Ucwaningo lwe-methylene blue elwa namagciwane nesikhunta: ukukhanya kuqhathaniswa nobumnyama

Abafundi ababili babona ibinzana elithi i-methylene blue elwa namagciwane bese becabanga izinto ezingafani. Omunye ucabanga uketshezi oluluhlaza okwesibhakabhaka olunoshevu obulala amagciwane lapho luwathinta. Omunye ucabanga inqubo lapho udayi nokukhanya okubomvu kuwabhubhisa. Yomibili le mibono ineqiniso elithile. Kodwa awukho ophelele, futhi ukuyixuba yikho okwenza izimangalo eziningi eziku-inthanethi ziphambuke.

Lo mehluko yiwona okuxoxwa ngawo kulokhu kubuyekezwa. I-methylene blue inobuthi obulinganiselwe ebumnyameni uma ukugxila kwayo kuphakeme ngokwanele. Ngokukhanya okubomvu iba into evuselelwa ukukhanya: imunca i-photon, ifinyelele esimweni se-triplet esikwazi ukusabela, bese iqhuba ukusabela kwamakhemikhali kwe-oxidation okulimaza izinto eziningi ngesikhathi esisodwa. Le ndlela evuselelwa ukukhanya inegama layo, ukwelapha amagciwane nge-photodynamic, kanye nemibandela yayo. Ukugxila kukadayi kukodwa akuyichazi. Ubude begagasi, umthamo wokukhanya, ukufukamela, umoya-mpilo, nesidalwa esihlolwayo kunquma umphumela.

Imibono emibili yokubulala

Qala ngokucabanga indlela esebenza ebumnyameni. I-methylene blue ingudayi we-phenothiazinium one-charge kagesi ephozithivu. Inamathela ezingxenyeni ezizungeze amaseli amagciwane, ingene kumaseli ngamazinga ahlukene, bese ibamba iqhaza ekusabeleni kwe-redox nasekubopheni amakhemikhali. Awekho ama-photon ahilelekile. Ukubulala kuyenzeka, kodwa kuphela ekugxileni okuhluka kakhulu kuye ngesidalwa, futhi indlela esebenza ngayo ayiqondiswanga entweni eyodwa kuphela.

Indlela ye-photodynamic yengeza ukukhanya. Ama-photon abomvu anobude begagasi obucishe bube ngu-630 kuya ku-680 nm ahambisana nebhendi yokumunca kadayi, kanti isiqongo se-monomer siseduze kuka-664 nm. Ukumunca kufaka udayi esimweni se-singlet esihlala isikhashana, bese udlulela esimweni se-triplet esihlala isikhathi eside kunaleso. Kusukela lapho kuvela amaqoqo amabili okusabela. Ekusabeleni kohlobo I, i-triplet idlulisela i-electron entweni okusabelwana nayo bese iqala izindlela zama-radical ezihlanganisa izinhlobo ze-superoxide ne-hydroxyl. Ekusabeleni kohlobo II, idlulisela amandla ngokuqondile kumoya-mpilo osesimweni samandla esiphansi bese yenza umoya-mpilo we-singlet. Zombili zingasebenza ngesikhathi esisodwa, futhi ukubiza udayi ngokuthi uyinto esebenza kuphela ngomoya-mpilo we-singlet kwenza indaba ibe lula ngokweqile. Ukubuyekezwa kobudlelwano phakathi kwesakhiwo nokusebenza kwezinto ze-phenothiazinium ezivuselelwa ukukhanya kukukhombisa lokhu ngokuqondile: phakathi kodayi abahlobene, ubuthi obubangelwa ukukhanya kumabhaktheriya abuzange buhambisane nenani lomoya-mpilo we-singlet okhiqizwayo kuphela, ngoba ukungena kukadayi, indawo ohlala kuyo, nokuncishiswa kwawo ube yisimo se-leuco esingasebenzi kwakubaluleke njengendlela yawo yokusebenzisana nokukhanya. Ukusabela kwamakhemikhali ngokukhanya okuchaza umthetho wobude begagasi kuchazwe ekubuyekezweni okuhambisana nalokhu okuphathelene nodayi nokukhanya okubomvu.

Umoya-mpilo we-singlet ubalulekile kwesinye isici esiwusizo. Usabela nama-biomolecule amaningi ngesikhathi esisodwa, ngakho ukuhlasela kwenzeka ngezindlela eziningi kunokuba kuqondiswe ku-enzyme eyodwa noma kuphrotheni eyodwa ebophayo. Incazelo eyisisekelo yokwelapha amagciwane ngamakhemikhali avuselelwa ukukhanya yakubeka lokhu njengesizathu sokuthi izindlela ezijwayelekile zokumelana nemithi zingabi nethonya elikhulu: iphrotheni eyengeziwe ebopha i-penicillin noma i-topoisomerase eguqukile ayivimbi ukuqhuma komoya-mpilo osabelayo. Lo mbono waqinisekiswa ekuhlolweni kwe-Staphylococcus aureus emelana ne-methicillin kanye nama-enterococci amelana ne-vancomycin, njengoba ithebula elingezansi likhombisa. Lokhu akusho ukuthi ukumelana akunakwenzeka, kodwa kusho ukuthi kusebenza ngendlela ehlukile. Amaphampu akhipha izinto kumaseli kanye nezivikelo ezilwa ne-oxidation zisengaguqula umthamo odingekayo, ikakhulukazi esikhuntwini.

Ithebula lezidalwa namamodeli ocwaningo

Leli thebula liyingxenye eyinhloko yalesi sihloko. Inani ngalinye lihambisana nenqubo ethile: udayi ofanayo, kodwa umthamo wokukhanya noma ukufukamela okuhlukile, kunikeza inani elihlukile. Ungathathi i-MBC, i-MLC, ne-MIC njengokungashintshaniswa. Lapha i-MBC ne-MLC kusho ukuthi akukho okusindile lapho kudluliselwa kwenye indawo yokukhulisa, kanti i-MIC isho ukuvinjelwa kokukhula. Imibandela yokukhanya ibhalwe ekungeneni ngakunye.

Isidalwa nemodeli	Umphumela ebumnyameni	Umphumela ngokukhanya	Imibandela
S. aureus NCTC 6571, amaseli antanta ngokukhululeka oketshezini	MBC 5 uM	MBC 5 uM	Ithebula lokubuyekeza, ukukhanya okumhlophe 12 J per cm2
S. aureus NCTC 6571, amaseli antanta ngokukhululeka oketshezini	MLC 2.5 uM	MLC 1 uM	Inqubo yocwaningo lokuqala yango-1997, 1.7 mW per cm2 isikhathi esingu-1 hour, cishe 6.3 J per cm2, inani lamagciwane okuqalwa ngawo eliseduze kuka-10^6 CFU per mL, njengoba kubikwe ocwaningweni lokuqala lwe-phenothiazinium ebulala amabhaktheriya ngokukhanya
MRSA EMRSA-15, amaseli antanta ngokukhululeka	MBC 80 uM	MBC 20 uM	Ithebula lokubuyekeza, cishe 12 J per cm2, ukwanda kokuzwela ngokuphindwe kane
MRSA EMRSA-16, amaseli antanta ngokukhululeka	MBC 40 uM	MBC 10 uM	Inqubo efanayo nale engenhla
E. coli NCTC 9001, amaseli antanta ngokukhululeka	MBC 500 uM	MBC 100 uM	Inqubo efanayo, ukwanda kokuzwela ngokuphindwe kahlanu
P. aeruginosa NCTC 10622, amaseli antanta ngokukhululeka	MBC 500 uM	MBC 25 uM	Inqubo efanayo, ukwanda kokuzwela ngokuphindwe amashumi amabili
E. faecalis NCTC 775, amaseli antanta ngokukhululeka	MLC 100 uM	MLC 100 uM	Inqubo yango-1997 ku-6.3 J per cm2, akukho ukuthuthuka kulolu hlobo lwamagciwane nakulo mthamo
E. faecium emelana ne-vancomycin, amaseli antanta ngokukhululeka	MBC 400 uM	MBC 400 uM	Ithebula lokubuyekeza, akukho ukuthuthuka ngodayi oyisisekelo, kuyilapho izinguqulo zawo ezinamaqembu e-methyl nezixwaya amanzi kakhulu zenze ngcono, ngokusho kocwaningo lwe-enterococcus emelana ne-vancomycin
Candida albicans, amaseli antanta ngokukhululeka	MIC ngaphezu kuka-12.79 ug per mL	MIC 12.79, 6.39, no-1.59 ug per mL ku-5, 10, no-15 J per cm2	Ukufukamela kokuqala ebumnyameni isikhathi esingu-30 minute, i-LED ebomvu engu-600 kuya ku-650 nm enesiqongo ku-635 nm, ngokusho kocwaningo lokusabela emthamo lapho i-methylene blue ilwa nesikhunta ngokukhanya
C. albicans YEM14 iqhathaniswa nenguqulo ekhiqiza ngokweqile amaphampu akhipha izinto, ukubulawa ngokubalwa kwe-CFU	Akusebenzi lapha	Ukubulawa okucishe kube ngu-5 log kohlobo lokuqala, cishe akukho ukubulawa kwenguqulo ekhiqiza ngokweqile i-CDR1/CDR2	Udayi ongu-100 uM, ukufukamela isikhathi esingu-30 minute, 660 nm ku-60 J per cm2, ngokusho kocwaningo lwemiphumela yamaphampu akhipha izinto ekubulaleni kwe-methylene blue ngokukhanya
Imicu ye-Trichophyton rubrum iqhathaniswa nezinhlamvu zayo	Akusebenzi lapha	MIC yemicu cishe 6.3 ug per mL, yezinhlamvu cishe 2.0 ug per mL; ukuvinjelwa okuphelele kwe-CFU kwakudinga 5 ug per mL kanye no-100 J per cm2 emicwini, kuqhathaniswa no-1.25 ug per mL kanye no-40 J per cm2 ezinhlamvwini	Inqubo ye-photodynamic engu-635 nm, ngokusho kocwaningo lokuzwela kwezigaba zokukhula kwesikhunta se-dermatophyte
I-biofilm evuthiwe yezinhlobo ezingu-33 ngaphansi kwezinsini, idivayisi ye-Calgary, usuku 6	MB iyodwa, ukwehla okungaphezu kuka-80 percent enanini eliphelele	MB kanye ne-LED engu-660 nm, ukwehla okucishe kube ngu-55 percent emsebenzini we-metabolism, ngaphandle kokushintsha okubanzi kokwakheka komphakathi	Udayi ongu-0.01 percent (100 ug per mL), 660 nm cishe ku-330 mW per cm2 isikhathi esingu-5 minutes, ukukhula ngaphandle komoya-mpilo, ngokusho kocwaningo lwe-biofilm yomlomo yezinhlobo eziningi olwenziwa ngaphandle komzimba

Kugqama amaphethini amathathu. Okokuqala, ama-staphylococci e-Gram-positive afa emazingeni aphansi e-micromolar, kuyilapho amabhaktheriya amise okwezinduku e-Gram-negative edinga udayi ocishe uphindwe kayikhulu ebumnyameni, ngoba ulwelwesi lwangaphandle lwama-Gram-negative lukhawulela ukungena. Ukukhanya kunciphisa lo mehluko kodwa akuwuqedi. Okwesibili, ukusebenza kwe-methylene blue ekulweni nesikhunta kungokoqobo kodwa kuncike emibandeleni: ukubulala kwayo i-Candida kukhula kakhulu ngokwanda komthamo wokukhanya, futhi ukushintsha isici esisodwa esibonakalayo, njengokukhiqizwa ngokweqile kwamaphampu akhipha izinto, kungaqeda ukubulawa okungu-5 log ngomthamo ofanayo. Okwesithathu, i-biofilm evuthiwe iziphatha ngokuhlukile kumaseli antanta oketshezini; yilapho umugqa we-biofilm yomlomo ungowakhona.

Amabhaktheriya: lapho ukukhanya kusiza kakhulu, nalapho kungasizi khona

Ama-staphylococci yiwona azwela kakhulu. Ngisho namanani asethebula lokubuyekeza e-MRSA ebhebhetheka ngezinga lobhubhane alingana noma aphansi kunawento yokuqhathanisa eyi-vancomycin lapho kusetshenziswa ukukhanya, kanti izinguqulo ezinamaqembu amabili e-methyl zenze kangcono nakakhulu ngoba ukwengezwa kwe-methyl kwandise ukuthanda amafutha nokukhiqizwa komoya-mpilo we-singlet, ngesikhathi kuvimbela ukuncishiswa zibe yisimo se-leuco. Lo mniningwane wamakhemikhali ubalulekile: ukhombisa ukuthi ukubulala kuwumphumela wokufinyeleliswa kukadayi kanye nokusebenzisana kwawo nokukhanya, hhayi ukusebenzisana nokukhanya kukodwa.

Izidalwa ze-Gram-negative zikhombisa uhlangothi lokufinyeleliswa kukadayi. I-Pseudomonas yayidinga 500 uM ebumnyameni kanye no-25 uM ngokukhanya, ukuthuthuka ngokuphindwe amashumi amabili, kodwa inani langaphansi kokukhanya lisengaphezulu kakhulu kwebanga lama-staphylococci. I-charge ephozithivu isiza udayi ukuthi usondele kulwelwesi lwangaphandle, kodwa ukungena kuseyisithiyo. Ukwakheka kwengxube, isikhathi sokufukamela, nokukhethwa kwenguqulo kadayi kushintsha la manani kakhulu kunokuphinda udayi ofanayo ngomthamo ophezulu.

Ama-enterococci aletha isexwayiso. Ezinhlelweni ezindala, i-E. faecalis ayizange ibonise inzuzo yokukhanya nhlobo, kanti i-E. faecium emelana ne-vancomycin yayidinga i-molecule ehlukile kunokukhanya okwengeziwe. Noma ubani ocaphuna i-MBC eyodwa ye-methylene blue njengenenani elingaguquki lokulwa nawo wonke amabhaktheriya ufunda ucwaningo ngendlela engafanele. Alikho inani elinjalo elingaguquki. Kukhona kuphela amanani aqondene nohlobo oluthile, indawo yokukhulisa, inani lamagciwane okuqalwa ngawo, ukufukamela, ubude begagasi, kanye namandla okukhanya endaweni ngayinye. Indlela yokufunda la manani ngaphandle kokuzikhohlisa ichazwe emhlahlandleleni wokuhumusha amajika okubulala.

Isikhunta: siyazwela, kodwa isigaba sokukhula namaphampu kuyakunquma

Izindonga zamaseli esikhunta ezakhiwe ngama-glucan, ama-mannan, ne-chitin zenza inkinga yokungena ehlukile kweyamabhaktheriya, futhi isikhunta sinezela nezindlela zokuzivikela ezisebenzayo. Umphumela wamaphampu akhipha izinto we-Candida ongenhla uyisibonelo esicace kakhulu: udayi ofanayo, ukukhanya okufanayo, indawo yokukhulisa efanayo, kodwa inguqulo ekhiqiza ngokweqile yasinda kuyilapho uhlobo lokuqala lwalahlekelwa cishe ngu-99.999 percent wamayunithi akha amakoloni. Ama-dermatophyte anezela umehluko wezigaba zokukhula. Izinhlamvu zifa kalula kakhulu kunemicu, ngakho inqubo eqeda izinhlamvu ingashiya izingcezu zemicu zisaphila ngomthamo ofanayo.

Yingakho imibiko yokulwa nesikhunta nge-methylene blue idinga ukunemba okufanayo ezimweni zokukhanya njengemibiko yamabhaktheriya. Ucwaningo lomtholampilo lwe-photodynamic yezinzipho nesikhumba lwaluhlanganisa udayi ongu-1 kuya ku-2 percent nokukhanya okubomvu okulinganisiwe, ngamandla okukhanya endaweni ngayinye achaziwe kanye nokulandelelwa kwemiphumela, futhi lwaluhlukanisa ukubukeka okubonwa emtholampilo nokuqedwa kwesikhunta okuqinisekiswe ngokuhlolwa. Lezo zivivinyo zibuyekezwe esifingqweni sobufakazi be-photodynamic yesikhunta sezinzipho nesikhumba. Ukugcoba udayi ohlanjululiwe endaweni ethintekile ngokukhanya kwegumbi kuphinda umbala kodwa kushiye inqubo ngaphandle.

Ukuhlolwa kwe-biofilm: imodeli yomlomo yezinhlobo ezingu-33

Ucwaningo lwango-2024 lwe-biofilm yomlomo yezinhlobo eziningi lufanelwe ukunakwa ngoba luhlola isimo esinzima kunesilula. Izinhlobo zomlomo ezingamashumi amathathu nantathu, ezihlanganisa i-Porphyromonas gingivalis, i-Tannerella forsythia, i-Aggregatibacter actinomycetemcomitans, i-Fusobacterium, nama-streptococci omlomo, zakhula ngaphandle komoya-mpilo zaba yi-biofilm evuthiwe ngosuku 6. Zafakwa udayi ongu-0.01 percent kanye ne-LED engu-660 nm isikhathi esingu-5 minutes, ngamandla aphezulu okukhanya afikayo endaweni. Kokubili udayi uwodwa nokukhanya kukodwa kwehlisa kakhulu amanani aphelele, kanti ukuhlanganiswa kwakho kwehlisa umsebenzi we-metabolism cishe ngesigamu, kodwa umphakathi awuzange ushintshe ukwakheka kwawo futhi i-biofilm ayizange iqedwe zonke izinto eziphilayo.

Lona umphumela wangempela onomkhawulo obekwe ngokwethembeka. Usekela ukwelapha nge-photodynamic njengendlela eyengeziwe yasendaweni ephaketheni lezinsini elihlanzwe, hhayi njengoketshezi lokugeza umlomo olwelapha lodwa i-periodontitis ehlonzwe ngokuhlolwa. Lokhu kusiletha egebeni elivamile.

Kungani ukubulala elabhorethri kungalingani nokwelapha

I-MBC yamaseli antanta ngokukhululeka ibeka amaseli afanayo kudayi ofanayo nasekukhanyeni okufanayo endaweni yokukhulisa enomoya-mpilo. Indawo ethintekile emzimbeni ayinikezi lutho lwalokho. Ulwelwesi lwe-biofilm lubambezela ukungena, izimo ezifana nezamaseli aphikelela ephila ziyakwazi ukubekezelela i-oxidation, igazi noketshezi oluphuma ezicutshini kumunca ukukhanya, ukwakheka kwezicubu kushiya izithunzi, futhi amaphakethe anomoya-mpilo omncane anciphisa ukusebenza kokusabela kohlobo II ngoba, ngokwencazelo yawo, umoya-mpilo we-singlet udinga umoya-mpilo oyimolecule. Ukutheleleka okuxubile kwengeza izinhlobo ezihlukene ngokungena kukadayi nangokuzwela, ngakho inani lohlobo olulodwa olusetshenziswa njengereferensi alikwazi ukumela yonke indawo.

Isibonelo somlomo sicacisa leli gebe. Ukunciphisa i-Porphyromonas noma ama-streptococci esitsheni selabhorethri, noma ukwehlisa ngehhafu i-biofilm ekhuliswe elabhorethri, akufakazeli ukuthi uketshezi oluluhlaza okwesibhakabhaka lokugeza umlomo lwelapha i-periodontitis, luqeda ukutheleleka ngaphakathi kwezinyo, noma luqeda amabhaktheriya egazini. Ukunakekelwa kwezinsini emtholampilo kuncike ekuphazamiseni nasekususeni i-biofilm ngezindlela zemishini kanye nokuphatha ukwakheka kwephakethe lezinsini, kanti ukwelapha ngokukhanya kufundwa njengendlela eyengeziwe esetshenziswa endaweni ethile. Ukugxila kukadayi okubulala embotsheni yesitsha sokuhlola akunabo ubudlelwano obungaguquki nokugxila okufinyelela kumabhaktheriya ngaphansi komugqa wezinsini, futhi ukukhanya kwegumbi akuwona umthamo ongu-660 nm. Imithamo, ukuphepha, nemikhawulo yokusebenzisana kwezinye izinto kunoma yikuphi ukusetshenziswa emlonyeni kufanele kunqunywe nodokotela, njengoba kuchazwe ereferensini yokuphepha nokusebenzisana.

Okungashiwo yilobu bufakazi

Abusona isimangalo sokubulala amagciwane ekhaya. Amathebula okubulala amabhaktheriya asebenzisa izinhlobo ezichaziwe, izindawo zokukhulisa, izikhathi zokuthintana, amabanga okukhanya, namandla okukhanya endaweni ngayinye. Awasho lutho ngezindawo zasekhishini, izikhathi zokuthintana ezidingekayo lapho kunokungcola okuvela ezintweni eziphilayo, noma izindinganiso zokubulala amagciwane ezibekwe yimithetho.

Futhi akusho ukuthi kungathatha indawo yemithi elwa namabhaktheriya esebenza kuwo wonke umzimba. Ukusebenza okuvamile kwe-photodynamic kudinga udayi, ukukhanya okumunnyiwe, nomoya-mpilo endaweni efanayo ngesikhathi esifanayo. Ukukhanya akufinyeleli ezindaweni ezijulile noma lapho ukutheleleka sekusabalele khona ngendlela umuthi ogwinyiwe ofinyelela ngayo egazini. Ucwaningo lubeka le ndlela njengendlela yokubhekana nokutheleleka okusendaweni efinyelelekayo noma njengendlela eyengeziwe, njengoba nje u-Wainwright achaza ukubulala amagciwane endaweni ethile kusukela ekuqaleni.

Isifingqo esifanele sinomkhawulo omncane kodwa sisathakazelisa. I-methylene blue inokusebenza okulinganiselwe kokulwa namagciwane ebumnyameni, kanti ukukhanya okubomvu kuvula umphumela omkhulu kakhulu we-photodynamic kumabhaktheriya amaningi nasesikhuntwini, ngokwehluka okukhulu phakathi kokukhanya nobumnyama okufana no-500 kuya ku-25 uM kwe-Pseudomonas kanye no-80 kuya ku-20 uM kwe-MRSA ebhebhetheka ngezinga lobhubhane, kanye nokungaphumeleli okufundisayo kuma-enterococci nase-Candida enamaphampu akhipha izinto asebenzayo. Ukuguqula lokhu kube ukusetshenziswa kwangempela kulawulwa ukufinyeleliswa kukadayi, indlela i-biofilm esebenza ngayo, nomoya-mpilo ngendlela elawulwa ngayo ukugxila kukadayi okubhaliwe. Gcina le mibandela icacile, bese amanani eba nencazelo. Yisuse, bese engasho lutho.