I-methylene blue ngokuchasene nee-biofilm: oko kulinganiswa zizifundo zaselabhoratri ezisebenzisa ukukhanya

I-biofilm ayisosihlwele seeseli ezidada kulwelo. Sisakhiwo: iiseli ezincamathele kumphezulu, ezifakwe kwisakhiwo ezizenzela sona ngeepolysaccharide, iiproteni ne-DNA engaphandle kweeseli, apho umlinganiselo weoksijini nezondlo utshintsha ukusuka ngaphandle ukuya ngaphakathi. Eso sakhiwo sisizathu sokuba ikhompawundi ebulala iibhaktheriya ezidadayo ngemizuzu ingaphumeleli xa isetyenziswa kwezo ntsholongwane zikhuliswe njenge-biofilm. Naliphi na ibango nge-methylene blue nee-biofilm kufuneka litolikwe kuqwalaselwa eso sakhiwo, kuba sona simisela ukuba ingakanani idayi efikelela kwiiseli nokuba kungakanani ukukhanya okufikelela kwidayi.

Olu hlaziyo lugubungela imithombo emithathu kuphela. Omnye unika imvelaphi. Emibini ziimvavanyo zaselabhoratri. Ukwahlula ezo ndima kungundoqo wale ndlela, kwaye kuthintela eyona mpazamo ixhaphakileyo kwesi sihloko: ukucaphula isiphumo esifunyenwe kwisitya selabhoratri ngokungathi sibubungqina bokunyanga usulelo emntwini.

Umthombo wemvelaphi umalunga nekhemistri, hayi ukubulala

Owona mdala kule mithombo mithathu luhlaziyo oluchazayo lokuphuhliswa kwezinto ze-phenothiazinium ezenza isenzo ngokukhanya, olwapapashwa ngo-2005. Lulandela i-methylene blue ukusuka kwidayi yenkulungwane ye-19 ukuya kwikhompawundi ephambili yophando kunyango lomhlaza lwe-photodynamic, yaza kamva yasetyenziselwa ichemotherapy ye-photodynamic echasene neentsholongwane. Igalelo lalo yinkcazelo yonxulumano phakathi kwesakhiwo nomsebenzi: ukongezwa kwamaqela e-methyl kwi-chromophore kwandisa ukutyekela kwayo kumafutha, imveliso yeoksijini ekwimeko ye-singlet, namandla okubulala iibhaktheriya ngokukhanya, ngoxa kucothisa ukuncitshiswa kwayo ukuya kwimo ye-leuco engasebenzi kangako. Likwachaza ngokucacileyo ingxaki ekule nkalo. Uninzi lweengxelo lwasebenzisa iidayi eziqhelekileyo zorhwebo, kungaphuhliswa kakhulu ezinye iikhompawundi ezivela kuzo.

Sebenzisa olo hlaziyo kwinxalenye yokuqonda kwakho echaza indlela oku kusebenza ngayo: ukukhanya kufaka idayi kwimeko yamandla aphezulu, imeko ye-triplet isebenzisana neoksijini ngokudluliselwa kwamandla kwe-Type II ukuze yenze ioksijini ye-singlet, okanye ngokudluliselwa kwee-electron kwe-Type I ukuze yenze iiradical. Olu hlaselo luneendlela ezininzi lusisizathu sokuba iindlela eziqhelekileyo zokuxhathisa zingasebenzi kakuhle ekubulaleni nge-photodynamic. Musa ukulucaphula njengobungqina bokuncipha okulinganiswa nge-log. Aluzange lulinganise oko. I-photochemistry ebanzi exhasa loo ndlela yokusebenza ichazwe kwingqokelela yobungqina bonyango lwe-photodynamic, kwaye imigaqo yobude bamaza emisela ukuba isibane esithile sinokuyiqhuba na ichazwe kwinqaku elithi indlela i-methylene blue efunxa ngayo ukukhanya okubomvu.

Iimvavanyo ezimbini zee-biofilm, iintlobo ezimbini ezahlukeneyo zeziphumo

Omabini amaphepha ophando avavanya isenzo se-methylene blue se-photodynamic kwii-biofilm zomlomo ku-660 nm, kwaye phantse akukho nto yimbi avumelana ngayo. Amaqela eentsholongwane ahlukileyo, ubudala obahlukileyo, imixube yedayi eyahlukileyo, izixa zokukhanya ezahlukileyo, kwaye ngaphezu kwako konke, imilinganiselo yeziphumo eyahlukileyo. Lo mahluko ngundoqo weli nqaku, kwaye ukwaboniswa kwitheyibhile yothelekiso engezantsi elungiselelwe eli nqaku.

Olokuqala sisifundo se-in vitro se-biofilm evuthiweyo eneentlobo ezininzi engaphantsi kweentsini, esapapashwa ngo-2024. Iintlobo ezingamashumi amathathu anesithathu zakhuliswa kunye kwizikhonkwane ze-polystyrene kwisixhobo se-Calgary Biofilm Device, zondliwa phantsi kweemeko ezingenayo ioksijini. Isiphakathi sokuzikhulisa satshintshwa emva kweeyure eziyi-72, kwaza ukukhula kwaqhubeka ukuya kwiintsuku eziyi-7 ngokwesicwangciso. Unyango lwaluyi-methylene blue engafakwanga kwisithwali, ene-0.01 percent, kunye ne-LED ebomvu eyi-660 nm malunga ne-330 mW nge-square cm nganye, imizuzu eyi-5 kumgama oyi-2 mm. Xa oku kuphindaphindwa, kuphuma malunga ne-99 J nge-square cm nganye. Uyilo lovavanyo lwahlula ngokucacileyo iimeko ezine ukuze kuhlolwe impembelelo yento nganye neyokudityaniswa kwazo: iqela lolawulo elinganyangwanga, idayi ngaphandle kokukhanya, ukukhanya ngaphandle kwedayi, nedayi kunye nokukhanya.

Olwesibini sisifundo se-in vitro se-biofilm yomlomo yeentsholongwane ezincamathela kuqala, esapapashwa ngo-2021. Iintlobo ezine ze-Streptococcus zakhuliswa kunye kangangeeyure eziyi-24 kwiipleyiti ezinemingxuma eyi-96 nge-sucrose eyi-1 percent. Into eyayisebenza ngokukhanya yayiyi-methylene blue edityaniswe neenanoparticle ze-beta cyclodextrin ku-4.65 micromolar, umxube wokuhambisa idayi owenzelwe ukuyidlulisa kwizithintelo ze-biofilm. Yomibini imithombo yokukhanya yayibomvu ku-660 nm, kwaye ukukhanya kwasetyenziswa imizuzwana eyi-90, kodwa izixa zahluka kakhulu: i-320 J nge-square cm nganye kwiqela lelaser ne-8.1 J nge-square cm nganye kwiqela le-LED. Amaqela olawulo ayequka idayi ngaphandle kokukhanya, ilaser kuphela, i-LED kuphela, isisombululo setyuwa njengolawulo olungenayo into esebenzayo, ne-chlorhexidine eyi-0.2 percent njengomgangatho wokuthelekisa isenzo esichasene neentsholongwane.

Iqela leentlobo eziyi-33 ezikhule iintsuku eziyi-7 ngaphandle kweoksijini ne-biofilm yeentlobo eziyi-4 ezincamathela kuqala, ekhule iiyure eziyi-24, ziinkqubo ezahlukeneyo zokulinganisa. Amanani alandelayo akanakudityaniswa ukuze kubalwe umndilili.

Oko isifundo ngasinye sikulinganisileyo ngokwenene

Kwisifundo se-biofilm evuthiweyo, imilinganiselo yeziphumo yayikukusebenza kwemetabolism ngokuncitshiswa kwe-TTC, okufundwe ku-485 nm, nobuninzi beebhaktheriya ngokudityaniswa kwe-DNA ne-DNA ngendlela ye-checkerboard kusetyenziswa iiprobe zeentlobo eziyi-33. Umda wokufumanisa wawumalunga neeseli eziyi-10,000. Akuzange kubalwe iiseli eziphilayo ngokuzikhulisa kwiipleyiti, akuzange kulinganiswe ubunzima bezinto eziphilayo nge-crystal violet, akuzange kuthathwe imifanekiso yokwahlula eziphilayo kwezifileyo, kwaye akuzange kulandelwe ukukhula kwakhona. I-LED iyodwa yacutha ukusebenza kwemetabolism malunga ne-50 percent xa kuthelekiswa namaqela angakhanyiswanga nge-LED, yaye idayi kunye ne-LED yakucutha malunga ne-55 percent. Ngokwezibalo, iqela ledayi iyodwa lalikwiqela elinye nolawulo olunganyangwanga. Kumanani afunyenwe ngokwe-DNA, idayi iyodwa, i-LED iyodwa, nedayi kunye ne-LED zonke zazingaphantsi kolawulo ngokomahluko obalulekileyo ngokwezibalo, kodwa kwakungekho mahluko ubalulekileyo phakathi kwazo zontathu. Ababhali basichaza ngqo isigqibo: unyango aluzange luyitshabalalise i-biofilm esele imile, incamathele, nequlethe iintlobo ezininzi zeentsholongwane; i-biofilm evuthiweyo yezicubu ezingqonge amazinyo ifuna ukuphazanyiswa kwesakhiwo ngendlela yomatshini, i-photodynamic ibe sisincedisi kuphela, ukuba iyasetyenziswa.

Kwisifundo seentsholongwane ezincamathela kuqala, umlinganiselo wesiphumo wawuziintsholongwane eziphilayo ezafunyanwa ngokuzikhulisa kwiziphakathi ezikhetha iintlobo ezithile, zaza iziphumo zaboniswa kwisikali selogarithm. Iqela ledayi ngaphandle kokukhanya alizange lahluke kwiqela lesisombululo setyuwa. Omabini amaqela asebenze ngokukhanya, ilaser kunye nedayi efakwe kwisithwali ne-LED kunye nedayi efakwe kwisithwali, acutha amanani eentsholongwane eziphilayo ukuya kutsho malunga ne-4 log xa kuthelekiswa nolawulo olungenayo into esebenzayo. Oko kukuncipha ngokuphindwe nge-10,000, okanye nge-99.99 percent, kuloo mlinganiselo, kwaye akukho nalinye kula maqela lahluka ngokwezibalo kwi-chlorhexidine. Amaxabiso okukhanya kuphela nawedayi ebumnyameni kuphela awazange axelwe njengamanani e-log azimeleyo anokuthenjwa. Ngoko isishwankathelo esithembekileyo sesi sixhaswa liphepha: idayi efakwe kwisithwali neyavuselelwa kukukhanya yavelisa ukuncipha, idayi ebumnyameni ayizange ikwenze oko, kwaye ubungakanani bokuncipha babulingana nomgangatho wento ebulala iintsholongwane kule modeli ye-biofilm eselula.

Akukho nalinye iphepha elalinganisa ukukhula kwakhona emva konyango, ubunzima bezinto eziphilayo obushiyekileyo, okanye ubume besakhiwo esingqonge iiseli. Ukuncipha kwe-CFU akutsho nto ngesakhiwo esishiyekileyo. Ukuhla kwemetabolism akutsho nto ngeentsholongwane ezisindileyo. Inani elifunyenwe nge-DNA alinakuxela ukuba zeziphi eziphilayo.

Uthelekiso lweendlela zovavanyo nemilinganiselo yeziphumo

Le theyibhile sisixhobo sothelekiso seli nqaku. Funda ngokunqamlezayo kumqolo ukuze ubone oko isifundo esinye sikungqinayo, uze ufunde usihla kwikholam ukuze ubone imibuzo engaphendulwayo nasisiphi kwezi zifundo.

Imihluko emithathu ifuna ukugxininiswa. Okokuqala, kwimodeli evuthiweyo, iqela lokukhanya kuphela lichaza phantse yonke impembelelo kwimetabolism, ngoko inani le-55 percent alibonisi ukubulawa okubangelwa yidayi. Okwesibini, kwimodeli eselula, umxube awusebenzi ebumnyameni kwaye impembelelo ibonakala kuphela xa kukho ukukhanya, nto leyo eluphawu lokuvuselelwa kukukhanya. Okwesithathu, ukuhla nge-80 percent kumanani afunyenwe nge-DNA kusondele ngokwezibalo kwi-0.7 log kuphela ukuba loo manani ibingamanani eentsholongwane eziphilayo. Akunjalo, ngoko akufanele kuze kucatshulwe njengokubulawa okulinganiswa nge-log. Umxholo onxulumene nee-biofilm zamazinyo uqokelelwe kuphando lwee-biofilm zomlomo nolwamazinyo, kwaye indlela ngokubanzi yokwahlula iimodeli, iindlela zokuhambisa izinto, nemilinganiselo yeziphumo kwizifundo ezahlukeneyo ichazwe ngokweenkcukacha kwisikhokelo seendlela zaselabhoratri.

Kutheni izifundo zee-biofilm zifuna imigaqo yazo yokutolikwa

Izinto ezine eziguquguqukayo zimisela ukuba isiphumo se-methylene blue kwi-biofilm sithetha nantoni na ngaphaya kwepleyiti esifunyenwe kuyo.

Ubudala be-biofilm nesakhiwo esingqonge iiseli ziza kuqala. Ii-biofilm ezindala zijiyile ngakumbi, zineebhondi ezininzi ezidibanisa isakhiwo sazo, kwaye kunzima ngakumbi ukuba idayi ingene kuzo. Ababhali bango-2021 bachaza ukungena kwedayi nokusasazeka kakubi kokukhanya kwi-biofilm ecwangcisiweyo njengezona zithintelo ziphambili. Iindlela zokusebenzisa izithwali, njengokufaka idayi kwi-cyclodextrin, zizama ukusombulula kanye eso sithintelo. Yiloo nto umxube uyinto eguquguqukayo kuvavanyo, kungekhona nje inkcukacha encinane.

Ukubalwa kwezixa zokukhanya kuza okwesibini. Amaxabiso amandla okukhanya afunyanwa ngumphezulu apha asuka kwi-8.1 ukuya kwi-320 J nge-square cm nganye kubude bamaza obufanayo, umahluko ophantse ube ngamashumi amane ophindaphindo. Amandla okukhanya afika ngexesha elithile asuka kwi-90 mW ukuya kwii-W ezininzi nge-square cm nganye. Ubude bamaza obufanayo abuthethi ukuba ukukhanya okufunyenweyo kuyafana, kwaye akukho nasinye isifundo esitshintsha isixa ngendlela ecwangciswe ngokwaneleyo ukuze sichaze igophe lonxulumano phakathi kwesixa nesiphumo.

Amaqela olawulo aza okwesithathu. Isifundo esingenalo iqela ledayi ebumnyameni asinakukhupha ukuba idayi inetyhefu ngokwayo, kwaye isifundo esingenalo iqela lokukhanya kuphela asinakuthi nayiphi na impembelelo ibangelwa kukuvuselelwa kukukhanya. Omabini amaphepha aquka omabini la maqela, nto leyo ewomelezayo. Iphepha le-biofilm evuthiweyo libonisa ukuba kutheni iqela lokukhanya kuphela libalulekile: ngaphandle kwalo, isiphumo semetabolism besiya kutolikwa njengempembelelo yedayi.

Imilinganiselo yeziphumo iza okwesine. Ukuphila, imetabolism, ubuninzi be-DNA, ubunzima bezinto eziphilayo, isakhiwo, nokukhula kwakhona ngamabango amathandathu ahlukeneyo. Akukho phepha apha lilinganisa ngaphezu kwezimbini. Ngokukodwa, ukukhula kwakhona akukho naliphi na elikulinganisayo, ngoko ukuqhubeka kwempembelelo akulinganiswanga, kunokuba kungqiniwe.

Oko oku kungakungqiniyo malunga nokunyanga usulelo

Akukho nalinye kula malungiselelo linokujikeleza kwegazi, iiseli zokuzikhusela komzimba, ukuhamba kwamathe, ukutshintshwa kolwelo olukumsantsa weentsini, okanye ukususwa kwezinto ezincamatheleyo ngendlela yomatshini. Uxinaniso lwedayi kumngxuma wepleyiti alufani noxinaniso kwizicubu emva kokuhanjiswa kwayo emntwini nangayiphi na indlela. Ukukhanyiswa kube kanye imizuzu eyi-5 okanye imizuzwana eyi-90 kwe-biofilm eselula, okanye nevuthiweyo, yaselabhoratri akuyonkqubo epheleleyo yonyango. Ababhali besifundo se-biofilm evuthiweyo bayakutsho oku ngokwabo xa bechaza i-photodynamic njengesincedisi esinokubakho ekususweni ngendlela yomatshini, kunokuba ibe yindlela ezimeleyo yokuyitshabalalisa ngokupheleleyo.

Isishwankathelo esichanekileyo simxinwa kwaye sihlala sixhaswa bubungqina. I-methylene blue evuselelwe kukukhanya inokucutha imetabolism nobuninzi obufunyenwe nge-DNA kwiqela leentsholongwane zomlomo elivuthiweyo laselabhoratri ngaphandle kokulitshitshabalalisa, ibe ukukhanya kuphela kwenza uninzi lomsebenzi wokucutha imetabolism kolo lungiselelo. Idayi ethwelwe yi-cyclodextrin nevuselelwe kukukhanya inokucutha amanani eentsholongwane eziphilayo ngamazinga elogarithm kwi-biofilm ye-Streptococcus eselula yaselabhoratri, ukuya kumlinganiselo othelekiseka nowe-chlorhexidine kwakuloo modeli. Ukuba naluphi na kwezi ziphumo lunokusebenza kusulelo emntwini kuxhomekeke ekuhanjisweni kwento, ekufikeleleni kokukhanya, kwioksijini, kubume bomzimba, nakwimilinganiselo yeziphumo zekliniki ezi mvavanyo ezingazange ziyilelwe ukuyilinganisa.