Uphando lwemethylene blue lokulwa neentsholongwane nokungunda: ukukhanya xa kuthelekiswa nobumnyama

Abafundi ababini babona ibinzana elithi imethylene blue yokulwa neentsholongwane baze bacinge ngezinto ezahlukileyo. Omnye ucinga ngolwelo oluluhlaza okwesibhakabhaka olutyhefa iintsholongwane xa ludibana nazo. Omnye ucinga ngenkqubo apho idayi kunye nokukhanya okubomvu kuzitshabalalisa. Zombini ezi ngcinga zinenxalenye yenyaniso. Akukho nanye enika umfanekiso opheleleyo, yaye ukuzibhidanisa kulapho uninzi lwamabango akwi-intanethi luphambuka khona.

Lo mahluko ngowona mxholo wolu phononongo. Imethylene blue inobuthi obulinganiselweyo ebumnyameni xa kukho uxinaniso olwaneleyo. Xa kukho ukukhanya okubomvu, iba yinto eyenza ukusabela kuvuselelwe kukukhanya: ifunxa ifotoni, ifikelele kwisimo se-triplet esisabelayo, ize iqhube ukusabela kwemichiza nge-oxidation eyonakalisa iithagethi ezininzi ngaxeshanye. Le nkqubo ivuselelwa kukukhanya inegama layo, unyango lwe-photodynamic lokulwa neentsholongwane, kunye neemeko zayo. Uxinaniso lodwa aluyichazi. Ubude bamaza, umthamo wokukhanya, ixesha lokufukama, ioksijini, nohlobo lwentsholongwane zimisela isiphumo.

Iindlela ezimbini zokubulala

Cinga kuqala ngendlela yasebumnyameni. Imethylene blue yidayi ye-phenothiazinium enentlawulo yombane eyiphozithivu. Incamathela kwizigqubuthelo zeentsholongwane, ingene kwiiseli ngemilinganiselo eyahlukahlukeneyo, ize ibandakanyeke ekusabeleni kwe-redox nasekubopheleleni kwemichiza. Akukho fotoni zibandakanyekayo. Ukubulala kuyenzeka, kodwa kuphela kuxinaniso olwahluka kakhulu ngokohlobo lwentsholongwane, yaye indlela esebenza ngayo ayijoliswanga kwithagethi enye ecacileyo.

Indlela ye-photodynamic yongeza ukukhanya. Iifotoni ezibomvu ezimalunga ne-630 ukuya kwi-680 nm zingqamana noluhlu lwamaza afunxwa yidayi, incopho yokufunxa kwe-monomer ikufuphi ne-664 nm. Ukufunxa kutyhalela idayi kwisimo se-singlet esihlala ixesha elifutshane, ize emva koko ibe kwisimo se-triplet esihlala ixesha elide. Ukusuka apho kulandela iintlobo ezimbini zokusabela. Ekusabeleni kohlobo I, i-triplet idlulisela i-elektroni kwisubstrate ize iqalise iindlela zeeradikhali ezibandakanya i-superoxide neentlobo ze-hydroxyl. Ekusabeleni kohlobo II, idlulisela amandla ngokuthe ngqo kwioksijini ekwisimo sayo esisiseko ize ivelise ioksijini ye-singlet. Zombini zinokwenzeka ngaxeshanye, yaye ukubiza idayi ngokuba sisixhobo seoksijini ye-singlet kuphela kukwenza lula kakhulu. Uphononongo lobudlelwane phakathi kolwakhiwo nokusebenza kwezinto ze-phenothiazinium ezivuselelwa kukukhanya lubonisa oku ngokuthe ngqo: phakathi kweedayi ezinxulumeneyo, ubuthi obuvuselelwa kukukhanya kwiibhaktheriya abuzange buhambelane nomthamo weoksijini ye-singlet ovelisiweyo kuphela, kuba ukufunxwa, indawo efikelela kuyo idayi, nokuncitshiswa kwayo ibe yifomu ye-leuco engasebenziyo zazibaluleke ngokulinganayo neempawu zayo zefiziksi yokukhanya. Ukusabela kwemichiza okubangelwa kukukhanya okusemva komgaqo wobude bamaza kuchazwe kuphononongo oluhambisana nolu lwedayi kunye nokukhanya okubomvu.

Ioksijini ye-singlet ingundoqo kolunye uphawu oluluncedo. Isabela neemolekyuli ezininzi zezinto eziphilayo ngaxeshanye, ngoko uhlaselo luchaphazela izinto ezininzi endaweni yokujolisa kwi-enzyme enye okanye kwiproteni ebophelelayo. Inkcazelo esisiseko yonyango lwemichiza lwe-photodynamic lokulwa neentsholongwane yachaza oku njengesizathu sokuba iipateni eziqhelekileyo zokuxhathisa zingabi nefuthe elikhulu: iproteni eyongezelelweyo ebophelela ipenicillin okanye i-topoisomerase etshintshileyo ayikuthinteli ukugqabhuka kweoksijini esabelayo. Le ngqiqo yangqineka kwiimvavanyo ze-Staphylococcus aureus exhathisa imethicillin nee-enterococci ezixhathisa ivancomycin, njengoko itheyibhile engezantsi ibonisa. Oku akuthethi ukuba ukuxhathisa akunakwenzeka, kodwa ukuba kusebenza ngendlela eyahlukileyo. Iimpompo zokukhupha izinto kunye neendlela zokuzikhusela kwi-oxidation zisenokutshintsha umthamo ofunekayo, ingakumbi ekungundeni.

Itheyibhile yeentlobo zeentsholongwane neemodeli zophando

Le theyibhile yeyona nto iphambili kweli nqaku. Ixabiso ngalinye lixhomekeke kwinkqubo yovavanyo ethile: idayi efanayo, umthamo wokukhanya okanye ixesha lokufukama elahlukileyo, nenani elahlukileyo. Musa ukuthatha i-MBC, i-MLC, ne-MIC njengezithetha into enye. I-MBC ne-MLC apha zithetha ukuba akukho zisindileyo xa ziphinde zakhuliswa kwenye indawo yokukhulisela, ngoxa i-MIC ithetha ukuthintelwa kokukhula. Iimeko zokukhanya zihamba nongeniso ngalunye.

Uhlobo lwentsholongwane nemodeli	Isiphumo ebumnyameni	Isiphumo ekukhanyeni	Iimeko
S. aureus NCTC 6571, ukunqunyanyiswa kweeseli ezidadayo	MBC 5 uM	MBC 5 uM	Itheyibhile yophononongo, ukukhanya okumhlophe 12 J per cm2
S. aureus NCTC 6571, ukunqunyanyiswa kweeseli ezidadayo	MLC 2.5 uM	MLC 1 uM	Inkqubo yovavanyo yokuqala yowe-1997, 1.7 mW per cm2 kangangeyure e-1, malunga ne-6.3 J per cm2, isixa seentsholongwane esifakwe ekuqaleni sikufuphi ne-10^6 CFU per mL, njengoko kuchaziwe kuphononongo lokuqala lokubulawa kweebhaktheriya yi-phenothiazinium ngokukhanya
MRSA EMRSA-15, iiseli ezidadayo	MBC 80 uM	MBC 20 uM	Itheyibhile yophononongo, malunga ne-12 J per cm2, ukwanda kokuchaphazeleka ngokuphindwe kane
MRSA EMRSA-16, iiseli ezidadayo	MBC 40 uM	MBC 10 uM	Inkqubo yovavanyo efanayo nale ingasentla
E. coli NCTC 9001, iiseli ezidadayo	MBC 500 uM	MBC 100 uM	Inkqubo yovavanyo efanayo, ukwanda kokuchaphazeleka ngokuphindwe kahlanu
P. aeruginosa NCTC 10622, iiseli ezidadayo	MBC 500 uM	MBC 25 uM	Inkqubo yovavanyo efanayo, ukwanda kokuchaphazeleka ngokuphindwe ngamashumi amabini
E. faecalis NCTC 775, iiseli ezidadayo	MLC 100 uM	MLC 100 uM	Inkqubo yovavanyo yowe-1997 kwi-6.3 J per cm2, akukho kuphucuka kolu hlobo nakulo mthamo
E. faecium exhathisa ivancomycin, iiseli ezidadayo	MBC 400 uM	MBC 400 uM	Itheyibhile yophononongo, akukho kuphucuka ngedayi esisiseko, ngoxa izinto eziphuma kuyo ezongezelelwe amaqela e-methyl nezichasa amanzi ngakumbi zaphucula, ngokwephononongo lwe-enterococcus exhathisa ivancomycin
Candida albicans, iiseli ezidadayo	MIC ngaphezu kwe-12.79 ug per mL	MIC 12.79, 6.39, kunye ne-1.59 ug per mL kwi-5, 10, kunye ne-15 J per cm2	Ukufukama kwangaphambili ebumnyameni imizuzu engama-30, i-LED ebomvu 600 ukuya kwi-650 nm enencopho kwi-635 nm, ngokwephononongo lobudlelwane phakathi komthamo nesiphumo sesenzo se-photodynamic semethylene blue sokulwa nokungunda
C. albicans YEM14 xa ithelekiswa nohlobo oluvelisa iimpompo zokukhupha ezigqithiseleyo, ukubulawa okulinganiswa nge-CFU	Ayisebenzi apha	Ukubulawa kohlobo olungumzali malunga ne-5 log, ukubulawa kohlobo oluvelisa i-CDR1/CDR2 ngokugqithiseleyo kusondele kuziro	Idayi 100 uM, ukufukama imizuzu engama-30, 660 nm kwi-60 J per cm2, ngokwephononongo lweempembelelo zeempompo zokukhupha ekubulaleni kwemethylene blue okuvuselelwa kukukhanya
Imisonto ye-Trichophyton rubrum xa ithelekiswa nembewu yayo	Ayisebenzi apha	MIC yemisonto malunga ne-6.3 ug per mL, eyembewu malunga ne-2.0 ug per mL; ukuthintela ngokupheleleyo i-CFU kwakufuna i-5 ug per mL kunye ne-100 J per cm2 kwimisonto xa kuthelekiswa ne-1.25 ug per mL kunye ne-40 J per cm2 kwimbewu	Inkqubo ye-photodynamic ye-635 nm, ngokwephononongo lokuchaphazeleka kwe-dermatophyte ngokwenqanaba lokukhula
Ibhayofilm evuthiweyo engaphantsi kweentsini yeentlobo ezingama-33, isixhobo seCalgary, usuku 6	MB yodwa, ukuhla okungaphezu kwe-80 pesenti kwinani lilonke	MB kunye ne-LED ye-660 nm, ukuhla malunga ne-55 pesenti ekusebenzeni kwemetabolism, akukho kulungelelaniswa ngokutsha koluntu lweentsholongwane ngokubanzi	Idayi 0.01 pesenti (100 ug per mL), 660 nm malunga ne-330 mW per cm2 imizuzu emi-5, ukukhula ngaphandle kweoksijini, ngokwephononongo lwebhayofilm yomlomo yeentlobo ezininzi olwenziwe ngaphandle komzimba

Iipateni ezintathu ziyabonakala. Okokuqala, ii-staphylococci ze-Gram-positive zifa kumanqanaba asezantsi e-micromolar, ngoxa iibhaktheriya ezimile okweentonga ze-Gram-negative zifuna malunga nedayi ephindwe kalikhulu ebumnyameni, kuba inwebu yangaphandle yee-Gram-negative ithintela ukungena. Ukukhanya kuyawunciphisa lo mahluko kodwa akuwuphelisi. Okwesibini, isenzo semethylene blue sokulwa nokungunda siyinyani kodwa sixhomekeke kwiimeko: ukubulawa kwe-Candida yimethylene blue kuphucuka kakhulu ngokwanda komthamo wokukhanya, yaye utshintsho olunye kwiimpawu zayo, olufana nokuveliswa ngokugqithiseleyo kweempompo zokukhupha, lunokuphelisa ukubulawa kwe-5 log kumthamo ofanayo. Okwesithathu, ibhayofilm evuthiweyo iziphatha ngokwahlukileyo kwiiseli ezinqunyanyiswe kulwelo, nto leyo eboniswa ngumqolo webhayofilm yomlomo.

Iibhaktheriya: apho ukukhanya kunceda kakhulu, nalapho kungancediyo

Ii-staphylococci zezona zichaphazeleka lula. Nkqu namanani akwithetibhile yophononongo eMRSA yobhubhane ayalingana okanye abe ngaphantsi kwalawo evancomycin esetyenziswe kuthelekiso xa kusetyenziswa ukukhanya, yaye izinto eziphuma kwidayi ezongezelelwe amaqela amabini e-methyl zenze ngcono nangakumbi kuba ukongezwa kwe-methyl kwandisa ukuthanda kwazo amafutha nemveliso yeoksijini ye-singlet, ngoxa zithintela ukuncitshiswa zibe yifomu ye-leuco. Loo nkcukacha yemichiza ibalulekile: ibonisa ukuba ukubulala sisiphumo sokufikelelwa kwithagethi kunye neempawu zefiziksi yokukhanya, kungekhona ezokukhanya kuphela.

Iintsholongwane ze-Gram-negative zibonisa indima yokufikelela kwidayi kwithagethi. I-Pseudomonas yafuna i-500 uM ebumnyameni ne-25 uM ekukhanyeni, ukuphucuka okuphindwe ngamashumi amabini, kodwa ixabiso lasekukhanyeni lisengaphezulu kakhulu koluhlu lwee-staphylococci. Intlawulo yombane eyiphozithivu inceda idayi isondele kwinwebu yangaphandle, kodwa ukungena kuyo kusengumqobo. Indlela umxube olungiselelwe ngayo, ixesha lokufukama, nokukhethwa kwento ephuma kwidayi zitshintsha la manani ngaphezu kokuphinda kusetyenziswe idayi efanayo ngomthamo ophezulu.

Ii-enterococci zinika isilumkiso. Kwiinkqubo zovavanyo ezindala, i-E. faecalis ayizange ibonise nzuzo yokukhanya kwaphela, yaye i-E. faecium exhathisa ivancomycin yayifuna imolekyuli eyahlukileyo endaweni yokukhanya okungakumbi. Nabani na ocaphula i-MBC enye yemethylene blue njengexabiso elisisigxina lokulwa neebhaktheriya ngokubanzi uyifunda ngendlela ephosakeleyo imithombo yophando. Alikho ixabiso elinjalo elisisigxina. Kukho kuphela amaxabiso axhomekeke kuhlobo oluthile, indawo yokukhulisela, isixa seentsholongwane ezifakwe ekuqaleni, ixesha lokufukama, ubude bamaza, kunye namandla okukhanya ngeyunithi yendawo. Indlela yokufunda loo maxabiso ngaphandle kokuzikhohlisa ichazwe kwisikhokelo sokutolika iigrafu zokubulala.

Ukungunda: kuyachaphazeleka, kodwa inqanaba lokukhula neempompo ziyagqiba

Iindonga zokungunda ezenziwe nge-glucan, i-mannan, ne-chitin zenza ingxaki yokufunxwa eyahlukileyo kweyeebhaktheriya, yaye ukungunda kongeza iindlela ezisebenzayo zokuzikhusela. Isiphumo seempompo zokukhupha ze-Candida esingasentla sesona sibonisa oku ngokucacileyo: idayi efanayo, ukukhanya okufanayo, indawo yokukhulisela efanayo, kodwa uhlobo oluvelisa iimpompo ngokugqithiseleyo lwasinda ngoxa uhlobo olungumzali lwalahlekelwa malunga ne-99.999 pesenti yeeyunithi ezenza amakholoni. Ii-dermatophyte zongeza umba wenqanaba lokukhula. Imbewu ifa lula kakhulu kunemisonto, ngoko inkqubo esusa imbewu isenokushiya amaqhekeza emisonto ephila kumthamo ofanayo.

Kungenxa yoko iingxelo zemethylene blue zokulwa nokungunda zifuna ukuchazwa ngokungqongqo kweemeko zokukhanya, njengezo zeebhaktheriya. Izifundo zonyango lwe-photodynamic lwezikhonkwane nolusu zadibanisa idayi eyi-1 ukuya kwi-2 pesenti nokukhanya okubomvu okulinganisiweyo, kumandla okukhanya ngeyunithi yendawo achaziweyo, kunye nokulandelelwa kweziphumo, zaza zahlula inkangeleko yezonyango ekususweni kokungunda okungqinwe luvavanyo. Ezo mvavanyo ziphononongwa kwisishwankathelo sobungqina be-photodynamic bokungunda kwezikhonkwane nolusu. Ukuqaba idayi exutyiweyo kwindawo eyonakeleyo phantsi kokukhanya kwegumbi kuphinda kuvelise umbala kodwa kushiye ngaphandle inkqubo.

Uvavanyo lwebhayofilm: imodeli yomlomo yeentlobo ezingama-33

Uphononongo lwebhayofilm yomlomo yeentlobo ezininzi lowama-2024 lufuna ingqalelo kuba luvavanya imeko enzima kunelula. Iintlobo zomlomo ezingamashumi amathathu anesithathu, kuquka i-Porphyromonas gingivalis, i-Tannerella forsythia, i-Aggregatibacter actinomycetemcomitans, i-Fusobacterium, nee-streptococci zomlomo, zakhula ngaphandle kweoksijini zaba yibhayofilm evuthiweyo ngosuku 6. Unyango lwaluyidayi eyi-0.01 pesenti kunye ne-LED ye-660 nm imizuzu emi-5 kumandla aphezulu okukhanya afika kumphezulu. Zombini idayi yodwa nokukhanya kuphela zanciphisa kakhulu amanani eentsholongwane ewonke, yaye ukudityaniswa kwazo kwanciphisa ukusebenza kwemetabolism malunga nesiqingatha, kodwa uluntu lweentsholongwane aluzange lulungelelaniswe ngokutsha yaye ibhayofilm ayizange icocwe ide ingabi nazintsholongwane.

Eso sisiphumo sokwenene esinomda ocacileyo. Sixhasa unyango lwe-photodynamic njengoncediso lwendawo ethile kwipokotho yeentsini ecociweyo, kungekhona njengesihlambululi somlomo esinyanga ngokwaso isifo sokudumba kwezicubu ezixhasa amazinyo esifunyaniswe luvavanyo. Oku kusizisa kumsantsa oqhelekileyo.

Kutheni ukubulala elabhoratri kungalingani nonyango

Uvavanyo lwe-MBC lweeseli ezidadayo lubeka iiseli ezifanayo kwidayi efanayo nakukukhanya okufanayo kwindawo yokukhulisela eneoksijini. Indawo eyonakeleyo emzimbeni ayinayo nayiphi na kwezi meko. Ulwakhiwo olujikeleze iiseli lucothisa ukungena, iimeko ezifana nezokuhlala ixesha elide zisinda ziyayinyamezela i-oxidation, igazi nolwelo oluphuma kwindawo eyonakeleyo zifunxa ukukhanya, ukumila kwezicubu kushiya izithunzi, yaye iipokotho ezineoksijini encinane zinciphisa ukusabela kohlobo II kuba ioksijini ye-singlet ifuna ioksijini yemolekyuli ngokwenkcazelo yayo. Usulelo oluxubeneyo longeza iintlobo ezahlukileyo ekufunxeni nasekuchaphazelekeni, ngoko ixabiso lohlobo olunye olusetyenziswa njengereferensi alinako ukumela yonke indawo.

Umzekelo womlomo uwenza lo msantsa ucace. Ukunciphisa i-Porphyromonas okanye ii-streptococci kwisitya, okanye ukuthomalalisa ibhayofilm ekhuliswe elabhoratri ngesiqingatha, akuqinisekisi ukuba isihlambululi somlomo esiluhlaza okwesibhakabhaka sinyanga ukudumba kwezicubu ezixhasa amazinyo, siphilisa usulelo olungaphakathi ezinyo, okanye sisusa iibhaktheriya egazini. Unyango lwezicubu ezixhasa amazinyo luxhomekeke ekuphazamiseni nasekususeni ibhayofilm ngeendlela zoomatshini, kunye nokulawulwa kolwakhiwo lwepokotho yeentsini, ngoxa unyango olusekelwe ekukhanyeni luphando njengesincedisi sendawo ethile. Uxinaniso lwedayi olubulala kumngxunyana wesitya sovavanyo alunabudlelwane obusisigxina noxinaniso olufikelela kwiibhaktheriya ezingaphantsi komgca weentsini, yaye ukukhanya kwegumbi akungomthamo we-660 nm. Imida yomthamo, ukhuseleko, nokusebenzisana nayo nayiphi na into xa isetyenziswa emlonyeni ifuna ukukhokelwa yingcali yezonyango, njengoko kucacisiwe kwireferensi yokhuseleko nokusebenzisana.

Into obungayithethiyo obu bungqina

Abulobango lokubulala iintsholongwane ekhaya. Iitheyibhile zokubulawa kweebhaktheriya zisebenzisa iintlobo ezichaziweyo, iindawo zokukhulisela, amaxesha okudibana, uluhlu lwamaza okukhanya, namandla okukhanya ngeyunithi yendawo. Azithethi nto ngemiphezulu yasekhitshini, amaxesha okudibana afunekayo xa kukho ukungcola kwezinto eziphilayo, okanye imigangatho yokubulala iintsholongwane ebekwe ngabalawuli.

Kananjalo abubungqina bokuthatha indawo yeyeza lokubulala iibhaktheriya elisebenza kuwo wonke umzimba. Isenzo esiqhelekileyo se-photodynamic sifuna idayi, ukukhanya okufunxiweyo, neoksijini kwindawo enye ngexesha elinye. Ukukhanya akufikeleli kwiindawo ezinzulu okanye ezisasazekileyo ngendlela iyeza eliginyiweyo elifikelela ngayo egazini. Imithombo yophando ibeka le ndlela njengelungele usulelo lwendawo ethile efikelelekayo okanye njengesincedisi, kanye njengoko uWainwright wachaza ukubulala iintsholongwane kwindawo ethile kwasekuqaleni.

Isishwankathelo esifanelekileyo sinomda omncinane kodwa sisengumdla. Imethylene blue inomsebenzi olinganiselweyo wokulwa neentsholongwane ebumnyameni, yaye ukukhanya okubomvu kuvulela isiphumo se-photodynamic esikhulu kakhulu kwiibhaktheriya ezininzi nakukungunda, kukho umahluko omkhulu phakathi kokukhanya nobumnyama onjengokusuka kwi-500 ukuya kwi-25 uM kwi-Pseudomonas nakwi-80 ukuya kwi-20 uM kwiMRSA yobhubhane, kunye nokungaphumeleli okufundisayo kwii-enterococci nakwi-Candida enobuchule bokukhupha idayi. Ukusetyenziswa kwezi ziphumo ngaphandle kwelabhoratri kulawulwa kukufikelelwa kwithagethi, indlela ibhayofilm esebenza ngayo, neoksijini, ngokulinganayo noxinaniso lwedayi olubhaliweyo. Gcina ezo meko zicacile ukuze amanani abe nentsingiselo. Zisuse, aze amanani angabi nantsingiselo.