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ʻO ka ulu ʻana o nā microbial i loko o nā ʻeha e hōʻike pinepine ʻia ma ke ʻano he biofilms, kahi e hoʻopilikia ai i ka ho'ōla ʻana a he paʻakikī ke hoʻopau. Ke koi nei nā ʻaʻahu kālā hou e hakakā i nā maʻi ʻeha, akā ʻaʻole ʻike nui ʻia ko lākou pono antibiofilm a me nā hopena ho'ōla kūʻokoʻa i ka maʻi. Ma ka hoʻohana ʻana i nā hiʻohiʻona biofilm in vitro a me in vivo o Staphylococcus aureus lāua ʻo Pseudomonas aeruginosa, hōʻike mākou i ka pono o nā ʻaʻahu hana ion Ag1+; Nā ʻaʻahu Ag1+ e loaʻa ana ka ethylenediaminetetraacetic acid a me benzethonium chloride (Ag1+/EDTA/BC), a me nā ʻaʻahu e loaʻa ana ka nitrate kālā (Ag oxysalts). , e hana ana i nā ion Ag1+, Ag2+ a me Ag3+ e hakakā i ka biofilm ʻeha a me kona hopena i ka ho'ōla ʻana. He liʻiliʻi nā hopena o nā ʻaʻahu Ag1+ i ka biofilm ʻeha ma vitro a me nā ʻiole (C57BL/6j). I ka hoʻohālikelike ʻana, ua hoʻemi nui nā paʻakai Ag oxygenated a me nā ʻaʻahu Ag1+/EDTA/BC i ka helu o nā bacteria ola i loko o nā biofilms in vitro a ua hōʻike i ka emi nui ʻana o nā ʻāpana bacteria a me EPS i loko o nā biofilms ʻeha ʻiole. He ʻokoʻa nā hopena o kēia mau kāhiko ʻana i ka ho'ōla ʻana o nā ʻeha i loaʻa i ka biofilm a me nā ʻeha i loaʻa ʻole i ka biofilm, me nā kāhiko paʻakai oxygenated i loaʻa nā hopena maikaʻi aʻe ma ka reepithelialization, ka nui o ka ʻeha, a me ka mumū i hoʻohālikelike ʻia me nā lāʻau lapaʻau kaohi a me nā kāhiko kālā ʻē aʻe. Hiki i nā ʻano physicochemical like ʻole o nā kāhiko kālā ke loaʻa nā hopena like ʻole ma ka biofilm ʻeha a me ka ho'ōla ʻana, a pono e noʻonoʻo ʻia kēia i ke koho ʻana i kahi kāhiko no ka mālama ʻana i nā ʻeha i loaʻa i ka biofilm.
Ua wehewehe ʻia nā ʻeha mau loa ʻo ia hoʻi "nā ʻeha i hiki ʻole ke holomua ma o nā pae maʻamau o ka ho'ōla ʻana ma ke ʻano maʻamau a me ka manawa kūpono" 1. Hoʻokumu nā ʻeha mau loa i kahi kaumaha noʻonoʻo, pilikanaka a me ka hoʻokele waiwai no nā maʻi a me ka ʻōnaehana mālama ola. Ua manaʻo ʻia ka hoʻolilo kālā makahiki a ka NHS no ka mālama ʻana i nā ʻeha a me nā maʻi pili i ka £8.3 biliona ma 2017–182. He pilikia koʻikoʻi nō hoʻi nā ʻeha mau loa ma ʻAmelika Hui Pū ʻIa i kēia manawa, me ka manaʻo o Medicare i ke kumukūʻai makahiki o ka mālama ʻana i nā maʻi me nā ʻeha ma $28.1–$96.8 biliona3.
ʻO ka maʻi kahi kumu nui e pale ai i ka ho'ōla ʻana o ka ʻeha. Hōʻike pinepine ʻia nā maʻi ma ke ʻano he biofilms, aia ma 78% o nā ʻeha mau loa ʻaʻole e ho'ōla. Hoʻokumu ʻia nā biofilms i ka wā e hoʻopili mau ʻia ai nā microorganism i nā ʻili, e like me nā ʻili ʻeha, a hiki ke hōʻuluʻulu e hana i nā kaiāulu hana polymer extracellular (EPS). Pili ka biofilm ʻeha me ka hoʻonui ʻia o ka pane mumū e alakaʻi ana i ka hōʻino ʻana o nā ʻiʻo, hiki ke hoʻopaneʻe a pale paha i ka ho'ōla4. ʻO ka hoʻonui ʻia o ka hōʻino ʻana o nā ʻiʻo ma muli paha o ka hoʻonui ʻia o ka hana o nā matrix metalloproteinases, collagenase, elastase a me nā reactive oxygen species5. Eia kekahi, ʻo nā cell mumū a me nā biofilms he mau mea hoʻohana kiʻekiʻe o ka oxygen a no laila hiki ke hoʻoulu i ka hypoxia o nā ʻiʻo kūloko, e hoʻopau ana i nā cell o ka oxygen koʻikoʻi e pono ai no ka hoʻoponopono ʻana i nā ʻiʻo6.
He kūpaʻa loa nā biofilms makua i nā mea antimicrobial, e koi ana i nā hoʻolālā ikaika e kāohi i nā maʻi biofilm, e like me ka mālama mechanical a ukali ʻia e ka mālama antimicrobial kūpono. No ka mea hiki i nā biofilms ke hana hou koke, hiki i nā antimicrobial kūpono ke hōʻemi i ka pilikia o ka hana hou ʻana ma hope o ka debridement ʻoki kino7.
Hoʻohana nui ʻia ke kālā i nā ʻaʻahu antimicrobial a hoʻohana pinepine ʻia ma ke ʻano he lāʻau lapaʻau mua no nā ʻeha i loaʻa i ka maʻi mau. Nui nā ʻaʻahu kālā i loaʻa ma ke kālepa, kēlā me kēia me kahi ʻano kālā like ʻole, ka nui, a me ka matrix kumu. Ua alakaʻi ka holomua i nā ʻaʻahu lima kālā i ka hoʻomohala ʻana o nā ʻaʻahu lima kālā hou. ʻO ke ʻano metala o ke kālā (Ag0) he inert; No ka hoʻokō ʻana i ka pono antimicrobial, pono ia e nalowale i kahi electron e hana i ke kālā ionic (Ag1+). Loaʻa i nā ʻaʻahu kālā kuʻuna nā hui kālā a i ʻole ke kālā metala, i ka wā e hōʻike ʻia ai i ka wai, e palaho e hana i nā ion Ag1+. Hoʻopili kēia mau ion Ag1+ me ka cell bacteria, e wehe ana i nā electrons mai nā ʻāpana kūkulu a i ʻole nā kaʻina hana koʻikoʻi e pono ai no ke ola. Ua alakaʻi ka ʻenehana patent i ka hoʻomohala ʻana o kahi hui kālā hou, ʻo Ag oxysalts (silver nitrate, Ag7NO11), i hoʻokomo ʻia i loko o nā ʻaʻahu ʻeha. ʻAʻole like me ke kālā kuʻuna, ʻo ka palaho ʻana o nā paʻakai oxygen e hana i nā kūlana kālā me ka valence kiʻekiʻe (Ag1+, Ag2+ a me Ag3+). Ua hōʻike ʻia nā haʻawina in vitro he ʻoi aku ka maikaʻi o nā haʻahaʻa haʻahaʻa o nā paʻakai kālā oxygenated ma mua o ke kālā ion hoʻokahi (Ag1+) e kūʻē i nā bacteria pathogenic e like me Pseudomonas aeruginosa, Staphylococcus aureus a me Escherichia coli8,9. ʻO kekahi ʻano hou o ke kāhiko kālā e komo pū ana me nā mea hoʻohui hou, ʻo ia hoʻi ka ethylenediaminetetraacetic acid (EDTA) a me benzethonium chloride (BC), i hōʻike ʻia e kuhikuhi i ka biofilm EPS a pēlā e hoʻonui ai i ke komo ʻana o ke kālā i loko o ka biofilm. Hāʻawi kēia mau ʻenehana kālā hou i nā ala hou e kuhikuhi ai i nā biofilms ʻeha. Eia nō naʻe, he mea nui ka hopena o kēia mau antimicrobial ma ke kaiapuni ʻeha a me ka ho'ōla kūʻokoʻa i ka maʻi e hōʻoia ʻaʻole lākou e hana i kahi kaiapuni ʻeha maikaʻi ʻole a hoʻopaneʻe paha i ka ho'ōla ʻana. Ua hōʻike ʻia nā hopohopo e pili ana i ka cytotoxicity kālā in vitro me kekahi mau kāhiko kālā10,11. Eia nō naʻe, ʻaʻole i unuhi ʻia ka cytotoxicity in vitro i loko o ka toxicity in vivo, a ua hōʻike kekahi mau kāhiko Ag1+ i kahi ʻano palekana maikaʻi12.
Maanei, ua noiʻi mākou i ka pono o nā ʻaʻahu carboxymethylcellulose e loaʻa ana nā ʻano kālā hou e kūʻē i ka biofilm ʻeha in vitro a in vivo. Eia kekahi, ua loiloi ʻia nā hopena o kēia mau ʻaʻahu i nā pane pale a me ka ho'ōla kūʻokoʻa o ka maʻi.
Ua loaʻa nā ʻaʻahu a pau i hoʻohana ʻia ma ke kālepa. ʻO 3M Kerracel Gel Fiber Dressing (3M, Knutsford, UK) he ʻaʻahu fiber gel 100% carboxymethylcellulose (CMC) non-antimicrobial i hoʻohana ʻia ma ke ʻano he ʻaʻahu hoʻomalu ma kēia haʻawina. Ua loiloi ʻia ʻekolu mau ʻaʻahu kālā CMC antimicrobial, ʻo ia hoʻi ka ʻaʻahu 3M Kerracel Ag (3M, Knutsford, UK), nona ka 1.7 wt%. paʻakai kālā oxygenated (Ag7NO11) i loko o nā ion kālā valence kiʻekiʻe (Ag1+, Ag2+ a me Ag3+). I ka wā o ka decomposition o Ag7NO11, ua hoʻokumu ʻia nā ion Ag1+, Ag2+ a me Ag3+ ma ka lakio o 1:2:4. ʻO ka ʻaʻahu Aquacel Ag Extra e loaʻa ana he 1.2% silver chloride (Ag1+) (ConvaTec, Deeside, UK) 13 a me ka ʻaʻahu Aquacel Ag + Extra e loaʻa ana he 1.2% silver chloride (Ag1+), EDTA a me benzethonium chloride (ConvaTec, Deeside, UK) 14.
ʻO nā ʻano i hoʻohana ʻia ma kēia noiʻi ʻo Pseudomonas aeruginosa NCTC 10781 (Public Health England, Salisbury) a me Staphylococcus aureus NCTC 6571 (Public Health England, Salisbury).
Ua ulu ʻia nā bacteria i ka pō i loko o ka wai Muller-Hinton (Oxoid, Altrincham, UK). A laila ua hoʻoheheʻe ʻia ka moʻomeheu i ka pō 1:100 i loko o ka wai Mueller-Hinton a ua hoʻopili ʻia he 200 µl ma luna o nā membrane cyclopore Whatman sterile 0.2 µm (Whatman plc, Maidstone, UK) ma luna o nā papa agar Mueller-Hinton (Sigma-Aldrich Company Ltd, Kent, Great Britain). ) Ka hoʻokumu ʻana o ka biofilm colonial ma 37°C no 24 mau hola. Ua hoʻāʻo ʻia kēia mau biofilms colonial no ka hoʻemi logarithmic.
E ʻokiʻoki i ke kapa komo i loko o nā ʻāpana huinahā 3 cm2 a hoʻomaʻū mua me ka wai deionized sterile. E kau i ke kāʻei ma luna o ka biofilm o ke kolone ma ka papa agar. Ua wehe ʻia kēlā me kēia 24 ha o ka biofilm, a ua helu ʻia nā bacteria ola i loko o ka biofilm (CFU/ml) ma o ka dilution serial (10−1 a 10−7) i loko o ka Day-Angle neutralization broth (Merck-Millipore). Ma hope o 24 mau hola o ka incubation ma 37°C, ua hana ʻia nā helu papa maʻamau ma nā papa agar Mueller-Hinton. Ua hana ʻia kēlā me kēia lāʻau a me ka manawa i ʻekolu mau manawa, a ua hana hou ʻia nā helu papa no kēlā me kēia dilution.
Loaʻa ka ʻili ʻōpū puaʻa mai nā puaʻa wahine Nui Keʻokeʻo i loko o 15 mau minuke o ka pepehi ʻana e like me nā kūlana hoʻokuʻu aku o ka ʻAhahui ʻEulopa. Ua kahi ʻia ka ʻili a hoʻomaʻemaʻe ʻia me nā ʻūlū waiʻona, a laila ua hoʻopaʻahau ʻia ma -80°C no 24 mau hola e hoʻopau i ka ʻili. Ma hope o ka hoʻoheheʻe ʻana, ua holoi ʻia nā ʻāpana ʻili 1 cm2 ʻekolu manawa me PBS, 0.6% sodium hypochlorite, a me 70% ethanol no 20 mau minuke i kēlā me kēia manawa. Ma mua o ka wehe ʻana i ka epidermis, e wehe i kekahi ethanol i koe ma ka holoi ʻana i 3 mau manawa i loko o ka PBS sterile. Ua hoʻoulu ʻia ka ʻili i loko o kahi pā 6-well me kahi membrane nylon 0.45-μm-mānoanoa (Merck-Millipore) ma luna a me 3 mau pads absorbent (Merck-Millipore) e loaʻa ana he 3 ml fetal bovine serum (Sigma) i hoʻohui ʻia me 10% Dulbecco's modified Eagle. Medium (Dulbecco's Modified Eagle Medium - Aldrich Ltd.).
Ua ulu ʻia nā biofilms kolone e like me ka mea i wehewehe ʻia no nā haʻawina hōʻike biofilm. Ma hope o ka hoʻoulu ʻana i ka biofilm ma ka membrane no 72 mau hola, ua hoʻopili ʻia ka biofilm i ka ʻili o ka ʻili me ka hoʻohana ʻana i kahi loop inoculation sterile a ua wehe ʻia ka membrane. A laila ua hoʻoulu ʻia ka biofilm ma ka dermis o ka puaʻa no 24 mau hola hou ma 37°C e ʻae i ka biofilm e oʻo a pili i ka ʻili o ka puaʻa. Ma hope o ka oʻo ʻana a me ka hoʻopili ʻana o ka biofilm, ua hoʻopili pololei ʻia kahi ʻaʻahu 1.5 cm2, i hoʻomaʻū mua ʻia me ka wai distilled sterile, i ka ʻili o ka ʻili a hoʻoulu ʻia ma 37°C no 24 mau hola. Ua ʻike ʻia nā bacteria ola ma ke kala ʻana ma ka hoʻopili like ʻana i ka reagent ola cell PrestoBlue (Invitrogen, Life Technologies, Paisley, UK) i ka ʻili apical o kēlā me kēia explant a hoʻoulu iā ia no 5 mau minuke. E hoʻohana i ke kāmela kikohoʻe Leica DFC425 e hopu koke i nā kiʻi ma ka microscope Leica MZ8. Ua helu ʻia nā wahi i kala ʻia i ka poni me ka hoʻohana ʻana i ka polokalamu Image Pro mana 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (mediacy.com)). Ua hana ʻia ka microscopy electron scanning e like me ka mea i wehewehe ʻia ma lalo nei.
Ua hoʻoheheʻe ʻia nā bacteria i ulu i ka pō me ka helu 1:100 i loko o ka wai Mueller-Hinton. Ua hoʻohui ʻia he 200 μl o ka moʻomeheu i ka membrane cyclopore Whatman sterile 0.2 μm (Whatman, Maidstone, UK) a ua kau ʻia ma luna o ka agar Mueller-Hinton. Ua hoʻomoʻa ʻia nā papa Biofilm ma 37°C no 72 mau hola e ʻae i ka hoʻokumu ʻana o ka biofilm makua.
Ma hope o 3 mau lā o ka oʻo ʻana o ka biofilm, ua kau pololei ʻia kahi kāʻei huinaha 3 cm2 ma luna o ka biofilm a hoʻomoʻa ʻia ma 37°C no 24 mau hola. Ma hope o ka wehe ʻana i ke kāʻei mai ka ʻili biofilm, ua hoʻohui ʻia he 1 ml o PrestoBlue Cell Viability Reagent (Invitrogen, Waltham, MA) i ka ʻili o kēlā me kēia biofilm no 20 kekona. Ua hoʻomaloʻo ʻia nā ʻili ma mua o ka hoʻopaʻa ʻia ʻana o nā loli kala me ka hoʻohana ʻana i kahi kāmela kikohoʻe Nikon D2300 (Nikon UK Ltd., Kingston, UK).
E hoʻomākaukau i nā moʻomeheu i ka pō ma luna o ka agar Mueller-Hinton, e hoʻoili i nā kolone pākahi i 10 ml Mueller-Hinton broth a hoʻoulu ma luna o kahi shaker ma 37°C (100 rpm). Ma hope o ka hoʻoulu ʻana i ka pō, ua hoʻoheheʻe ʻia ka moʻomeheu 1:100 i loko o ka Mueller-Hinton broth a ua ʻike ʻia he 300 µl ma luna o ka membrane cyclopore Whatman circular 0.2 µm (Whatman International, Maidstone, UK) ma luna o ka Mueller-Hinton agar a hoʻoulu ʻia ma 37°C i loko o 72 mau hola. . Ua hoʻopili ʻia ka biofilm makua i ka ʻeha e like me ka mea i wehewehe ʻia ma lalo nei.
Ua hoʻokō ʻia nā hana āpau me nā holoholona ma ke Kulanui o Manchester ma lalo o kahi laikini papahana i ʻāpono ʻia e ke Keʻena o ka Pono Holoholona a me ka Loiloi Pono (P8721BD27) a e like me nā alakaʻi i paʻi ʻia e ke Keʻena Home ma lalo o ka ASPA i hoʻoponopono hou ʻia i ka makahiki 2012. Ua pili nā mea kākau āpau i nā alakaʻi hōʻea. Ua hoʻohana ʻia nā ʻiole C57BL/6j ʻewalu pule (Envigo, Oxon, UK) no nā haʻawina in vivo āpau. Ua hoʻomaʻemaʻe ʻia nā ʻiole me ka isoflurane (Piramal Critical Care Ltd, West Drayton, UK) a ua kahi ʻia ko lākou mau ʻili dorsal a hoʻomaʻemaʻe ʻia. A laila ua hāʻawi ʻia kēlā me kēia ʻiole i kahi ʻeha excisional 2 × 6 mm me ka hoʻohana ʻana i kahi punch biopsy Stiefel (Schuco International, Hertfordshire, UK). No nā ʻeha i loaʻa i ka biofilm, e hoʻopili i ka biofilm colonial 72-hola i ulu ʻia ma ka membrane e like me ka mea i wehewehe ʻia ma luna i ka papa dermal o ka ʻeha me ka hoʻohana ʻana i kahi loop inoculation sterile ma hope koke o ka ʻeha a hoʻolei i ka membrane. Hoʻomaʻū mua ʻia hoʻokahi kenimika kuea o ka ʻaʻahu me ka wai sterile e mālama i kahi ʻeha pulu. Ua hoʻopili pololei ʻia nā ʻaʻahu i kēlā me kēia ʻeha a uhi ʻia me ka ʻili 3M Tegaderm (3M, Bracknell, UK) a me ka mea hoʻopili wai Mastisol (Eloquest Healthcare, Ferndale, MI) i hoʻopili ʻia a puni nā kihi e hāʻawi i ka hoʻopili hou aku. Ua lawelawe ʻia ʻo Buprenorphine (Animalcare, York, UK) ma kahi nui o 0.1 mg/kg ma ke ʻano he analgesic. E ʻoki i nā ʻiole i ʻekolu lā ma hope o ka ʻeha me ka hoʻohana ʻana i ke ʻano Schedule 1 a wehe, ʻoki i ka hapalua, a mālama i ka wahi ʻeha e like me ka mea e pono ai.
Ua hana ʻia ke kala ʻana o Hematoxylin (ThermoFisher Scientific) a me ka eosin (ThermoFisher Scientific) e like me ke kaʻina hana a ka mea hana. Ua helu ʻia ka ʻāpana ʻeha a me ka reepithelialization me ka hoʻohana ʻana i ka polokalamu Image Pro mana 10 (Media Cybernetics Inc, Rockville, MD).
Ua hoʻokahe ʻia nā ʻāpana ʻiʻo i loko o ka xylene (ThermoFisher Scientific, Loughborough, UK), hoʻomaʻū hou ʻia me ka 100-50% ethanol graded, a hoʻokomo pōkole ʻia i loko o ka wai deionized (ThermoFisher Scientific). Ua hana ʻia ka Immunohistochemistry me ka hoʻohana ʻana i ka VectaStain Elite ABC PK-6104 kit (Vector Laboratories, Burlingame, CA) e like me ke kaʻina hana a ka mea hana. Ua hoʻoheheʻe ʻia nā antibodies mua i nā neutrophils NIMP-R14 (Thermofisher Scientific) a me nā macrophages Ms CD107b Pure M3/84 (BD Biosciences, Wokingham, UK) i ka 1:100 i loko o ka hopena blocking a hoʻohui ʻia i ka ʻili ʻoki, a ukali ʻia e 2 antibodies Anti-, VectaStain ABC a me Vector Nova Red Peroxidase (HRP) substrate kit (Vector Laboratories, Burlingame, CA) a ua counterstained me ka hematoxylin. Ua kiʻi ʻia nā kiʻi me ka hoʻohana ʻana i kahi microscope Olympus BX43 a me kahi kāmela kikohoʻe Olympus DP73 (Olympus, Southend-on-Sea, UK).
Ua hoʻopaʻa ʻia nā laʻana ʻili i loko o 2.5% glutaraldehyde a me 4% formaldehyde i loko o 0.1 M HEPES (pH 7.4) no 24 mau hola ma 4°C. Ua hoʻomaloʻo ʻia nā laʻana me ka hoʻohana ʻana i ka ethanol i hoʻokaʻawale ʻia a ua hoʻomaloʻo ʻia i loko o CO2 me ka hoʻohana ʻana i kahi mea hoʻomaloʻo kiko koʻikoʻi Quorum K850 (Quorum Technologies Ltd, Loughton, UK) a ua uhi ʻia me ka hui gula-palladium me ka hoʻohana ʻana i kahi ʻōnaehana hoʻokuʻu mini sputterer/glow Quorum SC7620. Ua kiʻi ʻia nā laʻana me ka hoʻohana ʻana i kahi microscope electron scanning FEI Quanta 250 (ThermoFisher Scientific) e ʻike i ke kikowaena o ka ʻeha.
Ua hoʻopili ʻia ʻo Toto-1 iodide (2 μM) ma luna o ka ʻili ʻeha o ka ʻiole i ʻoki ʻia a hoʻomoʻa ʻia no 5 min ma 37 °C (ThermoFisher Scientific) a mālama ʻia me Syto-60 (10 μM) ma 37 °C (ThermoFisher Scientific). Ua hana ʻia nā kiʻi Z-stack 15-minuke me ka hoʻohana ʻana i kahi Leica TCS SP8.
Ua hoʻopaʻa ʻia a kālailai ʻia nā ʻikepili hana hou olaola a me nā ʻenehana me ka hoʻohana ʻana i ka polokalamu Graphpad Prism V9 (GraphPad Software, La Jolla, CA). Ua hoʻohana ʻia ka loiloi hoʻokahi ala o ka ʻokoʻa me nā hoʻohālikelike he nui me ka hoʻohana ʻana i ka hoʻāʻo post hoc a Dunnett e hoʻāʻo no nā ʻokoʻa ma waena o kēlā me kēia lāʻau a me ka ʻaʻahu hoʻomalu non-antimicrobial. Ua manaʻo ʻia he koʻikoʻi ka waiwai p <0.05.
Ua loiloi mua ʻia ka pono o nā ʻaʻahu fibrous gel kālā e kūʻē i nā kolone biofilm o Staphylococcus aureus a me Pseudomonas aeruginosa in vitro. Loaʻa i nā ʻaʻahu kālā nā ʻano like ʻole o ke kālā: hana nā ʻaʻahu kālā kuʻuna i nā ion Ag1+; hiki i nā ʻaʻahu kālā, hiki ke hana i nā ion Ag1+ ma hope o ka hoʻohui ʻana o EDTA/BC, ke luku i ka matrix biofilm a hōʻike i ka bacteria i ke kālā ma lalo o ka hopena antibacterial o ke kālā. ions15 a me nā ʻaʻahu e loaʻa ana nā paʻakai Ag oxygenated e hana i nā ion Ag1+, Ag2+ a me Ag3+. Ua hoʻohālikelike ʻia kona pono me kahi ʻaʻahu hoʻomalu non-antimicrobial i hana ʻia mai nā fibers gelled. Ua loiloi ʻia nā bacteria ola i koe i loko o ka biofilm i kēlā me kēia 24 hola no 8 mau lā (Kiʻi 1). I ka lā 5, ua hoʻihoʻi hou ʻia ka biofilm me 3.85 × 105S. Staphylococcus aureus a i ʻole 1.22 × 105P. aeruginosa e loiloi i ka hoʻōla ʻana o ka biofilm. Ke hoʻohālikelike ʻia me nā ʻaʻahu hoʻomalu ʻole antimicrobial, ʻaʻole i loaʻa ka hopena liʻiliʻi o nā ʻaʻahu Ag1+ i ke ola bacteria ma nā biofilms Staphylococcus aureus a me Pseudomonas aeruginosa ma luna o 5 mau lā. I ka hoʻohālikelike ʻana, ʻo nā ʻaʻahu e loaʻa ana ka oxygenated Ag a me Ag1+ + EDTA/BC salts ua kūpono i ka pepehi ʻana i nā bacteria i loko o ka biofilm i loko o 5 mau lā. Ma hope o ka hoʻopili hou ʻia ʻana me nā bacteria planktonic i ka lā 5, ʻaʻohe hoʻihoʻi ʻana o ka biofilm i ʻike ʻia (Kiʻi 1).
Ka helu ʻana o nā bacteria ola i loko o nā biofilms Staphylococcus aureus a me Pseudomonas aeruginosa ma hope o ka mālama ʻana me nā ʻaʻahu kālā. Ua mālama ʻia nā kolone Biofilm o Staphylococcus aureus a me Pseudomonas aeruginosa me nā ʻaʻahu kālā a i ʻole nā ʻaʻahu hoʻomalu non-antimicrobial, a ua hoʻoholo ʻia ka helu o nā bacteria ola i koe i kēlā me kēia 24 hola. Ma hope o 5 mau lā, ua hoʻihoʻi hou ʻia ka biofilm me 3.85 × 105S. Staphylococcus aureus a i ʻole 1.22 × 105P. Ua hoʻokumu ʻia nā kolone o ka bacterioplankton Pseudomonas aeruginosa pākahi e loiloi i ka hoʻōla ʻana o ka biofilm. Hōʻike nā kiʻi i ka awelika +/- hewa maʻamau.
No ka ʻike ʻana i ka hopena o nā ʻaʻahu kālā i ke ola o ka biofilm, ua hoʻopili ʻia nā ʻaʻahu i nā biofilm makua i ulu ʻia ma ka ʻili puaʻa ex vivo. Ma hope o 24 mau hola, wehe ʻia ke ʻaʻahu a hoʻoluʻu ʻia ka biofilm me kahi dye reactive polū, kahi i hoʻololi ʻia e nā bacteria ola i kahi kala poni. ʻO nā biofilms i mālama ʻia me nā ʻaʻahu kaohi he poni, e hōʻike ana i ke alo o nā bacteria ola i loko o ka biofilm (Kiʻi 2A). I ka hoʻohālikelike ʻana, ʻo ka biofilm i mālama ʻia me ke ʻaʻahu Ag oxysols he polū nui ia, e hōʻike ana he bacteria ola ʻole nā bacteria i koe ma ka ʻili o ka ʻili o ka puaʻa (Kiʻi 2B). Ua ʻike ʻia ke kala polū a me ka poni i hui pū ʻia i nā biofilms i mālama ʻia me nā ʻaʻahu Ag1+, e hōʻike ana i ke alo o nā bacteria ola a me nā bacteria ola ʻole i loko o ka biofilm (Kiʻi 2C), ʻoiai ʻo nā ʻaʻahu EDTA/BC i loaʻa iā Ag1+ he polū nui me kekahi mau kiko poni. e hōʻike ana i nā wahi i hoʻopilikia ʻole ʻia e ke ʻaʻahu kālā (Kiʻi 2D). Ua hōʻike ka helu ʻana o nā wahi hana (poni) a me nā wahi hana ʻole (polū) he 75% ka hana o ka ʻāpana kaohi (Kiʻi 2E). Ua hana like nā ʻaʻahu Ag1+ + EDTA/BC me nā ʻaʻahu paʻakai Ag oxygenated, me nā helu ola o 13% a me 14%, kēlā me kēia. Ua hōʻemi pū ka ʻaʻahu Ag1+ i ke ola bacteria ma 21%. A laila ua nānā ʻia kēia mau biofilms me ka hoʻohana ʻana i ka microscopy electron scanning (SEM). Ma hope o ka mālama ʻana me ka ʻaʻahu hoʻomalu a me ka ʻaʻahu Ag1+, ua ʻike ʻia kahi papa o Pseudomonas aeruginosa e uhi ana i ka ʻili puaʻa (Kiʻi 2F, H), ʻoiai ma hope o ka mālama ʻana me ka ʻaʻahu Ag1+, ua loaʻa kekahi mau cell bacteria a ua loaʻa kekahi mau cell bacteria ma lalo. Hiki ke noʻonoʻo ʻia nā olonā collagen ʻo ia ke ʻano o ka ʻiʻo o ka ʻili puaʻa (Kiʻi 2G). Ma hope o ka mālama ʻana me ka ʻaʻahu Ag1+ + EDTA/BC, ua ʻike ʻia nā plaque bacteria a me nā plaque fiber collagen ma lalo (Kiʻi 2I).
ʻIke ʻia o ka biofilm Pseudomonas aeruginosa ma hope o ka mālama ʻana i ke kāhiko kālā. (A–D) Ua ʻike ʻia ke ola ola bacteria i loko o nā biofilm Pseudomonas aeruginosa i ulu ʻia ma ka ʻili puaʻa me ka hoʻohana ʻana i ka pena ola PrestoBlue 24 mau hola ma hope o ka mālama ʻana me nā kāhiko kālā a i ʻole nā kāhiko hoʻomalu non-antimicrobial. He poni nā bacteria ola, he polū nā bacteria ola ʻole a he uliuli ka ʻili puaʻa. (E) Ka helu ʻana o nā biofilm Pseudomonas aeruginosa i ulu ʻia ma ka ʻili puaʻa (kiko poni) me ka hoʻohana ʻana i ka microscopy electron scanning Image Pro mana 10 (FI) a mālama ʻia me kahi kāhiko kālā a i ʻole kahi kāhiko hoʻomalu non-antimicrobial no 24 mau hola. ʻO ka pā unahi SEM = 5 µm. (J–M) Ua ulu nā biofilm Colonial ma nā kānana a ua hoʻoluʻu ʻia me ka pena reactive PrestoBlue ma hope o 24 h o ka incubation me nā kāhiko kālā.
No ka hoʻoholo ʻana inā pili ka pilina kokoke ma waena o nā ʻaʻahu a me nā biofilms i ka pono o nā ʻaʻahu, ua mālama ʻia nā biofilms colonial i kau ʻia ma luna o kahi pālahalaha me nā ʻaʻahu no 24 mau hola a laila ua hoʻoluʻu ʻia me nā dyes reactive. He ʻulaʻula ʻeleʻele ke kala o ka biofilm i mālama ʻole ʻia (Kiʻi 2J). I ka hoʻohālikelike ʻana i nā biofilms i mālama ʻia me nā ʻaʻahu i loaʻa nā paʻakai Ag oxygenated (Kiʻi 2K), ua hōʻike nā biofilms i mālama ʻia me nā ʻaʻahu i loaʻa iā Ag1+ a i ʻole Ag1+ + EDTA/BC i nā kaula o ka waihoʻoluʻu ʻulaʻula (Kiʻi 2L,M). Hōʻike kēia kala ʻulaʻula i ke alo o nā bacteria ola a pili pū me ka wahi suture i loko o ke ʻaʻahu. Hana kēia mau wahi i humuhumu ʻia i nā hakahaka make e hiki ai i nā bacteria i loko o ka biofilm ke ola.
No ka loiloi ʻana i ka pono o nā ʻaʻahu kālā i loko o ke kino ola, ua mālama ʻia nā ʻeha ʻokiʻoki piha o nā ʻiole i loaʻa i ka maʻi oʻo S. aureus a me P. aeruginosa biofilms me nā ʻaʻahu hoʻomalu nonantimicrobial a i ʻole nā ʻaʻahu kālā. Ma hope o 3 mau lā o ka mālama ʻana, ua hōʻike ka nānā ʻana i ke kiʻi macroscopic i nā nui ʻeha liʻiliʻi i ka wā i mālama ʻia ai me nā ʻaʻahu paʻakai oxygenated i hoʻohālikelike ʻia me nā ʻaʻahu hoʻomalu non-antimicrobial a me nā ʻaʻahu kālā ʻē aʻe (Kiʻi 3A-H). No ka hōʻoia ʻana i kēia mau nānā ʻana, ua ʻohi ʻia nā ʻeha a ua helu ʻia ka ʻāpana ʻeha a me ka reepithelialization ma nā ʻāpana ʻiʻo hematoxylin a me ka eosin me ka hoʻohana ʻana i ka polokalamu image pro mana 10 (Kiʻi 3I-L).
ʻO ka hopena o nā kāʻei kālā ma ka ʻili o ka ʻeha a me ka hoʻihoʻi hou ʻana o nā ʻeha i loaʻa i nā biofilms. (A–H) Nā ʻāpana liʻiliʻi i loaʻa i nā biofilms o Pseudomonas aeruginosa (A–D) a me Staphylococcus aureus (E–H) ma hope o ʻekolu lā o ka mālama ʻana me kahi kāʻei hoʻomalu nonantimicrobial, kahi kāʻei paʻakai Ag oxygenated, kahi kāʻei Ag1+, a me kahi kāʻei Ag1+. Nā kiʻi macroscopic hōʻike. nā ʻeha o nā ʻiole me ka kāʻei Ag1+ + EDTA/BC. (IL) Hōʻike i ka maʻi Pseudomonas aeruginosa, nā ʻāpana histological i hoʻoluʻu ʻia me ka hematoxylin a me ka eosin, i hoʻohana ʻia e helu i ka ʻāpana ʻeha a me ka hoʻoulu hou ʻana o ka epithelial. Ka helu ʻana o ka ʻāpana ʻeha (M, O) a me ka pakeneka reepithelialization (N, P) o nā ʻeha i loaʻa i nā biofilms Pseudomonas aeruginosa (M, N) a me Staphylococcus aureus (O, P) (no kēlā me kēia hui mālama n = 12). Hōʻike nā kiʻi i ka awelika +/- hewa maʻamau. * ʻo ia hoʻi ʻo p = < 0.05 ** ʻo ia hoʻi ʻo p = < 0.01; ʻO ke anakahi Macroscopic = 2.5 mm, ʻo ke anakahi histological = 500 µm.
Ua hōʻike ʻia ka helu ʻana o ka ʻāpana ʻeha i nā ʻeha i loaʻa i ka biofilm Pseudomonas aeruginosa (Kiʻi 3M) he awelika ka nui o ka ʻeha o nā ʻeha i mālama ʻia me Ag oxysalts he 2.5 mm2, ʻoiai ʻo ka ʻaʻahu hoʻomalu non-antimicrobial he awelika ka nui o ka ʻeha o 3.1 mm2, ʻaʻole ia he ʻoiaʻiʻo. ua hōʻea i ke koʻikoʻi helu (Kiʻi 3M). p = 0.423). ʻAʻole i hōʻike nā ʻeha i mālama ʻia me Ag1+ a i ʻole Ag1+ + EDTA/BC i ka emi ʻana o ka ʻāpana ʻeha (3.1 mm2 a me 3.6 mm2, kēlā me kēia). ʻO ka mālama ʻana me ka ʻaʻahu paʻakai Ag oxygenated i hoʻolaha i ka re-epithelialization i kahi nui ma mua o ka ʻaʻahu hoʻomalu non-antimicrobial (34% a me 15%, kēlā me kēia; p = 0.029) a me Ag1+ a i ʻole Ag1+ + EDTA/BC (10% a me 11%) (Kiʻi 3N). . , kēlā me kēia).
Ua ʻike ʻia nā ʻano like ma ka wahi ʻeha a me ka hoʻoulu hou ʻana o ka epithelial i nā ʻeha i loaʻa i nā biofilms S. aureus (Kiʻi 3O). Ua hōʻemi nā ʻaʻahu i loaʻa nā paʻakai kālā oxygenated i ka wahi ʻeha (2.0 mm2) ma 23% i hoʻohālikelike ʻia me ke kāʻei antimicrobial kaohi (2.6 mm2), ʻoiai ʻaʻole koʻikoʻi kēia hōʻemi (p = 0.304) (Kiʻi 3O). Eia kekahi, ua emi iki ka wahi ʻeha ma ka hui mālama Ag1+ (2.4 mm2), ʻoiai ʻo ka ʻeha i mālama ʻia me ke kāʻei Ag1+ + EDTA/BC ʻaʻole i hōʻemi i ka wahi ʻeha (2.9 mm2). Ua hoʻolaha pū nā paʻakai oxygen o Ag i ka re-epithelialization o nā ʻeha i loaʻa i ka biofilm S. aureus (31%) i kahi nui ma mua o nā mea i mālama ʻia me nā kāʻei kaohi non-antimicrobial (12%, p = 0.003) (Kiʻi 3P). Ua hōʻike ʻia nā pae o ka hana hou ʻana o ka epithelial e like me ka mana hoʻomalu ʻo Ag1+ (16%, p = 0.903) a me ka hana ʻana o Ag+1 + EDTA/BC (14%, p = 0.965).
No ka ʻike ʻana i ka hopena o nā ʻaʻahu kālā ma ka matrix biofilm, ua hana ʻia ke kala ʻana o Toto 1 a me Syto 60 iodide (Kiʻi 4). ʻO Toto 1 iodide kahi dye impermeable cell hiki ke hoʻohana ʻia e ʻike pololei i nā waikawa nucleic extracellular, ka mea i nui i ka EPS o nā biofilms. ʻO Syto 60 kahi dye permeable cell i hoʻohana ʻia ma ke ʻano he counterstain16. Ua hōʻike ʻia nā nānā ʻana o Toto 1 a me Syto 60 iodide i loko o nā ʻeha i hoʻokomo ʻia me nā biofilms o Pseudomonas aeruginosa (Kiʻi 4A-D) a me Staphylococcus aureus (Kiʻi 4I-L) ma hope o 3 mau lā o ka mālama ʻana i ke kapa komo, ua emi nui ka EPS i loko o ka biofilm. loaʻa nā paʻakai oxygenated Ag a me Ag1+ + EDTA/BC. ʻO nā ʻaʻahu Ag1+ me ka ʻole o nā ʻāpana antibiofilm hou i hoʻemi nui i ka DNA cell-free i loko o nā ʻeha i hoʻokomo ʻia me Pseudomonas aeruginosa akā ʻaʻole i kūpono i nā ʻeha i hoʻokomo ʻia me Staphylococcus aureus.
ʻO ke kiʻi ʻana i loko o ke kino o ka biofilm ʻeha ma hope o 3 mau lā o ka mālama ʻana me nā ʻaʻahu kaohi a i ʻole nā ʻaʻahu kālā. Nā kiʻi confocal o Pseudomonas aeruginosa (A–D) a me Staphylococcus aureus (I–L) i hoʻoluʻu ʻia me Toto 1 (ʻōmaʻomaʻo) e ʻike i nā waikawa nucleic extracellular, kahi ʻāpana o nā polymers biofilm extracellular. No ka hoʻoluʻu ʻana i nā waikawa nucleic intracellular, e hoʻohana iā Syto 60 (ʻulaʻula). nā waikawa. P. Ke nānā ʻana i ka microscopy electron o nā ʻeha i loaʻa i ka maʻi Pseudomonas aeruginosa (E–H) a me Staphylococcus aureus (M–P) biofilms ma hope o 3 mau lā o ka mālama ʻana me nā ʻaʻahu kaohi a me ke kālā. Ka pā unahi SEM = 5 µm. Ka pā unahi kiʻi confocal = 50 µm.
Ua hōʻike ʻia ka microscopy electron scanning ua emi nui ka bacteria i loko o nā ʻiole i hoʻopili ʻia me nā kolone biofilm o Pseudomonas aeruginosa (Kiʻi 4E-H) a me Staphylococcus aureus (Kiʻi 4M-P) ma hope o 3 mau lā o ka mālama ʻana me nā ʻaʻahu kālā āpau.
No ka loiloi ʻana i ka hopena o nā ʻaʻahu kālā ma ka mumū ʻeha i loko o nā ʻiole i loaʻa i ka biofilm, ua hoʻoluʻu ʻia nā ʻāpana o nā ʻeha i loaʻa i ka biofilm i mālama ʻia me ka kaohi a i ʻole nā ʻaʻahu kālā no 3 mau lā me ka hoʻohana ʻana i nā antibodies kūikawā no nā neutrophils a me nā macrophages. Hoʻoholo nui o nā neutrophils a me nā macrophages i loko. ʻiʻo granulation. Kiʻi 5). Ua hōʻemi nā ʻaʻahu kālā āpau i ka helu o nā neutrophils a me nā macrophages i nā ʻeha i loaʻa i ka Pseudomonas aeruginosa i hoʻohālikelike ʻia me nā ʻaʻahu hoʻomalu non-antimicrobial ma hope o ʻekolu lā o ka mālama ʻana. Eia nō naʻe, ʻo ka mālama ʻana me ka ʻaʻahu paʻakai kālā oxygenated i hopena i ka emi nui ʻana o nā neutrophils (p = <0.0001) a me nā macrophages (p = <0.0001) i hoʻohālikelike ʻia me nā ʻaʻahu kālā ʻē aʻe i hoʻāʻo ʻia (Kiʻi 5I,J). ʻOiai ʻoi aku ka hopena o Ag1+ + EDTA/BC i ka biofilm ʻeha, ua hoʻemi ia i nā pae neutrophil a me macrophage i kahi liʻiliʻi ma mua o ka ʻaʻahu Ag1+. Ua ʻike ʻia nā ʻeha kaulike i loaʻa i ka biofilm S. aureus ma hope o ke kāhiko ʻana me Ag (p = <0.0001), Ag1+ (p = 0.0008) a me Ag1++ EDTA/BC (p = 0.0043) oxisols i hoʻohālikelike ʻia me ka mana . Ua ʻike ʻia nā ʻano like no ka neutropenia. kāʻei (Kiʻi 5K). Eia naʻe, ʻo ka ʻaʻahu paʻakai Ag oxygenated wale nō i hōʻike i ka emi nui ʻana o ka helu o nā macrophages i loko o ka ʻiʻo granulation i hoʻohālikelike ʻia me ka mana i nā ʻeha i loaʻa i ka biofilms S. aureus (p = 0.0339) (Kiʻi 5L).
Ua helu ʻia nā Neutrophils a me nā macrophages i loko o nā ʻeha i loaʻa i ka maʻi Pseudomonas aeruginosa a me Staphylococcus aureus biofilms ma hope o 3 mau lā o ka mālama ʻana me ka mana antimicrobial ʻole a i ʻole nā ʻaʻahu kālā. Ua helu ʻia nā Neutrophils (AD) a me nā macrophages (EH) i loko o nā ʻāpana ʻiʻo i hoʻoluʻu ʻia me nā antibodies kikoʻī no nā neutrophils a i ʻole macrophages. Ka helu ʻana o nā neutrophils (I a me K) a me nā macrophages (J a me L) i nā ʻeha i loaʻa i ka maʻi Pseudomonas aeruginosa (I a me J) a me Staphylococcus aureus (K & L) biofilms. N = 12 no kēlā me kēia hui. Hōʻike nā kiʻi i ka awelika +/- hewa maʻamau, nā waiwai koʻikoʻi i hoʻohālikelike ʻia me ka ʻaʻahu mana antibacterial ʻole, * ʻo ia hoʻi p = < 0.05, ** ʻo ia hoʻi p = < 0.01; *** ʻo ia hoʻi p = < 0.001; hōʻike p = < 0.0001).
A laila ua loiloi mākou i ka hopena o nā ʻaʻahu kālā i ka ho'ōla kūʻokoʻa o ka maʻi. Ua mālama ʻia nā ʻeha excisional i loaʻa ʻole ka maʻi me kahi ʻaʻahu hoʻomalu non-antimicrobial a i ʻole kahi ʻaʻahu kālā no 3 mau lā (Kiʻi 6). Ma waena o nā ʻaʻahu kālā i hoʻāʻo ʻia, ʻo nā ʻeha wale nō i mālama ʻia me ka ʻaʻahu paʻakai oxygenated i ʻike ʻia he liʻiliʻi ma nā kiʻi macroscopic ma mua o nā ʻeha i mālama ʻia me ka mana (Kiʻi 6A-D). Ua hōʻike ka helu ʻana o ka ʻāpana ʻeha me ka hoʻohana ʻana i ka loiloi histological ʻo ka awelika o ka ʻāpana ʻeha ma hope o ka mālama ʻana me ka ʻaʻahu Ag oxysols he 2.35 mm2 i hoʻohālikelike ʻia me 2.96 mm2 no nā ʻeha i mālama ʻia me ka hui hoʻomalu, akā ʻaʻole i hiki kēia ʻokoʻa i ka helu koʻikoʻi (p = 0.488) (Kiʻi 6I). I ka hoʻohālikelike ʻana, ʻaʻohe emi ʻana o ka ʻāpana ʻeha i ʻike ʻia ma hope o ka mālama ʻana me Ag1+ (3.38 mm2, p = 0.757) a i ʻole Ag1+ + EDTA/BC (4.18 mm2, p = 0.054) i hoʻohālikelike ʻia me ka hui hoʻomalu. Ua ʻike ʻia ka hoʻonui ʻia ʻana o ka hana hou ʻana o ka epithelial me ka ʻaʻahu Ag oxysol i hoʻohālikelike ʻia me ka hui kaohi (30% vs. 22%, kēlā me kēia), ʻoiai ʻaʻole i hiki kēia i ke koʻikoʻi (p = 0.067), he mea koʻikoʻi loa kēia a hōʻoia i nā hopena i hala. ʻO ka ʻaʻahu me nā oxysols e paipai i ka re-epithelialization. -Epithelization o nā ʻeha i loaʻa ʻole ka maʻi17. I ka hoʻohālikelike ʻana, ʻaʻohe hopena o ka mālama ʻana me Ag1+ a i ʻole Ag1+ + EDTA/BC dressings a i ʻole hōʻike i ka emi ʻana o ka re-epithelialization i hoʻohālikelike ʻia me ka kaohi.
Ka hopena o ke kāʻei ʻeha kālā i ka ho'ōla ʻana o ka ʻeha i nā ʻiole i loaʻa ʻole ka maʻi me ka resection piha. (AD) Nā kiʻi macroscopic hōʻike o nā ʻeha ma hope o ʻekolu mau lā o ka mālama ʻana me kahi kāʻei hoʻomalu non-antimicrobial a me kahi kāʻei kālā. (EH) Nā ʻāpana ʻeha hōʻike i hoʻoluʻu ʻia me ka hematoxylin a me ka eosin. Ua helu ʻia ka helu ʻana o ka ʻāpana ʻeha (I) a me ka pakeneka o ka reepithelialization (J) mai nā ʻāpana histological ma ke kikowaena o ka ʻeha me ka hoʻohana ʻana i ka polokalamu loiloi kiʻi (n = 11-12 no kēlā me kēia hui mālama). Hōʻike nā kiʻi i ka awelika +/- hewa maʻamau. * ʻo ia hoʻi p = <0.05.
He mōʻaukala lōʻihi ko ke kālā no ka hoʻohana ʻana ma ke ʻano he lāʻau antimicrobial i ka ho'ōla ʻana i nā ʻeha, akā ʻo nā ʻano hana like ʻole a me nā ʻano lawe ʻana e hopena i nā ʻokoʻa i ka pono antimicrobial 18. Eia kekahi, ʻaʻole maopopo loa nā waiwai antibiofilm o nā ʻōnaehana lawe kālā kikoʻī. ʻOiai he kūpono ka pane ʻana o ka pale kino o ka mea hoʻokipa e kūʻē i ka bacteria planktonic, ʻaʻole ia he maikaʻi loa i nā biofilms19. Hoʻopili koke ʻia nā bacteria planktonic e nā macrophages, akā i loko o nā biofilms, hoʻopilikia hou nā pūnaewele i hōʻuluʻulu ʻia ma ka palena ʻana i ka pane a ka mea hoʻokipa i ka nui e hiki ai i nā pūnaewele pale ke hana i ka apoptosis a hoʻokuʻu i nā mea proinflammatory e hoʻonui i ka pane pale20. Ua ʻike ʻia hiki i kekahi mau leukocytes ke komo i loko o nā biofilms21 akā ʻaʻole hiki ke phagocytose i nā bacteria ke hoʻopilikia ʻia kēia pale22. Pono e hoʻohana ʻia kahi ala holistic e kākoʻo i ka pane ʻana o ka pale kino o ka mea hoʻokipa e kūʻē i ka maʻi biofilm ʻeha. Hiki i ka debridement ʻeha ke hoʻopilikia kino i ka biofilm a wehe i ka hapa nui o ka bioburden, akā ʻaʻole paha e kūpono ka pane ʻana o ka pale kino o ka mea hoʻokipa e kūʻē i ke koena o ka biofilm, ʻoiai inā ua hoʻopilikia ʻia ka pane ʻana o ka pale kino o ka mea hoʻokipa. No laila, hiki i nā lāʻau lapaʻau antimicrobial e like me nā ʻaʻahu kālā ke kākoʻo i ka pane pale o ka mea hoʻokipa a hoʻopau i nā maʻi biofilm. ʻO ka haku mele ʻana, ka noʻonoʻo ʻana, ka solubility, a me ka substrate lawe ke hoʻohuli i ka pono antimicrobial o ke kālā. I nā makahiki i hala iho nei, ua ʻoi aku ka maikaʻi o ka holomua o ka ʻenehana hana kālā i kēia mau ʻaʻahu9,23. I ka holomua ʻana o ka ʻenehana ʻaʻahu kālā, he mea nui e hoʻomaopopo i ka pono o kēia mau ʻaʻahu i ka kaohi ʻana i ka maʻi ʻeha a, ʻoi aku ka nui, ka hopena o kēia mau ʻano kālā ikaika i ke kaiapuni ʻeha a me ka ho'ōla ʻana.
Ma kēia haʻawina, ua hoʻohālikelike mākou i ka pono o ʻelua mau ʻaʻahu kālā holomua me nā ʻaʻahu kālā maʻamau e hana ana i nā ion Ag1+ e kūʻē i nā biofilms me ka hoʻohana ʻana i nā hiʻohiʻona in vitro a me in vivo. Ua loiloi pū mākou i ka hopena o kēia mau ʻaʻahu ma ke kaiapuni ʻeha a me ka ho'ōla kūʻokoʻa o ka maʻi. No ka hōʻemi ʻana i ka mana o ka matrix lawe ʻana, ua hana ʻia nā ʻaʻahu kālā a pau i hoʻāʻo ʻia me ka carboxymethylcellulose.
ʻO kā mākou loiloi mua ʻana o kēia mau ʻaʻahu kālā e kūʻē i nā biofilms kolone o Pseudomonas aeruginosa lāua ʻo Staphylococcus aureus e hōʻike ana, ʻaʻole like me nā ʻaʻahu Ag1+ kuʻuna, ʻelua mau ʻaʻahu kālā holomua, ʻo Ag1+ + EDTA/BC a me nā paʻakai Ag oxygenated, he pono ma 5. Ke pepehi pono nei i ka bacteria biofilm i loko o kekahi mau lā. Eia kekahi, pale kēia mau ʻaʻahu i ka hana hou ʻana o ka biofilm ma hope o ka hōʻike pinepine ʻana i nā bacteria planktonic. Loaʻa i ka ʻaʻahu Ag1+ ka chloride kālā, ka hui kālā like a me ka matrix kumu e like me Ag1+ + EDTA/BC, a he hopena palena ʻole i ka ola bacteria i loko o ka biofilm i ka manawa like. ʻO ka ʻike ʻana he ʻoi aku ka maikaʻi o ka ʻaʻahu Ag1+ + EDTA/BC e kūʻē i ka biofilm ma mua o ka ʻaʻahu Ag1+ i loaʻa i ka matrix like a me kahi hui kālā e kākoʻo ana i ka manaʻo e pono ana nā mea hoʻohui hou e hoʻonui i ka pono o ka chloride kālā e kūʻē i ka biofilm, e like me ka mea i hōʻike ʻia ma kahi ʻē aʻe15. Kākoʻo kēia mau hopena i ka manaʻo e pāʻani ana ʻo BC a me EDTA i kahi kuleana hou e kōkua ana i ka pono o ke kāhiko ʻana a ʻo ka nele o kēia ʻāpana i nā kāhiko Ag1+ paha i kōkua i ka hōʻike ʻole ʻana i ka pono in vitro. Ua ʻike mākou ua hōʻike nā kāhiko paʻakai Ag oxygenated e hana ana i nā ion Ag2+ a me Ag3+ i ka pono antibacterial ikaika ma mua o Ag1+ a ma nā pae like me Ag1+ + EDTA/BC. Eia naʻe, ma muli o ke kiʻekiʻe o ka hiki ke redox, ʻaʻole maopopo pehea ka lōʻihi o ka hana a me ka pono o nā ion Ag3+ e kūʻē i nā biofilms ʻeha a no laila e pono ai ke aʻo hou ʻia. Eia kekahi, he nui nā kāhiko like ʻole e hana ana i nā ion Ag1+ i hoʻāʻo ʻole ʻia ma kēia haʻawina. Hana ʻia kēia mau kāhiko me nā hui kālā like ʻole, nā ʻano, a me nā matrices kumu, kahi e hoʻohuli ai i ka lawe ʻana o nā ion Ag1+ a me ko lākou pono e kūʻē i nā biofilms. He mea kūpono hoʻi ke hoʻomaopopo ʻana he nui nā ʻano hoʻohālike in vitro a me in vivo i hoʻohana ʻia e loiloi i ka pono o nā kāhiko ʻeha e kūʻē i nā biofilms. ʻO ke ʻano o ke kumu hoʻohālike i hoʻohana ʻia, a me ka paʻakai a me ka protein o ka media i hoʻohana ʻia i kēia mau hiʻohiʻona, e hoʻohuli i ka pono o ke kāhiko ʻana. I loko o kā mākou kumu hoʻohālike in vivo, ua ʻae mākou i ka biofilm e oʻo i loko o ka vitro a laila hoʻoili iā ia i ka ʻili o ka ʻeha. He kūpono ka pane pale ʻana o ka ʻiole kikowaena e kūʻē i ka bacteria planktonic i hoʻopili ʻia i ka ʻeha, a laila e hana ana i kahi biofilm i ka wā e ho'ōla ai ka ʻeha. ʻO ka hoʻohui ʻana o ka biofilm makua i kahi ʻeha e kaupalena i ka pono o ka pane pale ʻana o ka kikowaena i ka hoʻokumu ʻana o ka biofilm ma ka ʻae ʻana i ka biofilm makua e hoʻokumu iā ia iho i loko o ka ʻeha ma mua o ka hoʻomaka ʻana o ka hoʻōla. No laila, hiki i kā mākou kumu hoʻohālike iā mākou ke loiloi i ka pono o nā ʻaʻahu antimicrobial ma nā biofilm makua ma mua o ka hoʻomaka ʻana o nā ʻeha e ho'ōla.
Ua ʻike pū mākou ua hoʻopilikia ke kūpono o ke kāhiko ʻana i ka pono o nā kāhiko kālā ma nā biofilms i ulu ʻia ma vitro a me ka ʻili puaʻa. Manaʻo ʻia ka pili kokoke ʻana me ka ʻeha he mea koʻikoʻi no ka pono antimicrobial o ke kāhiko ʻana24,25. ʻO nā kāhiko e loaʻa ana nā paʻakai Ag oxygenated i pili kokoke me nā biofilms makua, e hopena ana i ka emi nui ʻana o ka nui o nā bacteria ola i loko o ka biofilm ma hope o 24 mau hola. I ka hoʻohālikelike ʻana, i ka wā i mālama ʻia ai me nā kāhiko Ag1+ a me Ag1+ + EDTA/BC, ua koe ka nui o nā bacteria ola. Loaʻa i kēia mau kāhiko nā sutures ma ka lōʻihi holoʻokoʻa o ke kāhiko ʻana, kahi e hana ai i nā hakahaka make e pale ai i ka pili kokoke me ka biofilm. I kā mākou mau haʻawina in vitro, ua pale kēia mau wahi pili ʻole i ka pepehi ʻana i nā bacteria ola i loko o ka biofilm. Ua loiloi mākou i ke ola bacteria ma hope o 24 mau hola o ka mālama ʻana; I ka hala ʻana o ka manawa, i ka wā e piha ai ke kāhiko ʻana, hiki ke emi ka hakahaka make, e hōʻemi ana i ka wahi no kēia mau bacteria ola. Eia naʻe, hōʻike kēia i ke koʻikoʻi o ka haku mele ʻana o ke kāhiko ʻana, ʻaʻole wale ke ʻano kālā i loko o ke kāhiko ʻana.
ʻOiai he mea pono nā haʻawina in vitro no ka hoʻohālikelike ʻana i ka pono o nā ʻenehana kālā like ʻole, he mea nui hoʻi e hoʻomaopopo i nā hopena o kēia mau mea hoʻonani ma nā biofilms in vivo, kahi e hāʻawi ai nā ʻiʻo kikowaena a me nā pane pale i ka pono o nā mea hoʻonani e kūʻē i nā biofilms. Ua ʻike ʻia ka hopena o kēia mau mea hoʻonani ma nā biofilms ʻeha me ka hoʻohana ʻana i ka microscopy electron scanning a me ka EPS staining o ka biofilm me ka hoʻohana ʻana i nā dyes DNA intracellular a me extracellular. Ua ʻike mākou ma hope o 3 mau lā o ka mālama ʻana, ua kūpono nā mea hoʻonani āpau i ka hōʻemi ʻana i ka DNA cell-free i nā ʻeha i loaʻa i ka biofilm, akā ʻaʻole i kūpono ka mea hoʻonani Ag1+ i nā ʻeha i loaʻa i ka Staphylococcus aureus. Ua hōʻike pū ka microscopy electron scanning he emi nui ka bacteria i loaʻa i nā ʻeha i mālama ʻia me nā mea hoʻonani kālā, ʻoiai ua ʻoi aku ka ʻike ʻia o kēia me ka mea hoʻonani paʻakai Ag oxygenated a me ka mea hoʻonani Ag1+ + EDTA/BC i hoʻohālikelike ʻia me ka mea hoʻonani Ag1+. Hōʻike kēia mau ʻikepili he ʻokoʻa nā hopena o nā mea hoʻonani kālā i hoʻāʻo ʻia ma ke ʻano biofilm, akā ʻaʻohe o nā mea hoʻonani kālā i hiki ke hoʻopau i ka biofilm, e kākoʻo ana i ka pono no kahi ala holoʻokoʻa i ka mālama ʻana i nā maʻi biofilm ʻeha; ka hoʻohana ʻana i nā ʻaʻahu lima kālā. Hoʻomaka ʻia ka mālama ʻana e ka debridement kino e wehe i ka hapa nui o ka biofilm.
ʻO nā ʻeha mau loa e noho pinepine ana i kahi ʻano mumū nui, me nā hunaola mumū keu e waiho ana i loko o ka ʻiʻo ʻeha no kahi manawa lōʻihi, e hōʻino ana i nā ʻiʻo a hoʻopau i ka oxygen e pono ai no ka metabolism cellular maikaʻi a me ka hana i loko o ka ʻeha26. Hoʻonui nā biofilms i kēia ʻano ʻeha ʻino ma o ka hoʻopilikia maikaʻi ʻole ʻana i ka ho'ōla ʻana ma nā ʻano like ʻole, me ka pale ʻana i ka hoʻonui ʻia ʻana o nā cell a me ka neʻe ʻana a me ka hoʻoulu ʻana o nā cytokines proinflammatory27. I ka lilo ʻana o nā ʻaʻahu kālā i mea ʻoi aku ka maikaʻi, he mea nui e hoʻomaopopo i ka hopena o lākou i ke kaiapuni ʻeha a me ka ho'ōla ʻana.
ʻO ka mea hoihoi, ʻoiai ua hoʻopilikia nā ʻaʻahu kālā āpau i ka haku mele biofilm, ʻo nā ʻaʻahu paʻakai kālā oxygenated wale nō i hoʻonui i ka re-epithelialization o kēia mau ʻeha i loaʻa i ka maʻi. Kākoʻo kēia mau ʻikepili i kā mākou mau ʻike mua17 a me nā mea a Kalan et al. (2017)28, nāna i hōʻike i nā ʻano palekana maikaʻi a me nā ʻona o nā paʻakai kālā oxygenated, ʻoiai ʻo nā haʻahaʻa haʻahaʻa o ke kālā he kūpono ia e kūʻē i nā biofilms.
Hōʻike kā mākou noiʻi o kēia manawa i nā ʻokoʻa o ka ʻenehana kālā ma waena o nā ʻaʻahu kālā antimicrobial a me ka hopena o kēia ʻenehana ma ke kaiapuni ʻeha a me ka ho'ōla kūʻokoʻa o ka maʻi. Eia nō naʻe, ʻokoʻa kēia mau hopena mai nā noiʻi mua e hōʻike ana ua hoʻomaikaʻi ka ʻaʻahu Ag1+ + EDTA/BC i nā palena ho'ōla o nā pepeiao rabbit i ʻeha i loko o ke kino. Eia nō naʻe, hiki ke kumu o kēia i nā ʻokoʻa o nā hiʻohiʻona holoholona, nā manawa ana, a me nā ʻano noi bacteria29. I kēia hihia, ua lawe ʻia nā ana ʻeha 12 mau lā ma hope o ka ʻeha e ʻae i nā mea hana o ka ʻaʻahu e hana ma ka biofilm i kahi manawa lōʻihi. Kākoʻo ʻia kēia e kahi noiʻi i hōʻike i ka nui o nā ʻeha wāwae i loaʻa i ka maʻi i mālama ʻia me Ag1+ + EDTA/BC i hoʻonui mua ʻia ma hope o hoʻokahi pule o ka mālama ʻana, a laila ma luna o 3 mau pule e hiki mai ana o ka mālama ʻana me Ag1+ + EDTA/BC a i loko o 4 mau pule o ka hoʻohana ʻana i nā lāʻau lapaʻau ʻole. Nā ʻaʻahu CMC e hōʻemi i ka nui o nā ʻeha30.
Ua hōʻike mua ʻia kekahi mau ʻano a me nā ʻano o ke kālā he cytotoxic in vitro 11, akā ʻaʻole i unuhi mau ʻia kēia mau hopena in vitro i nā hopena maikaʻi ʻole i vivo. Eia kekahi, ʻo ka holomua ʻana i ka ʻenehana kālā a me ka hoʻomaopopo maikaʻi ʻana i nā hui kālā a me nā ʻano i loko o nā ʻaʻahu ua alakaʻi i ka hoʻomohala ʻana o nā ʻaʻahu kālā palekana a maikaʻi hoʻi. Eia naʻe, i ka holomua ʻana o ka ʻenehana ʻaʻahu kālā, he mea nui e hoʻomaopopo i ka hopena o kēia mau ʻaʻahu ma ke kaiapuni ʻeha31,32,33. Ua hōʻike mua ʻia ʻo ka piʻi ʻana o ka nui o ka re-epithelialization e pili ana i ka nui o ka nui o nā macrophages anti-inflammatory M2 i hoʻohālikelike ʻia me ka phenotype pro-inflammatory M1. Ua ʻike ʻia kēia ma kahi kumu hoʻohālike ʻiole ma mua kahi i hoʻohālikelike ʻia ai nā ʻaʻahu ʻeha hydrogel kālā me ka sulfadiazine kālā a me nā hydrogels non-antimicrobial34.
Hiki i nā ʻeha mau ke hōʻike i ka mumū nui a ua ʻike ʻia ʻo ke alo o nā neutrophils keu he mea hōʻino paha i ka ho'ōla ʻana o ka ʻeha35. Ma kahi noiʻi i nā ʻiole i hoʻopau ʻia e ka neutrophil, ua hoʻopaneʻe ka alo o nā neutrophils i ka reepithelialization. ʻO ke alo o nā neutrophils keu e alakaʻi ai i nā pae kiʻekiʻe o nā protease a me nā ʻano oxygen reactive, e like me ka superoxide a me ka hydrogen peroxide, e pili ana me nā ʻeha mau a lohi ka ho'ōla37,38. Pēlā nō, ʻo ka hoʻonui ʻana i nā helu macrophage, inā ʻaʻole i kāohi ʻia, hiki ke alakaʻi i ka hoʻopaneʻe ʻana o ka hoʻōla ʻana o ka ʻeha39. He mea nui loa kēia hoʻonui inā ʻaʻole hiki i nā macrophages ke hoʻololi mai kahi phenotype pro-inflammatory i kahi phenotype pro-healing, e hopena ana i nā ʻeha i hiki ʻole ke puka i waho o ka pae mumū o ka ho'ōla40. Ua ʻike mākou i ka emi ʻana o nā neutrophils a me nā macrophages i nā ʻeha i loaʻa i ka biofilm ma hope o 3 mau lā o ka mālama ʻana me nā ʻaʻahu kālā āpau, akā ʻoi aku ka ʻike ʻia o ka emi ʻana me nā ʻaʻahu paʻakai oxygenated. ʻO kēia emi ʻana paha he hopena pololei ia o ka pane pale i ke kālā, kahi pane i ka emi ʻana o ka bioburden, a i ʻole ka ʻeha ma kahi pae hope o ka ho'ōla ʻana a no laila ua hoʻemi ʻia nā cell pale i loko o ka ʻeha. ʻO ka hōʻemi ʻana i ka helu o nā hunaola mumū i loko o ka ʻeha e mālama paha i kahi ʻano e kūpono ai i ka ho'ōla ʻana o ka ʻeha. ʻAʻole maopopo ke ʻano o ka hana ʻana o ke ʻano o ka hoʻoulu ʻana o Ag oxysalts i ka ho'ōla kūʻokoʻa i ka maʻi, akā ʻo ka hiki o Ag oxysalts ke hana i ka oxygen a luku i nā pae ʻino o ka hydrogen peroxide, kahi mea hoʻoponopono o ka mumū, e wehewehe paha i kēia a pono e noiʻi hou aku17.
ʻO nā ʻeha maʻi i hoʻōla ʻole ʻia he pilikia ia no nā kauka a me nā maʻi. ʻOiai he nui nā ʻaʻahu e koi ana i ka pono antimicrobial, ʻaʻole kākaʻikahi ka noiʻi ʻana i nā kumu koʻikoʻi ʻē aʻe e hoʻopilikia ana i ka microenvironment ʻeha. Hōʻike kēia haʻawina he ʻokoʻa ka pono antimicrobial o nā ʻenehana kālā like ʻole a, ʻo ka mea nui, nā hopena like ʻole ma ke kaiapuni ʻeha a me ka ho'ōla ʻana, kūʻokoʻa i ka maʻi. ʻOiai ʻo kēia mau haʻawina in vitro a me in vivo e hōʻike ana i ka pono o kēia mau ʻaʻahu i ka mālama ʻana i nā maʻi ʻeha a me ka hoʻolaha ʻana i ka ho'ōla ʻana, pono nā hoʻokolohua i hoʻomalu ʻia e loiloi i ka pono o kēia mau ʻaʻahu i loko o ke keʻena lapaʻau.
Loaʻa nā ʻikepili i hoʻohana ʻia a/a i ʻole i kālailai ʻia i ka wā o ke aʻo ʻana o kēia manawa mai ka mea kākau pili ma ke noi kūpono.
Ka manawa hoʻouna: Iulai-15-2024