I-methylene blue ngokumelene nama-biofilm: lokho okukalwa izifundo zaselabhorethri ezisebenzisa ukukhanya

I-biofilm ayilona iqoqo lamangqamuzana antanta oketshezini. Iyisakhiwo: amangqamuzana anamathele endaweni ethile, agxile kunethiwekhi azakhela yona yama-polysaccharide, amaprotheni, ne-DNA engaphandle kwamangqamuzana, kanti amazinga omoya-mpilo nezakhamzimba ayashintsha ukusuka ngaphandle kuya ngaphakathi. Leso sakhiwo siyisizathu sokuthi ikhemikhali ebulala amabhaktheriya antantayo ngemizuzu ingahluleka ngokumelene nalezo zinhlobo ezifanayo lapho zikhuliswe njenge-biofilm. Noma yisiphi isimangalo nge-methylene blue nama-biofilm kufanele sihunyushwe kubhekwe leso sakhiwo, ngoba inethiwekhi inquma ukuthi ungakanani udayi ofinyelela emangqamuzaneni nokuthi kungakanani ukukhanya okufinyelela kudayi.

Lokhu kubuyekezwa kuhlanganisa imithombo emithathu kuphela. Omunye unikeza ulwazi oluyisisekelo. Emibili iyizivivinyo zaselabhorethri. Ukuhlukanisa lezo zindima kuyindlela yonke yokubuyekeza, futhi kuvimbela iphutha elivame kakhulu kulesi sihloko: ukucaphuna umphumela wesitsha sokuhlola njengokungathi uwubufakazi bokwelapha ukutheleleka kumuntu.

Umthombo wolwazi oluyisisekelo ukhuluma ngamakhemikhali, hhayi ngokubulala

Omdala kunayo yonke kule mithombo emithathu ukubuyekezwa okulandisayo kokuthuthukiswa kwezinto ze-phenothiazinium ezizwela ukukhanya okwashicilelwa ngo-2005. Kulandelela i-methylene blue kusukela ekubeni udayi wekhulu le-19 kuya ekubeni ikhemikhali eyisisekelo sokuthuthukisa ukwelashwa komdlavuza ngokukhanya, futhi kamuva nokwelashwa kwamagciwane ngamakhemikhali asebenza ngokukhanya. Umnikelo wako uchaza ubudlelwano phakathi kwesakhiwo nokusebenza: ukufakwa kwamaqembu e-methyl ku-chromophore kwandisa ukuthambekela kokuhlangana namafutha, ukukhiqizwa komoya-mpilo osesimweni se-singlet, namandla okubulala amabhaktheriya ngokukhanya, kuyilapho kubambezela ukuguquka ngokwehliswa kube uhlobo lwe-leuco olungasebenzi kangako. Kuphinde kuyisho ngokucacile inkinga yalo mkhakha. Imibiko eminingi yayisebenzisa odayi abavamile abathengiswayo, kanti izinhlobo ezithuthukisiwe ezisuselwa kubo zazincane uma kuqhathaniswa.

Sebenzisa lokho kubuyekezwa engxenyeni echaza indlela yokusebenza ekuqondeni kwakho: ukukhanya kwenza udayi ube sesimweni samandla aphezulu, isimo se-triplet sisebenzisana nomoya-mpilo ngokudluliswa kwamandla kwe-Type II ukuze kwakheke umoya-mpilo osesimweni se-singlet, noma ngokudluliswa kwama-electron kwe-Type I ukuze kwakheke ama-radical. Lokho kuhlasela ngezindlela eziningi yisizathu sokuthi izindlela ezivamile zokumelana zingasebenzi kahle ngokumelene nokubulala ngokukhanya. Ungakucaphuni njengobufakazi bokuncipha kwe-log. Akuzange kukale lokho. Amakhemikhali okukhanya abanzi asekela le ndlela achazwe eqoqweni lobufakazi bokwelashwa ngokukhanya, kanti imithetho yobude begagasi enquma ukuthi isibani esithile singayiqalisa yini ichazwe kokuthi i-methylene blue ikumunca kanjani ukukhanya okubomvu.

Izivivinyo ezimbili zama-biofilm, izindlela ezimbili ezihlukene zokuchaza imiphumela

Womabili amaphepha ezivivinyo ahlola ukusebenza kwe-methylene blue ngokukhanya kuma-biofilm omlomo ku-660 nm, kodwa cishe akukho okunye afana ngakho. Amaqoqo ezinhlobo ezihlukene, ubudala obuhlukene, ukwakheka kukadayi okuhlukene, amanani okukhanya ahlukene, futhi ngaphezu kwakho konke, imiphumela ekalwayo ehlukene. Lowo mehluko uyiphuzu lalesi sihloko, futhi uyisisekelo sethebula lokuqhathanisa elingezansi.

Esokuqala isifundo se-in vitro se-biofilm evuthiwe enezinhlobo eziningi engaphansi kwezinsini esashicilelwa ngo-2024. Izinhlobo ezingamashumi amathathu nantathu zakhuliswa ndawonye ezikhonkwaneni ze-polystyrene ku-Calgary Biofilm Device, zanikezwa izakhamzimba ngaphandle komoya-mpilo, kwashintshwa uketshezi lokukhulisa ngemva kwamahora angu-72, futhi ukukhula kwaqhubeka kwaze kwaba yizinsuku eziyi-7 ezazihlosiwe. Ukwelashwa kwakuyi-methylene blue engaboshiwe ku-0.01 percent, kusetshenziswa i-LED ebomvu engu-660 nm ngamandla acishe abe ngu-330 mW per square cm imizuzu engu-5 ebangeni elingu-2 mm, okuthi uma kubalwa kube cishe ngu-99 J per square cm. Uhlelo lokuhlola lwaluhlukanise ngokucacile izinhlanganisela ezine: iqembu lokuqhathanisa elingelashwanga, udayi ngaphandle kokukhanya, ukukhanya ngaphandle kukadayi, kanye nodayi nokukhanya.

Esesibili isifundo se-in vitro se-biofilm yomlomo yezinhlobo eziqala ukuhlala endaweni esashicilelwa ngo-2021. Izinhlobo ezine ze-Streptococcus zakhuliswa ndawonye amahora angu-24 emapuletini anemigodi engu-96, kusetshenziswa i-sucrose engu-1 percent. Into ezwela ukukhanya kwakuyi-methylene blue ehlanganiswe nama-nanoparticle e-beta cyclodextrin ku-4.65 micromolar, okuwukwakheka kokuyithwala okwakuhloswe ukudlulisa udayi ezithiyweni ze-biofilm. Yomibili imithombo yokukhanya yayibomvu ku-660 nm, nokuchayeka kwamasekhondi angu-90, kodwa amanani okukhanya ayehluke kakhulu: 320 J per square cm eqenjini le-laser no-8.1 J per square cm eqenjini le-LED. Amaqembu okuqhathanisa ayehlanganisa udayi ngaphandle kokukhanya, i-laser kuphela, i-LED kuphela, isixazululo sikasawoti njengeqembu lokuqhathanisa elingasebenzi, kanye ne-chlorhexidine engu-0.2 percent njengesilinganiso sokuqhathanisa ukusebenza ngokumelene namagciwane.

Iqoqo lezinhlobo ezingu-33 elinezinsuku eziyi-7 elikhule ngaphandle komoya-mpilo ne-biofilm yezinhlobo ezi-4 ezifika kuqala enamahora angu-24 kuyizinhlelo zokukala ezihlukene. Izibalo ezilandelayo azinakuhlanganiswa ukuze kutholakale isilinganiso esimaphakathi.

Lokho isifundo ngasinye esakukala ngempela

Esifundweni se-biofilm evuthiwe, imiphumela eyakalwa kwakuwukusebenza kwemetabolism ngokwehliswa kwe-TTC okufundwa ku-485 nm, kanye nobuningi bamabhaktheriya ngokuhlanganiswa kwe-DNA ne-DNA ngendlela ye-checkerboard kusetshenziswa izihlonzi zezinhlobo ezingu-33, okunomkhawulo wokuthola osondele kumangqamuzana angu-10,000. Akuzange kubalwe amangqamuzana aphilayo ngokuwakhulisa emapuletini, kukalwe ubuningi bezinto eziphilayo nge-crystal violet, kuthathwe izithombe ezihlukanisa aphilayo nafile, noma kulandelelwe ukukhula kabusha. I-LED kuphela yehlisa ukusebenza kwemetabolism cishe ngo-50 percent uma kuqhathaniswa namaqembu angazange athole i-LED, kanti udayi ne-LED bakwehlisa cishe ngo-55 percent. Ngokwezibalo, udayi kuphela wawuseqenjini elifanayo neqembu lokuqhathanisa elingelashwanga. Ezibalweni ezisuselwa ku-DNA, udayi kuphela, i-LED kuphela, kanye nodayi ne-LED konke kwakuphansi kakhulu kuneqembu lokuqhathanisa, ngaphandle komehluko obalulekile ngokwezibalo phakathi kwakho kokuthathu. Ababhali bawusho ngokuqondile umphumela: izindlela zokwelapha azizange ziyibhubhise i-biofilm enamagciwane amaningi eyayisivele yakhekile futhi inamathele, futhi i-biofilm evuthiwe ezungeze amazinyo idinga ukuphazanyiswa kwesakhiwo ngezindlela zemishini, ukusetshenziswa kokukhanya kube yisengezo kuphela.

Esifundweni sezinhlobo ezifika kuqala, umphumela okaliwe kwakuyizilwanyana ezincane eziphilayo ezatholakala ngokukhuliswa ezindaweni zokukhulisa ezikhetha izinhlobo ezithile, zivezwe ngesikali se-logarithm. Iqembu likadayi ngaphandle kokukhanya alizange lehluke kwesixazululo sikasawoti. Womabili amaqembu asebenzisa ukukhanya ukwenza udayi usebenze, i-laser nod ayi ofakwe ngaphakathi kwezithwali kanye ne-LED nod ayi ofakwe ngaphakathi kwezithwali, anciphisa izibalo zamangqamuzana aphilayo ngokufinyelela cishe ku-4 log uma kuqhathaniswa neqembu lokuqhathanisa elingasebenzi. Lokhu kwakuwukuncipha ngokuphindwe ngo-10,000 noma ngo-99.99 percent kuleso silinganiso, futhi alikho kula maqembu elalehluke ngokwezibalo ku-chlorhexidine. Amanani okukhanya kuphela nawodayi ngaphandle kokukhanya awazange abikwe njengezinombolo ze-log ezihlukene nezithembekile. Ngakho isifinyezo esithembekile yileso esisekelwa yiphepha: udayi ofakwe ngaphakathi kwezithwali nowenziwe wasebenza ngokukhanya wabangela ukuncipha, udayi ngaphandle kokukhanya awukwenzanga lokho, futhi ubukhulu bomphumela babufana nesilinganiso sesibulali-magciwane kule modeli ye-biofilm esencane.

Alikho iphepha elakala ukukhula kabusha ngemva kokwelashwa, ubuningi bezinto eziphilayo ezisele, noma isakhiwo senethiwekhi. Ukuncipha kwe-CFU akusho lutho ngenethiwekhi esele. Ukwehla kwemetabolism akusho lutho ngalawo asindile. Isibalo se-DNA asikwazi ukusho ukuthi yimaphi aphilayo.

Ukuqhathaniswa kwezindlela zokuhlola nemiphumela ekalwayo

Leli thebula liyithuluzi lokuqhathanisa lalesi sihloko. Funda uvundlile emgqeni ukuze ubone ukuthi isifundo esisodwa sifakazelani, bese ufunda wehle ngekholomu ukuze ubone ukuthi yimiphi imibuzo engaphendulwa yilesi nalesiya sifundo.

Ukuqhathanisa okuthathu kudinga ukugcizelelwa. Okokuqala, emodelini evuthiwe iqembu lokukhanya kuphela lichaza cishe wonke umphumela kwimetabolism, ngakho inani elingu-55 percent alisho ukubulala okubangelwa udayi. Okwesibili, emodelini esencane ukwakheka kukadayi ngaphandle kokukhanya akusebenzi, futhi umphumela uvela kuphela lapho kunokukhanya, okuwuphawu lokwenziwa kusebenze ngokukhanya. Okwesithathu, ukwehla ngo-80 percent ezibalweni ezisuselwa ku-DNA kusondele ngokwezibalo ku-0.7 log kuphela uma okukalwayo bekuyizibalo zamangqamuzana aphilayo. Kodwa akunjalo, ngakho akufanele neze kucashunwe njengokubulala okulinganiswe nge-log. Ulwazi oluhlobene nama-biofilm amazinyo luqoqwe ocwaningweni lwama-biofilm omlomo namazinyo, kanti indlela ejwayelekile yokuhlukanisa amamodeli, izindlela zokufakwa, nemiphumela ekalwayo ezifundweni ezihlukene ichazwe ngokuningiliziwe kumhlahlandlela wezindlela zaselabhorethri.

Kungani izifundo zama-biofilm zidinga imithetho yazo yokufundwa

Izinto ezine eziguquguqukayo zinquma ukuthi umphumela we-methylene blue ku-biofilm unencazelo yini ngaphandle kwepuleti lawo.

Ubudala be-biofilm nenethiwekhi yayo kuza kuqala. Ama-biofilm amadala ajiyile, anezixhumanisi eziningi phakathi kwezingxenye zawo, futhi kunzima kakhulu ukuba udayi ungene kuwo. Ababhali bango-2021 babala ukungena kukadayi nokungasabalali kahle kokukhanya ku-biofilm ehlelekile njengezithiyo eziyinhloko. Izindlela zezithwali ezifana nokufakwa ngaphakathi kwe-cyclodextrin ziyimizamo yokubhekana ngqo naleso sithiyo. Yingakho ukwakheka kuyinto eguquguqukayo esivivinyweni, hhayi imininingwane encane nje.

Ukubalwa kwenani lokukhanya kuza okwesibili. Amanani amandla okukhanya atholwa indawo ngayinye lapha asukela ku-8.1 kuya ku-320 J per square cm ngobude begagasi obufanayo, umehluko ocishe uphindwe ngamashumi amane, kanti amandla okukhanya afikayo asukela ku-90 mW kuya kuma-W amaningana per square cm. Ubude begagasi obufanayo abusho ukuchayeka okufanayo, futhi alikho kulezi zifundo eliguqula inani lokukhanya ngendlela ehleleke ngokwanele ukuze kwakhiwe ijika lobudlelwano.

Amaqembu okuqhathanisa eza okwesithathu. Isifundo esingenalo iqembu likadayi ngaphandle kokukhanya asikwazi ukukhipha ukuthi udayi unobuthi ngokwawo, kanti isifundo esingenalo iqembu lokukhanya kuphela asikwazi ukuthi umphumela ubangelwa ukwenziwa kusebenze ngokukhanya. Womabili amaphepha awafaka womabili la maqembu, okuyinto enhle ngawo. Iphepha le-biofilm evuthiwe libonisa ukuthi kungani iqembu lokukhanya kuphela libalulekile: ngaphandle kwalo, umphumela wemetabolism ubuzohunyushwa njengomphumela kadayi.

Imiphumela ekalwayo iza okwesine. Ukuphila kwamangqamuzana, imetabolism, ubuningi be-DNA, ubuningi bezinto eziphilayo, isakhiwo, nokukhula kabusha kuyizimangalo eziyisithupha ezihlukene. Alikho iphepha lapha elikala ngaphezu kwezimbili. Ikakhulukazi ukukhula kabusha akukho kuzo zonke lezi zifundo, ngakho ukuqhubeka komphumela akukalwanga, kunokuba kube kufakazelwe.

Lokhu okungakufakazeli ngokwelapha ukutheleleka

Azikho kulezi zinhlelo zokuhlola ezinokujikeleza kwegazi, amangqamuzana amasosha omzimba, ukugeleza kwamathe, ukuvuselelwa koketshezi olusemiseleni yezinsini, noma ukususwa kwezinto ngezindlela zemishini. Ukugxila kukadayi emgodini wepuleti akukhona ukugxila kwawo esicutshini ngemva kwanoma iyiphi indlela yokuwufaka kumuntu. Ukuchayeka okukodwa kwemizuzu engu-5 noma amasekhondi angu-90 kwe-biofilm esencane noma ngisho evuthiwe yaselabhorethri akulona uhlelo lokwelashwa. Ababhali be-biofilm evuthiwe bakusho lokhu ngokwabo ngokuchaza ukusetshenziswa kokukhanya njengesengezo esingase sisize ekususweni ngezindlela zemishini, hhayi njengendlela yokuyiqeda ngokuphelele esetshenziswa yodwa.

Isifinyezo esifanele silinganiselwe futhi sihlala siyiqiniso. I-methylene blue eyenziwe yasebenza ngokukhanya ingacindezela imetabolism nobuningi obususelwa ku-DNA eqoqweni elivuthiwe lezilwanyana ezincane zomlomo zaselabhorethri ngaphandle kokulibhubhisa, kanti ukukhanya kuphela kwenza ingxenye enkulu yomsebenzi wokunciphisa imetabolism kuleyo nhlelo. Udayi othwalwa yi-cyclodextrin nowenziwe wasebenza ngokukhanya unganciphisa izibalo zamangqamuzana aphilayo ngamazinga amaningana okuphindeka ngeshumi ku-biofilm esencane ye-streptococci yaselabhorethri, ngezinga eliqhathaniseka ne-chlorhexidine kuleyo modeli efanayo. Ukuthi noma yikuphi kwalokhu okubonwayo kuyasebenza yini ekuthelelekeni kumuntu kuncike ekufinyeleleni kukadayi, ekufinyeleleni kokukhanya, emoyeni-mpilo, ekwakhekeni komzimba, nasemiphumeleni yomtholampilo lezi zivivinyo ezazingaklanyelwe ukuyikala.