama-bacterial kanye nama-collagen fiber ama-collagen fiber abonakala (Umdwebo 2i).
Ukubona ngeso lengqondo i-pseudomonas aeruginosa biofilm ngemuva kokwelashwa kokugqoka kwesiliva. . Amagciwane abukhoma anamagciwane apinki, angasebenzi kahle kanye nesikhumba sengulube aluhlaza okwesibhakabhaka. . I-Sem Scale Bar = 5 μm. (J-M) amakoloni biofilms akhula ezihlungi futhi anamabala odayi wokusebenza kwe-prestoblue ngemuva kokufakwa kwe-24 h yokufakwa ngezingubo zesiliva.
Ukuthola ukuthi ukuxhumana okusondelene phakathi kokugqoka kanye ne-biofilms kuthinte ukusebenza kwezingubo, ama-biofilms amakoloni abekwe endaweni ephansi aphathwa ngokugqoka amahora angama-24 abese enamanzi asebenzayo. I-biofilm engalashwa yayipinki emnyama ngombala (umfanekiso 2J). Ngokuphambene ne-biofilms iphathwe ngokugqoka okuqukethe usawoti we-ag ag (umdwebo 2k), i-biofilms ephethwe ngokugqoka okuqukethe i-AG1 + noma i-AG1 + BC ikhombise amabhendi epinki esitengeli (isithombe 2l, m). Lo mbala wepinki ukhombisa ukuba khona kwamagciwane asebenzayo futhi kuhlotshaniswa nendawo yokusungula ngaphakathi kokugqoka. Lezi zindawo ezithungelwe zidala izikhala ezifile ezivumela amagciwane ngaphakathi kwe-biofilm ukuze asinde.
Ukuhlola ukusebenza kwezingubo zesiliva eVivo, ukuqina okugcwele amanxeba amagundane angenwe nguS. Aureus noP. Aeruginosa biofilms aphathwe ngezingubo zokulawulwa kweziliva noma ukugqoka kwesiliva. Ngemuva kwezinsuku ezi-3 zokwelashwa, ukuhlaziywa kwezithombe ze-macroscopic kubonise osayizi abancane besilonda lapho belashwa ngokugqoka kukasawoti oxyged uma kuqhathaniswa nokugqoka okungekho oku-antimicrobial okuqukethwe kanye nezinye izingubo zesiliva (Umdwebo 3a-H). Ukuqinisekisa lokhu okubukwayo, amanxeba avunwa futhi alimala indawo kanye ne-peepikelizelization kwahlelwa ngezigaba ze-hematoxylin kanye ne-eosin-stized izicubu usebenzisa i-Image Pro Software Version 10 (Umdwebo 3i-L).
Umphumela wokugqoka kwesiliva endaweni yenxeba nasekuvuseleleni kabusha amanxeba atheleleke nge-biofilms. . ukugqoka. Izithombe ze-macroscopic. Amanxeba amagundane anokugqoka kwe-AG1 + + edta / BC. (Il) Ukutheleleka okumele i-Pseudomonas Aeruginosa, izingxenye zomlando ezinamabala nge-hematoxylin ne-eosin, zisetshenziselwa ukwanda indawo yendawo kanye nokuvuselelwa kwe-epithelial. Ukuqwashiswa kwendawo ye-WNINTS (M, O) kanye namaphesenti we-penductiveelifelization (n, p) amanxeba atheleleke nge-pseudomonas aeruginosa (m, n) ne-staphylococcus aureus (O, p) biofilms (O, P) Biofilms (O, P) Biofilms (Pear Treatment Group N 12 12). Amagrafu akhombisa kusho ukuthi: * Kusho p = <0.05 ** kusho p = <0.01; Isikali seMacroscopic = 2,5 mm, isikali se-Histolocal = 500 μm.
Ukuthambekelwa kwendawo yokulimala emanxebeni angenwe yi-pseudomonas aeruginosa biofilm (Umdwebo 3m) kubonise ukuthi amanxeba aphathwa ngamanxeba angu-2,5 mm2, kanti ukugqokwa kwezilindi okuphakathi kwe-3.1 mm2, okungeyona kuyiqiniso. ukufinyelela okubonakalayo kwezibalo (Umdwebo 3m). p = 0.423). Amanxeba aphathwe nge-AG1 + noma i-AG1 + + Edta / BC akhombisa ukuthi akukho ukuncishiswa endaweni yokulimala (3.1 mm2 no-3.6 mm2, ngokulandelana). Ukwelashwa ngotshani kasawoti we-AG kukhushulwe kabusha okuvuselelwa kabusha ngezinga elikhulu ukwedlula ukugqoka okungekho emthethweni (34% no-15%, ngokulandelana: Umdwebo 3n). . , ngokulandelana).
Amathrendi afanayo enxebeni le-Inlingle's Area kanye ne-Epithelial Regeneration babonwa emanxebeni angenwe yi-S. Aureus Biofilms (Umdwebo 3o). Ukugqoka okuqukethe usawoti wesiliva okuxhunyiwe kuncishisiwe indawo yenxeba (2.0 mm2) ngo-23% uma kuqhathaniswa nokugqoka okungalawulwa kokulawula (2.6 mm2), yize lokhu kuncishiswa bekungabalulekile (p = 0.304). Ngaphezu kwalokho, indawo yokulimala eqenjini le-AG1 + yancishiswa kancane (2.4 MM2), ngenkathi isilonda siphathwa nge-AG1 + + Edta / bc ukugqoka akuzange kunciphise indawo yenxeba (2.9 mm2). Usawoti we-oksijini we-AG futhi wagqugquzela kabusha amanxeba atheleleke nge-S. Aureus Biofilm (31%) ngezinga elikhulu kunalawo aphathwe izingubo zokulawula ezingezona eze-antimicrobial (12%, p = 0.003) (Umdwebo 3p). I-AG1 + Ukugqoka (16%, p = 0.903) kanye ne-AG + 1 + Edta / BC ukugqoka (14%, p = 0.965) kukhombise amazinga we-epithelial asport.
Ukubona ngeso lengqondo umphumela wokugqoka kwesiliva kuma-matrix we-biofilm, i-toto 1 ne-syto 60 i-iodide stain yenziwa (Fig. 4). Udayi we-toto 1 uwudayi ongenakufinyeleleka esitokisini esingasetshenziswa ukuze ngeso lengqondo ngokunemba ama-nucleic acid, anamaphakathi kuma-eps we-biofilms. I-Syto 60 yidayi evunyelwe eseli esetshenzisiwe njenge-Countrestain16. Ukubonwa kwe-Toto 1 ne-SYTO 60 Iodide emanxebeni atholwe ngama-biofilms we-pseudomonas aeruginosa (Umdwebo 4i-d) noStaphylococcus aureus (isithombe 4i-d) noStaphylococcus aureus, ama-eps nge-biofilm ancishiswe kakhulu. equkethe usawoti oxygenated ag ne-AG1 + + Edta / BC. Izakhiwo ze-AG1 + ngaphandle kwezinto ezingezekile ze-antibiiofilm zinciphise kakhulu ama-DNA angenamakhalekhukhwini ngamanxeba afakwe amanxeba e-aeruginosa kodwa aphumelela kakhulu emanxebeni ane-areus.
E-Vivo ukucabanga nge-biofilm ngemuva kwezinsuku ezi-3 zokwelashwa ngezingubo zokulawula noma zesiliva. Izithombe ezivumayo ze-pseudomonas aeruginosa (A-D) kanye ne-staphylococcus aureus (i-l) zinamatshe nge-toto 1 (eluhlaza) ukubona ngeso lengqondo ama-nucleil acid ama-aucleils, ingxenye yama-polymers e-externacellular aymembers. Ukubeka amabala ama-nucleellar acid, sebenzisa i-syto 60 (ebomvu). ama-acid. P. Ukuskena i-elektroni microscopy yamanxeba atheleleke nge-pseudomonas aeruginosa (e-h) kanye ne-staphylococcus aureus (m-p) biofilms (m-p) biofilms (m-p) biofilms (m-p) biofilms (m-p) biofilms ngemuva kwezinsuku ezi-3 zokuphuza. I-Sem Scale Bar = 5 μm. Ibha yokucabanga evumayo = 50 μm.
Ukuskena i-elektroni
Ukuhlola umphumela wokugqoka kwesiliva ekuvuvukeni kwesilimo kumagundane atheleleke ngegciwane, izingxenye zamanxeba okutheleleka aphathwe ngokulawulwa kwama-antibophils asetshenziswa ama-antibophils nama-macrophage. Ukunqunywa kwenani lama-neutrophils kanye nama-macrophage ngaphakathi. izicubu zomzimba. Umdwebo 5). Zonke izigqoko zesiliva zinciphise inani lama-neutrophils nama-macrophages emanxebeni angenwe yi-pseudomonas aeruginosa uma kuqhathaniswa nokugqoka okungekho emthethweni kwezokwelapha ngemuva kwezinsuku ezintathu zokwelashwa. Kodwa-ke, ukwelashwa ngokugqokwa usawoti wesiliva okuxhunyiwe kwaholela ekunciphiseni okukhulu kuma-neutrophils (p = <0.0001) kanye nama-macrophages (p = <0.0001) uma kuqhathaniswa nezinye izingubo zesiliva ezihloliwe (Umdwebo 5i, j). Yize i-AG1 + + Edta / BC yaba nomthelela omkhulu kwinxeba le-biofilm, inciphise amazinga we-neutrophil kanye nama-macrophage ngezinga elincane kakhulu kunokugqoka okuphansi kwe-AG1 +. Amanxeba alinganiselayo atheleleke ngo-S. Aureus Biofilm nawo abhekwe ngemuva kokugqoka nge-AG (P = <0.0001) kanye ne-AG1 ++ edta / BC (P = 0.000043) Izitayela ezifanayo zibhekwa ne-neutropenia. I-bandage (Fig. 5k). Kodwa-ke, ukugqokwa kukasawoti oxygedged ag kubonise ukwehliswa okukhulu kwenani lama-macrophages ezicubu ze-granution uma kuqhathaniswa nokulawulwa kwamanxeba angenwe yi-S. Aures biofilms (P = 0.0339) (Umdwebo 5l).
Ama-neutrophils kanye nama-macrophage ahlukaniswe emanxebaneni atheleleke nge-pseudomonas aeruginosa ne-staphylococcus aureus biofilms ngemuva kwezinsuku ezi-3 zokulawulwa noma ukugqoka kwesiliva. I-Neutrophils (isikhangiso) kanye nama-macrophages (eh) ahlukaniswe ngezigaba zezicubu ezinama-antibodies aqondile ama-neutrophils noma ama-macrophage. Ukuqwashiswa kwama-neutrophils (i nok) kanye nama-macrophages (J no-l) emanxebeni angenwe yi-pseudomonas aeruginosa (i no j) kanye ne-staphylococcus aureus (k & l) biofilms. N = 12 iqembu ngalinye. Amagrafu akhombisa ukuthi asho ukuthini- *** kusho p = <0.001; kukhombisa p = <0.0001).
Sibe sesihlola umphumela wezingubo zesiliva ekuphulukeni kwezifo ezizimele. Amanxeba edlula angenagciwane athelelekile aphathwa ngokugqoka okulawulwa yi-antimicrobial antimicrobial noma ukugqoka kwesiliva izinsuku ezi-3 (Umdwebo 6). Phakathi kwezingubo zesiliva ezihlolwe, kuphela amanxeba aphethwe ngusawoti oxyged oxyged avele emincane ezithombeni zeMacroscopic ngaphandle kwamanxeba aphathwa ngokulawulwa (umfanekiso we-6a-D). Ukuqanjwa kwendawo yokulimala kusetshenziswa Ukuhlaziywa kwe-Histological kukhombisa ukuthi indawo yokulimala esele ngemuva kokwelashwa nge-AL Oxysols egqoke amanxeba aphathwe ngeqembu lokulawula, kepha lo mehluko awufinyelelanga ukubaluleka kwezibalo (P = 0.488) (Fig . 6i). Ngokuphambene nalokho, akukho ukwehliswa kwelensi yeLwindi okwabonwa ngemuva kokwelashwa nge-AG1 + (3.38 MM2, P = 0.757) noma i-AG1 + MM2, p = 0.054) kuqhathaniswa neqembu lokulawula. Ukwanda kwe-epithelial kwaqashelwa nge-AG Oxysol ukugqoka uma kuqhathaniswa neqembu elilawulayo (30% vs. 22%, ngokulandelana), yize lokhu kungafinyeleli ukubaluleka (P = 0.067), lokhu kubaluleke kakhulu futhi kuqinisekisa imiphumela edlule. Ukugqoka nge-Oxysols kukhuthaza ukuphindaphinda kwe-epithelization. -Infensization of amanxeba angafakwanga17. Ngokuphambene nalokho, ukwelashwa nge-AG1 + noma i-AG1 + + EDTA / BC yokugqoka bekungenamthelela noma kuboniswe ukwehla kwe-epithelialization okuqhathaniswa nokulawula.
Umphumela wokugqoka isilonda sesiliva ekuphulukeni kwamanxeba kumagundane angavikelekile ngokuqalwa okuphelele. . (EH) izingxenye ezimele izilonda ezinamatshe nge-hematoxylin ne-eosin. Ukuncishiswa kwendawo yokulingisa (i) namaphesenti e-repeepikelialization (j) kubalwe kusuka ezingxenyeni zomlando eziphakathi nendawo kwesilonda kusetshenziswa isoftware yokuhlaziywa kwezithombe (n = 11-16 ngeqembu ngalinye lokwelashwa). Amagrafu akhombisa kusho ukuthi: * Kusho P = <0.05.
Isiliva linomlando omude wokusebenzisa njengokwelashwa kwe-antimicrobial ekuphulukeni kwezilonda, kepha izindlela eziningi ezahlukahlukene nezindlela zokulethwa kungaholela ekungafani kokusebenza ngempumelelo kwe-antimicrobial 18. Ngaphezu kwalokho, izakhiwo ze-antibioFilm zezinhlelo ezithile zesiliva ukulethwa aziqondakali ngokuphelele. Yize impendulo yokusingathwa kokuphendula isebenza ngempumelelo ekulweni namagciwane e-planktonic, ngokuvamile ayisebenza kangako ngokulwa ne-biofilms19. Ama-Planktonic Bacteria atholakala kalula ama-macrophages, kepha ngaphakathi kwama-biofilms, amangqamuzana ahlanganisiwe afaka izinkinga ezengeziwe ngokukhawulela ukuphendula komzimba kuze kufike lapho amangqamuzana e-apoptosis futhi akhulule izici zokuvuselela ama-apop. Kubonile ukuthi amanye ama-leukocytes angangena e-biofilms21 kepha awakwazi ukwenza amagciwane e-phagocytose uma lokhu kuvikela kuphinde kwehlelwe22. Indlela ephelele kufanele isetshenziselwe ukusekela impendulo yokuvikelwa kokuvikezela kokutheleleka nge-biofilmm. Ukudonswa kwamanxeba kungaphazamisa ngokomzimba i-biofilm futhi ususe iningi lama-biofilden, kepha ukusabela kokuvikelwa kokuvikelwa komzimba kungenzeka kungaphumeleli ngokumelene ne-biofilm, ikakhulukazi uma impendulo yomgcini yokuzivikela emzimbeni isengozini. Ngakho-ke, izindlela zokwelapha eziphilayo ezinjengezingubo zesiliva zingasekela umgcini ziphendule futhi ziqede ukutheleleka nge-biofilm. Ukwakheka, ukugxilisa ingqondo, ukusebenzisa kanye nokulethwa kokubeletha kungathonya ukusebenza kwe-antimicrobial kwesiliva. Eminyakeni yamuva nje, intuthuko ebukhwini bokulungiswa kwesiliva benze lezi zingubo zokugqoka ziphumelele ngokwengeziwe9,23. Njengobuchwepheshe bokugqoka isiliva buthuthuka, kubalulekile ukuqonda ukusebenza kwalokhu kugqoka ekunikezeni ukutheleleka kwesilonda futhi, okubaluleke kakhulu, umthelela walezi zinhlobo ezinamandla zesiliva nasekuphulukeni.
Kulolu cwaningo, siqhathanisa ukusebenza kwezingubo ezimbili ezithuthukile zesiliva ezinezingubo ezijwayelekile zesiliva ezikhiqiza i-AG1 + i-Ion ngokumelene ne-Biofilms zisebenzisa okuhlukile ku-vitro nasemodeli ye-vivo. Siphinde sahlola umphumela walezi zingubo zokugqoka emvelweni yenxha yenxemba nasekuthelelekeni kwezifo ezizimele. Ukunciphisa ithonya le-matrix yokulethwa, zonke izigqoko zesiliva ezihlolwe zakhiwa i-carboxymethylcelcellellulose.
Ukuhlolwa kwethu kokuqala kwalezi zingubo zesiliva ezimelene nama-biofilms amakoloni ama-aeruudomonas aeruginosa kanye ne-staphylococcus aureus kukhombisa lokho, ukugqoka okungafani nosawoti we-AG1 +, kuyasebenza ngempumelelo ngosawoti we-ag, kusebenza ngempumelelo ngo-5. Ngokuphumelelayo ukubulala amagciwane e-biofilm ngaphakathi izinsuku ezimbalwa. Ngaphezu kwalokho, lezi zingubo zokugqoka zivimbela ukwakheka kabusha kwe-biofilm ekuchayweni okuphindaphindiwe kumagciwane we-planktonic. Ukugqoka kwe-AG1 + kwakuqukethe isiliva chloride, i-matrix efanayo yesiliva kanye ne-matrix yasekuqaleni njenge-AG1 + + edta / Bc, futhi yayinomphumela olinganiselwe ekusebenzeni kwamagciwane ngaphakathi kwesikhathi esifanayo. Ukuqashelwa ukuthi ukugqoka kwe-AG1 + + EDTA / BC kwasebenza ngempumelelo kangaka ngokumelene ne-biofilm kunesigqoko se-AG1 + esihlanganisa i-matrix efanayo nengxenye yesiliva kusekela umbono wokuthi izithako ezengeziwe zidingeka ukwandisa ukusebenza nge-biofilm, njengoba sekubikiwe kwenye indawo15. Le miphumela isekela umbono wokuthi uBc no-Edta babamba iqhaza elengeziwe lokufaka isandla ekugqokeni ukugqoka okuphelele nokuthi ukungabikho kwalesi sakhi ekugqokeni kwa-AG1 + kungenzeka kube nomthelela ekwehlulekeni ukukhombisa ukusebenza kahle kwe-vitro. Sithole ukuthi ukugqoka kukasawoti ag ag ukukhiqiza i-AG2 + ne-AG3 + I-Ion kukhombisa ukusebenza nge-antibacterial ukwedlula i-AG1 + nasemazingeni afanayo ne-AG1 + + Edta / BC. Kodwa-ke, ngenxa yekhono eliphakeme kakhulu, akukacaci ukuthi isikhathi esingakanani se-AG3 + i-Ion sihlala sisebenza futhi sisebenza ngempumelelo ekunqandeni ama-biofilms ngakho-ke kufanele sifunde ukutadisha okwengeziwe. Ngaphezu kwalokho, kunezingubo eziningi ezahlukene ezakhiqiza i-AG1 + i-Ion ezingahloliwe kulolu cwaningo. Lezi zingubo zihlanganiswa zezinhlanganisela zesiliva ezahlukahlukene, ukugxila, kanye nezisekelo zamatric, ezingathonya ukulethwa kwama-ag1 + ama-ion kanye nokusebenza kwawo ngokumelene ne-biofilms. Kuhle futhi ukuqaphela ukuthi kunezinto eziningi ezihlukile ku-vitro nakumodeli we-vivo ezisetshenziselwa ukuhlola ukusebenza kwezingubo zokugqoka ngama-biofilms. Uhlobo lwemodeli olusetshenzisiwe, kanye nokuqukethwe kukasawoti kanye namaprotheni ezindaba ezisetshenziswe kulezi zinhlobo, kuzothonya ukusebenza kokugqoka. E-Vivo Model yethu, savumela i-biofilm ukuthi ivuthwe e-vitro yabe isidlulisela endaweni ye-dermal yesilonda. Impendulo yokuzivikela kwegundane isebenza ngempumelelo ekulweni namagciwane e-planktonic asetshenziswa esilondeni, ngaleyo ndlela akha i-biofilm njengoba isilonda siphulukisa. Ukungezelelwa kwe-biofilm evuthiwe esilimazeni kukhawulela ukusebenza kahle kokuphendula kokutholwa kokuzivikela ekwakhekeni kwe-biofilm ngokuvumela i-biofilm evuthiwe ngokwayo ngaphakathi kwesilonda ngaphambi kokuphulukiswa kungaqala. Ngakho-ke, imodeli yethu isivumela ukuthi sihlaziye ukusebenza kwezinto zokugqoka ezivuthiwe kuma-biofilms avuthiwe ngaphambi kokuba amanxeba aqale ukwelapha.
Siphinde sathola ukuthi ukugqoka kufanelekile kuthonya ukusebenza ngempumelelo kwezingubo zesiliva ku-biofilms esekhulile nesikhumba se-porcine. Ukuxhumana okusondelene nesilonda kubhekwa njengokubalulekile ekusebenzeni kahle kwe-antimicrobial kwe-dressing24,25. Ukugqoka okuqukethe usawoti we-ag oxygedated ag bekusondelene nama-biofilms avuthiwe, okuholela ekunciphiseni okukhulu kwinani lamagciwane asebenzayo ngaphakathi kwama-biofilm ngemuva kwamahora angama-24. Ngokuphambene nalokho, lapho ephathwa nge-AG1 + kanye ne-AG1 + + Edta / BC yokugqoka, izinombolo ezibalulekile zamagciwane asebenzayo asala. Lezi zingubo ziqukethe ama-suture kulo lonke ubude bokugqoka, okudala izikhala ezifile ezivimbela ukuxhumana eduze ne-biofilm. Kuzo ezifundweni zethu ze-vitro, lezi zindawo ezingaxhumana nazo zivimbela ukubulawa kwamagciwane asebenzayo ngaphakathi kwe-biofilm. Sihlole ukusebenza ngegciwane ngemuva kwamahora angama-24 wokwelashwa; Ngokuhamba kwesikhathi, njengoba ukugqoka kugcwele ngokwengeziwe, kungahle kube khona isikhala esincane esincane, kunciphisa le ndawo yalawa magciwane asebenzayo. Kodwa-ke, lokhu kuqokomisa ukubaluleka kokwakheka kokugqoka, hhayi nje uhlobo lwesiliva ekugqokeni.
Ngenkathi esezifundweni ze-vitro zilusizo ngokuqhathanisa ukusebenza kobuchwepheshe obuhlukile besiliva, kubalulekile futhi ukuqonda imiphumela yalezi zingubo zokugqoka kuma-bivo, lapho izimpendulo zomsingathi kanye nezimpendulo zokuzivikela zinomthelela ekugqokeni kwezingubo zokugqoka ezibhekene ne-biofilms. Umphumela walezi zingubo ezigqokwa amanxeba e-biofilms waqashelwa kusetshenziswa ukuskena ama-elektroni ama-microscopy kanye ne-ePS ye-biofilm esebenzisa udayi lwe-DNA we-intracellular ne-externacelar. Sithole ukuthi ngemuva kwezinsuku ezi-3 zokwelashwa, konke ukugqoka kwakusebenza ngempumelelo ekunciphiseni i-DNA engenamakhalekhukhwini emanxebeni athelelekile, kepha ukugqoka kwa-AG1 + bekungasebenzi kahle emanxebeni atheleleke eStaphylococcus aureus. Ukuskena i-elektroni microscopy kubonise ukuthi amagciwane amancane kakhulu ayekhona emanxebeni aphathwe ngezingubo zesiliva, yize lokhu kubhekwe kakhulu ngokugqoka usawoti we-ag kanye ne-ag1 + + edta / BC yokugqoka uma kuqhathaniswa nokugqoka kwe-AG1 +. Le mininingwane ikhombisa ukuthi ukugqoka kwesiliva okuhlolwe kwakunamazinga ahlukahlukene onomthelela esakhiweni se-biofilm, kepha azikho izigqoko zesiliva ezakwazi ukuqeda i-biofilm, zisekela isidingo sendlela ephelele yokwelashwa kwezifo ze-biofilm; Ukusetshenziswa kwe-welver armands. Ukwelashwa kwandulelwa ukungcola ngokomzimba ukususa iningi le-biofilm.
Amanxeba angapheli avame esimweni sokuvuvukala okunzima, okunamangqamuzana okuvuvukala ngokweqile asele ezicubu zensizwa isikhathi eside, okwenza ukulimala kwezicubu kanye nokunciphisa umoya-mpilo odingekayo nge-metabolism yeselula. Ama-Biofilms akhulisa le ndawo elinamanxeba elinobutha ngokuthinta kabi ukuphulukiswa ngezindlela ezahlukahlukene, kufaka phakathi ukuvimbela ukwanda kwamaseli nokufuduka kanye nokwenza kusebenze kwe-proinflammatorator cytokines27. Njengoba ukugqoka kwesiliva kusebenza kakhulu, kubalulekile ukuqonda umthelela abanawo emvelweni yenxeba nokuphulukiswa.
Kuyathakazelisa ukuthi yize zonke izigqoko zesiliva zithinte ukwakheka kwe-biofilm, kuphela ukugqoka kukasawoti okuxhumeke komoya okotshani kwandise kabusha kwalawa manxeba athelelekile. Le mininingwane isekela ukutholwa kwethu kwangaphambilini17 naleyo yeKalan et al. .
Ucwaningo lwethu lwamanje luqokomisa umehluko kubuchwepheshe besiliva phakathi kokugqoka kwesiliva okulwa nokudlalwa kanye nomthelela walobu buchwepheshe emvelweni wenxeba kanye nokutheleleka ngokutheleleka. Kodwa-ke, le miphumela ihlukile ezifundweni zangaphambilini ezibonisa ukuthi amapharamitha we-AG1 + + Edta / BC athuthukisiwe athuthukisiwe ezindlebe zonogwaja ezilimele eVivo. Kodwa-ke, lokhu kungahle kube ngenxa yokwehluka kumamodeli wezilwane, izikhathi zokulinganisa, kanye nezindlela zohlelo lokusebenza lwe-bacterial29. Kulokhu, izilinganiso zokulimala zithathwe ezinsukwini eziyi-12 ngemuva kokulimala ukuvumela izithako ezisebenzayo zokugqoka ukuze zisebenze nge-biofilm esikhathini eside. Lokhu kusekelwa isifundo esibonisa ukuthi izilonda ezitheleleke emtholampilo ziphathwe nge-AG1 + + edta / BC ekuqaleni zikhuphuke ngosayizi ngemuva kwesonto elilodwa lokwelashwa, bese kuthi amasonto amathathu alandelayo ekwelashweni nge-AG1 + + Edta / BC futhi kungakapheli amasonto amane ukusetshenziswa kwama-non-antimicrobials. izidakamizwa. Ukugqoka kwe-CMC ukunciphisa usayizi we-Ulcers30.
Amafomu athile nokugxila kwesiliva kunakhonjiswe ngaphambili ukuba yi-cytotoxic eVitro 11, kepha lezi ngemiphumela ye-vitro azihlali zihumusha zibe yimiphumela emibi eVivo. Ngaphezu kwalokho, ukuthuthuka kobuchwepheshe besiliva nokuqonda okungcono kwamakhompiyutha wesiliva nokugxila kokugqoka kuholele ekuthuthukisweni kwezingubo eziningi zesiliva eziphephile nezisebenzayo. Kodwa-ke, njengoba ubuchwepheshe bokugqoka besiliva buthuthuka, kubalulekile ukuqonda umthelela walezi zingubo zokugqoka kwinhlangano yenyane31,32,33. Phambilini bekubikwa ukuthi izinga elikhuphukayo le-epithelialization kabusha lihambelana nengxenye eyandayo ye-anti-inflammatory m2 macrophages uma iqhathaniswa ne-pro-inflammatory m1 phenotype. Lokhu kwaphawulwa kwimodeli yegundane edlule lapho kuqhathaniswa nezilonge zesiliva ze-hydrogel kuqhathaniswa nesiliva sulfadiazine nama-non-antimicrobial hydrogels34.
Amanxeba angapheli angabonisa ukuvuvukala ngokweqile futhi sekugcinwe ukuthi ukuba khona kwama-neutrophils ngokweqile kungalimaza ukulungiswa kwe-Healing35. Ocwaningweni kumagundane aseNeutrophil-ancipha, ukuba khona kwama-neutrophils kubambezele kabusha kabusha. Ukuba khona kwama-neutrophils ngokweqile kuholela emazingeni aphezulu we-proteries kanye nezinhlobo ze-oxygen ezisebenzayo, njenge-superoxide ne-hydrogen peroxide, ezihambisana namanxeba angapheli ahamba kancane aphulukisa37,38. Ngokunjalo, ukwanda kwezinombolo ze-macrophage, uma kungalawulwa, kungaholela ekubambezelweni kokupholisa amanxeba. Lokhu kunyuka kubaluleke kakhulu uma ama-macrophages engakwazi ukuguqukela kusuka ku-phenotype okuvuvukala kwi-phenotype ye-pro-aphulukisa, okuholela emanxebeni ehluleka ukuphuma esigabeni sokuvuvukala se-Healing40. Sibonile ukwehla kwama-neutrophils kanye nama-macrophages emanxebeni atheleleke - atheleleke ngemuva kwezinsuku ezi-3 zokwelashwa nazo zonke izingubo zesiliva, kepha ukwehla kwabikwa kakhulu ngokugqoka kukasawoti oxygen. Lokhu kuncipha kungaba umphumela oqondile wokuphendula izivikeli esivikelweni, noma ukuncipha kwe-bioburden, noma isilonda esisesigabeni sokuphulukisa kamuva futhi ngenxa yalokho amangqamuzana omzimba asenxeleni. Ukunciphisa inani lamaseli wokuvuvukala esilondeni kungagcina indawo efanelekayo yokuthola ukuphulukiswa. Indlela yokusebenza kwe-O OXEsAlds egqugquzela ukwelashwa ngezifo izifo ngokungethelelekile ayicaci, kepha amandla esikhumba agysogen ukukhiqiza umoya-mpilo futhi aqede amazinga alimazayo we-hydrogen peroxide, angakuchaza lokhu futhi kudinga okwengeziwe17.
Amanxeba angenacala angapholi aphulukisa abhekana nenkinga kubo bobabili odokotela neziguli. Yize ukugqoka okuningi kudinga ukusebenza kwe-antimicrobial, ucwaningo aluvamile ukugxila kwezinye izinto ezibalulekile ezithonya inxeba ama-microenvelo. Lolu cwaningo lukhombisa ukuthi ubuchwepheshe besiliva obuhlukile bunokusebenza okuhlukile kwe-antimicrobial futhi, okubalulekile, imiphumela ehlukile kwimvelo yenxeba nasekuphulukeni, ezimele ngokutheleleka. Yize lezi eVitro nase-Vivo izifundo zibonisa ukusebenza kwalezi zingubo zokugqoka ekwelapheni izifo zezilonda kanye nokuthuthukisa ukwelashwa, kudingeka izivivinyo ezilawulwa ngokungahleliwe ukuze kuhlolwe ukusebenza kwalezi zingubo.
Ama-datasets asetshenzisiwe futhi / noma ahlaziye phakathi nocwaningo lwamanje ayatholakala kumlobi ohambelana nesicelo esifanele.
Isikhathi Sokuposa: Jul-15-2024