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Kukura kwemicrobial mumaronda kunowanzo kuratidzira seBiofilms, uko kunovhiringidza nekurapa uye kwakaoma kutsauka. New Silver Sirthings inoti kurwisa ronda renjodzi, asi avo antibiofilm yekubudirira uye hutachiona-hwakazvimirira-hwekuporesa mhedzisiro kazhinji hazviwanzo kuzivikanwa. Uchishandisa muVitro uye muVivo Biofilm Models of Staphylococcus Aureus Aeruginosa, isu tinozivisa kushanda kwekubudirira kweAG1 + majira eAG1. AG1 + nguo dzaive nemapfekero ane etherynediamailasetraacetic acloride (ag1 + / edta / bc), uye kupfeka zvine sirivheri nitrate (AG oxysalts). , inoburitsa AG1 +, AG2 + uye AG3 + INI INS kurwadzisa biofilm uye zvarinoita pakuporesa. AG1 + nguo dzakave nemitengo miviri pane ronda biofilm mu vitro uye mumakonzo (c57bl / 6j). Mukupesana, Oxynated ag Salts uye Ag1 + / edta / BC Drivings zvakanyanya kuderedzwa muBiafro muVitro uye EPS Zvikamu uye EPS Zvikamu muBouse Ronda Biofils. Izvi zvipfeko zvaive nemhedzisiro dzakasiyana-siyana pakurapa kweBiofilm-yakatapukirwa uye isina-biofilm-yemigwagwa yemugodhi ine zvakawanda zvinobatsira pakurekodha, size size, uye kuzvimba kwakaenzaniswa nekukanganisa kurapa uye zvimwe zvipfeko zvesirivheri. Iyo yakasiyana physicochemical zvivakwa zvesirivha zvesirivha zvinogona kuve nemhedzisiro dzakasiyana pane ronda biofilm uye kupora, uye izvi zvinofanirwa kutariswa, uye izvi zvinofanirwa kutariswa kana uchisarudza kupfeka maronda ebiofilm-isina hutachiona.
Maronda okusingaperi anotsanangurwa se "maronda anotadza kufambira mberi kuburikidza nematanho ekurapa nenzira yakarongeka uye nenguva" 1. Maronda ekusingaperi anogadzira psychological, munharaunda uye ehupfumi mutoro kune varwere uye hutano hwehutano. Peji yegore negore ichishandiswa pakurapa maronda uye yakabatana comorbidities inofungidzirwa ku £ 8,3 bhiriyoni muna 2017-182. Maronda anguva refu ari zvakare dambudziko iri muUnited States, aine Medicare anofungidzira mutengo wegore rose wekurapa varwere aine maronda ari pamadhora 281- $ 96,8 bhiriyoni 3.
Utachiona chinhu chikuru chinodzivirira kurwadza kwekurwadziwa. Utachiona hunowanzo kuratidzwa seBiofilms, iyo iripo muna 78% yeasina-kurapa maronda. Biofilms fomu apo microorganiss isingadzivisiki yakabatanidzwa kune maficha, senge shuwa zvevarwi, uye inogona kuunganidzira kuumba yekuwedzera polymer (ePS) -Ukiti nharaunda. Ronda biofilm inosanganiswa nemhinduro yekuwedzera inowedzera inotungamira kune tishu kukuvara, iyo inogona kunonoka kana kudzivirira kupora4. Kuwedzera kweTishu kukuvara kunogona kuve kwakakonzerwa nechikamu chekuwedzera chiitiko cheMatrix metalloparteteines, collagenase, elastase uye inoshanda okisijeni mitezo5. Uyezve, masero anozvimba uye biofilms ari kumusoro kwevatengi veokisijeni uye saka anogona kukonzera nzeve dzemunharaunda hypoxia, ichipedza masero eokisijeni inodiwa kuti ishande tishu
Biofilms yakakura inodzivirira zvakanyanya kune antimicrobial vamiriri, inoda nzira dzehasha dzekudzora hutachiona hwehutachiona, dzakadai sekurapa kwemuchina kwakateverwa nekubata kwakaringana kurapa. Nekuti Biofilms inogona kukurumidza kudzora, inoshanda antimicrobials inogona kuderedza njodzi yekuumbwa mushure mekuvhiya mushure mekuvhiyiwa kwemadhiri7.
Sirivheri iri kuwedzera kushandiswa mune antimicrobial nguo uye inowanzoshandiswa seyambiro-yekutanga kurapwa kwemaronda asina kutapukirwa. Pane zvakawanda zvekushambadzira zviratidzo zvekutengesa, chimwe nechimwe chine chakasiyana yesirivheri, kusangana, uye base matrix. Kufambira mberi muArtirst Iyo simbi yesimbi yesirivheri (AG0) is inert; Kuti uwane antimicrobial kushanda, inofanira kurasikirwa nemagetsi kuti aumbe Ionic sirivheri (AG1 +). Zvinyoro zvesirivha sirivha zvine zvesimbi misheni kana simbi sirivha iyo, kana ichiburitswa nemvura, enda kuumba AG1 + ion. Aya AG1 + ion anogadzirisa neiyo bharo rechirwere, achibvisa magetsi kubva pazvinhu zvemukati kana maitiro ekutsoropodza anodikanwa kuti apone. Tekinoroji tekinoroji yakatungamira kukuvuka kweiyo sirivha nyowani, AG oxysalts (sirivheri nitrate, AG7no11), iyo inosanganisirwa muzvipfeko zveruvhu. Kusiyana sirivheri yechinyakare, kuora kweiyo oksijeni-ine salts inobereka nyika dzesirivheri neEAG1 +, AG2 + naG3 +). MuVitro Ongororo dzakaratidzira kuti dzakaderera dzekuratidzira sirivha sirivha inobudirira kupfuura imwe sirivheri (AG1 +) kubva pseaudenic bactia, staphyloccus auseus uye escherichia coli8,9. Imwe mhando nyowani yekupfeka sirivheri inosanganisira zvimwe zvinongedzo, kureva ethyleentianisetraactraactracetic aclide (BC), izvo zvinonzi zvinyorwe kune biofilm eps uye nekudaro vanowedzera kupinda kwesirivheri mune biofilm. Aya matsva etsiva matekinoroji anopa nzira itsva dzekutarisira vanga biofilms. Nekudaro, kukanganisika kweaya antimicrobials munzvimbo dzemazhinji uye hutachiona-dzakazvimirira kurapa kwakakosha kuve nechokwadi chekuti ivo vasingagadzi nharaunda isina kunaka kana kuti ichinonoka kupora. Kunetseka nezve mu vitro sirivheri sirivha yakataurwa nezvakangoita sirivheri. Nekudaro, mu vitro cytototicity haisati yashanduka muVivo yeVivo yeVivo yeVivo yeVivo yeAG1 yaAg1 yakaratidza yakanaka yekuchengetedza nhoroondo12.
Pano, takatarisa kushanda kweCarboxyMetheselmellose kupfekedzwa kupfeka zvine chekuita nekuvhenekera mafomu ekurwisa vitro uye mu vivo. Uye zvakare, mhedzisiro yekupfeka uku pamhinduro dzekudzivirira uye kupora kwakazvimirira kwekutapukirwa kwakaongororwa.
Kupfeka kwese kwakashandiswa kwaive kuwanika. 3m Kerracen Gel Filer Kupfeka (3m, Knutsford, UK) is a Antimicrobial 100% CarboxyMelloseLellose Matatu Antimacrobial CMC Silver akapfeka, kureva 3M Kerracen ag kupfeka (3m, Knutsford, UK), iyo ine 1.7 wt%. Oxygenated Silver Munyu (AG7no11) muValence Sirlenti Sirtions Ions (AG1 +, AG2 + naG3 +). Munguva yekuenda kwechiitiko cheAG7no11, AG1 +, AG2 + uye AG3 + Ions dzinoumbwa mune chiyero che1: 2: 4. Aquacel AG yakawedzera kupfeka zvine 1,2% sirivha chloride (AG1 +) (Deesidec, leadal +
Marara anoshandiswa muchidzidzo ichi aive Pseudomsonas Aeruginosa Nctc 10781 (Peabhari Hutano hweEngland, Salisbury) uye StaphylocoCuccus Nctc 6571
Buttia yakakurirwa husiku mu muler-hinton muto (Oxoid, Altrincham, UK). Tsika yekupfuura yehukuru yakangoerekana yadyarwa ). ) Colonial biofilm fomu pa37 ° C kwemaawa makumi maviri nemana. Aya makoko ekoloni akaedzwa kuti logarithmic shrkage.
Cheka kupfekwa kuita matatu cm2 masaga zvidimbu uye pre-moisten ine zvisina kujairika madhiri emvura. Isa bandage pamusoro peiyo biofilm yekoloni pane iyo agar ndiro. Chese chose 24 ha e biofilm chakabviswa, uye chinowanikwa bota mukati meiyo biofilm (CFU / mL) zvakavezwa ne serial dilution (10-1 kusvika 10-7) muzuva-angle kusarerekera broth (merck-milipre). Mushure memaawa makumi maviri nemana ekukwana kuCocubation pa 37 ° C, yakajairika mapuranga akaitwa paMueller-Hinton Agar Plates. Imwe neimwe yekurapa uye nguva yenguva yakaitwa mune tsaona, uye plate kuverenga kwakadzokororwa kune imwe neimwe dhifeni.
Nguruve dumbu ganda rinowanikwa kubva kune vakadzi hombe chena nguruve mukati memaminetsi gumi nemashanu ekuuraya zvinoenderana neEuropean Union Export Export Export Lies Ganda rakaveurwa uye rakanatswa nedoro rinopukuta, rinobva romesa pa -80 ° C kwemaawa makumi maviri nemana kuti aongorore ganda. Mushure meTying, 1 CM2 zvidimbu zvegirazi zvakashambwa katatu nePBS, 0.6% Sodium hypochlorite, uye 70% ethanol kwemaminetsi makumi maviri nguva imwe neimwe nguva. Usati wabvisa iyo epidermis, bvisa chero yasara ethanol nekushambidza katatu nguva mune zvisina tsvina pbs. Ganda rakange rakabatanidzwa mune 6-zvakanaka ndiro ne1.45-μM-milki plon membrane (mlik-milipore) ine 3 ml fetal bovine serum (Sigma) yakawedzera ne10% Dulbecco's Yakagadziridzwa Gondo. Pakati (Dulbecco's Modified Eagle Medium - Aldrich Ltd.).
Colonial biofilms yakakurirwa sezvakatsanangurwa kwe biofilm kufumurwa zvidzidzo. Mushure meCustomer Iyo Biofilm paMesemberi kwemaawa makumi manomwe neshanu, iyo biofilm yakaiswa kune iyo ganda pamusoro pekusvibisa loop uye membrane yakabviswa. Iyo biofilm yakabva yanyudzwa pane iyo inguruve dermis yemaawa makumi maviri nemana pa37 ° C kubvumira iyo biofilm yekukura uye inonamatira kune ganda renguruve. Pashure pokunge biofilm yakanga yakura uye yakasungirirwa, prepty yekupfeka, yakasarudzika nemvura yakasvibira yakadzika, yakaiswa zvakananga kuganda kumusoro uye yakaiswa pa37 ° C kwemaawa makumi maviri nemana. Hutachiona hwehukama hwakaonekwa nekusagadzikana nekushandisa prestoblue cell serbs (engitrogen Shandisa leica dfc425 Digital kamera kuenda kuIntery Kutora mifananidzo pane leica Mz8 microscope. Nzvimbo dzine mavara mavara Kuongorora electron microscopy yakaitwa sekutsanangurwa pazasi.
Hutachiona hwehukuru hwakatanga husiku hwakafambiswa 1: 100 muMueller-Hinton muto. 200 μL yetsika yakawedzerwa kune yakasviba 0.2 μM whomyman cyclopore membrane (Whatman, MAIDStone, UK) uye yakarongedzwa paMoueller-Hinton Agar. Mahwendefa ebioflm akaiswa muvhare na37 ° C kweawa makumi manomwe nemasere ekutendera akura biofilm fomu.
Mushure memazuva matatu eBiofilm Maturation, A 3 CM2 Square Bandage yakaiswa zvakananga pane biofilm uye yakavezwa pa37 ° C kwemaawa makumi maviri nemana. Mushure mekubvisa bandage kubva kune biofilm pamusoro, 1 ml yePrestoblue cell viagent Reagent (envitrogen Nzvimbo dzakaomeswa dzisina kuomeswa usati washandura ruvara zvakanyorwa uchishandisa Nikon D2300 Digital Kamera (NIKON UK LTD., Kingston, UK).
Gadzirira Tsika dzehukama paMueller-Hinton Agar, Chinjana Colonies kusvika gumi ML Mueller-Hinton muto uye incubate pane shahker na37 ° C (100 RPM). Mushure mehusiku hwehusiku, tsika yakanyorwa . . Iyo yakakura biofilm yakaiswa kune ronda sezvakatsanangurwa pazasi.
Zvese zvinoshanda nemhuka zvakaitwa kuYunivhesiti yeManchester pasi pechirongwa cheReful Licence Yakabvumidzwa neHofisi Yemhuka Nedheni Yakaburitswa neHofisi Yemusha pasi pe2012 APPA. Vanyori vese vanoomerera kumitungamiri anosvika. Makumi masere-vhiki Makonzo ainyanyoka neIsomlurane (Piramal akarwadziswa neLTD, West Drayton, UK) uye nzvimbo dzadzo dzeDorsal dzakaveurwa uye dzakacheneswa. Imwe neimwe mbeveve yakabva yapihwa 2 × 6 mm stionsal ronda uchishandisa stofel bipsy punch (Schuco International, Hertfordshire, UK). Kune maronda e biofilm-atachiona, isa makumi manomwe nematanhatu-awa coloni. Chikamu chimwe chekirekita chekupfeka chakanyorova chakanyoroveswa nemvura isina kusvibirira kuchengetedza nharaunda yakashata. Kupfekerwa kwaishandiswa zvakananga kune imwe neimwe ronda uye yakafukidzwa ne 3M tegaderm film (3m, bracktell yehutano hwekuzvidza (eloquest healthcare, ferndale, mi) yakaiswa mumipendero yekupa mamwe kuwedzeredza. Buprenorphine (AnimalCare, York, UK) yakapihwa kuiswa kwe1.1 mg / kg se analgesic. Cull makonzo mazuva matatu mushure mekukuvara uchishandisa iyo nzira yehurongwa 1 uye bvisa, halve, uye chengetedza iyo nzvimbo yenguruve sezvinodiwa.
Hematoxylin (Thermofisher Science) uye Eosin (Thermofisher Science) Stain's akaitwa zvinoenderana neiyo protocol. Nzvimbo yemakundu uye reepithelization yaive denderedzwa uchishandisa Image Pro software yeSoftware 10 (Media Cybernetics Inc, Rockville, MD).
Zvikamu zveTisue zvakasikwa muXylene (Thermofisher Science, Loughborough, Ryddrated ne100-50% yakanyorwa nemvura yemvura (iyo yesaiti yemvura (thermofisher sainzi). Immunohistochemistry yakaitwa uchishandisa vertiastin Elite ABC PK-6104 Kit (Vector Laboratories, Burlingame, CA's Producturetr's protocol. Misheni yekutanga kuNitrophils Nimp-R14 (Themmofisher Science) uye Macrophages Ms ABC uye Vector Nova Red Peroxidase (HRP) Substrate Kit (Vector Laboratories, Burlingame, CA) uye akaverengeka neHematolelin. Mifananidzo yakawanikwa uchishandisa Olympus bx43 microscope uye olympus dp73 dialmpus kamera (Olympus, Southend-paGungwa, UK).
Samples yeganda yakatemwa mu2,5% glutaraldehyde uye 4% formaldehyde mu 0,1 m ph 7.4) kwemaawa makumi maviri nemana pa4 ° C. Sampuli dzakasvibiswa uchishandisa Graded Ethanol uye akaomeswa muCO2 ichishandisa Quorum K80logies Ltd Fungidziro dzakafungidzirwa kushandisa iyo FEI QuAnta 250 kuongorora Electron microscope (Thermofisher Science) kuti uone pfungwa yepakati yeiyo ronda.
Toto-1 idode (2 μm) yakaiswa kune yakakwiridzirwa mbevha nzvimbo uye yakagadziriswa kwe5 min pa37 ° C (Tormofisher Science) na 37 ° C (themmofishser Scificy). 15-Minute Z-Stack Mifananidzo yakagadzirwa uchishandisa leica tc8.
Biological uye technical replicate data yaive yakaongororwa uye yakaongororwa uchishandisa graphpad prism v9 software (graphpad software, la golla, ca). Imwe-nzira yekuongorora kusiyanisa nekuenzanisa kweDunnett's post Hoc bvunzo yakashandiswa kuyedza nemisiyano pakati pekurapwa kwega kwega uye iyo is antimicrobial kudzora. A P kukosha <0.05 yaionekwa seyakakosha.
Kubudirira kwesirivheri gel zvipfeko zvekupfeka zvakatanga kuongororwa neBiofilm colonies ye staphylococcus Aureus uye pseudomonas aeruginosa mu vitro. Kupfeka sirivheri kune mafomu akasiyana esirivheri: Zvinyoro zvesirivha sirivheri zvinoburitsa AG1 + ions; Sirivhi nguo, iyo inogona kuburitsa ag1 + ion mushure meEdta / BC, inogona kuparadza iyo biofilm matrix uye kufumura mabhakitiriya kune sirivheri pasi pesirivheri yesirivheri. Ion15 uye kupfeka kuchinyora okisigenated ag munyu unoburitsa AG1 +, AG2 + uye AG2 + ion. Kunoshanda kwayo kwakaenzaniswa neasina-antimicrobial kudzora kupfeka yakagadzirwa kubva pamabwe akakomberedzwa. Kuramba uchiita bactia inoshandisika mukati meiyo biofilm yakaongororwa maawa ese makumi maviri nemana kwemazuva masere (Mufananidzo 1). Pazuva rechishanu, biofilm rakaburitswa na 3.85 × 105s. Staphylococcus Aureus kana 1.22 × 105P. Aeruginosa kuti aongorore biofilm kupora. Kuenzaniswa ne-artimicrobial kutonga, majekisi aGO AG1 aive nemhedzisiro yakaderera pane bacterial viabilcuccus aureus auruginosa biofinsa biofoms pamusoro pemazuva mashanu. Mukupesana, kupfeka kunemukara okisixgenated aG uye ag1 + Edta / BC Salets yaive inoshanda mukuuraya mabhakitiriya mukati meiyo biofilm mukati memazuva mashanu. Mushure mekudzokororwa kudzokororwa nePlanktonic Bactia pazuva 5, hapana kudzoreredzwa kweiyo biofilm yakacherechedzwa (Fig. 1).
Kurehwa kweBhaleabhasi anoshanda muStaphylococccus Aureus uye Pseudomson A Aeruginosa Biofilms mushure mekurapa nejira ririvha. Biofilm colonies ye staphylococcuccus uye pseudomsonas aeruginosa akarapwa nezvipfeko zvesirivha kana zvisiri-artimicrobial controcings, uye huwandu hwehutachiona hunodzora hwasara hwakasara hwaive hwakatsanangurwa maawa makumi maviri nemana. Mushure memazuva mashanu, biofilm yakakomborerwa ne 3.85 × 105s. Staphylococcus Aureus kana 1.22 × 105P. Colonies yeBacerioplankton Pseudomsonas Aeruginosa yakaumbwa mumwe nemumwe kuti uongorore biofilm kupora. Graphs inoratidza zvinoreva +/-yakajairika chikanganiso.
Kuti uone mhedzisiro yemhedzisiro yezvipfeko zvesirivheri pane biofilm budilly, kupfeka kwaishandiswa kune biofilms yakakura pane porcine skin ex vivo. Mushure meawa maawa makumi maviri nemana, kupfeka kunobviswa uye biofilm yakasvibiswa nebhuruu rekodhi, iyo iri metabolized ne bactia yekurarama kune ruvara rwepinki. Biofilms yakarapwa nemapfekero ekutonga aive pink, kuratidza kuvapo kwemabhakisi anoshanda mukati meiyo biofilm (Mufananidzo 2a). Mukupesana, biofilm yakarapwa neAG oxyshsols kupfeka yaive yebhuruu, zvichiratidza kuti iyo bhairasi yakasara pamusoro peganda renguruve risina basa (Mufananidzo 2B). Yakavhenganiswa bhuruu uye yepinki yakaonekwa muBiofilms yakarapwa neAg1 + kuratidza nzvimbo dzisina kukanganiswa nekupfeka sirivheri (nhamba 2d). Kurehwa kweiyo inoshanda (pink) uye isingashande (yebhuruu) nzvimbo dzakaratidza kuti chigamba chaive chechiitiko ichizoshanda 78% (Mufananidzo 2e). AG1 + + Edta / BC Draphiro yakaitwa zvakafanana kune oxygenated AG mungozi wemunyu, nemitero ye13% uye 14%, zvichiteerana. Iyo AG1 + kupfeka + kupfeka zvakare yakaderedzwa cabical viability ne 21%. Idzi biofilms dzakazoonekwa dzichishandisa scanning electron microscopy (SEM). After treatment with the control dressing and the Ag1+ dressing, a layer of Pseudomonas aeruginosa was observed covering the porcine skin (Figure 2F,H), whereas after treatment with the Ag1+ dressing, few bacterial cells were found and few bacterial cells were found underneath. Colagen Fibers inogona kutariswa seye tishu chimiro cheganda re porcine (Mufananidzo 2G). Mushure mekurapwa neAG1 + + Edta / BC kupfeka, mabhata ezvipfeko uye pasi peColagen Fiber Plubes aionekwa (Mufananidzo 2I).
Visualization yePseudomonas aeruginosa biofilm mushure mekurapwa kwesirivheri. . Kurarama mabhakitiriya ari pink, asiri-anobatsira mabhakitiriya uye ganda renguruve nderebhuruu. . Sem sero saizi bar = 5 μm. .
Kuti uone kana padhuze nepamusoro pekupfekedzwa nepamhepo uye biofilms yakakanganisa zvakanyanya zvekupfekedza Iyo isina kutapwa biofilm yaive yakasviba pink muvara (Mufananidzo 2j). Mukupesana neBiofilms akabata neCraps Iyi pink claping inoratidza kuvapo kwehutsinye hwehutachiona uye hunobatanidzwa nenzvimbo yerudzi mukati mekupfeka. Iyi sewebhu-munzvimbo dzinogadzira nzvimbo dzakafa dzinobvumira mabhakitiriya mukati meiyo biofilm kuti rirarame.
Kuti aongorore kushanda kwekupfekedza kwesirivheri muVivo, kwakazara maronda emakonzo ane hutachiona Mushure memazuva matatu ekurapa, macroscopic jawu ongororo yakaratidza madiki madiki pakaitwa nekupfekedzwa kwemunyu kwemunyu kunoenzaniswa nemapfekero asiri ekupfeka uye zvimwe zvipfeko zvesirivha (Mufananidzo 3a-h). Kuti uone izvi zviono, maronda aya akakodzwa nzvimbo uye ronda uye reepithelialis pane hematoxixylin zvikamu uye eosin-yakasvibiswa tissue zvikamu uchishandisa image pro software 10 (Mufananidzo 3I-L).
Mhedzisiro yezvipfeko zvesirivheri pane ronda pamusoro uye zvakare-epithelialization yemaronda ane hutachiona ne biofilms. (A-h) masero madiki ane biofilms ePseudomonas (a-d) uye staphylocccucc Control Control, AG1 AG1 kupfeka, uye AG1 + kupfeka. Mumiriri wecroscopic mifananidzo. Maronda emakonzo ane AG1 + Edta / BC kupfeka. . Kuregedzwa kwenzvimbo yemakumbo (m, m, o) uye muzana reepithelialization (N, p) yemaronda ane pseududomonas (o, p) biofilms (pane biofilms (pane biofilms (pane biofilms (pane kurapwa kweboka n = 12). Graphs inoratidza zvinoreva +/-yakajairika chikanganiso. * zvinoreva p = <0.05 ** zvinoreva p = <0.01; Macroscopic chiyero = 2,5 mm, his hamuzano = 500 μm.
Kuregedzwa kwenzvimbo yemaronda mumaronda ane pseudomsonas aerudulinosa biofinosa biofinosa bioficm Chokwadi. yakasvika kukosha kwehuwandu (Mufananidzo 3m). p = 0.423). Maronda akarapwa neAG1 + kana AG1 + + edta / BC akarishandisa pasina kuderedzwa kwenzvimbo yevaraya (3.1 mm2 uye 3.6 mm2, zvichiteerana). Kurapa neOxygenated AG Munyu Wemunyu Akasimudzirwa RE-EpitHelialization kune iyo Yakakura Kupfuura Kupfeka Kupfeka (34% uye ne1.029) uye Ag1 + Edta / BC (10% uye 11%) ( Mufananidzo 3n). . , zvichiteerana).
Maitiro akafanana muRonda Area uye Epithelial Kuzoratidzwa kwaonekwa mumaronda ane hutachiona neS. Aureus Biofilms (Mufananidzo 3o). Kupfekedzwa kupfeka kune oxygenated sirivha yakaderedzwa nzvimbo (2.0 mm2) ne20% kana ichienzaniswa neyekutora Uye zvakare, iyo nzvimbo yemavanga muAg1 + boka rekurapa raideredzwa zvishoma Oxygen Salts yeAG zvakare yakakurudzirwa zvakare ye-Epithelialization yerwiro dzinotakura neSauselm (31%) kusvika kune isina-kurwisa AG1 + kupfeka (16%, p = 0.903) uye ag + edta / bc kupfeka (14%, P =
Kuti uone mhedzisiro yemhedzisiro yezvipfeko zvesirivheri pane biofilm matrix, toto 1 uye syto 60 iodside staining (Fig. 4). Toto 1 iodide is sero-isinganyatsoita dhayi inogona kushandiswa kunyatsoona Exclatellular lecleic acids, iyo yakawanda mumaps of biofilms. Syto 60 is sero inobvumirwa dhayi inoshandiswa seyakaisa nyaya1. Kucherechedzwa kwetoto 1 uye Syto 60 Iodide mumaronda akasungwa neBiofinomonas (Mufananidzo 4a-D) zvakaratidza kuti mushure memazuva matatu ekupfekedza, EPS muBiofilm yakaderedzwa zvakanyanya. Inayo okisixgenated Salts aG uye AG1 + Edta / BC. AG1 + kupfeka pasina zvimwe zvinowedzera zveantibiofilm zvikamu zvakanyanya kuderedza cell-Mahara eDNA mumaronda akasungwa nePseudomson
Mu vivo kufungidzira kwevarairidzo biofilm mushure memazuva matatu ekurapa nekutonga kana sirivheri. Confocal mifananidzo yePseuginosa (A-D) uye staphylococcus Aureus (i-l) yakasvibiswa neyakaona Acclecucular Kusunga mubatanidzwa wechiitiko, shandisa syto 60 (tsvuku). acids. P. Scaning Electron microscopy yemaronda ane pseudomonas aeruginosa (e-h) uye staphylococcus aureus (m-p) biofilms mushure memazuva matatu ekurapa uye sirivheri. Sem sero saizi bar = 5 μm. Confocal kufungidzira chiyero bar = 50 μm.
Scaning Electron Microscopy yakaratidza kuti makonzo akasungwa ne biofilm colonies yePsegusonas Aureus (Mufananidzo 4m-h) zvakave nemaronda mashoma mushure memazuva matatu ekurapa neese sirivheri.
Kuti aongorore mhedzisiro yezvipfeko zvesirivha peronda muBiofilm-yakatapukirwa-madhiri ezvekutapudza kwemazuva matatu. Kutsunga kwehuwandu hweNetrophils uye macrophagy mukati. granulation tishu. Mufananidzo 5). Mapfekero ese eSirl akaderedza huwandu hwetrophils uye macrophalls mumaronda ane hutachiona eruginosa akaenzaniswa neyekutora mfekero ekutonga mushure memazuva matatu ekurapa. Nekudaro, kurapwa neoxygenated sirivha sirivha yakakonzera kuderedzwa kukuru muNeatrophils (p = <0. 0. 0. 0.00001) Kunyangwe ag1 + + edta / BC aive aine mari yakakura pakuvhuna biofilm, yakaderedza neutrophil naMacrophage mazinga mashoma pane kuderera pane iyo AG1 kupfeka kwaAg1. Maronda ane mwero ane hutachiona naSauseus biofilm akaonekwawo mushure mekupfeka neAG (P = <0.0 edta / bc (p = 0.003 Mafambiro akafanana anoonekwa neutropenia. bandage (fig. 5k). Nekudaro, chete oxygenated ag munyu wakaratidza kukosha kwakakura muhuwandu hweMacrophaes muGranuse tishu mumaronda ane hutachiona
Neutrophils uye MacProphes zvaive nehurombo hutachiona nePseudomonas Aeruginosa uye staphylocccus Aureus Biofilms mushure memazuva matatu ekurapa neasina-antimicrobial kudzora kana kupfeka sirivheri. Neutrophils (Ad) neMacrophages (EH) zvaive zvakabatanidzwa muzvikamu zve tishu zvakarongedzwa nemanyodhi akananga kune neutrophils kana macrophills kana macrophies. Kurehwa kweNeutrophils (i naK) neMacrophages (j uye l) mumaronda ane pseududomonas (i uye staphylocccus aureus (k & l) biofilms. N = 12 paboka. Graphs Ratidza zvinoreva +/-standard chikanganiso, kukosha kwakakosha kunoenzaniswa neye antibacterial control kupfeka, * zvinoreva p = <0.05, ** zvinoreva p = <0.01; *** zvinoreva p = <0.001; inoratidza p = <0.0001).
Isu takabva taongorora mhedzisiro yezvipfeko zvesirivheri pane hutachiona. Maronda asiri ehutsinye akabatwa neasina-antimicrobial kudzora kupfeka kana kupfeka sirivheri kwemazuva matatu (Mufananidzo 6). Pakati pezvipfeko zvesirivheri zvinoongororwa, chete maronda akabatwa neyekupfeka kwemunyu wemunyu akaonekwa zvidiki pamacroscopic mifananidzo pane maronda akarapwa nehurombo (Mufananidzo 6a-D). Kuregedzwa kwenzvimbo yemakumbo uchishandisa Hunoshuwa kuongorora kwakaratidza kuti nzvimbo yevarwere vakurumwa mushure mekurapwa neAg Onsers . 6Ini). Mukupesana, hapana kuderedzwa kwenzvimbo yemavanga kucherechedzwa mushure mekurapwa mushure mekurapwa neAG1 + (3.38 mm2, 0,797) kana edta / bc (4.18 mm2, p = Kuwedzera Epithelial Kuzivisa neAG OXYYSL Kupfeka Kwekudzora Boka (30% vs. 22%, izvi hazvina kusvika), izvi zvakakosha uye zvichisimbisa mibairo yapfuura. Kupfeka neoyshysolss inosimudzira re-Epithelialization. -Pitarization yemiwande isina kutapukirwa17. Mukupesana, kurapwa neAG1 + kana AG1 + Edta / BC Draphing yaive isina mhedzisiro kana kuratidza kuderera kwe-Epithelialization kuenzanisa nekudzora.
Mhedzisiro yesironda ruva rekupfeka paronda kupora mumakonzo asina kutapurira. . . Kuregedzwa kwenzvimbo yevaraya (i) uye peresenti yeReepitHelialization (J) yakaverengerwa kubva muzvikamu zvehunyanzvi paMitero yekuongorora kwemifananidzo (n = 11-12 pane kurapwa kweboka). Graphs inoratidza zvinoreva +/-yakajairika chikanganiso. * zvinoreva p = <0.05.
Sirivheri ine nhoroondo refu yekushandiswa seyakareba kurapa murima kupora, asi mafambiro akawanda akasiyana uye nzira dzekutakura dzinogona kukonzera kusawirirana muAntimicrobial Elesses. Uyezve, iyo antibiofilm zvivakwa zvetsiva zvakananga Sirivha masisitimu haana kunyatsonzwisiswa. Kunyangwe iyo yekurova yemuviri yekudzivirira ine chekuita nehurongwa hwekupokana nePlishktonic Bactia, kazhinji hazvibatsiri kurwisa biofilms19. Buttia rePlanktonic dzinogona kurongwa Izvo zvakaonekwa kuti mamwe leukocytes anogona kupinza biofilms21 asi akatadza kuPhagocytose Bactia kana ichi chidziviriro ichi chakamisikidzwa22. Maitiro eHolistic anofanirwa kushandiswa kutsigira nhambo yehondo yekudzivirira pamusoro pevanga biofilm utachiona. Kuzvivara kwemarara kunogona kukanganisa biofilm uye bvisa yakawanda ye biobelched, asi iyo yekushambadzira yekushomeka inogona kusashanda kune yakasara biofilm, kunyanya kana iyo yekushambadzira maumine mhinduro inokanganiswa. Saka, kurapa kweanotamba kwakadai sokupfeka sirivheri kunogona kutsigira nhambo yehondo yekudzivirira uye kubvisa hutachiona hweBiofilm. Kuumbwa, kusimbisa, Solubility, uye kuendesa Kukurukurirana kunogona kukanganisa antimicrobial kushanda kwe sirivheri. Mumakore achangopfuura, kufambira mberi musungwa rekugadzirisa sirivha kwakaita kuti zvipfeko izvi zvikure zvakanaka9,23. Sevhu ririvha rinofambira mberi-sirivha rinofambira mberi pakukura kwezvakapfeka izvi mukudzora zvemawere uye, zvakanyanya kukosha, kukanganiswa kwemhando idzi sirivha pakatemaruva uye kupora.
Muchidzidzo ichi, takafananidza nekubudirira kwevakapfeka zvipfeko zviviri zvepamberi nezvipfeko zvesirivha zvirema zvinogadzira AG1 + INS SIL1 Isu takaongororawo mhedzisiro yezvipfeko izvi pane zvavanga zvakatipoteredza uye hutachiona-hwakazvimiririra kupora. Kuti uderedze pesvedzero yeiyo Matrix Matrix, zvese zvipfeko zvesirivha zvinoongororwa zvakaumbwa zvearboxymethcylosellose.
Yedu yekutanga kuongororwa kweiyi zvipfeko zvesirivha kubva kuPseudomonas Aeruginosa uye StaffEccus AG1 + Edta / BC SEAFEGED AG SALTS, inobata ne5. Zvinoshanda zvinouraya biofilm bacteria mukati mazuva mashoma. Pamusoro pezvo, idzi zvipfeko dzinodzivirira kuumbwa kwe biofilm pane kudzokorora kuratidzwa kune planktonic mabhakitiriya. Iyo AG1 + yekupfeka iri ine sirivha chloride, iyo sirivheri yakaenzana uye base matrix se ag1 + edta / bc, uye yaive nemhedzisiro yekubatira bota mukati meiyo biofilm pane imwe nguva. Iko kucherechedza uko AG1 + + Edta / BC Dressing yaive inoshanda neBiofilm pane iyo Matrix inodikanwa kuti uwedzere kushanda kweiyo sirivheri kumwe kunhu15. Mhedzisiro iyi inotsigira iyo pfungwa yekuti Bc naEdta play chiitiko chekuwedzera kupfekedza kushanda uye kusavapo kwechikamu ichi muAg1 + kupfeka kunogona kunge kwakakonzera kutadza kuratidza muVitro kushanda kwevitro. Takaona kuti oxygenated ag salt prassing kuchigadzira AG2 + uye AG3 + IIS yakaratidza yakasimba Antibacterial Aug1 + uye pamatanho akafanana neAG1 + Edta / BC. Nekudaro, nekuda kwekukwira kweRedox kugona, hakuna kujeka kuti ag3 + long3 + ingoita nguva yakareba sei inoshanda uye inoshanda pamusoro pevhu biofilms uye saka inofanirwa kuenderera mberi nekudzidza. Pamusoro pezvo, pane akawanda madhirowa akasiyana anogovera AG1 + ions isina kuedzwa muchidzidzo ichi. Izvi zvipfeko zvinoumbwa nemakomisheni akasiyana-siyana, zvivakwa, uye zvigadziko zvemateki, izvo zvinogona kukanganisa kutumirwa kweAG1 + iS uye kushanda kwavo kune biofilms. Zvakakoshawo kuziva kuti kune vazhinji vakasiyana muVitro uye muVivo Modiels vaishandisa kuongorora kushanda kwekurwara kwekurwara kurwisa biofilms. Rudzi rwemuenzaniso rwakashandiswa, pamwe nemunyu uye mapuroteni ezvinyorwa zvinoshandiswa mumhando idzi, achapesvedzera kushanda kwekupfeka. Mudu muVivo Model, takabvumira iyo biofilm yekukura mu vitro uye yakabva yaendesa kune iyo denmal pamusoro penjodzi. Iyo inogamuchira mbeva yemumhepo yekushomeka ndeyekuti inoshanda nePlanktonic Bacsertia yakashandiswa kune vanga, nekudaro vachiumba biofilm sezvo iyo ronda renjodzi. Kuwedzera kweBiofilm kwakakura kune ronda rinodzikisira kushanda kweiyo yekushambadzira yekudzivirira kweBiofilm yekuumbwa nekubvumira iyo Biofilm yekuisa pachayo kuti isati yasimudzwa. Nekudaro, muenzaniso wedu unotibvumidza kuongorora kushanda kwekushandisa kwehutongi hwekupfekedza kwakakura pane dzimba dzakura dzimba dzakatarisana nemirwere inotanga kupora.
Isu takawanawo kuti kupfeka kwakakwenenzverwa kwakapesvedzera kugona kwezvipfeko zvesirivha zviri muVitro-Goofms uye Ganda rePorcine. Kusangana padhuze ne ronda kunoonekwa kwakashata kune iyo Intimicrobial Kubudirira kwekupfeka24,25. Kupfekerwa kune Oxygenated ag Salts vaive padhuze nepedyo neBiofilms vakakura, zvichikonzera kuderedzwa kwakakura muhuwandu hwemabhakisi anoshanda mukati meiyo biofilm mushure maawa makumi maviri nemana. Kusiyana neizvi, kana kurapwa neAG1 + uye AG1 + Edta / BC Driving, nhamba dzakakosha dzemaoko anobatsira akashanda. Izvi zvipfeko zvine chekunyepa padhuze nehurefu hwese hwekupfeka, izvo zvinogadzira nzvimbo dzakafa dzinotadzisa kusangana neiyo biofilm. Muzvidzidzo zvedu muVitro, nzvimbo idzi dzisiri-dzekutaurirana dzakatadzisa kuuraya kwebhakitiriya rinoshandika mukati meiyo biofilm. Takaongorora butti bacterial viability mushure meawa makumi maviri nemana ekurapwa; Nekufamba kwenguva, sezvo kupfeka kunowedzera kugutsurwa, panogona kunge paine nzvimbo isina kufa, ichidzora nzvimbo yeiyi mabhakitiriya anoshanda. Nekudaro, izvi zvinoratidzira kukosha kwekuumbwa kwekupfeka, kwete mhando yesirivheri mukupfeka.
Ndichiri muVitro Ongororo dzinobatsira pakuenzanisa kushanda kwesirivheri sirivheri, zvakakoshawo kunzwisisa zvinopfekedzwa zveBiofms muBiofms uye mhinduro dzisingamiriri Mhedzisiro yeaya majira pane vanga biofils yakaonekwa uchishandisa kuongorora Equiroscopy elekroscopy uye ePs Takaona kuti mushure memazuva matatu ekurapa, zvese zvipfeko zvaive zvichishanda mukudzora cell-emahara mu biofilm-hutachiona-chakarukwa + chipfeko chaive chisiri kushanda mumaronda eAureus-hutachiona. Kuongorora Electron Microscopy kwakaratidzawo kuti mabhakitiriya akakosha aive aripo mumaronda akabatwa nezvipfeko zvesirivheri, + AG1 (+ edta / bc kupfeka kunoenzaniswa neAG1 + kupfeka. Idzi data dzinoratidza kuti zvipfeko zvesirivheri zvakaverengerwa zvakakanganisa madhigirii ezvemhando yepamusoro, asi hapana chimwe chezvipfeko chesirivha chakakwanisa kudzima biofilm, kutsigira kudiwa kwenzira yekubatwa nehutachiona hwehupfumi hweronda. kushandiswa kwesirivhare. Kurapwa kunotangirwa nekushomeka kwemuviri kubvisa yakawanda yeiyo biofilm.
Maronda erambadza anowanzove ari mumamiriro ekuzvimba zvakanyanya, aine masero akawandisa akasununguka akasarudzika kwenguva yakareba, achikonzera okisijeni inodiwa kune anoshanda metabolus uye inoshanda muhwonda. Biofilms inowedzera iyi nzvimbo ine njodzi nekukasira kukanganisa kurapa munzira dzakasiyana siyana, kusanganisira kudzivirira kwesero kuwedzera uye kutama uye kuiswa kweProinflamiritsi Cytokugara Cytokutorarati Cytokines27. Sezvo zvipfeko zvesirivheri zvinoshanda zvinobudirira, zvakakosha kuti unzwisise zvakaringana zvavanazvo pakatovirisa nzvimbo uye kupora.
Sezvineiwo, kunyangwe zvipfeko zvese zvesirivha zvakakanganisa kuumbwa kweBixym Idzi data ritsigira yedu yapfuura kuwana kuwana17 uye avo veKalan et al. .
Kudzidza kwedu zvazvino kunosimbisa misoro yematongerwo enyika pakati pemapfekero eantimucrobial sirivheri uye kukanganiswa kwehunyanzvi uhwu pakatarisana nenhamo. Nekudaro, izvi zvinobuda zvakasiyana nezvidzidzo zvepamberi zvinoratidza kuti AG1 + Edta / BC kupfeka kwakavandudza maratidziro ezvakakuvadzwa nenzeve dzakakuvara mu vivo. Nekudaro, izvi zvinogona kuve nekuda kwemisiyano mumhando yemhuka, kuyerwa nguva, uye mabharari ekushandisa nzira29. Mune ino kesi, zviyero zveruvanga zvakatorwa mazuva gumi nemaviri mushure mekukuvara kubvumira izvo zvinoshanda zvinongedzo zvekupfeka kuti uite pane iyo biofilm kwenguva yakareba. Izvi zvinotsigirwa nechidzidzo chakaratidza kuti chirwere chehurombo chakabatwa neAL1 + Edta / BC SEVEL LEATE BEATE NEG1 (+ Edta / BC uye Mukati Makumi mana e kushandiswa kwe asiri antimicrobials. zvinodhaka. CMC kupfeka kuderedza saizi yehutanho30.
Mamwe mafomu uye esirivheri zvakamboratidzwa kuve cytotoxic muVitro 11, asi izvi mumhedzisiro yeVitro haigone kushandurwa kuita zvakaipa mu vivo. Mukuwedzera, kufambira mberi kwesirivheri tekinoroji uye nzwisiso iri nani yezvinyorwa zvesirivheri uye kutarisisa kwakapfeka kwakakwenenzverwa kwakakura kukuvadzwa kwevakawanda vakachengeteka uye vanoshanda sirivha. Zvisinei, sesirivheri yekupfeka tekinoroji inofambira mberi, zvakakosha kuti unzwisise kukanganiswa kweaya majira pakatarisana nenzvimbo Izvo zvakambotaurwa kuti kuwanda kwemari yekudzokorora kunoenderana neyekuwedzera kuwanda kweAnti-Farlammatory M2 Macrophags Izvi zvakacherechedzwa mune yadarika mbeva modhiyo uko Sirl Hydrogel Wand crapy yakafananidzwa ne Silver Sulfadiaazine uye isiri-antimicrobial hydrogels34.
Maronda ekusingaperi anogona kuratidza kuzvimba kwakanyanya uye zvakacherechedzwa kuti kuvapo kwesimba rakawandisa kungave kwakashata kukuvadza kuporesa35. Mukudzidza mumakonzo-akatsemuka-akapfeka mbeva, kuvapo kweNetrophils kunonoka kuora. Kuvapo kwekunyanyisa neutrophils kunotungamira kune yakakwira mapoka emhando dzekupedza uye sosi yemaokisijeni, senge superoxide uye hydrogen peroxide, iyo inosanganiswa nemaronda anosereya uye anononoka kupora uye anononoka kupora. Saizvozvowo, kuwedzera muMacrophage nhamba, kana kusingadzoreki, kunogona kutungamira kunonoka kuvanda kuporesa39. Kuwedzera uku kwakanyanya kukosha kana Macrophages kutadza kushandurwa kubva kuPro-Farlamiture phenotype kune pro-kupora phenotype, zvichikonzera maronda ari kutadza kubuda chikamu chekuporesa40. Takaona kuderera kweNetrophils uye macrophills maronda eBiofilm mushure memazuva matatu ekurapa nezvipfeko zvese zvesirivha, asi kuderera kwacho kwacho kwacho kwacho kwacho kwacho kwacho kwakataurwa nekupfekedza kwemugodhi wemugunwe. Kuderera uku kungave kwakananga kwemhinduro yekudzivirirwa kune sirivheri, mhinduro yekuderedza Biobelden, kana kuti ronda riri mune imwe nhanho yekupora uye saka masero emagetsi ari muonda acho ari kuderedzwa. Kuderedza huwandu hwemasero anozvimba mune ronda anogona kuchengetedza nharaunda yakakodzera kupora. Mashandiro echiitiko chekuita kuti AG obysalts inosimudzira kurapwa isingabudisike, asi kugona kweiyo hydsagen peroxide, murevereri wekuzvimba, anogona kutsanangura izvi uye zvinoda kuenderera mberi nekufunda17.
Maronda asina kurapa anotapukira anoisa dambudziko kune vanachiremba uye varwere. Kunyangwe akawanda macressing akawanda achiti antimicrobial kushanda, kutsvagisa hakuwanzo kutarisa pane zvimwe zvinhu zvakakosha zvinokanganisa iyo micronvironment. Ichi chidzidzo chinoratidza kuti kwakasiyana sirivha tekinoroji kushanda kwakasiyana uye, zvakakosha, mhedzisiro dzakasiyana-siyana pamatare mazhinji uye kupora, kwakazvimiririra. Kunyangwe izvi muVito zvidzidzo uye muVivo zvidzidzo zvinoratidza kushanda kwekupfeka uku kurapa hutachiona uye kusimudzira miedzo yekupora, yakasarudzika inodzorwa inodiwa kuti uongorore kugona kweiyi machira.
Iyo madheti anoshandiswa uye / kana kuongororwa panguva yazvino yekudzidza inowanikwa kubva kune inoenderana munyori pane chikumbiro chine musoro.
Kutumira Nguva: Jul-15-2024