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Girman ƙwayoyin cuta a cikin raunuka sau da yawa yana bayyana kansa a matsayin biofilms, wanda ke tsoma baki ga warkarwa kuma yana da wahalar kawar da shi. Sabbin kayan shafa na azurfa suna da'awar yaƙi da cututtukan raunuka, amma ingancinsu na maganin rigakafi da tasirin warkarwa mai zaman kansa ba a san su ba gabaɗaya. Ta amfani da samfuran biofilm na Staphylococcus aureus da Pseudomonas aeruginosa a cikin vitro da in vivo, muna ba da rahoton ingancin kayan shafa na Ag1+ ion; kayan shafa na Ag1+ waɗanda ke ɗauke da ethylenediaminetetraacetic acid da benzetonium chloride (Ag1+/EDTA/BC), da kayan shafa waɗanda ke ɗauke da silver nitrate (Ag oxysalts). , waɗanda ke samar da ions na Ag1+, Ag2+ da Ag3+ don yaƙi da biofilm na rauni da tasirinsa akan warkarwa. Kayan shafa na Ag1+ ba su da tasiri sosai akan biofilm na rauni a cikin vitro da a cikin beraye (C57BL/6j). Sabanin haka, gishirin Ag mai iskar oxygen da miya na Ag1+/EDTA/BC sun rage yawan ƙwayoyin cuta masu rai a cikin biofilms a cikin vitro kuma sun nuna raguwa mai yawa a cikin abubuwan da ke cikin ƙwayoyin cuta da EPS a cikin biofilms na raunin beraye. Waɗannan miya suna da tasiri daban-daban akan warkar da raunukan da suka kamu da biofilm da waɗanda ba su kamu da biofilm ba, tare da miya na gishiri mai iskar oxygen yana da ƙarin tasiri mai amfani akan sake farfadowa, girman rauni, da kumburi idan aka kwatanta da magungunan sarrafawa da sauran miya na azurfa. Daban-daban halayen sinadarai na miya na azurfa na iya samun tasiri daban-daban akan biofilm na rauni da warkarwa, kuma ya kamata a yi la'akari da wannan lokacin zaɓar miya don maganin raunukan da suka kamu da biofilm.
An bayyana raunuka masu tsanani a matsayin "raunuka waɗanda suka kasa ci gaba ta hanyar matakan warkarwa na yau da kullun cikin tsari da kuma kan lokaci" 1. Raunuka masu tsanani suna haifar da nauyi na tunani, zamantakewa da tattalin arziki ga marasa lafiya da tsarin kiwon lafiya. An kiyasta kashe kuɗin da NHS ke kashewa a kowace shekara kan maganin raunuka da sauran cututtuka masu alaƙa da su ya kai £8.3 biliyan a 2017–182. Raunuka masu tsanani suma matsala ce mai matuƙar muhimmanci a Amurka, inda Medicare ta kiyasta farashin shekara-shekara na kula da marasa lafiya da raunuka a kan $28.1–$96.8 biliyan3.
Kamuwa da cuta babban abu ne da ke hana warkar da rauni. Kamuwa da cuta galibi yana bayyana a matsayin biofilms, wanda ke cikin kashi 78% na raunukan da ba sa warkarwa na yau da kullun. Biofilms suna samuwa ne lokacin da ƙananan ƙwayoyin cuta suka makale a saman fata ba tare da juyawa ba, kamar saman rauni, kuma suna iya taruwa don samar da al'ummomin da ke samar da polymer (EPS). Biofilm ɗin rauni yana da alaƙa da ƙaruwar amsawar kumburi wanda ke haifar da lalacewar nama, wanda zai iya jinkirta ko hana warkarwa4. Ƙara lalacewar nama na iya faruwa ne saboda ƙaruwar ayyukan matrix metalloproteinases, collagenase, elastase da nau'in oxygen mai amsawa5. Bugu da ƙari, ƙwayoyin kumburi da biofilms kansu suna yawan amfani da iskar oxygen kuma saboda haka suna iya haifar da hypoxia na nama na gida, suna rage ƙwayoyin oxygen mai mahimmanci da ake buƙata don ingantaccen gyaran nama6.
Manyan halittun biofilms suna da matuƙar juriya ga magungunan kashe ƙwayoyin cuta, suna buƙatar dabarun yaƙi don magance cututtukan biofilm, kamar maganin injiniya sannan kuma maganin kashe ƙwayoyin cuta mai inganci. Saboda biofilms na iya sake farfaɗowa cikin sauri, magungunan kashe ƙwayoyin cuta masu inganci na iya rage haɗarin sake farfaɗowa bayan tiyatar cire su.
Ana ƙara amfani da azurfa a cikin kayan maye na ƙwayoyin cuta kuma galibi ana amfani da shi azaman magani na farko don raunuka masu kamuwa da cuta na yau da kullun. Akwai kayan miya na azurfa da yawa da ake samu a kasuwa, kowannensu yana ɗauke da nau'ikan kayan azurfa daban-daban, maida hankali, da ma'aunin tushe. Ci gaba a cikin kayan miya na azurfa ya haifar da haɓaka sabbin kayan miya na azurfa. Siffar ƙarfe ta azurfa (Ag0) ba ta da ƙarfi; Don cimma tasirin maganin rigakafi, dole ne ya rasa electron don samar da azurfar ionic (Ag1+). Kayan miya na azurfa na gargajiya suna ɗauke da mahaɗan azurfa ko azurfa na ƙarfe waɗanda, idan aka fallasa su ga ruwa, suka ruɓe don samar da ions na Ag1+. Waɗannan ions na Ag1+ suna amsawa da ƙwayar cuta, suna cire electrons daga abubuwan gini ko mahimman hanyoyin da ake buƙata don rayuwa. Fasaha mai lasisi ta haifar da haɓaka sabon mahaɗan azurfa, Ag oxysalts (silver nitrate, Ag7NO11), wanda aka haɗa a cikin kayan miya na rauni. Ba kamar azurfa na gargajiya ba, rugujewar gishirin da ke ɗauke da iskar oxygen yana samar da yanayin azurfa tare da babban ƙarfin kuzari (Ag1+, Ag2+ da Ag3+). Nazarin In vitro ya nuna cewa ƙarancin yawan gishirin azurfa mai iskar oxygen ya fi inganci fiye da azurfar ion guda ɗaya (Ag1+) akan ƙwayoyin cuta masu cutarwa kamar Pseudomonas aeruginosa, Staphylococcus aureus da Escherichia coli8,9. Wani sabon nau'in miya na azurfa ya haɗa da ƙarin sinadarai, wato ethylenediaminetetraacetic acid (EDTA) da benzetonium chloride (BC), waɗanda aka ruwaito suna kai hari ga biofilm EPS kuma ta haka suna ƙara shigar azurfa cikin biofilm. Waɗannan sabbin fasahohin azurfa suna ba da sabbin hanyoyin kai hari ga biofilm na rauni. Duk da haka, tasirin waɗannan magungunan kashe ƙwayoyin cuta akan yanayin rauni da warkarwa mara kamuwa da cuta yana da mahimmanci don tabbatar da cewa ba sa haifar da mummunan yanayin rauni ko jinkirta warkarwa. An ba da rahoton damuwa game da gubar azurfa a cikin vitro tare da miya da yawa na azurfa10,11. Duk da haka, gubar guba a cikin vitro ba ta fassara zuwa guba a cikin in vivo ba, kuma miya da yawa na Ag1+ sun nuna kyakkyawan bayanin aminci12.
A nan, mun binciki ingancin kayan miya na carboxymethylcellulose waɗanda ke ɗauke da sabbin hanyoyin samar da azurfa don magance raunin da ke cikin jini a cikin vitro da kuma a cikin jiki. Bugu da ƙari, an tantance tasirin waɗannan kayan miya akan martanin garkuwar jiki da warkarwa ba tare da la'akari da kamuwa da cuta ba.
Duk kayan miya da aka yi amfani da su an same su a kasuwa. 3M Kerracel Gel Fiber Dressing (3M, Knutsford, UK) wani kayan miya ne na gel fiber wanda ba ya kashe ƙwayoyin cuta 100% carboxymethylcellulose (CMC) wanda aka yi amfani da shi azaman kayan miya mai sarrafawa a cikin wannan binciken. An kimanta kayan miya na azurfa guda uku na CMC masu maganin ƙwayoyin cuta, wato 3M Kerracel Ag dressing (3M, Knutsford, UK), wanda ya ƙunshi 1.7 wt% na gishirin azurfa mai iskar oxygen (Ag7NO11) a cikin ions na azurfa masu ƙarfi (Ag1+, Ag2+ da Ag3+). A lokacin ruɓewar Ag7NO11, an samar da ions na Ag1+, Ag2+ da Ag3+ a cikin rabo na 1:2:4. Miyar Aquacel Ag Karin miyar da ke ɗauke da 1.2% azurfa chloride (Ag1+) (ConvaTec, Deeside, UK) 13 da kuma Aquacel Ag + Karin miyar da ke ɗauke da 1.2% azurfa chloride (Ag1+), EDTA da benzetonium chloride (ConvaTec, Deeside, UK) 14.
Nau'ikan da aka yi amfani da su a cikin wannan binciken sune Pseudomonas aeruginosa NCTC 10781 (Jami'ar Lafiyar Jama'a ta Ingila, Salisbury) da Staphylococcus aureus NCTC 6571 (Jami'ar Lafiyar Jama'a ta Ingila, Salisbury).
An shuka ƙwayoyin cuta cikin dare ɗaya a cikin ruwan Muller-Hinton (Oxoid, Altrincham, UK). Daga nan aka narkar da ruwan da aka yi amfani da shi a cikin dare 1:100 a cikin ruwan Mueller-Hinton sannan aka shafa 200 µl a kan membranes na Whatman cyclopore mai 0.2 µm (Whatman plc, Maidstone, UK) a kan faranti na agar na Mueller-Hinton (Sigma-Aldrich Company Ltd, Kent, Great Britain).) Tsarin biofilm na mulkin mallaka a zafin jiki na 37°C na tsawon awanni 24. An gwada waɗannan biofilm na mulkin mallaka don rage girman logarithmic.
A yanka miyar zuwa murabba'i 3 cm2 sannan a jika ta da ruwa mai tsafta. A sanya bandeji a kan biofilm ɗin yankin a kan farantin agar. An cire kowace hekta 24 na biofilm, kuma an auna ƙwayoyin cuta masu rai a cikin biofilm ɗin (CFU/ml) ta hanyar dilution na serial (10−1 zuwa 10−7) a cikin ruwan da ke rage kusurwar Rana (Merck-Millipore). Bayan awanni 24 na shiryawa a zafin jiki na 37°C, an yi ƙididdigar faranti na yau da kullun akan faranti na Mueller-Hinton. An yi kowane magani da lokacin da aka ƙayyade sau uku, kuma an maimaita ƙididdigar faranti ga kowane dilution.
Ana samun fatar cikin alade daga mata manyan aladu fari cikin mintuna 15 bayan an yanka ta bisa ga ƙa'idodin fitar da kaya na Tarayyar Turai. An aske fatar kuma an tsaftace ta da goge-goge na barasa, sannan a daskare ta a -80°C na tsawon awanni 24 don cire fata daga fata. Bayan narkewa, an wanke sassan fata 1 cm2 sau uku da PBS, 0.6% sodium hypochlorite, da 70% ethanol na tsawon mintuna 20 a kowane lokaci. Kafin cire epidermis, cire duk wani ethanol da ya rage ta hanyar wanke sau 3 a cikin PBS mara tsafta. An dasa fatar a cikin faranti mai rijiya 6 tare da membrane na nailan mai kauri 0.45-μm (Merck-Millipore) a saman da kuma kushin sha 3 (Merck-Millipore) wanda ke ɗauke da sinadarin sabulun shanu na tayi 3 ml (Sigma) wanda aka ƙara masa 10% na Dulbecco's modified Eagle. Medium (Dulbecco's Modified Eagle Medium – Aldrich Ltd.).
An noma fina-finan halittu na mulkin mallaka kamar yadda aka bayyana don nazarin fallasa fina-finan halittu. Bayan an yi wa fim ɗin biofilm ɗin da ke kan membrane na tsawon awanni 72, an shafa fim ɗin biofilm ɗin a saman fata ta amfani da madauri mai hana zubar jini sannan aka cire membrane ɗin. Daga nan aka sanya fim ɗin biofilm ɗin a kan fatar alade na ƙarin awanni 24 a zafin jiki na 37°C don ba da damar fim ɗin biofilm ɗin ya girma ya manne da fatar alade. Bayan fim ɗin biofilm ɗin ya girma kuma aka haɗa shi, an shafa wani miya mai tsawon santimita 1.5, wanda aka jika shi da ruwa mai narkewa, kai tsaye a saman fata kuma aka sanya shi a zafin jiki na 37°C na tsawon awanni 24. An hango ƙwayoyin cuta masu rai ta hanyar yin tabo ta hanyar shafa reagent na ƙwayoyin PrestoBlue (Invitrogen, Life Technologies, Paisley, UK) a saman kowane tsiro sannan a saka shi na mintuna 5. Yi amfani da kyamarar dijital ta Leica DFC425 don ɗaukar hotuna nan take akan na'urar hangen nesa ta Leica MZ8. An auna yankunan da aka yi wa launin ruwan hoda ta amfani da manhajar Image Pro sigar 10 (Media Cybernetics Inc, Rockville, MD Image-Pro (mediacy.com)). An yi amfani da na'urar daukar hoton lantarki kamar yadda aka bayyana a ƙasa.
An narkar da ƙwayoyin cuta da aka noma cikin dare ɗaya da rabi a cikin ruwan Mueller-Hinton. An ƙara μl 200 na al'ada a cikin membrane na Whatman cyclopore mai 0.2 μm (Whatman, Maidstone, UK) mai tsafta sannan aka shafa a kan agar na Mueller-Hinton. An sanya faranti na Biofilm a zafin digiri 37 na Celsius na tsawon awanni 72 don ba da damar samar da biofilm ɗin da ya girma.
Bayan kwana 3 na balaga da fim ɗin biofilm, an sanya bandeji mai girman santimita 3 a kan fim ɗin biofilm ɗin kai tsaye sannan aka saka shi a zafin jiki na digiri 37 na Celsius na tsawon awanni 24. Bayan cire bandeji daga saman fim ɗin biofilm ɗin, an ƙara 1 ml na PrestoBlue Cell Viability Reagent (Invitrogen, Waltham, MA) a saman kowane fim ɗin biofilm na tsawon daƙiƙa 20. An busar da saman kafin a yi rikodin canjin launi ta amfani da kyamarar dijital ta Nikon D2300 (Nikon UK Ltd., Kingston, UK).
A shirya al'adun dare a kan agar Mueller-Hinton, a mayar da al'adun daban-daban zuwa ruwan Mueller-Hinton mai 10 ml sannan a saka a cikin abin shaker a zafin digiri 37 Celsius (100 rpm). Bayan an dafa shi cikin dare, an narkar da al'adun da 1:100 a cikin ruwan Mueller-Hinton kuma an ga 300 µl a kan membrane na Whatman cyclopore mai 0.2 µm (Whatman International, Maidstone, UK) a kan agar Mueller-Hinton kuma a saka a zafin digiri 37 cikin awanni 72. An shafa al'adun da suka girma a kan raunin kamar yadda aka bayyana a ƙasa.
An gudanar da dukkan ayyukan da dabbobi a Jami'ar Manchester a ƙarƙashin lasisin aikin da Ofishin Kula da Jin Dadin Dabbobi da Bitar Ɗabi'a (P8721BD27) ya amince da shi kuma bisa ga jagororin da Ofishin Harkokin Cikin Gida ya buga a ƙarƙashin ASPA da aka sake duba a shekarar 2012. Duk marubutan sun bi ƙa'idodin isowa. An yi amfani da berayen C57BL/6j masu makonni takwas (Envigo, Oxon, UK) don duk nazarin in vivo. An yi wa berayen maganin sa barci da isoflurane (Piramal Critical Care Ltd, West Drayton, UK) kuma an aske saman bayansu kuma an tsaftace su. Sannan aka ba wa kowane bera raunin cirewa mai girman 2 × 6 mm ta amfani da naushin Stiefel biopsy (Schuco International, Hertfordshire, UK). Don raunukan da suka kamu da biofilm, a shafa biofilm na mulkin mallaka na awanni 72 da aka shuka a kan membrane kamar yadda aka bayyana a sama a kan fatar jikin raunin ta amfani da madauri na allurar rigakafi nan da nan bayan rauni sannan a jefar da membrane ɗin. Ana jiƙa santimita murabba'in ɗaya na miya da ruwa mai tsafta don kula da yanayin rauni mai danshi. An shafa kayan miya kai tsaye a kan kowace rauni kuma an rufe ta da fim ɗin 3M Tegaderm (3M, Bracknell, UK) da manne mai ruwa na Mastisol (Eloquest Healthcare, Ferndale, MI) da aka shafa a gefuna don samar da ƙarin mannewa. An ba da Buprenorphine (Animalcare, York, UK) a cikin adadin 0.1 mg/kg a matsayin maganin rage zafi. A cire beraye kwana uku bayan rauni ta amfani da hanyar Jadawali na 1 sannan a cire, a raba su biyu, sannan a adana yankin raunin kamar yadda ake buƙata.
An yi fenti na Hematoxylin (ThermoFisher Scientific) da eosin (ThermoFisher Scientific) bisa ga ka'idar masana'anta. An auna yankin rauni da sake farfaɗo da shi ta amfani da software na Image Pro version 10 (Media Cybernetics Inc, Rockville, MD).
An cire sassan nama a cikin xylene (ThermoFisher Scientific, Loughborough, UK), an sake sanya shi cikin ruwa da ethanol mai daraja 100-50%, sannan aka nutsar da shi na ɗan lokaci a cikin ruwan da aka cire ionized (ThermoFisher Scientific). An yi amfani da kayan aikin VectaStain Elite ABC PK-6104 (Vector Laboratories, Burlingame, CA) bisa ga ka'idar masana'anta. An narkar da manyan ƙwayoyin rigakafi ga neutrophils NIMP-R14 (Thermofisher Scientific) da macrophages Ms CD107b Pure M3/84 (BD Biosciences, Wokingham, UK) da ruwa mai toshewa 1:100 sannan aka ƙara su a saman da aka yanke, sannan aka sake haɗa su da kayan aikin antibodies guda biyu na Anti-, VectaStain ABC da Vector Nova Red Peroxidase (HRP) (Vector Laboratories, Burlingame, CA) sannan aka maye gurbinsu da hematoxylin. An samo hotunan ta amfani da na'urar hangen nesa ta Olympus BX43 da kyamarar dijital ta Olympus DP73 (Olympus, Southend-on-Sea, UK).
An gyara samfuran fata a cikin 2.5% glutaraldehyde da 4% formaldehyde a cikin 0.1 M HEPES (pH 7.4) na tsawon awanni 24 a zafin jiki na 4°C. An cire samfuran ta amfani da ethanol mai daraja kuma an busar da su a cikin CO2 ta amfani da na'urar busar da Quorum K850 mai mahimmanci (Quorum Technologies Ltd, Loughton, UK) kuma an shafa sputter da aka shafa da zinare-palladium ta amfani da tsarin Quorum SC7620 mini sputterer/glow discharge. An ɗauki hotunan samfuran ta amfani da na'urar hangen nesa ta FEI Quanta 250 scanning electron microscope (ThermoFisher Scientific) don ganin tsakiyar wurin raunin.
An shafa Toto-1 iodide (2 μM) a saman raunin linzamin kwamfuta da aka cire sannan aka saka shi a cikin injin na tsawon minti 5 a zafin digiri 37 (ThermoFisher Scientific) sannan aka yi masa magani da Syto-60 (10 μM) a zafin digiri 37 (ThermoFisher Scientific). An ƙirƙiri hotunan Z-stack na minti 15 ta amfani da Leica TCS SP8.
An tattara bayanai na kwafi na halittu da fasaha kuma an yi nazari a kansu ta amfani da manhajar Graphpad Prism V9 (GraphPad Software, La Jolla, CA). An yi amfani da nazarin bambance-bambancen hanya ɗaya tare da kwatantawa da yawa ta amfani da gwajin bayan hoc na Dunnett don gwada bambance-bambance tsakanin kowace magani da miyar da ba ta maganin ƙwayoyin cuta ba. An ɗauki ƙimar p <0.05 a matsayin mai mahimmanci.
An fara tantance ingancin miyar gel ɗin azurfa mai siffar fibrous a kan ƙwayoyin halittar Staphylococcus aureus da Pseudomonas aeruginosa a cikin vitro. Miyar azurfa tana ɗauke da dabarun azurfa daban-daban: miyar azurfa ta gargajiya tana samar da ions na Ag1+; miyar azurfa, waɗanda za su iya samar da ions na Ag1+ bayan ƙara EDTA/BC, na iya lalata matrix na biofilm kuma su fallasa ƙwayoyin cuta ga azurfa a ƙarƙashin tasirin maganin kashe ƙwayoyin cuta na azurfa. ions15 da miyar da ke ɗauke da gishirin Ag mai iskar oxygen waɗanda ke samar da ions na Ag1+, Ag2+ da Ag3+. An kwatanta ingancinsa da miyar da ba ta maganin ƙwayoyin cuta ba wadda aka yi da zare masu gelled. An tantance sauran ƙwayoyin cuta masu rai a cikin biofilm ɗin kowane sa'o'i 24 na tsawon kwanaki 8 (Hoto na 1). A rana ta 5, an sake haɗa biofilm ɗin da 3.85 × 105S. Staphylococcus aureus ko 1.22×105P. aeruginosa don tantance murmurewa na biofilm. Idan aka kwatanta da miyar da ba ta maganin ƙwayoyin cuta ba, miyar Ag1+ ba ta da tasiri sosai kan wanzuwar ƙwayoyin cuta a cikin fina-finan Staphylococcus aureus da Pseudomonas aeruginosa na tsawon kwanaki 5. Sabanin haka, miyar da ke ɗauke da gishirin Ag1+ + EDTA/BC mai iskar oxygen ta yi tasiri wajen kashe ƙwayoyin cuta a cikin fim ɗin biofilm cikin kwanaki 5. Bayan an yi wa ƙwayoyin cuta na planktonic allura akai-akai a rana ta 5, ba a ga wani abu da ya dawo da fim ɗin biofilm ɗin ba (Hoto na 1).
Ƙididdige ƙwayoyin cuta masu rai a cikin ƙwayoyin halittar Staphylococcus aureus da Pseudomonas aeruginosa bayan an yi musu magani da kayan shafa na azurfa. An yi wa ƙwayoyin halittar Staphylococcus aureus da Pseudomonas aeruginosa magani da kayan shafa na azurfa ko kayan shafa marasa maganin ƙwayoyin cuta, kuma an ƙayyade adadin ƙwayoyin halittar da suka rage a kowace awa 24. Bayan kwana 5, an sake haɗa sinadarin biofilm ɗin da 3.85×105S. Staphylococcus aureus ko 1.22×105P. An samar da ƙwayoyin halittar bacterioplankton Pseudomonas aeruginosa daban-daban don tantance murmurewa daga ƙwayoyin halittar biofilm. Zane-zane suna nuna matsakaicin kuskuren +/- na yau da kullun.
Domin ganin tasirin kayan shafa na azurfa akan wanzuwar fim ɗin biofilm, an shafa kayan shafa a kan manyan fina-finan biofilm da aka shuka a fatar alade. Bayan awanni 24, an cire kayan shafa kuma an yi wa fim ɗin biofilm fenti da launin shuɗi mai amsawa, wanda ƙwayoyin cuta masu rai ke narkewa zuwa launin ruwan hoda. Hotunan biofilm da aka yi wa magani da kayan shafa na sarrafawa ruwan hoda ne, wanda ke nuna kasancewar ƙwayoyin cuta masu rai a cikin fim ɗin biofilm (Hoto na 2A). Sabanin haka, fim ɗin biofilm da aka yi wa magani da kayan shafa na Ag oxysols galibi shuɗi ne, yana nuna cewa sauran ƙwayoyin cuta a saman fatar alade ƙwayoyin cuta ne marasa rai (Hoto na 2B). An lura da launukan shuɗi da ruwan hoda masu gauraya a cikin fina-finan biofilm da aka yi wa magani da kayan shafa na Ag1+, wanda ke nuna kasancewar ƙwayoyin cuta masu rai da marasa rai a cikin fim ɗin biofilm (Hoto na 2C), yayin da kayan shafa na EDTA/BC da ke ɗauke da Ag1+ galibi shuɗi ne tare da wasu tabo masu ruwan hoda. yana nuna wuraren da kayan shafa na azurfa ba su shafa ba (Hoto na 2D). Ƙididdigar wuraren aiki (ruwan hoda) da marasa aiki (shuɗi) ya nuna cewa facin sarrafawa yana aiki da kashi 75% (Hoto na 2E). An yi miyar Ag1+ + EDTA/BC kamar miyar gishirin Ag mai iskar oxygen, tare da ƙimar rayuwa ta 13% da 14%, bi da bi. Miyar Ag1+ kuma ta rage yuwuwar ƙwayoyin cuta da 21%. Daga nan aka lura da waɗannan fina-finan biofilms ta amfani da na'urar daukar hoto ta electron microscopy (SEM). Bayan an yi magani da miyar sarrafawa da miyar Ag1+, an ga wani Layer na Pseudomonas aeruginosa yana rufe fatar alade (Hoto na 2F,H), yayin da bayan an yi magani da miyar Ag1+, an sami ƙwayoyin cuta kaɗan kuma an sami ƙwayoyin cuta kaɗan a ƙasa. Za a iya ɗaukar zare na Collagen a matsayin tsarin nama na fatar alade (Hoto na 2G). Bayan an yi magani da miyar Ag1+ + EDTA/BC, an ga plaques na ƙwayoyin cuta da plaques na fiber na collagen a ƙarƙashinsu (Hoto na 2I).
Kwaikwayon Pseudomonas aeruginosa biofilm bayan maganin azurfa. (A–D) An yi amfani da rini na kwayoyin cuta a cikin fim ɗin Pseudomonas aeruginosa biofilms da aka girma a kan fatar alade ta amfani da rini na PrestoBlue awanni 24 bayan an yi amfani da rini na azurfa ko kuma rini na hana ƙwayoyin cuta. Kwayoyin cuta masu rai suna da ruwan hoda, ba sa rayuwa kuma fatar alade shuɗi ce. (E) Ƙididdige fim ɗin Pseudomonas aeruginosa biofilms da aka girma a kan fatar alade (tabo mai ruwan hoda) ta amfani da na'urar daukar hoto ta electron microscopy Image Pro version 10 (FI) kuma an yi musu magani da rini na azurfa ko rini na hana ƙwayoyin cuta na tsawon awanni 24. SEM sikelin bar = 5 µm. (J–M) Fim ɗin halittu na mulkin mallaka sun girma akan matattara kuma an yi musu fenti da rini mai amsawa na PrestoBlue bayan awanni 24 na haɗuwa da rini na azurfa.
Domin tantance ko kusancin da ke tsakanin kayan miya da kuma kayan biofilms ya shafi tasirin kayan miya, an yi wa kayan biofilms na mulkin mallaka da aka sanya a kan wani wuri mai faɗi magani da kayan miya na tsawon awanni 24 sannan aka yi musu fenti da rini mai amsawa. Kayan biofilm ɗin da ba a yi wa magani ba launinsu ruwan hoda ne mai duhu (Hoto na 2J). Sabanin kayan biofilms da aka yi wa magani da kayan miya da ke ɗauke da gishirin Ag mai iskar oxygen (Hoto na 2K), kayan biofilms da aka yi wa magani da kayan miya da ke ɗauke da Ag1+ ko Ag1+ + EDTA/BC sun nuna madaurin launin ruwan hoda (Hoto na 2L,M). Wannan launin ruwan hoda yana nuna kasancewar ƙwayoyin cuta masu rai kuma yana da alaƙa da yankin dinki a cikin kayan miya. Waɗannan wuraren da aka dinka suna ƙirƙirar wurare matattun da ke ba da damar ƙwayoyin cuta a cikin kayan biofilm su rayu.
Domin tantance ingancin kayan shafa na azurfa a cikin jiki, an yi wa beraye masu kauri da aka cire da suka kamu da S. aureus da P. aeruginosa biofilms magani da kayan shafa na azurfa ko kayan shafa na azurfa. Bayan kwana 3 na magani, nazarin hotunan macroscopic ya nuna ƙananan girman raunuka lokacin da aka yi musu magani da kayan shafa na gishiri mai iskar oxygen idan aka kwatanta da kayan shafa na azurfa marasa maganin ƙwayoyin cuta da sauran kayan shafa na azurfa (Hoto na 3A-H). Don tabbatar da waɗannan abubuwan da aka lura, an girbe raunuka kuma an auna yankin rauni kuma an auna reepithelialization akan sassan nama da aka shafa da hematoxylin da eosin ta amfani da software na image pro sigar 10 (Hoto na 3I-L).
Tasirin kayan shafa na azurfa akan saman rauni da sake farfaɗo da raunukan da suka kamu da biofilms. (A–H) Ƙananan ƙwayoyin da suka kamu da biofilms na Pseudomonas aeruginosa (A–D) da Staphylococcus aureus (E–H) bayan kwana uku na magani da kayan shafa marasa maganin ƙwayoyin cuta, kayan shafa na gishirin Ag mai iskar oxygen, kayan shafa na Ag1+, da kayan shafa na Ag1+. Hotunan macroscopic masu wakiltar. raunukan beraye da kayan shafa na Ag1+ + EDTA/BC. (IL) Wakili kamuwa da cutar Pseudomonas aeruginosa, sassan histological da aka shafa da hematoxylin da eosin, ana amfani da su don auna yankin rauni da sake farfaɗo da epithelial. Ƙididdige yankin rauni (M, O) da kashi na sake farfaɗo da epithelialization (N, P) na raunukan da suka kamu da Pseudomonas aeruginosa (M, N) da Staphylococcus aureus (O, P) biofilms (ga kowane rukunin magani n = 12). Zane-zane suna nuna matsakaicin kuskuren +/- na yau da kullun. * yana nufin p = < 0.05 ** yana nufin p = < 0.01; Sikelin macroscopic = 2.5 mm, sikelin histological = 500 µm.
Ƙididdige yankin rauni a cikin raunukan da suka kamu da cutar Pseudomonas aeruginosa biofilm (Hoto na 3M) ya nuna cewa raunukan da aka yi wa magani da Ag oxysalts suna da matsakaicin girman rauni na 2.5 mm2, yayin da miya wadda ba ta maganin ƙwayoyin cuta ba tana da matsakaicin girman rauni na 3.1 mm2, wanda ba gaskiya ba ne. ya kai ga mahimmancin ƙididdiga (Hoto na 3M). p = 0.423). Raunukan da aka yi wa magani da Ag1+ ko Ag1+ + EDTA/BC ba su nuna raguwar yankin rauni ba (3.1 mm2 da 3.6 mm2, bi da bi). Jiyya da miya mai gishirin Ag mai iskar oxygen ya haɓaka sake-epithelialization zuwa mafi girma fiye da miya wadda ba ta maganin ƙwayoyin cuta ba (34% da 15%, bi da bi; p = 0.029) da Ag1+ ko Ag1+ + EDTA/BC (10% da 11%) (Hoto na 3N). . , bi da bi).
An lura da irin wannan yanayin a yankin rauni da kuma sake farfaɗowar epithelial a cikin raunukan da suka kamu da S. aureus biofilms (Hoto na 3O). Kayan miya da ke ɗauke da gishirin azurfa mai iskar oxygen sun rage yankin rauni (2.0 mm2) da kashi 23% idan aka kwatanta da kayan miya marasa maganin ƙwayoyin cuta (2.6 mm2), kodayake wannan raguwar ba ta da mahimmanci (p = 0.304) (Hoto na 3O). Bugu da ƙari, an ɗan rage yankin rauni a cikin rukunin maganin Ag1+ (2.4 mm2), yayin da raunin da aka yi wa magani da kayan miya na Ag1+ + EDTA/BC bai rage yankin rauni ba (2.9 mm2). Gishirin iskar oxygen na Ag kuma ya haɓaka sake farfaɗo da raunukan da suka kamu da S. aureus biofilm (31%) zuwa mafi girma fiye da waɗanda aka yi wa magani da kayan miya marasa maganin ƙwayoyin cuta (12%, p = 0.003) (Hoto na 3P). Miyar Ag1+ (16%, p = 0.903) da miyar Ag+1 + EDTA/BC (14%, p = 0.965) sun nuna matakan sake farfaɗo da epithelial kamar sarrafawa.
Domin ganin tasirin miyar azurfa akan matrix na biofilm, an yi amfani da fenti na iodide na Toto 1 da Syto 60 (Hoto na 4). Iodide na Toto 1 rini ne wanda ba ya shiga cikin tantanin halitta wanda za a iya amfani da shi don ganin daidai nucleic acid na extracellular, waɗanda ke da yawa a cikin EPS na biofilms. Syto 60 rini ne mai shiga cikin tantanin halitta wanda ake amfani da shi azaman maganin hana tabo16. Lura da Toto 1 da Syto 60 iodide a cikin raunuka da aka yi wa allurar biofilms na Pseudomonas aeruginosa (Hoto na 4A-D) da Staphylococcus aureus (Hoto na 4I-L) ya nuna cewa bayan kwana 3 na maganin miyar, EPS a cikin biofilm ya ragu sosai. yana ɗauke da gishirin oxygen Ag da Ag1+ + EDTA/BC. Miyar Ag1+ ba tare da ƙarin abubuwan hana ƙwayoyin cuta ba ta rage DNA mara tantanin halitta sosai a cikin raunuka da aka yi wa allurar Pseudomonas aeruginosa amma ba su da tasiri sosai a cikin raunuka da aka yi wa allurar Staphylococcus aureus.
Hoton fim ɗin raunin da aka yi a cikin jiki bayan kwana 3 na magani da maganin sarrafawa ko na azurfa. Hotunan Confocal na Pseudomonas aeruginosa (A–D) da Staphylococcus aureus (I–L) an yi musu fenti da Toto 1 (kore) don ganin ƙwayoyin nucleic na extracellular, wani ɓangare na polymers na biofilm na extracellular. Don yin fenti da ƙwayoyin nucleic na intracellular, yi amfani da Syto 60 (ja). acid. P. Duba na'urar daukar hoto ta electron na raunukan da suka kamu da Pseudomonas aeruginosa (E–H) da Staphylococcus aureus (M–P) biofilms bayan kwana 3 na magani da maganin sarrafawa da na azurfa. Scal bar = 5 µm. Confocal imageing ma'aunin bar = 50 µm.
Binciken na'urar daukar hoton lantarki ya nuna cewa beraye da aka yi musu allurar rigakafin ƙwayoyin halittar Pseudomonas aeruginosa (Hoto na 4E-H) da Staphylococcus aureus (Hoto na 4M-P) suna da ƙananan ƙwayoyin cuta a cikin raunukansu bayan kwana 3 na magani da duk wani maganin azurfa.
Domin tantance tasirin miyar azurfa akan kumburin rauni a cikin beraye masu kamuwa da biofilm, an yi wa sassan raunukan da suka kamu da biofilm magani da maganin sarrafawa ko na azurfa na tsawon kwanaki 3 fenti ta hanyar amfani da ƙwayoyin rigakafi na musamman ga neutrophils da macrophages. Ƙimar adadi na neutrophils da macrophages a ciki. kyallen granulation. Hoto na 5). Duk miyar azurfa ta rage adadin neutrophils da macrophages a cikin raunukan da suka kamu da Pseudomonas aeruginosa idan aka kwatanta da miyar da ba ta maganin rigakafi ba bayan kwana uku na magani. Duk da haka, magani da miyar gishirin azurfa mai iskar oxygen ya haifar da raguwar neutrophils (p = <0.0001) da macrophages (p = <0.0001) idan aka kwatanta da sauran miyar azurfa da aka gwada (Hoto na 5I,J). Kodayake Ag1+ + EDTA/BC yana da tasiri mafi girma akan biofilm na rauni, ya rage matakan neutrophil da macrophage zuwa ƙasa da miyar Ag1+. An kuma lura da raunuka masu matsakaicin kamuwa da cutar S. aureus biofilm bayan an yi musu ado da Ag (p = <0.0001), Ag1+ (p = 0.0008) da Ag1++ EDTA/BC (p = 0.0043) oxisols idan aka kwatanta da sarrafawa. An lura da irin wannan yanayin ga bandeji na neutropenia. (Hoto na 5K). Duk da haka, kawai miyar gishirin Ag mai iskar oxygen ta nuna raguwa mai yawa a yawan macrophages a cikin kyallen granulation idan aka kwatanta da sarrafawa a cikin raunukan da suka kamu da cutar S. aureus biofilms (p = 0.0339) (Hoto na 5L).
An auna ƙwayoyin neutrophils da macrophages a cikin raunukan da suka kamu da cutar Pseudomonas aeruginosa da Staphylococcus aureus biofilms bayan kwana 3 na magani da magungunan hana ƙwayoyin cuta ko kuma na azurfa. An auna ƙwayoyin neutrophils (AD) da macrophages (EH) a cikin sassan nama da aka yi wa fenti da ƙwayoyin rigakafi na musamman ga ƙwayoyin neutrophils ko macrophages. Adadin ƙwayoyin neutrophils (I da K) da macrophages (J da L) a cikin raunukan da suka kamu da cutar Pseudomonas aeruginosa (I da J) da Staphylococcus aureus (K & L) biofilms. N = 12 ga kowace ƙungiya. Zane-zane suna nuna matsakaicin +/- kuskuren yau da kullun, ƙimar mahimmanci idan aka kwatanta da maganin hana ƙwayoyin cuta, * yana nufin p = < 0.05, ** yana nufin p = < 0.01; *** yana nufin p = < 0.001; yana nuna p = <0.0001).
Sai muka tantance tasirin miyar azurfa akan warkarwa mara kamuwa da cuta. An yi wa raunukan da ba su kamu da cutar magani da miyar da ba ta da ƙwayoyin cuta ko miyar azurfa na tsawon kwana 3 (Hoto na 6). Daga cikin miyar azurfa da aka gwada, raunukan da aka yi wa magani da miyar gishiri mai iskar oxygen kawai sun bayyana ƙarami akan hotunan macroscopic fiye da raunukan da aka yi wa magani da magani (Hoto na 6A-D). Ƙididdige yankin rauni ta amfani da nazarin histological ya nuna cewa matsakaicin yankin rauni bayan an yi masa magani da miyar Ag oxysols shine 2.35 mm2 idan aka kwatanta da 2.96 mm2 ga raunukan da aka yi wa magani da ƙungiyar kulawa, amma wannan bambancin bai kai ga mahimmancin ƙididdiga ba (p = 0.488) (Hoto na 6I). Akasin haka, ba a lura da raguwar yankin rauni ba bayan an yi masa magani da miyar Ag1+ (3.38 mm2, p = 0.757) ko Ag1+ + EDTA/BC (4.18 mm2, p = 0.054) idan aka kwatanta da ƙungiyar kulawa. An lura da ƙaruwar sake farfaɗowar epithelial tare da miyar Ag oxysol idan aka kwatanta da ƙungiyar kulawa (30% vs. 22%, bi da bi), kodayake wannan bai kai ga mahimmanci ba (p = 0.067), wannan yana da matuƙar muhimmanci kuma yana tabbatar da sakamakon da ya gabata. Miyar da oxysols yana haɓaka sake farfaɗowa. -Epithelization na raunuka marasa kamuwa17. Sabanin haka, magani da miyar Ag1+ ko Ag1+ + EDTA/BC bai yi wani tasiri ba ko ya nuna raguwar sake farfaɗowa idan aka kwatanta da sarrafawa.
Tasirin miyar raunukan azurfa akan warkar da rauni a cikin beraye marasa kamuwa da cuta tare da cirewa gaba ɗaya. (AD) Hotunan macroscopic na raunuka bayan kwana uku na magani tare da miyar da ba ta maganin ƙwayoyin cuta ba da miyar azurfa. (EH) Waɗansu sassan raunuka da aka yi wa fenti da hematoxylin da eosin. An ƙididdige adadin yankin rauni (I) da kashi na sake epithelialization (J) daga sassan histological a tsakiyar raunin ta amfani da software na nazarin hoto (n = 11–12 ga kowace ƙungiyar magani). Zane-zane suna nuna matsakaicin +/- kuskuren yau da kullun. * yana nufin p = <0.05.
Silver yana da dogon tarihi na amfani da shi azaman maganin kashe ƙwayoyin cuta a warkar da raunuka, amma nau'ikan tsari daban-daban da hanyoyin bayarwa na iya haifar da bambance-bambance a cikin ingancin maganin kashe ƙwayoyin cuta 18. Bugu da ƙari, ba a fahimci kaddarorin maganin kashe ƙwayoyin cuta na takamaiman tsarin isar da azurfa ba. Kodayake amsawar garkuwar jiki mai masaukin baki yana da tasiri sosai akan ƙwayoyin cuta na planktonic, gabaɗaya ba shi da tasiri sosai akan biofilms19. Kwayoyin cuta na Planktonic ana iya yin phagocytose cikin sauƙi ta hanyar macrophages, amma a cikin biofilms, ƙwayoyin da aka tara suna haifar da ƙarin matsaloli ta hanyar iyakance martanin mai masaukin baki har zuwa lokacin da ƙwayoyin rigakafi za su iya fuskantar apoptosis da sakin abubuwan da ke haifar da kumburi don haɓaka amsawar garkuwar jiki20. An lura cewa wasu leukocytes na iya shiga biofilms21 amma ba za su iya phagocytose ƙwayoyin cuta da zarar an lalata wannan kariya22. Ya kamata a yi amfani da hanyar gabaɗaya don tallafawa martanin garkuwar jiki mai masaukin baki akan kamuwa da cutar biofilm ta rauni. Rage rauni na iya lalata biofilm ɗin jiki kuma ya cire yawancin nauyin bio, amma martanin garkuwar jiki mai masaukin baki na iya zama mara tasiri akan sauran biofilm, musamman idan martanin garkuwar jiki mai masaukin baki ya lalace. Saboda haka, magungunan kashe ƙwayoyin cuta kamar su kayan shafa na azurfa na iya tallafawa martanin garkuwar jiki na mai masaukin baki da kuma kawar da cututtukan biofilm. Haɗawa, tattarawa, narkewa, da kuma isar da sinadarai na iya yin tasiri ga ingancin maganin kashe ƙwayoyin cuta na azurfa. A cikin 'yan shekarun nan, ci gaban fasahar sarrafa azurfa ya sa waɗannan kayan shafa su fi tasiri9,23. Yayin da fasahar gyaran azurfa ke ci gaba, yana da mahimmanci a fahimci tasirin waɗannan kayan shafa wajen sarrafa kamuwa da raunuka, kuma mafi mahimmanci, tasirin waɗannan nau'ikan azurfa masu ƙarfi akan yanayin rauni da warkarwa.
A cikin wannan binciken, mun kwatanta tasirin kayan miya guda biyu na azurfa da kayan miya na azurfa na gargajiya waɗanda ke samar da ions na Ag1+ akan biofilms ta amfani da samfuran in vitro da in vivo daban-daban. Mun kuma tantance tasirin waɗannan kayan miya akan yanayin rauni da warkarwa ba tare da kamuwa da cuta ba. Don rage tasirin matrix na isarwa, duk kayan miya na azurfa da aka gwada an haɗa su da carboxymethylcellulose.
Bincikenmu na farko game da waɗannan miya na azurfa akan biofilms na mulkin mallaka na Pseudomonas aeruginosa da Staphylococcus aureus ya nuna cewa, ba kamar miya na Ag1+ na gargajiya ba, miya na azurfa guda biyu masu inganci, Ag1+ + EDTA/BC da gishirin Ag mai iskar oxygenated, suna da tasiri a 5. Suna kashe ƙwayoyin cuta na biofilm cikin 'yan kwanaki. Bugu da ƙari, waɗannan miya suna hana sake ƙirƙirar biofilm bayan an sake fallasa su ga ƙwayoyin cuta na planktonic. Miyar Ag1+ ta ƙunshi azurfa chloride, mahaɗin azurfa da matrix na tushe kamar Ag1+ + EDTA/BC, kuma tana da tasiri mai iyaka akan yuwuwar ƙwayoyin cuta a cikin biofilm a cikin lokaci ɗaya. Lura da cewa miya na Ag1+ + EDTA/BC ya fi tasiri akan biofilm fiye da miya na Ag1+ wanda ya ƙunshi matrix iri ɗaya da mahaɗin azurfa yana goyon bayan ra'ayin cewa ana buƙatar ƙarin sinadarai don ƙara tasirin azurfa chloride akan biofilm, kamar yadda aka ruwaito a wasu wurare15. Waɗannan sakamakon sun goyi bayan ra'ayin cewa BC da EDTA suna taka muhimmiyar rawa wajen ba da gudummawa ga ingancin miya gabaɗaya kuma rashin wannan sashi a cikin miya Ag1+ wataƙila ya taimaka wajen gazawar nuna ingancin in vitro. Mun gano cewa miya Ag gishiri mai iskar oxygen wanda ke samar da ions Ag2+ da Ag3+ ya nuna ingantaccen maganin kashe ƙwayoyin cuta fiye da Ag1+ kuma a matakan kama da Ag1+ + EDTA/BC. Duk da haka, saboda yawan ƙarfin redox, ba a san tsawon lokacin da ions Ag3+ ke aiki da tasiri akan biofilms na rauni ba don haka ya cancanci ƙarin bincike. Bugu da ƙari, akwai miya daban-daban da yawa waɗanda ke samar da ions Ag1+ waɗanda ba a gwada su ba a cikin wannan binciken. Waɗannan miya sun ƙunshi mahaɗan azurfa daban-daban, yawansu, da matrices na tushe, waɗanda zasu iya tasiri ga isar da ions Ag1+ da ingancinsu akan biofilms. Hakanan ya kamata a lura cewa akwai samfuran in vitro da in vivo da yawa da ake amfani da su don kimanta ingancin miya rauni akan biofilms. Nau'in samfurin da aka yi amfani da shi, da kuma abubuwan da ke cikin gishiri da furotin na kafofin watsa labarai da aka yi amfani da su a cikin waɗannan samfuran, zasu yi tasiri ga ingancin miya. A cikin samfurin in vivo ɗinmu, mun bar biofilm ɗin ya girma a cikin vitro sannan muka mayar da shi zuwa saman fatar raunin. Amsar garkuwar linzamin kwamfuta mai masaukin baki tana da tasiri sosai akan ƙwayoyin cuta na planktonic da aka shafa wa raunin, ta haka ne ke samar da biofilm yayin da raunin ke warkewa. Ƙara biofilm ɗin da ya girma a cikin rauni yana iyakance tasirin martanin garkuwar jiki ga samuwar biofilm ta hanyar barin biofilm ɗin da ya girma ya kafa kansa a cikin raunin kafin a fara warkarwa. Don haka, samfurinmu yana ba mu damar tantance tasirin miya mai kashe ƙwayoyin cuta akan biofilm ɗin da suka girma kafin raunuka su fara warkewa.
Mun kuma gano cewa daidaita suturar ta shafi tasirin kayan miya na azurfa akan biofilms da aka girma a cikin vitro da fatar alade. Ana ɗaukar kusanci da raunin a matsayin mahimmanci ga ingancin maganin rigakafi na kayan miya24,25. Kayan miya da ke ɗauke da gishirin Ag mai iskar oxygen suna da kusanci da biofilms masu girma, wanda ya haifar da raguwa mai yawa a cikin adadin ƙwayoyin cuta masu rai a cikin biofilm bayan awanni 24. Sabanin haka, lokacin da aka yi wa magani da kayan miya na Ag1+ da Ag1+ + EDTA/BC, adadi mai yawa na ƙwayoyin cuta masu rai sun rage. Waɗannan kayan miya suna ɗauke da dinki a duk tsawon kayan miya, wanda ke haifar da wuraren da ba su da rai waɗanda ke hana kusanci da biofilm. A cikin bincikenmu na in vitro, waɗannan wuraren da ba su da rai sun hana kashe ƙwayoyin cuta masu rai a cikin biofilm. Mun tantance yuwuwar ƙwayoyin cuta ne kawai bayan awanni 24 na magani; Bayan lokaci, yayin da kayan miya suka zama cikakke, ƙila a sami ƙarancin sarari mara rai, wanda ke rage yankin waɗannan ƙwayoyin cuta masu rai. Duk da haka, wannan yana nuna mahimmancin abun da ke cikin kayan miya, ba kawai nau'in azurfa a cikin kayan miya ba.
Duk da cewa nazarin in vitro yana da amfani wajen kwatanta tasirin fasahar azurfa daban-daban, yana da mahimmanci a fahimci tasirin waɗannan miya akan biofilms a cikin jiki, inda nama mai masaukin baki da martanin garkuwar jiki ke ba da gudummawa ga ingancin miya akan biofilms. An lura da tasirin waɗannan miya akan biofilms na rauni ta amfani da na'urar duba electron microscopy da kuma tabon EPS na biofilm ta amfani da rini na DNA na cikin jiki da na waje. Mun gano cewa bayan kwana 3 na magani, duk miya sun yi tasiri wajen rage DNA mara tantanin halitta a cikin raunukan da suka kamu da biofilm, amma miya Ag1+ bai yi tasiri sosai a cikin raunukan da suka kamu da Staphylococcus aureus ba. Duba microscopy na electron kuma ya nuna cewa ƙananan ƙwayoyin cuta ne ke cikin raunukan da aka yi wa miya da azurfa, kodayake wannan ya fi bayyana tare da miya gishirin Ag mai iskar oxygen da miya Ag1+ + EDTA/BC idan aka kwatanta da miya Ag1+. Waɗannan bayanai sun nuna cewa miya azurfa da aka gwada suna da tasiri daban-daban akan tsarin biofilm, amma babu ɗayan miya azurfa da ya sami damar kawar da biofilm, yana tallafawa buƙatar hanyar gaba ɗaya don magance cututtukan biofilm na rauni; amfani da madaurin azurfa. Ana yin magani kafin a cire mafi yawan fim ɗin biofilm ɗin.
Raunuka masu tsanani galibi suna cikin yanayi mai tsanani na kumburi, tare da ƙwayoyin kumburi da yawa da ke wanzuwa a cikin kyallen rauni na tsawon lokaci, suna haifar da lalacewar nama da kuma rage iskar oxygen da ake buƙata don ingantaccen metabolism da aiki a cikin raunin26. Biofilms suna ƙara ta'azzara wannan yanayin rauni mai ƙiyayya ta hanyar yin mummunan tasiri ga warkarwa ta hanyoyi daban-daban, gami da hana yaduwar ƙwayoyin halitta da ƙaura da kunna cytokines masu haifar da kumburi27. Yayin da kayan shafa na azurfa suka ƙara yin tasiri, yana da mahimmanci a fahimci tasirin da suke da shi akan yanayin rauni da warkarwa.
Abin sha'awa, duk da cewa duk wani kayan shafa na azurfa ya shafi abun da ke cikin biofilm, kayan shafa na gishirin azurfa mai iskar oxygen ne kawai ya ƙara sake haifar da waɗannan raunukan da suka kamu da cutar. Waɗannan bayanai sun goyi bayan bincikenmu na baya17 da na Kalan et al. (2017)28, wanda ya nuna ingantaccen aminci da guba na gishirin azurfa mai iskar oxygen, saboda ƙarancin yawan azurfa ya yi tasiri akan biofilms.
Bincikenmu na yanzu ya nuna bambance-bambancen da ke tsakanin kayan shafa na azurfa masu kashe ƙwayoyin cuta da tasirin wannan fasaha akan yanayin rauni da warkarwa ba tare da kamuwa da cuta ba. Duk da haka, waɗannan sakamakon sun bambanta da na baya da ke nuna cewa kayan shafa na Ag1+ + EDTA/BC sun inganta sigogin warkarwa na kunnuwan zomo da suka ji rauni a cikin jiki. Duk da haka, wannan na iya zama saboda bambance-bambance a cikin samfuran dabbobi, lokutan aunawa, da hanyoyin amfani da ƙwayoyin cuta29. A wannan yanayin, an ɗauki ma'aunin rauni kwanaki 12 bayan rauni don ba da damar sinadaran aiki na kayan shafa su yi aiki akan biofilm na tsawon lokaci. Wannan yana da goyan bayan wani bincike wanda ya nuna cewa gyambon ƙafa da suka kamu da cutar da aka yi wa magani da Ag1+ + EDTA/BC da farko sun ƙaru a girma bayan mako guda na magani, sannan kuma a cikin makonni 3 masu zuwa na magani da Ag1+ + EDTA/BC da kuma cikin makonni 4 na amfani da magungunan da ba sa kashe ƙwayoyin cuta. magunguna. Kayan shafa na CMC don rage girman gyambo30.
An riga an nuna cewa wasu siffofi da yawan azurfa suna da guba a cikin vitro 11, amma waɗannan sakamakon in vitro ba koyaushe suke haifar da mummunan sakamako a cikin jiki ba. Bugu da ƙari, ci gaban fasahar azurfa da fahimtar mahaɗan azurfa da yawan su a cikin miya sun haifar da haɓaka miya masu aminci da tasiri da yawa na azurfa. Duk da haka, yayin da fasahar miya ta azurfa ke ci gaba, yana da mahimmanci a fahimci tasirin waɗannan miya akan yanayin rauni31,32,33. An ruwaito a baya cewa ƙaruwar yawan sake-epithelialization ya yi daidai da ƙaruwar adadin macrophages masu hana kumburi na M2 idan aka kwatanta da yanayin M1 mai hana kumburi. An lura da wannan a cikin wani samfurin linzamin kwamfuta na baya inda aka kwatanta miya na rauni na azurfa hydrogel da sulfadiazine na azurfa da hydrogels marasa maganin ƙwayoyin cuta34.
Raunuka na yau da kullun na iya nuna kumburi mai yawa kuma an lura cewa kasancewar ƙwayoyin neutrophils masu yawa na iya zama illa ga warkar da rauni35. A cikin wani bincike da aka yi a kan beraye da suka ƙare da ƙwayoyin neutrophil, kasancewar ƙwayoyin neutrophils yana jinkirta sake farfaɗowa. Kasancewar ƙwayoyin neutrophils masu yawa yana haifar da yawan ƙwayoyin proteases da nau'ikan iskar oxygen masu amsawa, kamar su superoxide da hydrogen peroxide, waɗanda ke da alaƙa da raunuka na yau da kullun da na warkarwa a hankali37,38. Haka nan, ƙaruwar adadin ƙwayoyin macrophage, idan ba a sarrafa su ba, na iya haifar da jinkirin warkar da rauni39. Wannan ƙaruwar yana da mahimmanci musamman idan ƙwayoyin macrophages ba za su iya canzawa daga ƙwayar pro-inflammatory zuwa ƙwayar pro-warkarwa ba, wanda ke haifar da raunukan da ba su iya fita daga matakin kumburi na warkarwa40. Mun lura da raguwar ƙwayoyin neutrophils da macrophages a cikin raunukan da suka kamu da cutar biofilm bayan kwana 3 na magani da duk wani maganin azurfa, amma raguwar ta fi bayyana tare da maganin gishiri mai iskar oxygen. Wannan raguwar na iya zama sakamakon kai tsaye na martanin garkuwar jiki ga azurfa, martani ga raguwar nauyin halittu, ko raunin yana cikin matakin warkewa na gaba don haka rage ƙwayoyin rigakafi a cikin raunin. Rage adadin ƙwayoyin kumburi a cikin raunin na iya kiyaye yanayi mai dacewa don warkar da rauni. Hanyar aikin yadda Ag oxysalts ke haɓaka warkarwa mai zaman kanta ba ta da alaƙa da kamuwa da cuta ba a fayyace ba, amma ikon Ag oxysalts na samar da iskar oxygen da lalata matakan cutarwa na hydrogen peroxide, mai shiga tsakani na kumburi, na iya bayyana wannan kuma yana buƙatar ƙarin bincike17.
Raunuka masu kamuwa da cuta na dogon lokaci waɗanda ba sa warkarwa suna haifar da matsala ga likitoci da marasa lafiya. Duk da cewa yawancin kayan shafa suna da'awar ingancin maganin rigakafi, bincike ba kasafai yake mai da hankali kan wasu muhimman abubuwan da ke shafar yanayin rauni ba. Wannan binciken ya nuna cewa fasahohin azurfa daban-daban suna da tasirin maganin rigakafi daban-daban kuma, mahimmanci, tasirin daban-daban akan yanayin rauni da warkarwa, ba tare da la'akari da kamuwa da cuta ba. Duk da cewa waɗannan nazarin in vitro da in vivo sun nuna ingancin waɗannan kayan shafa wajen magance cututtukan rauni da haɓaka warkarwa, ana buƙatar gwaje-gwajen da aka sarrafa bazuwar don tantance ingancin waɗannan kayan shafa a asibitin.
Bayanan da aka yi amfani da su da/ko aka yi nazari a kansu a lokacin binciken na yanzu suna samuwa daga marubucin da ya dace bisa ga buƙata mai ma'ana.
Lokacin Saƙo: Yuli-15-2024