Imethylene blue nokuguga kwamathambo: kutheni iziphumo zeeseli zingakhange ziqikelele iziphumo kumathambo eempuku

Khawucinge ngekhompawundi ethomalalisa kanye iiseli ezitya ithambo, ivavanywa kwisitya selebhu ize isebenze kanye njengoko bekulindelwe. Ii-osteoclast ezimbalwa ezineenucleus ezininzi, ukusabela okucacileyo ngokobungakanani bedosi, nengcaciso enokukholeleka enxulumene neemitochondria. Emva koko le khompawundi inye inikwa iimpuku phantse lonke ixesha lobomi bazo njengabantu abadala, amathambo azo askenwa kumacala amathathu, kodwa akukho nto itshintshayo. Amathambo amadala akhangeleka emadala, enyangwe okanye enganyangwanga.

Leyo yinkcazelo ethembekileyo yophando olunye lowama-2024, kwaye kufanelekile ukuluqonda ngokuchanekileyo, kuba lumi kanye kwindawo apho uninzi lwamabango ngezongezelelo zokutya namathambo luqhekeka khona. Eli phepha lelikaPoudel noogxa bakhe, lipapashwe kwi-Aging ngoMatshi 2024, kwaye lifumaneka simahla, ngoko yonke imilinganiselo engezantsi inokuthelekiswa nomthombo. Esona siphumo salo siphambili kukungabikho kwempembelelo. Iziphumo ezinjalo azikokusilela. Ziimpendulo, kwaye esi siphumo siphendula kakuhle umbuzo othile: ngaba unyango lwexesha elide nge-antioxidant, imethylene blue okanye imitoquinone, esasazwa kuwo wonke umzimba luyawagcina amathambo eempuku ezigugayo? Hayi.

Ukuguga kwamathambo kubonakala njani kumanani

Ukulahleka kwamathambo ngenxa yobudala akuyonto enye, ngoko uphando aluzange lulinganise into enye kuphela. Besebenzisa i-micro-computed tomography, abaphandi baskena iindawo ezintathu zamathambo: ithambo eliqinileyo elingaphandle embindini wesiqu sefemur (isikhondo esixineneyo), ithambo elinesakhiwo somnatha kwi-metaphysis esekupheleni kwefemur kufutshane nedolo (umnatha ofana nesiponji kufutshane nelungu), kunye nethambo elinesakhiwo somnatha kumqolo wesihlanu wesinqe. Kwindawo nganye babhala imilinganiselo eqhelekileyo yesakhiwo esincinane sethambo. I-BV/TV ngumlinganiselo womthamo oyithambo kunokuba ube sisithuba somongo. I-BMD kukuxinana kwezimbiwa. I-Tb.Th, i-Tb.N, ne-Tb.Sp zichaza ubukhulu, inani, nomgama phakathi kwemicu yethambo lomnatha. Ubukhulu bethambo elingaphandle neendawo zalo xa lisikwa linqumle zichaza isikhondo.

Cinga ngethambo lomnatha njengesakhiwo senkxaso ngaphakathi kwindawo yokugcina impahla. Ukuguga kususa ezinye izixhaso ngokupheleleyo kuze kunciphise ubukhulu beziseleyo, ngoko umlinganiselo wesithuba esizaliswe sisakhiwo uyancipha, izixhaso eziseleyo ziba kude ngakumbi enye kwenye, kwaye isakhiwo siphela sithwala umthwalo omncinane. Ithambo elingaphandle lufana nodonga lwendawo yokugcina impahla ngokwayo, oluya lusiba bhityileyo. Unyango olukhusela amathambo kufanele lubonakale kula manani. Lolo luvavanyo olwasetyenziswa luphando.

Okwenzekayo kwisitya selebhu

Uvavanyo lweeseli lwalusebenzisa umongo othathwe kwiifemur nakwiitibia zeempuku ze-UM-HET3 ezineenyanga ezi-6 ukuya kwezi-7, uhlobo oluneentlobo ezahlukeneyo zemfuza olukhuliselwa uphando lokuguga. Kwavela iziphumo ezine, kwaye zalatha kumacala ahlukeneyo, ngoko ngasinye sifuna isivakalisi saso.

Okokuqala, iiseli zesiqu ze-mesenchymal, iqela eliquka iiseli eziza kwakha amathambo, zahlala ziphila kubungakanani bemethylene blue nemitoquinone obusuka kwi-0.125 ukuya kwi-0.5 micromolar kangangeeyure ezingama-48. Kwakungekho tyhefu, kungekho kukhuthazwa. Okwesibini, xa ezo seli zesiqu zakhuthazwa ukuba zibe zii-osteoblast kwindawo yokukhulisa ekhuthaza ukwakheka kwethambo, zaza zafakwa ibala lokubonisa i-alkaline phosphatase ngomhla we-18, akukho nanye kwezi khompawundi eyatshintsha iziphumo nakweyiphi inqanaba elavavanywayo. Icala lokwakha ithambo kwisitya alizange lisabele.

Okwesithathu, icala eliqhekeza ithambo lasabela. Iiseli zomongo ezandulela ezinye iiseli, ezikhuthazwe nge-M-CSF ne-RANKL, indlela eqhelekileyo yokwenza ii-osteoclast, zenza iiseli ezimbalwa ezineenucleus ezininzi ezibonisa i-TRAP njengoko ubungakanani bekhompawundi babusanda. Ukuncipha okucacileyo kwabonakala ngaphezu kwe-0.25 micromolar, oko kukuthi, kwelona nqanaba liphezulu lavavanywayo, i-0.5. Esi sesona siphumo esivelisa amawaka ezishwankathelo ezinethemba, kwaye siyinyani ngaphakathi kwemida yaso: ukudibana ngqo nangokuqhubekayo nekhompawundi kwiiseli ezikhuliswe elebhu kwanciphisa ukwakheka kwee-osteoclast ngendlela exhomekeke kwidosi.

Okwesine, ii-osteoblast ezivuthiweyo ezachanabeka kangangeeyure ezingama-48 zaza zavavanywa ngovavanyo loxinzelelo lweemitochondria azibonisanga lutshintsho lubalulekileyo ngokwezibalo ekusetyenzisweni kweoksijini okusisiseko okanye okuphezulu ngayo nayiphi na ikhompawundi. Kwi-0.5 micromolar zombini zabonakala zinciphisa ukuphefumla kweeseli kancinci, kodwa utshintsho aluzange lufikelele ekubalulekeni ngokwezibalo. Ngoko isitya selebhu asibonisanga kuphuculwa kokusebenza kweemitochondria kwiiseli ezakha ithambo okuhambisana nempembelelo yokuthintela ii-osteoclast. Indlela ecetywayo, ukuphuculwa kwamandla nge-khonkco lothutho lwee-elektroni kwiimitochondria, ayizange ivelise ukwanda okunokulinganiswa kokuphefumla kweeseli apho abaphandi babekukhangela khona.

Okwenzekayo kwiimpuku

Kwalandela iimvavanyo ezimbini zezilwanyana, ezazinobukhulu obahluke kakhulu.

Oluncinane lwasebenzisa iimazi ze-C57BL/6J ezizalwa ngokudibanisa izizalwane, uhlobo olusetyenziswa kakhulu kuphando lwebhayomedikhali. Ukuqala kwiinyanga ezili-18 ubudala, malunga nasekupheleni kobudala obuphakathi kwimpuku, isiqingatha sazo sasela amanzi ane-250 micromolar yemethylene blue kangangeenyanga ezi-6 okanye ezili-12, de zafikelela kwiinyanga ezingama-24 okanye ezingama-30. Iimazi ezinganyangwanga ezineenyanga ezili-18 zanika imilinganiselo yokuqala yokuthelekisa, kwaye iimazi ezineenyanga ezi-5 zabonisa ukuba amathambo aselula anjani. Ukuguga kwenza kanye into eyenziwa kukuguga. Kumqolo we-L5, umlinganiselo womthamo wethambo wehla malunga ne-35 pesenti, ukuxinana kwezimbiwa malunga ne-18 pesenti, kwaye inani lemicu yethambo lomnatha lehla laya malunga nesiqingatha, nokuba lwalukho unyango okanye lungekho. Ubukhulu bethambo elingaphandle behla malunga ne-10 pesenti phakathi kweenyanga ezili-18 nezingama-24, kwaye malunga ne-30 pesenti ngeenyanga ezingama-30. Imethylene blue ayizange itshintshe nalinye kula magophe.

Uvavanyo olukhulu lwenziwa kwi-Interventions Testing Program ye-National Institute on Aging, evavanya iikhompawundi ezinokuba luncedo kwiimpuku ze-UM-HET3 zeemazi nezamadoda kwiindawo ezintathu. Imethylene blue kwi-28 parts per million ekutyeni, nemitoquinone kwi-100 parts per million, nganye yaqalwa ekuqaleni kobudala obupheleleyo yaza yaqhubeka de zaba malunga neenyanga ezingama-22 ukuya kwezingama-24 ubudala. Kwaezo ndawo zintathu zamathambo zavavanywa nge-micro-CT. Itheyibhile epapashiweyo inika umlinganiselo ngamnye ngokwesini nangonyango, isebenzisa izibalo ezilungelelanisiweyo, kwaye ikholamu yonyango ifundeka ngokufanayo kuyo yonke indawo: akukho mpembelelo ibalulekileyo ngokwezibalo, akukho kudibana kwempembelelo yonyango nesini, naphi na. Ithambo elingaphandle okanye elomnatha, ifemur okanye umqolo, inkunzi okanye imazi, imethylene blue okanye imitoquinone. Amathambo aqhubeka eguga ngendlela elindelekileyo.

Itheyibhile ethelekisayo edibanisa amacala omabini

Le yintsika elisekelwe kuyo lonke eli nqaku. Umqolo ngamnye udibanisa oko isitya selebhu sakubonisayo noko isilwanyana sakubonisayo kwinkqubo enye yebhayoloji, kusetyenziswa imilinganiselo yophando ngokwayo.

Xa ifundwa umqolo ngomqolo, le theyibhile isusa ukungqubana okubonakala kukho. Yinye kuphela kwiinkqubo ezine zeeseli eyasabelayo, kwaye yasabela kuphela kufutshane nelona nqanaba liphezulu elavavanywayo, phantsi kokuchanabeka ngqo nangokuqhubekayo. Yonke enye into kwisitya selebhu yayisele ibonisa ukungabikho kwempembelelo. Isiphumo sesilwanyana ke asibonakali kangako njengokuphikisana, koko sibonakala njengesiqinisekiso: iziphumo ezintathu ezingenampembelelo kunye nempembelelo enye exhomekeke kwiimeko kwiiseli ezikhuliswe elebhu zaba kukungabikho kwempembelelo kuyo yonke into kwisilwanyana.

Isakhiwo samathambo asibobude bobomi

Kukho ukudideka okufanele icandelo lako, kuba imethylene blue inalo irekhodi lobude bobomi, kwaye kulula ukulifunda ngokungachanekanga njengerekhodi lamathambo. Kwiqela langaphambili le-Interventions Testing Program, imethylene blue kwidosi efanayo ekutyeni yandisa ubude bobomi kwipesentayili ye-90 kwiimazi ze-UM-HET3 malunga ne-6 pesenti. Le yayiyimpembelelo yokusinda ekupheleni kobomi, ngaphandle kotshintsho kubude bobomi obuphakathi nangaphandle kwempembelelo kwiinkunzi. Imitoquinone ayizange ibonise mpembelelo kubude bobomi nakwesiphi na isini. Loo manani okusinda avela kwiirekhodi zobude bobomi ezipapashiweyo zale nkqubo, hayi kwiphepha lamathambo.

Igalelo lephepha lamathambo kukugcina ezi ngxelo zahlukene. Ikhompawundi inokutshintsha ukusinda ekupheleni kobomi kwisini esinye ngaphandle kokugcina naluphi na uphawu olulinganisiweyo lwamathambo. Ukusinda kubala ukufa kuzo zonke izizathu; i-micro-CT ibala izixhaso kumqolo. Akukho mgaqo othi ungenelelo kufuneka lusebenze ngamathambo ukuze lwandise ubomi, kwaye olu phando lububungqina obuthe ngqo bokuba umqondiso wemethylene blue ekupheleni kobomi kwiimazi wawuhamba ngenye indlela. Nabani na ocaphula isiphumo sobude bobomi njengenkxaso yemethylene blue ne-osteoporosis uchitha imali esuka kwiakhawunti engafanelekanga. Indlela eqhelekileyo yokugcina ezi ngxelo zahlukene ichazwa kwinqaku elithi indlela yokufunda uphando ngemethylene blue.

Kutheni isitya selebhu singakhange siqikelele isiphumo kwimpuku

Iinkcazelo ezintlanu ziyahambelana nedatha, kwaye zinokwenzeka kunye.

Ukuchanabeka yeyokuqala. Iiseli ezikhuliswe elebhu zazintywiliselwe ngokuqhubekayo kubungakanani bekhompawundi obaziwayo. Iimpuku zatya ukutya okuxutywe needosi kangangeenyanga, kwaye uphando aluzange lulinganise amanqanaba ekhemikhali emongweni okanye nasiphi na isalathisi somonakalo owenziwa yioksideshini ethanjeni. Inqanaba elinciphisa ii-osteoclast xa lisetyenziswa ngqo lisenokungaze lifike kwiiseli ezandulelayo emongweni kuloo mlinganiselo. Iphepha lavavanya idosi enye kwikhompawundi nganye kwizilwanyana eziphilayo, ngoko indlela isilwanyana esisabela ngayo kwiidosi ezahlukeneyo isahleli ingaziwa.

Ikhemistri yeyesibini. Iintlobo zeoksijini ezisebenza kakhulu azihlali njengeqela elinye elixutywe kakuhle ngaphakathi kweseli. Zisebenza kwiindawo ezincinane ngamaxesha amafutshane, kwaye iinkqubo ze-redox kwiimitochondria, kwi-cytosol, nakwiinwebu zisebenza ngokuzimeleyo ngokuyinxenye. Imitoquinone yenzelwe ukuba iqokelelane kwiimitochondria, kanti imiqondiso yeoksideshini eqhuba ukwakheka kwee-osteoclast inokuvela kwenye indawo. Ukunciphisa iqela elinye akuyomfuneko ukuba kuvelise oko kufezwa kukuntywilisela yonke iseli ngokuqhubekayo.

Ukudityaniswa kweenkqubo kokwesithathu. Kwithambo eliphilayo, ukuqhekeka nokwakheka kudityanisiwe: ii-osteoclast kunye neeseli ezikufutshane nazo zikhupha imiqondiso etsala iiseli ezakha ithambo kwindawo esandul’ ukuqhekekwa. Ukunciphisa ukwakheka kwee-osteoclast akongezi ithambo elilinganayo nelo lingaqhekekanga, kuba kunokususa nemiqondiso ebiza abakhi. Isitya selebhu esineeseli ezandulelayo kuphela asinakubonisa le nkqubo yempendulo. Amathambo akanakuyifihla.

Umthwalo ngowesine. Ii-osteocyte zivakalelwa luxinzelelo lomatshini zize zilungelelanise ukwakhiwa ngokutsha kwethambo lonke. Njengoko izilwanyana ziguga zize zihambe kancinci, umqondiso womthwalo womatshini uyaphela, kwaye ukulahleka kuyalandela nokuba kukho impembelelo encinane yekhemikhali kuhlobo olunye lweseli. Ababhali ngokwabo bachaza umthambo nomthwalo okhuthaza ukwakheka njengethuba elinokuba nefuthe ngakumbi, indlela enobubele yokuthi amathambo aphulaphula ifiziksi ngakumbi kunekhemistri ye-redox.

Ixesha lokuqalisa lelesihlanu, kodwa kukho umda kule ngcaciso. Kuvavanyo lwe-C57BL/6J, unyango lwaqala kwiinyanga ezili-18, xa ilahleko enkulu yesakhiwo yayisele yenzekile, kwaye akukho nto ithintela ukuqhekeka kwethambo enokuphinda yakhe imicu yethambo lomnatha esele igqobhoziwe yaza yasuswa ngaphandle kokukhuthazwa okuhambelanayo kokwakheka. Kodwa ixesha lokuqalisa alinakuchaza yonke into, kuba ukunikwa kweedosi kwi-Interventions Testing Program kwaqala ekuqaleni kobudala obupheleleyo kodwa akuzange kutshintshe nto. Ukuqala emva kwexesha kuchaza inxalenye yovavanyo olunye. Akuchazi nto kolunye.

Oko kuthethwa koku ngamabango emethylene blue ne-osteoporosis

Kumfundi ophonononga imethylene blue ngempilo yamathambo okanye ukuguga kwamathambo, intsingiselo esebenzayo imfutshane. Obona bungqina bufumanekayo bovavanyo lwexesha elide kwiimpuku buchaza ukuba akukho nzuzo kumathambo evela ekusetyenzisweni okuqhubekayo kwemethylene blue okanye kwemitoquinone, nangona kukho umqondiso wokwenene wokunciphisa ii-osteoclast kwiiseli ezikhuliswe elebhu. Uncwadi lobude bobomi lunika imethylene blue impembelelo encinane ekupheleni kobomi kwiimazi kuphela, leyo olu phando lubonisa ukuba ayizange idlule ngamathambo. Akukho nanye kwezi ngxelo exhasa ukusetyenziswa kwayo ebantwini kwi-osteoporosis, kwaye akukho datha yamathambo abantu ingenayo kulo mfanekiso. Imibuzo ngeedosi nokhuseleko lweemveliso ebantwini yeyenqaku elithi ukhuseleko nemiphumo emibi yemethylene blue, hayi eyala mathambo empuku.

Isifundo esinzulu siphathelele ulandelelwano lobungqina. Isiphumo seeseli libango malunga noko ikhompawundi inokukwenza kuhlobo olunye lweseli phantsi kokuchanabeka okulawulwayo. Isiphumo samathambo libango malunga noko kwenziwa ziinyanga zokuchanabeka komzimba wonke kwilungu elidibanisa iintlobo ezimbini zeeseli phantsi kolawulo lomthwalo womatshini. Esokuqala yindawo apho iingqikelelo zizalelwa khona. Esesibini yindawo apho, ngokuphathelele ukuguga kwamathambo nezi antioxidant zimbini, le ngqikelelo yafela khona.