Enkosi ngokundwendwela i-Nature.com. Inguqulelo yesikhangeli oyisebenzisayo inenkxaso enyiniweyo yeCSS. Ngeziphumo ezingcono, sincoma ukusebenzisa isikhangeli esitsha (okanye ukuvala imo ehambelanayo kwi-Internet Explorer). Okwangoku, ukuqinisekisa inkxaso eqhubekayo, siya kubonisa isayithi ngaphandle kwezitayela kunye neJavaScript.
Ukusekwa kweemodeli zezilwanyana zotshintsho lwemodic (MC) sisiseko esibalulekileyo sokufunda iMC. Imivundla eMhlophe yaseNew Zealand engamashumi amahlanu anesine yahlulahlulwe yaba yi-sham-operation group, i-muscle implantation group (iqela le-ME) kunye neqela le-nucleus pulposus implantation (iqela le-NPE). Kwiqela le-NPE, i-disc ye-intervertebral yavezwa yindlela yokuhlinzwa ye-anterolateral lumbar kwaye inaliti yayisetyenziselwa ukuhlahlela umzimba we-L5 we-vertebral kufuphi neplate yokuphela. I-NP ikhutshwe kwi-L1 / 2 intervertebral disc ngesirinji kwaye ifakwe kuyo. Ukugaya umngxuma kwithambo le-subchondral. Iinkqubo zokuhlinzwa kunye neendlela zokugaya kwiqela lokufakelwa kwe-muscle kunye neqela le-sham-operation lalifana neqela le-NP yokufakelwa. Kwiqela le-ME, iqhekeza le-muscle lifakwe emngxunyeni, ngelixa kwiqela le-sham-operation, akukho nto ifakwe emngxunyeni. Emva kotyando, ukuskenwa kweMRI kunye novavanyo lwebhayoloji yemolekyuli lwenziwa. Isiginali kwiqela le-NPE litshintshile, kodwa kwakungekho tshintsho lucacileyo kwiqela le-sham-operation kunye neqela le-ME. Ukuqwalaselwa kwe-Histological kubonise ukuba ukwanda kwezicubu ezingaqhelekanga kubonwe kwindawo yokufakelwa, kwaye inkcazo ye-IL-4, IL-17 kunye ne-IFN-γ yonyuswa kwiqela le-NPE. Ukufakelwa kwe-NP kwithambo le-subchondral kunokwenza imodeli yezilwanyana ze-MC.
Iinguqu ze-Modic (MC) zizilonda ze-vertebral endplates kunye nomongo wethambo osondeleyo obonakalayo kwi-imaging resonance magnetic (MRI). Zixhaphake kakhulu kubantu abaneempawu ezinxulumeneyo1. Izifundo ezininzi ziye zagxininisa ukubaluleka kwe-MC ngenxa yokudibanisa kunye neentlungu ezisezantsi (LBP) 2,3. de Roos et al.4 kunye ne-Modic et al.5 ngokuzimeleyo kuqala bachaze iintlobo ezintathu ezahlukeneyo ze-subchondral signal ezingaqhelekanga kwi-vertebral bone umongo. Uhlobo lwe-Modic I utshintsho luyi-hypointense kwi-T1-weighted (T1W) ulandelelwano kunye ne-hyperintense kwi-T2-weighted (T2W) ulandelelwano. Esi silonda sityhila ii-endplates ze-fissure kunye nezicubu ezikufutshane ze-vascular granulation kumongo wethambo. Iinguqu ze-Modic ze-II zibonisa isignali ephezulu kuzo zombini i-T1W kunye ne-T2W ulandelelwano. Kolu hlobo lwesilonda, ukutshatyalaliswa kwe-endplate kunokufunyanwa, kunye nokutshintshwa kwamafutha e-histological yomongo wethambo okufutshane. Iinguqu ze-Modic ze-III zibonisa isignali ephantsi kwi-T1W kunye ne-T2W yokulandelelana. Izilonda ze-Sclerotic ezihambelana nee-endplates ziye zabonwa6. I-MC ithathwa njengesifo se-pathological of the spine kwaye ihambelana ngokusondeleyo nezifo ezininzi eziphazamisayo zomqolo7,8,9.
Ukuthathela ingqalelo idatha ekhoyo, izifundo ezininzi zinike ulwazi oluneenkcukacha kwi-etiology kunye neendlela ze-pathological ze-MC. UAlbert et al. ucebise ukuba i-MC ingabangelwa yi-disc herniation8. Hu et al. kubangelwa i-MC kwi-disc degeneration10. U-Kroc ucebise ingcamango "ye-disc rupture yangaphakathi," ethi ukuphindaphinda kwe-disc trauma kunokukhokelela kwii-microtears kwi-endplate. Emva kokubunjwa kwe-cleft, ukutshatyalaliswa kwe-endplate yi-nucleus pulposus (NP) kunokubangela impendulo ye-autoimmune, ekhokelela ngakumbi kuphuhliso lwe-MC11. UMa et al. wabelane ngombono ofanayo kwaye wachaza ukuba i-NP-induced autoimmunity idlala indima ebalulekileyo kwi-pathogenesis ye-MC12.
Iiseli ze-immune system, ngakumbi i-CD4 + T encedisa i-lymphocytes, idlala indima ebalulekileyo kwi-pathogenesis ye-autoimmunity13. I-subset ye-Th17 esandula kufunyanwa ivelisa i-proinflammatory cytokine IL-17, ikhuthaza ukubonakaliswa kwe-chemokine, kwaye ivuselela iiseli ze-T kwizitho ezonakalisiweyo ukuvelisa i-IFN-γ14. Iiseli ze-Th2 nazo zidlala indima ekhethekileyo kwi-pathogenesis yeempendulo ze-immune. Ukubonakaliswa kwe-IL-4 njengommeli we-Th2 iseli kunokukhokelela kwimiphumo enzima ye-immunopathological15.
Nangona uphando oluninzi lwezonyango lwenziwe kwi-MC16,17,18,19,20,21,22,23,24, kusekho ukunqongophala kwemizekelo yovavanyo lwezilwanyana ezifanelekileyo ezinokuxelisa inkqubo yeMC eyenzeka rhoqo ebantwini kwaye inokuba esetyenziselwa ukuphanda i-etiology okanye unyango olutsha olufana nonyango ekujoliswe kulo. Ukuza kuthi ga ngoku, zimbalwa iimodeli zezilwanyana ze-MC eziye zaxelwa ukuba zifunde iindlela ezisisiseko ze-pathological.
Ngokusekelwe kwithiyori ye-autoimmune ecetywayo ngu-Albert kunye no-Ma, olu pho nonongo luseke imodeli ye-MC yomvundla olula kunye nokuveliswa kwakhona ngokufaka i-NP ngokuzenzekelayo kufuphi nepleyiti yokuphela kwe-vertebral. Ezinye iinjongo kukuqwalasela iimpawu ze-histological zemizekelo yezilwanyana kunye nokuvavanya iindlela ezithile ze-NP kuphuhliso lwe-MC. Ukuza kuthi ga ngoku, sisebenzisa ubuchule obunje ngebhayoloji yemolekyuli, i-MRI, kunye nezifundo ze-histological ukufunda ukuqhubela phambili kwe-MC.
Imivundla emibini yafa ngokopha ngexesha lotyando, yaye imivundla emine yafa ngexesha lokuthomalalisa iintlungu ngexesha leMRI. Eminye imivundla engama-48 yasinda kwaye ayizange ibonise iimpawu zokuziphatha okanye zemithambo-luvo emva kotyando.
I-MRI ibonisa ukuba ubuninzi bomqondiso wezicubu ezifakwe kwimingxuma eyahlukeneyo ziyahluka. Ubungakanani bomqondiso we-L5 umzimba we-vertebral kwiqela le-NPE latshintsha ngokuthe ngcembe kwiiveki ze-12, i-16 kunye ne-20 emva kokufakwa (ulandelelwano lwe-T1W lubonise isignali ephantsi, kwaye ukulandelelana kwe-T2W kubonisa uphawu oluxubileyo kunye nomqondiso ophantsi) (umzobo 1C), ngelixa ukubonakala kwe-MRI kwamanye amaqela amabini eenxalenye ezifakelweyo zahlala zizinzile ngexesha elifanayo (Umfanekiso 1A, B).
(A) Ummeli we-MRIs olandelelanayo we-rabbit lumbar spine kwixesha le-3. Akukho zibonakaliso ezingaqhelekanga zifunyenwe kwimifanekiso yeqela le-sham-operation. (B) Iimpawu zeempawu zomzimba we-vertebral kwiqela le-ME zifana nezo zeqela le-sham-operation, kwaye akukho tshintsho oluphawulekayo lwesignali lubonwa kwindawo yokubethelela ngokuhamba kwexesha. (C) Kwiqela le-NPE, umqondiso ophantsi ubonakala ngokucacileyo kwi-T1W yokulandelelana, kwaye umxube odibeneyo kunye nomqondiso ophantsi ubonakala ngokucacileyo ngokulandelelana kwe-T2W. Ukususela kwiiveki ze-12 ukuya kwiiveki ze-20, izibonakaliso eziphezulu ze-sporadic ezijikeleze izibonakaliso eziphantsi kwi-T2W yokulandelelana kwehle.
I-hyperplasia yamathambo ecacileyo ingabonwa kwindawo yokufakelwa kwe-vertebral body kwiqela le-NPE, kunye ne-bone hyperplasia yenzeke ngokukhawuleza ukusuka kwi-12 ukuya kwiiveki ze-20 (umzobo we-2C) xa kuthelekiswa neqela le-NPE, akukho tshintsho oluphawulekayo olubonwayo kwi-vertebral eyimodeli. imizimba; Iqela le-Sham kunye neqela le-ME (Umfanekiso 2C) 2A, B).
(A) Umphezulu womzimba we-vertebral kwinxalenye edibeneyo ilula kakhulu, umgodi uphilisa kakuhle, kwaye akukho hyperplasia kwi-vertebral body. (B) Ubume bendawo emiliselweyo kwiqela le-ME iyafana naleyo ikwiqela lomsebenzi we-sham, kwaye akukho tshintsho lucacileyo kwinkangeleko yesiza esimiliselweyo ekuhambeni kwexesha. (C) I-Bone hyperplasia yenzeke kwindawo efakwe kwiqela le-NPE. I-hyperplasia yamathambo yanda ngokukhawuleza kwaye yanda nge-disc intervertebral ukuya kumzimba we-vertebral we-contralateral.
Uhlalutyo lwe-Histological lubonelela ngolwazi oluthe kratya malunga nokubunjwa kwamathambo. Umzobo wesi-3 ubonisa iifoto zamacandelo asemva kotyando angcoliswe yi-H&E. Kwiqela le-sham-operation, i-chondrocytes ihlelwe kakuhle kwaye akukho ukwanda kweeseli kufunyenwe (Umfanekiso 3A). Imeko kwiqela le-ME yayifana naleyo kwiqela le-sham-operation (Fig. 3B). Nangona kunjalo, kwiqela le-NPE, inani elikhulu le-chondrocytes kunye nokwanda kweeseli ze-NP-like zabonwa kwindawo yokufakelwa (umzobo 3C);
(A) I-Trabeculae ingabonwa kufuphi neplate yokuphela, i-chondrocytes ihlelwe ngokufanelekileyo kunye nobukhulu obufanayo beeseli kunye nokuma kwaye akukho ukwanda (amaxesha angama-40). (B) Imeko yendawo yokufakelwa kwiqela le-ME ifana neqela le-sham. I-Trabeculae kunye ne-chondrocytes ingabonwa, kodwa akukho kukhula okucacileyo kwindawo yokufakelwa (izihlandlo ezingama-40). (B) Kuyabonakala ukuba i-chondrocytes kunye neeseli ze-NP-ezifana ne-NP zikhula kakhulu, kwaye imilo kunye nobukhulu bee-chondrocytes azilingani (amaxesha angama-40).
Ukubonakaliswa kwe-interleukin 4 (IL-4) mRNA, interleukin 17 (IL-17) mRNA, kunye ne-interferon γ (IFN-γ) mRNA zabonwa kumaqela amabini e-NPE kunye ne-ME. Xa amanqanaba okuchazwa kwejene ekujoliswe kuyo afaniswa, ukubonakaliswa kofuzo lwe-IL-4, i-IL-17, kunye ne-IFN-γ yanda kakhulu kwiqela le-NPE xa kuthelekiswa neqela le-ME kunye neqela le-sham operation (Fig. 4) (P <0.05). Xa kuthelekiswa neqela lomsebenzi we-sham, amanqanaba okuvakalisa i-IL-4, i-IL-17, kunye ne-IFN-γ kwiqela le-ME landa nje kancinci kwaye alizange lifikelele utshintsho lwamanani (P> 0.05).
Inkcazo ye-mRNA ye-IL-4, IL-17 kunye ne-IFN-γ kwiqela le-NPE libonise umgangatho ophezulu kakhulu kunabo bakwiqela le-sham operation kunye neqela le-ME (P <0.05).
Ngokwahlukileyo, amanqanaba okuvakalisa kwiqela le-ME abonisanga mahluko ubalulekileyo (P> 0.05).
Uhlalutyo lwe-blot lwaseNtshona lwenziwa kusetyenziswa i-antibodies ekhoyo ngokurhweba ngokuchasene ne-IL-4 kunye ne-IL-17 ukuqinisekisa ipateni ye-mRNA etshintshileyo. Njengoko kuboniswe kwiMifanekiso 5A, B, xa kuthelekiswa neqela le-ME kunye neqela le-sham operation, amanqanaba eprotheyini ye-IL-4 kunye ne-IL-17 kwiqela le-NPE anyuswe kakhulu (P <0.05). Xa kuthelekiswa neqela lomsebenzi we-sham, amanqanaba eprotheyini ye-IL-4 kunye ne-IL-17 kwiqela le-ME nalo aliphumelelanga ukufikelela utshintsho olubalulekileyo (P> 0.05).
(A) Amanqanaba eprotheyini ye-IL-4 kunye ne-IL-17 kwiqela le-NPE laliphezulu kakhulu kuneqela le-ME kunye neqela le-placebo (P <0.05). (B) Ihistogram yeblothi yaseNtshona.
Ngenxa yenani eliqingqiweyo leisampulu zabantu ezifunyenwe ngexesha lotyando, izifundo ezicacileyo nezineenkcukacha malunga ne-pathogenesis ye-MC zinzima. Sizame ukuseka imodeli yezilwanyana ze-MC ukuze sifunde iindlela zayo ezinokubakho ze-pathological. Kwangaxeshanye, uvavanyo lweradiyoloji, uvandlakanyo lwe-histological kunye novavanyo lwebhayoloji yemolekyuli zasetyenziselwa ukulandela ikhosi ye-MC eyenziwa yi-NP autograft. Ngenxa yoko, imodeli yokufakelwa kwe-NP ibangele utshintsho oluthe ngcembe kwi-signal intensity ukusuka kwi-12-iveki ukuya kwii-20-ixesha lexesha leeveki (i-signal ephantsi exutywe ngokulandelelana kwe-T1W kunye nomqondiso ophantsi kwi-T2W ulandelelwano), ebonisa utshintsho lwezicubu, kunye ne-histological and molecular. uphononongo lwebhayoloji luqinisekise iziphumo zophononongo ngemitha.
Iziphumo zolu vavanyo zibonisa ukuba utshintsho olubonakalayo kunye ne-histological lwenzekile kwindawo yokwaphulwa komzimba we-vertebral kwiqela le-NPE. Ngelo xesha, ukubonakaliswa kwe-IL-4, i-IL-17 kunye ne-IFN-γ yofuzo, kunye ne-IL-4, i-IL-17 kunye ne-IFN-γ yabonwa, ebonisa ukuba ukuphulwa kwe-autologous nucleus pulposus tissue kwi-vertebral. Umzimba unokubangela uthotho lweempawu kunye notshintsho lwe-morphological. Kulula ukufumanisa ukuba iimpawu zeempawu zemizimba ye-vertebral yemodeli yezilwanyana (umqondiso ophantsi kwi-T1W ulandelelwano, umqondiso odibeneyo kunye nomqondiso ophantsi kwi-T2W ulandelelwano) zifana kakhulu neeseli ze-vertebral yomntu, kunye neempawu ze-MRI nazo. qinisekisa ukuqwalaselwa kwe-histology kunye ne-anatomy gross, oko kukuthi, utshintsho kwiiseli zomzimba we-vertebral ziqhubela phambili. Nangona impendulo yokuvuvukala ebangelwa yi-trauma enzima ingabonakala ngokukhawuleza emva kokugqabhuka, iziphumo ze-MRI zibonise ukuba utshintsho oluqhubekayo lwesignali lubonakala kwiiveki ze-12 emva kokuhlatywa kwaye luqhubekile ukuya kwiiveki ze-20 ngaphandle kweempawu zokubuyisela okanye ukuguqulwa kweenguqu ze-MRI. Ezi ziphumo zibonisa ukuba i-autologous vertebral NP yindlela ethembekileyo yokuseka i-MV eqhubekayo kwimivundla.
Le modeli yokugqobhoza ifuna isakhono esaneleyo, ixesha, kunye nomzamo wotyando. Kwiimvavanyo zokuqala, ukuchithwa okanye ukuvuselela ngokugqithiseleyo kwezakhiwo ze-ligamentous paravertebral kunokubangela ukubunjwa kwe-vertebral osteophytes. Ukunyamekela kufuneka kuthatyathwe ukungonakalisi okanye ukucaphukisa iidiski ezikufutshane. Ekubeni ubunzulu bokungena kufuneka balawulwe ukuze kufumaneke iziphumo ezihambelanayo kunye neziphindaphindayo, senze iplagi ngesandla ngokusika i-sheath yenaliti ye-3 mm ubude. Ukusebenzisa le plagi kuqinisekisa ubunzulu bokomba obufanayo kumzimba we-vertebral. Kwiimvavanyo zangaphambili, oogqirha abathathu be-orthopedic ababandakanyekayo ekusebenzeni bafumanisa iinaliti ze-16-gauge kulula ukusebenza kunye neenaliti ze-18-gauge okanye ezinye iindlela. Ukuthintela ukopha okugqithisileyo ngexesha lokomba, ukubamba inaliti imile okwethutyana kuya kubonelela umngxuma wokufaka ofanelekileyo ngakumbi, ebonisa ukuba iqondo elithile le-MC linokulawulwa ngolu hlobo.
Nangona uphando oluninzi lujolise kwi-MC, kuncinci okwaziwayo malunga ne-etiology kunye ne-pathogenesis ye-MC25,26,27. Ngokusekwe kwizifundo zethu zangaphambili, sifumanise ukuba inkqubo yokuzikhusela komzimba ngokuzenzekelayo idlala indima ephambili kwisenzeko nakuphuhliso lwe-MC12. Olu pho nonongo luhlolisise ukubonakaliswa kobungakanani be-IL-4, i-IL-17, kunye ne-IFN-γ, eyona ndlela iphambili yokuhlula iiseli ze-CD4 + emva kokuvuselela i-antigen. Kwisifundo sethu, xa kuthelekiswa neqela elibi, iqela le-NPE lalinokubonakaliswa okuphezulu kwe-IL-4, IL-17, kunye ne-IFN-γ, kwaye amanqanaba eprotheyini ye-IL-4 kunye ne-IL-17 nawo aphezulu.
Ngokonyango, i-IL-17 inkcazo ye-mRNA inyuswe kwiiseli ze-NP ezivela kwizigulane ezine-disc herniation28. Ukunyuka kwe-IL-4 kunye ne-IFN-γ amanqanaba okuvakalisa afunyenwe kwimodeli ye-disc ye-acute non-compressive disc xa kuthelekiswa nokulawula okunempilo29. I-IL-17 idlala indima ephambili ekudumbeni, ukulimala kwezicubu kwizifo ezizimelayo30 kunye nokwandisa impendulo yokuzivikela kwi-IFN-γ31. Ukuphuculwa kwe-IL-17-mediated tissue ukulimala kuye kwabikwa kwi-MRL / lpr mice32 kunye ne-autoimmunity-susceptible mice33. I-IL-4 inokuthintela ukubonakaliswa kwe-cytokines ye-proinflammatory (efana ne-IL-1β kunye ne-TNFα) kunye ne-macrophage activation34. Kwaxelwa ukuba intetho ye-mRNA ye-IL-4 yayihluke kwiqela le-NPE xa kuthelekiswa ne-IL-17 kunye ne-IFN-γ ngexesha elifanayo; Inkcazo ye-mRNA ye-IFN-γ kwiqela le-NPE yayiphezulu kakhulu kunokuba kwamanye amaqela. Ngoko ke, imveliso ye-IFN-γ ingaba ngumlamli wempendulo yokuvuvukala eyenziwa yi-NP intercalation. Ucwaningo luye lwabonisa ukuba i-IFN-γ iveliswa ziintlobo ezininzi zeeseli, kubandakanywa uhlobo olusebenzayo lwe-1 umncedisi we-T cells, iiseli ezibulalayo zendalo, kunye ne-macrophages35,36, kwaye i-cytokine ephambili ye-pro-inflammatory ekhuthaza iimpendulo ze-immune37.
Olu phononongo lubonisa ukuba impendulo ye-autoimmune inokubandakanyeka kwisehlo kunye nophuhliso lwe-MC. ULuoma et al. ifumene ukuba iimpawu zempawu ze-MC kunye ne-NP evelele zifana ne-MRI, kwaye zombini zibonisa uphawu oluphezulu kwi-T2W ukulandelelana38. Ezinye ii-cytokines ziye zaqinisekiswa ukuba zihambelana ngokusondeleyo nesiganeko se-MC, njenge-IL-139. UMa et al. icebise ukuba ukunyuka okanye ukuhla kwe-NP kunokuba nefuthe elikhulu ekwenzekeni nakuphuhliso lwe-MC12. I-Bobechko40 kunye ne-Herzbein et al.41 ibike ukuba i-NP yi-tissue ye-immunotolerant engenakungena kwi-vascular circulation ukususela ekuzalweni. Iiprotrusions ze-NP zazisa imizimba yangaphandle kunikezelo lwegazi, ngaloo ndlela ilamla i-autoimmune reactions yasekhaya42. Ukuphendulwa kwe-autoimmune kunokubangela inani elikhulu lezinto ezikhuselayo, kwaye xa ezi zinto zihlala zibonakaliswe kwizicubu, zingabangela utshintsho kwi-signing43. Kolu pho nonongo, ukugqithiswa kwe-IL-4, IL-17 kunye ne-IFN-γ zizinto eziqhelekileyo zokuzivikela, ezibonisa ngakumbi ubudlelwane obusondeleyo phakathi kwe-NP kunye ne-MCs44. Lo mzekelo wezilwanyana ulingisa kakuhle ukuphumelela kweNP kunye nokungena kwi-plate yokugqibela. Le nkqubo ibonise ngakumbi impembelelo ye-autoimmunity kwi-MC.
Njengoko kulindelekile, le modeli yezilwanyana isinika iqonga elinokwenzeka lokufunda iMC. Nangona kunjalo, le modeli isenayo imida: okokuqala, ngexesha lenqanaba lokujongwa kwezilwanyana, eminye imivundla yenqanaba eliphakathi kufuneka ixolelwe kuvavanyo lwe-histological and molecular biology, ke ezinye izilwanyana "ziyayeka ukusetyenziswa" ngokuhamba kwexesha. Okwesibini, nangona amaxesha amathathu ebekwe kolu phononongo, ngelishwa, senza kuphela uhlobo olulodwa lwe-MC (uhlobo lwe-Modic ndiyatshintsha), ngoko ke akwanelanga ukumela inkqubo yophuhliso lwesifo somntu, kwaye amaxesha amaninzi kufuneka abekwe. qaphela ngcono lonke utshintsho lwesignali. Okwesithathu, utshintsho kulwakhiwo lwethishu lunokubonakala ngokucacileyo ngokungcoliswa kwe-histological, kodwa ezinye iindlela ezikhethekileyo zinokubonisa ngcono utshintsho lwe-microstructural kule modeli. Ngokomzekelo, i-polarized light microscopy yayisetyenziselwa ukuhlalutya ukubunjwa kwe-fibrocartilage kwi-rabbit intervertebral discs45. Iziphumo zexesha elide ze-NP kwi-MC kunye ne-endplate zifuna uphando olongezelelweyo.
Imivundla emhlophe yaseNew Zealand engamashumi amahlanu anesine (ubunzima malunga ne-2.5-3 kg, iminyaka eyi-3-3.5 iinyanga) yahlulwa ngokungenamkhethe kwiqela le-sham operation, iqela lokufakelwa kwemisipha (iqela le-ME) kunye neqela lokufakelwa kweengcambu ze-nerve (iqela le-NPE). Zonke iinkqubo zokulinga zavunywa yiKomiti yezokuziphatha yeSibhedlele saseTianjin, kwaye iindlela zokuvavanya zenziwa ngokuhambelana nezikhokelo ezivunyiweyo.
Olunye uphuculo lwenziwe kwindlela yotyando ye-S. Sobajima 46. Umvundla ngamnye ubekwe kwindawo ye-lateral recumbency kunye ne-anterior surface of five consecutive lumbar intervertebral discs (IVDs) yavezwa ngokusebenzisa indlela ye-posterolateral retroperitoneal. Umvundla ngamnye wanikwa i-anesthesia jikelele (i-20% ye-urethane, i-5 ml / kg nge-ear vein). I-longitudinal skin incision yenziwa ukusuka kumgca ophantsi weembambo ukuya kwi-pelvic brim, i-2 cm ye-ventral ukuya kwi-paravertebral muscles. I-anterolateral spine echanekileyo ukusuka kwi-L1 ukuya kwi-L6 yavezwa ngokuchithwa okubukhali kunye nokuphazamiseka kwezicubu ezingaphantsi kwe-subcutaneous, izicubu ze-retroperitoneal, kunye nezihlunu (umzobo 6A). Inqanaba le-disc linqunywe ngokusebenzisa i-pelvic brim njenge-anatomical landmark ye-L5-L6 ye-disc level. Sebenzisa inaliti ye-16-gauge yokubhoboza umngxuma kufuphi neplate yokugqibela ye-L5 vertebra ukuya kubunzulu be-3 mm (Umfanekiso 6B). Sebenzisa i-5-ml yesirinji ukuze ufune i-autologous nucleus pulposus kwi-L1-L2 intervertebral disc (Umfanekiso 6C). Susa i-nucleus pulposus okanye isihlunu ngokweemfuno zeqela ngalinye. Emva kokuba umngxuma we-drill unzulu, i-sutures enokuthi ifakwe kwi-fascia enzulu, i-fascia engaphezulu kunye nolusu, unonophelo ukuba ungonakalisi izicubu ze-periosteal zomzimba we-vertebral ngexesha lotyando.
(A) I-disc ye-L5-L6 ibonakaliswe ngendlela ye-posterolateral retroperitoneal. (B) Sebenzisa inaliti ye-16-gauge ukumba umngxuma kufuphi ne-endplate ye-L5. (C) Ii-Autologous MF ziyavunwa.
I-anesthesia jikelele ilawulwa nge-20% ye-urethane (5 ml / kg) elawulwa nge-ear vein, kunye ne-lumbar spine radiographs iphindwe kwi-12, i-16, kunye ne-20 iiveki emva kokusebenza.
Imivundla yanikelwa nge-injection ye-intramuscular ye-ketamine (25.0 mg / kg) kunye ne-intravenous sodium pentobarbital (1.2 g / kg) kwi-12, i-16 kunye ne-20 iiveki emva kokuhlinzwa. Umqolo wonke wasuswa kuhlalutyo lwe-histological kwaye uhlalutyo lwangempela lwenziwa. Ukubhalwa kwe-quantitative reverse transcription (RT-qPCR) kunye ne-Western blotting yasetyenziselwa ukufumanisa utshintsho kwizinto zokuzivikela.
Iimviwo ze-MRI zenziwa kwimivundla usebenzisa i-3.0 T yemagnethi yekliniki (GE Medical Systems, Florence, SC) exhotywe nge-orthogonal i-coil receiver imilenze. Imivundla yathotywa i-anesthetized nge-20% ye-urethane (5 mL/kg) ngokusebenzisa umthambo wendlebe emva koko yabekwa phezu kwemagnethi kunye nommandla we-lumbar ogxile kwi-5-intshi ye-diameter ye-coil surface circular (GE Medical Systems). Imifanekiso ye-Coronal T2-weighted localizer (TR, 1445 ms; TE, 37 ms) yafunyanwa ukuze ichaze indawo ye-lumbar disc ukusuka kwi-L3-L4 ukuya kwi-L5-L6. I-Sagittal plane i-T2-weighted slices yafunyanwa ngezi zicwangciso zilandelayo: ukulandelelana ngokukhawuleza kwe-spin-echo kunye nexesha lokuphindaphinda (TR) ye-2200 ms kunye nexesha le-echo (TE) le-70 ms, i-matrix; intsimi ebonakalayo ye-260 kunye ne-stimuli esibhozo; Ubunzima bokusika bebuyi-2 mm, i-gap yayiyi-0.2 mm.
Emva kokuba ifoto yokugqibela ithathwe kwaye umvundla wokugqibela wabulawa, i-sham, i-muscle-embedded, kunye neediski ze-NP zasuswa ukuze kuhlolwe i-histological. Izicubu zilungiswe kwi-10% ye-neutral buffered formalin kwiveki ye-1, i-decalcified kunye ne-ethylenediaminetetraacetic acid, kunye ne-parafini icandelo. Iibhloko zezicubu zifakwe kwiparafini kwaye zisikwe kumacandelo e-sagittal (5 μm ubukhulu) kusetyenziswa i-microtome. Amacandelo afakwe i-hematoxylin kunye ne-eosin (H & E).
Emva kokuqokelela i-intervertebral discs ukusuka kwimvundla kwiqela ngalinye, i-RNA iyonke yachithwa kusetyenziswa ikholomu ye-UNIQ-10 (i-Shanghai Sangon Biotechnology Co., Ltd., China) ngokwemiyalelo yomenzi kunye ne-ImProm II reverse transcription system (Promega Inc. , Madison, WI, USA). Kwenziwe umva ukhuphelo.
I-RT-qPCR yenziwa ngokusebenzisa i-Prism 7300 (i-Applied Biosystems Inc., e-USA) kunye ne-SYBR Green Jump Qala i-Taq ReadyMix (i-Sigma-Aldrich, i-St. Louis, MO, USA) ngokwemiyalelo yomenzi. Umthamo wokuphendula we-PCR wawuyi-20 μl kwaye uqulethe i-1.5 μl ye-cDNA ehlanjululweyo kunye ne-0.2 μM ye-primer nganye. Iiprimers zenziwa ngu-OligoPerfect Designer (Invitrogen, Valencia, CA) kwaye zenziwe nguNanjing Golden Stewart Biotechnology Co., Ltd. (China) (Itheyibhile 1). Ezi meko zilandelayo zebhayisikile ezishushu zisetyenzisiwe: inyathelo lokuqala lokuvula i-polymerase kwi-94 ° C kwimizuzu emi-2, emva koko imijikelo ye-40 ye-15 s nganye kwi-94 ° C ye-template denaturation, i-annealing ye-1 min kwi-60 ° C, ukwandiswa, kunye ne-fluorescence. imilinganiselo yenziwe kwi-1 min kwi-72 ° C. Zonke iisampuli zandiswe kathathu kwaye ixabiso eliphakathi lisetyenziselwa uhlalutyo lwe-RT-qPCR. Idatha yokwandisa yahlaziywa kusetyenziswa i-FlexStation 3 (i-Molecular Devices, Sunnyvale, CA, USA). I-IL-4, i-IL-17, kunye ne-IFN-γ ye-gene expression yayiqhelekile kulawulo olungapheliyo (ACTB). Amanqanaba okuchaza azalanayo ekujoliswe kuko ku-mRNA abalwe kusetyenziswa indlela ye-2-ΔΔCT.
Iprotheyini epheleleyo ikhutshwe kwizicubu ezisebenzisa i-tissue homogenizer kwi-RIPA lysis buffer (equkethe i-protease kunye ne-phosphatase inhibitor cocktail) kwaye emva koko i-centrifuged kwi-13,000 rpm ye-20 min kwi-4 ° C ukususa i-debris yezicubu. I-micrograms ezingamashumi amahlanu zeprotheni zilayishwe ngomzila ngamnye, zihlulwe nge-10% ye-SDS-PAGE, kwaye emva koko idluliselwe kwi-membrane ye-PVDF. Ukuvinjwa kwenziwa kwi-5% yobisi olomileyo lwe-nonfat kwi-Tris-buffered saline (TBS) equkethe i-0.1% Phakathi kwe-20 kwi-1 h kwiqondo lokushisa. I-membrane ifakwe kunye ne-rabbit anti-decorin primary antibody (i-diluted 1:200; i-Boster, i-Wuhan, i-China) (i-diluted 1: 200; i-Bioss, i-Beijing, i-China) ubusuku bonke kwi-4 ° C kwaye iphendule ngeentsuku zesibini; kunye ne-antibody yesibini (ibhokhwe yokulwa nomvundla i-immunoglobulin G ngo-1:40,000 dilution) idityaniswe ne-horseradish peroxidase (Boster, Wuhan, China) iyure enye kwiqondo lobushushu begumbi. Iimpawu ze-blot zaseNtshona zifunyenwe ngokunyuka kwe-chemiluminescence kwi-membrane ye-chemiluminescent emva kwe-X-ray irradiation. Kuhlalutyo lwe-densitometric, ama-blots askenwa kwaye abalwa kusetyenziswa isoftware ye-BandScan kwaye iziphumo zachazwa njengomlinganiselo wokunganyangeki kwemfuza ekujoliswe kuyo kwi-tubulin immunoreactivity.
Ubalo lwamanani lwenziwe kusetyenziswa iphakheji yesoftware ye-SPSS16.0 (SPSS, USA). Idatha eqokelelwe ngexesha lophononongo ibonakaliswe njengentsingiselo ± ukuphambuka okusemgangathweni (ithetha ± SD) kwaye yahlalutya kusetyenziswa indlela enye yokulinganisa uhlalutyo lokuhluka (ANOVA) ukufumanisa umahluko phakathi kwamaqela amabini. I-P <0.05 ithathwa njengebaluleke kakhulu ngokwezibalo.
Ngaloo ndlela, ukusekwa kwemodeli yezilwanyana ze-MC ngokufakela i-NPs ye-autologous kwi-vertebral body kunye nokwenza i-macroanatomical observation, uhlalutyo lwe-MRI, uvavanyo lwe-histological kunye nohlalutyo lwebhayoloji ye-molecular ingaba sisixhobo esibalulekileyo sokuvavanya nokuqonda iindlela ze-MC zabantu kunye nokuphuhlisa unyango olutsha. ungenelelo.
Indlela yokucaphula eli nqaku: Han, C. et al. Imodeli yezilwanyana yeenguqu ze-Modic yasungulwa ngokufakela i-autologous nucleus pulposus kwithambo le-subchondral ye-lumbar spine. Sci. I-Rep. 6, 35102: 10.1038 / srep35102 (2016).
I-Weishaupt, D., Zanetti, M., Hodler, J., kunye ne-Boos, i-N. Magnetic resonance imaging of the lumbar spine: ukuxhaphaka kwe-disc herniation kunye nokugcinwa, ukunyanzeliswa kweengcambu ze-nerve, i-plate abnormalities, kunye ne-facet joint osteoarthritis kumavolontiya angabonakaliyo. . izinga. I-Radiology 209, 661-666, i-doi: 10.1148 / i-radiology.209.3.9844656 (1998).
Kjaer, P., Korsholm, L., Bendix, T., Sorensen, JS, kunye neLeboeuf-Eed, K. Modic utshintsho kunye nobudlelwane babo kwiziphumo zeklinikhi. I-European Spine Journal: ukupapashwa ngokusemthethweni kwe-European Spine Society, i-European Society of Spinal Deformity, kunye ne-European Society for Cervical Spine Research 15, 1312-1319, doi: 10.1007 / s00586-006-0185-x (2006).
Kuisma, M., et al. Utshintsho lwe-Modic kwi-endplates ye-lumbar vertebral: ukuxhaphaka kunye nokudibanisa neentlungu ezisezantsi kunye ne-sciatica kubasebenzi besilisa abaneminyaka ephakathi. Umqolo 32, 1116-1122, doi: 10.1097 / 01.brs.0000261561.12944.ff (2007).
de Roos, A., Kressel, H., Spritzer, K., kunye noDalinka, M. MRI yethambo lethambo litshintsha kufuphi neplate yokuphela kwisifo esibuhlungu se-lumbar spine. AJR. I-American Journal yeRadiology 149, 531-534, doi: 10.2214 / ajr.149.3.531 (1987).
I-Modic, MT, Steinberg, PM, Ross, JS, Masaryk, TJ, kunye neCarter, JR Isifo se-disc degenerative: ukuvavanywa kweenguqu ze-vertebral marrow ngeMRI. I-Radiology 166, 193-199, doi: 10.1148 / i-radiology.166.1.3336678 (1988).
I-Modic, MT, Masaryk, TJ, Ross, JS, kunye neCarter, JR Imaging of degenerative disc disease. I-Radiology 168, 177-186, doi: 10.1148 / radiology.168.1.3289089 (1988).
Jensen, TS, et al. Izibikezelo ze-neovertebral endplate (Modic) utshintsho lwesignali kuluntu jikelele. I-European Spine Journal: Ukupapashwa ngokusemthethweni kwe-European Spine Society, i-European Society of Spinal Deformity, kunye noMbutho waseYurophu woPhando lweCervical Spine, iCandelo 19, 129-135, doi: 10.1007 / s00586-009-1184-5 (2010).
UAlbert, HB kunye noMannisch, K. Modic utshintsho emva kwe-lumbar disc herniation. I-European Spine Journal : Ukupapashwa ngokusemthethweni kwe-European Spine Society, i-European Society of Spinal Deformity kunye ne-European Society for Cervical Spine Research 16, 977-982, doi: 10.1007 / s00586-007-0336-8 (2007).
I-Kerttula, L., Luoma, K., Vehmas, T., Gronblad, M., kunye ne-Kaapa, E. Uhlobo lwe-Modic I utshintsho lunokuqikelela ngokukhawuleza ukuguqulwa kwe-disc deformational disc degeneration: i-1-year prospective study. I-European Spine Journal 21, 1135-1142, doi: 10.1007 / s00586-012-2147-9 (2012).
Hu, ZJ, Zhao, FD, Fang, XQ and Fan, SW Modic utshintsho: izizathu ezinokwenzeka kunye negalelo kwi-lumbar disc degeneration. I-Hypotheses yezoNyango 73, 930-932, doi: 10.1016 / j.mehy.2009.06.038 (2009).
Krok, HV Internal disc rupture. Iingxaki zeDisc prolapse ngaphezulu kweminyaka engama-50. Umqolo (Phila Pa 1976) 11, 650-653 (1986).
Ixesha lokuposa: Dec-13-2024