Maƙala
Binciken shuɗin methylene kan kashe ƙwayoyin cuta: bakteriya, fungi, da kunnawa da haske
Masu karatu biyu suna ganin furucin shuɗin methylene mai kashe ƙwayoyin cuta, amma su yi tunanin abubuwa daban-daban. Ɗaya yana tunanin wani ruwa shuɗi da ke guba ga ƙwayoyin cuta da zarar ya taɓa su. Ɗayan yana tunanin wata hanya inda rini tare da jan haske ke lalata su. Dukkan tunanin biyu suna ɗauke da wani ɓangare na gaskiya. Babu wanda ya ba da cikakken bayani, kuma gauraya su shi ne inda yawancin iƙirarin yanar gizo ke kuskure.
Wannan bambanci shi ne batun wannan bita. Shuɗin methylene yana da ɗan tasirin guba a duhu idan maida hankalinsa ya isa. Tare da jan haske, yana zama sinadarin da ke tayar da aikin sinadarai ta haske: yana shan photon, ya kai ga yanayin triplet mai saurin yin martani, sannan ya haifar da aikin sinadarai na oxidation da ke lalata abubuwa da yawa lokaci guda. Wannan tsarin da haske ke kunnawa yana da sunansa, maganin photodynamic na kashe ƙwayoyin cuta, kuma yana da nasa sharuɗɗa. Maida hankali kaɗai ba ya ayyana shi. Tsawon zango, adadin haske, lokacin bari kafin gwaji, oxygen, da nau'in ƙwayar halitta ne ke tantance sakamakon.
Hanyoyin kashewa guda biyu
Da farko, a yi tunanin hanyar duhu. Shuɗin methylene rinin phenothiazinium ne mai cajin lantarki tabbatacce. Yana haɗuwa da suturar ƙwayoyin cuta, ya shiga ƙwayoyin zuwa matakai daban-daban, kuma ya shiga ayyukan sinadarai na redox da ɗaurewa. Babu photons a ciki. Ana samun kashewa, amma sai a maida hankali da ya bambanta sosai daga wata ƙwayar halitta zuwa wata, kuma hanyar aikin ba ta takaita ga abu guda ɗaya tak ba.
Hanyar photodynamic tana ƙara haske. Jan photons masu tsawon zango na kusan 630 zuwa 680 nm suna dacewa da zangon shan haske na rinin, inda kololuwar shan hasken monomer take kusa da 664 nm. Shan haske yana tura rinin zuwa yanayin singlet mai ɗan gajeren lokaci, sannan zuwa yanayin triplet mai ɗan tsawon lokaci. Daga nan, rukuni biyu na martanin sinadarai suna biyo baya. A aikin sinadarai na Type I, triplet yana miƙa electron ga wani sinadarin da yake yin martani da shi, ya fara hanyoyin radicals da suka haɗa da nau'ikan superoxide da hydroxyl. A aikin sinadarai na Type II, yana miƙa kuzari kai tsaye ga oxygen da ke yanayin kuzari na asali, ya samar da singlet oxygen. Duka biyun na iya faruwa lokaci guda, kuma kiran rinin sinadarin singlet oxygen kaɗai yana sauƙaƙa lamarin fiye da kima. Wata bita kan alaƙar tsarin sinadari da aikin sinadaran phenothiazinium masu tayar da martani da haske ta nuna wannan kai tsaye: a tsakanin rinuka masu alaƙa, gubar da haske ke haifarwa ga bakteriya ba ta bi adadin singlet oxygen kaɗai ba, domin shiga ƙwayoyin, wurin taruwa, da sauyawa ta reduction zuwa siffar leuco marar aiki suna da muhimmanci kamar halayen mu'amalar sinadarin da haske. An bayyana aikin sinadarai da haske da ke bayan ƙa'idar tsawon zango a bitar da ke tare da wannan kan rini da jan haske.
Singlet oxygen yana da muhimmanci ga wani hali mai amfani. Yana yin martani da biomolecules da yawa lokaci guda, don haka harin yana da fannoni da yawa maimakon nufin enzyme ɗaya ko furotin mai ɗaurewa. Wani bayani na farko kan amfani da sinadarai da haske wajen kashe ƙwayoyin cuta ya bayyana wannan a matsayin dalilin da ya sa tsarin juriya ga magungunan da aka saba amfani da su ba ya da tasiri sosai: ƙarin furotin mai ɗaure penicillin ko topoisomerase da ya samu sauyin gado ba ya hana fitowar oxygen mai saurin yin martani. Wannan tunani ya yi daidai da gwaje-gwajen Staphylococcus aureus mai juriya ga methicillin da enterococci masu juriya ga vancomycin, kamar yadda jadawalin da ke ƙasa ya nuna. Ba yana nufin juriya ba ta yiwuwa ba ne, sai dai tana aiki ta wata hanya. Famfunan fitar da sinadarai da kariyar antioxidants har yanzu na iya sauya adadin da ake buƙata, musamman a fungi.
Jadawalin ƙwayoyin halitta da tsarin bincike
Wannan jadawali shi ne babban ɓangaren bayanan wannan makala. Kowace ƙima ta danganta da takamaiman tsarin gwaji: rini ɗaya, amma adadin haske ko lokacin bari daban, yana ba da ƙima daban. Kada a ɗauki MBC, MLC, da MIC a matsayin abubuwa masu ma'ana ɗaya. MBC da MLC a nan suna nufin babu waɗanda suka tsira bayan sake renonsu, yayin da MIC ke nufin hana girma. An nuna yanayin haske ga kowace shigarwa.
| Ƙwayar halitta da tsarin gwaji | Sakamakon duhu | Sakamakon haske | Sharuɗɗa |
|---|---|---|---|
| S. aureus NCTC 6571, ƙwayoyi masu shawagi a ruwa | MBC 5 uM | MBC 5 uM | Jadawalin bita, farin haske 12 J per cm2 |
| S. aureus NCTC 6571, ƙwayoyi masu shawagi a ruwa | MLC 2.5 uM | MLC 1 uM | Tsarin gwaji na asali na 1997, 1.7 mW per cm2 na awa 1, kusan 6.3 J per cm2, adadin ƙwayoyin da aka fara da su kusa da 10^6 CFU per mL, kamar yadda aka ruwaito a binciken farko kan kashe bakteriya da phenothiazinium ta haske |
| MRSA EMRSA-15, ƙwayoyi masu shawagi a ruwa | MBC 80 uM | MBC 20 uM | Jadawalin bita, kusan 12 J per cm2, ƙaruwa ninki huɗu a sauƙin kashewa |
| MRSA EMRSA-16, ƙwayoyi masu shawagi a ruwa | MBC 40 uM | MBC 10 uM | Tsarin gwaji iri ɗaya da na sama |
| E. coli NCTC 9001, ƙwayoyi masu shawagi a ruwa | MBC 500 uM | MBC 100 uM | Tsarin gwaji iri ɗaya, ƙaruwa ninki biyar a sauƙin kashewa |
| P. aeruginosa NCTC 10622, ƙwayoyi masu shawagi a ruwa | MBC 500 uM | MBC 25 uM | Tsarin gwaji iri ɗaya, ƙaruwa ninki ashirin a sauƙin kashewa |
| E. faecalis NCTC 775, ƙwayoyi masu shawagi a ruwa | MLC 100 uM | MLC 100 uM | Tsarin gwaji na 1997 a 6.3 J per cm2, babu ingantuwar sakamako ga wannan nau'in da wannan adadi |
| E. faecium mai juriya ga vancomycin, ƙwayoyi masu shawagi a ruwa | MBC 400 uM | MBC 400 uM | Jadawalin bita, babu ingantuwar sakamako da rinin asali, amma sinadaran da aka samo daga gare shi masu ƙarin kauce wa ruwa da aka ƙara wa rukunin methyl sun inganta sakamakon, bisa ga binciken enterococcus mai juriya ga vancomycin |
| Candida albicans, ƙwayoyi masu shawagi a ruwa | MIC sama da 12.79 ug per mL | MIC 12.79, 6.39, da 1.59 ug per mL a 5, 10, da 15 J per cm2 | Barinsu a duhu na minti 30 kafin haskakawa, jan LED 600 zuwa 650 nm mai kololuwa a 635 nm, bisa ga binciken alaƙar adadi da sakamako na aikin shuɗin methylene da haske wajen kashe fungi |
| C. albicans YEM14 idan aka kwatanta da mai samar da famfunan fitarwa fiye da kima, kashewa bisa CFU | Bai shafa ba | Rage adadin nau'in asali da kusan 5 log, kashewa kusa da sifili ga mai samar da CDR1/CDR2 fiye da kima | Rini 100 uM, barinsu na minti 30, 660 nm a 60 J per cm2, bisa ga binciken tasirin famfunan fitarwa kan kashewa da shuɗin methylene ta haske |
| Zaren Trichophyton rubrum idan aka kwatanta da spores | Bai shafa ba | MIC na zaren kusan 6.3 ug per mL, na spores kusan 2.0 ug per mL; dakatar da CFU gaba ɗaya ya buƙaci 5 ug per mL tare da 100 J per cm2 ga zaren, idan aka kwatanta da 1.25 ug per mL tare da 40 J per cm2 ga spores | Tsarin gwajin photodynamic na 635 nm, bisa ga binciken sauƙin shafar dermatophytes a matakai daban-daban |
| Balagaggen biofilm mai nau'ikan 33 a ƙarƙashin dasashi, na'urar Calgary, rana ta 6 | MB kaɗai ya rage jimillar ƙidaya da sama da kashi 80 cikin ɗari | MB tare da LED na 660 nm ya rage ayyukan metabolism da kusan kashi 55 cikin ɗari, babu babban sauyi a tsarin al'ummar ƙwayoyin | Rini kashi 0.01 cikin ɗari (100 ug per mL), 660 nm a kusan 330 mW per cm2 na minti 5, girma ba tare da oxygen ba, bisa ga binciken biofilm na baki mai nau'ika da yawa a dakin gwaje-gwaje |
Abubuwa uku sun fito fili. Na farko, staphylococci masu Gram positive suna mutuwa a ƙananan matakan micromolar, yayin da bakteriya masu siffar sanda na Gram negative suke buƙatar rini kusan ninki ɗari a duhu, domin membrane na waje na Gram negative yana takaita shiga. Haske yana rage tazarar amma ba ya kawar da ita. Na biyu, aikin shuɗin methylene kan fungi yana faruwa da gaske amma yana dogara da sharuɗɗa: kashe Candida da shuɗin methylene yana ƙaruwa sosai da adadin haske, kuma sauyin siffar aiki guda ɗaya, kamar samar da famfunan fitarwa fiye da kima, na iya kawar da kashewar 5 log a adadi iri ɗaya. Na uku, balagaggen biofilm yana nuna halaye daban da ƙwayoyi masu shawagi a ruwa, kuma a nan ne layin biofilm na baki a jadawalin ya dace.
Bakteriya: inda haske ya fi taimakawa, da inda ba ya taimakawa
Staphylococci su ne masu sauƙin shafuwa. Ko ƙimomin jadawalin bita na MRSA mai haifar da annoba suna zama daidai ko ƙasa da na vancomycin da aka yi kwatanci da shi bayan an yi amfani da haske, kuma sinadaran da aka ƙara wa rukunin methyl biyu sun fi yin aiki, domin ƙara methyl ya ƙara son kitse da yawan singlet oxygen da ake samarwa, tare da jure sauyawa ta reduction zuwa siffar leuco. Wannan bayani na sinadarai yana da muhimmanci: yana nuna cewa kashewa sakamakon isar da sinadari ne tare da halayen mu'amalarsa da haske, ba mu'amalar da haske kaɗai ba.
Ƙwayoyin Gram negative suna nuna ɓangaren isar da sinadari. Pseudomonas ya buƙaci 500 uM a duhu da 25 uM tare da haske, wato ingantuwar sakamako ninki ashirin, amma ƙimar da aka samu tare da haske har yanzu tana sama sosai da zangon staphylococci. Cajin tabbatacce yana taimaka wa rinin ya kusanci membrane na waje, amma shiga ciki har yanzu shi ne abin da ke takaita aikin. Yadda aka haɗa sinadarin, lokacin bari, da zaɓin sinadarin da aka samo daga asali suna sauya waɗannan ƙimomi fiye da maimaita rini ɗaya a adadi mafi yawa.
Enterococci suna ba da dalilin yin taka-tsantsan. A tsoffin tsare-tsaren gwaji, E. faecalis bai nuna wata fa'idar haske ko kaɗan ba, kuma E. faecium mai juriya ga vancomycin ya buƙaci wani molecule daban maimakon ƙarin haske. Duk wanda ya ambaci MBC guda ɗaya na shuɗin methylene a matsayin ƙima tabbatacciya ta gama-gari ga bakteriya yana karanta wallafe-wallafen bincike ba daidai ba. Babu irin wannan ƙima tabbatacciya. Akwai ƙimomi ne kawai da suka danganta da takamaiman nau'i, muhallin reno, adadin ƙwayoyin da aka fara da su, lokacin bari, tsawon zango, da jimillar kuzarin haske a kowace faɗin wuri. An tattauna yadda za a karanta waɗannan ƙimomi ba tare da ruɗar da kai ba a jagorar fassara jadawalin layin kashe ƙwayoyin cuta.
Fungi: suna shafuwa, amma mataki da famfuna ne ke yanke hukunci
Bangon fungi da ya ƙunshi glucans, mannans, da chitin yana kawo matsalar shiga sinadari daban da ta bakteriya, kuma fungi suna da kariya mai aiki. Sakamakon famfunan fitarwa na Candida da ke sama shi ne misali mafi bayyana: rini iri ɗaya, haske iri ɗaya, muhallin reno iri ɗaya, amma mai samar da famfunan fiye da kima ya tsira, yayin da nau'in asali ya rasa kusan kashi 99.999 cikin ɗari na raka'o'in da ke iya samar da tarin ƙwayoyi. Dermatophytes suna ƙara bambancin matakin ci gaba. Spores suna mutuwa cikin sauƙi fiye da zaren fungi, don haka tsarin gwajin da ke kawar da spores zai iya barin gutsattsarin zaren da rai a adadi iri ɗaya.
Shi ya sa rahotannin shuɗin methylene kan fungi suke buƙatar kulawa da sharuɗɗan haske kamar na bakteriya. Binciken photodynamic na asibiti kan farce da fata ya haɗa rini kashi 1 zuwa 2 cikin ɗari da jan haske da aka auna a takamaiman jimillar kuzarin haske a kowace faɗin wuri, tare da bibiyar sakamako daga baya, kuma ya ware kamannin da ake gani a asibiti daga tabbacin kawar da fungi. An yi bitar waɗannan gwaje-gwajen a taƙaitaccen bayanin shaidar photodynamic kan fungi na farce da fata. Shafa rini mai rauni a wurin rauni ƙarƙashin hasken ɗaki yana maimaita launin ne amma ya bar hanyar aikin.
Gwajin biofilm: tsarin baki mai nau'ikan 33
Binciken biofilm na baki mai nau'ika da yawa na 2024 ya cancanci kulawa domin yana gwada yanayi mai wahala maimakon mai sauƙi. Nau'ikan ƙwayoyin baki talatin da uku, ciki har da Porphyromonas gingivalis, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, Fusobacterium, da streptococci na baki, sun girma ba tare da oxygen ba suka samar da balagaggen biofilm a rana ta 6. An yi amfani da rini kashi 0.01 cikin ɗari tare da LED na 660 nm na minti 5 a babban ƙarfin hasken da ke sauka a kowane faɗin wuri. Rini kaɗai da haske kaɗai duk sun rage jimillar ƙidaya sosai, kuma haɗinsu ya rage ayyukan metabolism da kusan rabi, amma tsarin al'ummar ƙwayoyin bai sauya ba, kuma ba a kawar da dukan ƙwayoyin biofilm ba.
Wannan tasiri ne na gaske da yake da iyaka bayyananniya. Yana goyon bayan amfani da photodynamic a matsayin ƙarin matakin magani a wani takamaiman aljihun dasashi da aka tsaftace, ba a matsayin ruwan kurkure baki da ke magance periodontitis da aka gano shi kaɗai ba. Wannan ya kai mu ga babban gibin.
Dalilin da ya sa kashewa a dakin gwaje-gwaje ba ta zama magani kai tsaye ba
Gwajin MBC na ƙwayoyi masu shawagi a ruwa yana fallasa ƙwayoyi iri ɗaya ga rini mai daidaito da haske mai daidaito a muhallin reno mai oxygen. Wurin rauni ba ya samar da ɗaya daga cikin waɗannan. Kayan da ke kewaye da ƙwayoyin suna rage saurin shiga, yanayi masu kama da na ƙwayoyin da ke dawwama suna jure oxidation, jini da ruwan da ke fita daga rauni suna shan haske, tsarin siffar nama yana barin inuwa, kuma aljihu masu ƙarancin oxygen suna rage aikin sinadarai na Type II domin singlet oxygen yana buƙatar oxygen na molecule bisa ma'anarsa. Cututtuka masu haɗuwar ƙwayoyin cuta suna ƙara nau'ikan da suka bambanta wajen shan sinadari da sauƙin shafuwa, don haka ƙimar nau'in gwaji guda ɗaya ba za ta iya wakiltar wurin ba.
Misalin baki yana fayyace wannan gibi. Rage Porphyromonas ko streptococci a kwanon gwaji, ko rage biofilm da aka reno da rabi, ba ya tabbatar da cewa ruwan kurkure baki shuɗi yana magance periodontitis, warkar da kamuwa da cuta a cikin haƙori, ko kawar da bakteriya daga jini. Kula da cututtukan dasashi a asibiti ya dogara da warwarewa da cire biofilm ta hanyar kayan aiki, tare da kula da tsarin aljihun dasashi, yayin da ake nazarin maganin da ya dogara da haske a matsayin ƙarin mataki a takamaiman wuri. Maida hankalin rinin da ke kashe ƙwayoyi a ƙaramin ramin gwaji ba shi da tabbatacciyar alaƙa da maida hankalin da ke kaiwa bakteriya ƙarƙashin iyakar dasashi, kuma hasken ɗaki ba adadin haske na 660 nm ba ne. Tantance adadin amfani, aminci, da iyakokin hulɗa da wasu sinadarai ga duk wani amfani a baki aikin ƙwararren mai kula da lafiya ne, kamar yadda aka bayyana a bayanin aminci da hulɗar sinadarai.
Abin da wannan shaida ba take nufi ba
Ba iƙirarin cewa wannan sinadarin kashe ƙwayoyin cuta ne na gida ba ne. Jadawalin kashe bakteriya suna amfani da takamaiman nau'ika, muhallin reno, lokutan taɓawa, zangon haske, da jimillar kuzarin haske a kowace faɗin wuri. Ba su bayyana komai game da saman abubuwan da ke cikin kicin, lokutan taɓawa da ake buƙata idan akwai datti na halitta, ko ƙa'idodin hukumomi na kashe ƙwayoyin cuta ba.
Haka kuma ba madadin maganin antibiotik da ke aiki a dukkan jiki ba ne. Aikin photodynamic da aka saba amfani da shi yana buƙatar rini, hasken da aka sha, da oxygen su kasance a wuri ɗaya lokaci guda. Haske ba ya kaiwa wurare masu zurfi ko wuraren da cuta ta bazu kamar yadda maganin da aka haɗiye ke kaiwa jini. Wallafe-wallafen bincike suna sanya wannan hanya don cuta a takamaiman wuri da ake iya kaiwa ko a matsayin ƙarin matakin magani, daidai da yadda Wainwright ya bayyana kashe ƙwayoyin cuta a takamaiman wuri tun farko.
Taƙaitaccen bayani mai adalci ya fi takaita, amma har yanzu yana da ban sha'awa. Shuɗin methylene yana da ɗan aikin kashe ƙwayoyin cuta a duhu, kuma jan haske yana ba da damar tasirin photodynamic mafi girma ga bakteriya da fungi da yawa, tare da manyan bambance-bambancen haske da duhu kamar 500 zuwa 25 uM ga Pseudomonas da 80 zuwa 20 uM ga MRSA mai haifar da annoba, tare da rashin nasara mai koyar da darasi a enterococci da Candida masu iya fitar da sinadari. Sauya sakamakon bincike zuwa amfani a zahiri yana dogara da isar da sinadari, yadda biofilm ke aiki, da oxygen kamar yadda yake dogara da maida hankalin rinin da aka bayyana. A kiyaye waɗannan sharuɗɗa a bayyane, lambobin za su kasance masu ma'ana. A cire su, lambobin ba za su nuna komai ba.