Na gode da ziyartar Nature.com. Sigar burauzar da kuke amfani da ita tana da iyakacin tallafin CSS. Don sakamako mafi kyau, muna ba da shawarar yin amfani da sabon burauza (ko kashe yanayin dacewa a cikin Internet Explorer). A halin yanzu, don tabbatar da ci gaba da tallafi, za mu nuna rukunin yanar gizon ba tare da salo da JavaScript ba.
Kafa samfuran dabbobi na canjin modic (MC) muhimmin tushe ne don nazarin MC. Hamsin da huɗu New Zealand White zomaye aka raba zuwa sham-aiki kungiyar, tsoka implantation kungiyar (ME kungiyar) da tsakiya pulposus implantation kungiyar (NPE kungiyar). A cikin ƙungiyar NPE, an fallasa diski na intervertebral ta hanyar aikin tiyata na lumbar na gaba kuma an yi amfani da allura don huda jikin L5 na kashin baya kusa da farantin ƙarshen. An ciro NP daga faifan intervertebral L1/2 ta sirinji kuma an yi masa allura. Hako rami a cikin kashin subchondral. Hanyoyin aikin tiyata da hanyoyin hakowa a cikin ƙungiyar ƙwayar tsoka da ƙungiyar sham-aiki sun kasance daidai da waɗanda ke cikin ƙungiyar NP. A cikin ƙungiyar ME, an sanya wani yanki na tsoka a cikin rami, yayin da a cikin ƙungiyar sham-aiki, ba a sanya wani abu a cikin rami ba. Bayan aikin, an yi gwajin MRI da gwajin kwayoyin halitta. Sigina a cikin ƙungiyar NPE ta canza, amma babu wani canji na sigina a cikin ƙungiyar sham-aiki da ƙungiyar ME. Kulawar tarihi ya nuna cewa an cika yaduwar ƙwayar nama a cikin shafin da ba shi da ƙarfi, da kuma furcin Il-4, IF-17 da IFN-γ ya ƙaru a cikin NPE kungiyar. Shigar da NP a cikin ƙashin subchondral zai iya samar da samfurin dabba na MC.
Canje-canje na Modic (MC) sune raunuka na ƙarshen ƙarshen kashin baya da marrow kusa da ƙasusuwan da ake iya gani akan hoton maganadisu (MRI). Suna da yawa a cikin mutane masu alaƙa da alamun alaƙa1. Yawancin karatu sun jaddada mahimmancin MC saboda haɗuwa da ƙananan ciwon baya (LBP) 2,3. De Roos et al.4 da modic et al.5 da farko da aka fara bayyana nau'ikan sigogi guda uku daban-daban a cikin bargo na pertebral kashi barna. Canje-canjen nau'in Modic I sune hypointense akan jerin T1-nauyin (T1W) da hyperintense akan jerin T2-nauyin (T2W). Wannan raunin yana bayyana fissure ƙusoshin da ke kusa da granulation na jijiyoyi a cikin kasusuwa. Canje-canjen nau'in Modic na II yana nuna babban sigina akan duka jerin T1W da T2W. A cikin irin wannan nau'in, ana iya samun lalacewa ta ƙarshe, da kuma maye gurbin kitse na tarihi na marrow na kusa. Canje-canjen nau'in Modic na III yana nuna ƙaramin sigina a cikin jerin T1W da T2W. An lura da raunuka na sclerotic daidai da faranti na ƙarshe6. MC ana la'akari da cutar cututtukan cututtuka na kashin baya kuma yana da alaƙa da alaƙa da yawancin cututtukan cututtuka na kashin baya7,8,9.
Yin la'akari da bayanan da ake samuwa, bincike da yawa sun ba da cikakkun bayanai game da ilimin etiology da hanyoyin ilimin cututtuka na MC. Albert et al. An ba da shawarar cewa MC na iya zama sanadin cutar ta diski8. Hu et al. dangana MC zuwa matsanancin degeneration na diski10. Kroc ya ba da shawarar manufar "katsewar diski na ciki," wanda ya ce maimaita raunin diski na iya haifar da microtears a cikin ƙarshen. Bayan samuwar tsagewar, lalatawar ƙarshen ta tsakiya pulposus (NP) na iya haifar da amsawar autoimmune, wanda ke ƙara haifar da haɓakar MC11. Ma et al. raba irin wannan ra'ayi kuma ya ruwaito cewa NP-induced autoimmunity yana taka muhimmiyar rawa a cikin pathogenesis na MC12.
Kwayoyin tsarin rigakafi, musamman CD4+ T masu taimakawa lymphocytes, suna taka muhimmiyar rawa a cikin pathogenesis na autoimmunity13. Ƙwararren Th17 da aka gano kwanan nan yana samar da cytokine proinflammatory IL-17, yana inganta maganganun chemokine, kuma yana ƙarfafa ƙwayoyin T a cikin gabobin da suka lalace don samar da IFN-γ14. Kwayoyin Th2 kuma suna taka muhimmiyar rawa a cikin pathogenesis na amsawar rigakafi. Bayyana IL-4 a matsayin wakili na Th2 cell zai iya haifar da mummunan sakamako na immunopathological15.
Kodayake an gudanar da karatun asibiti da yawa akan MC16,17,18,19,20,21,22,23,24, har yanzu akwai ƙarancin samfuran gwaji na dabba masu dacewa waɗanda zasu iya kwaikwayi tsarin MC da ke faruwa akai-akai a cikin mutane kuma yana iya zama. ana amfani da su don bincika ilimin etiology ko sababbin jiyya kamar maganin da aka yi niyya. Ya zuwa yau, ƴan nau'ikan dabbobi na MC ne kawai aka ba da rahoton yin nazarin hanyoyin cututtukan cututtukan da ke ciki.
Dangane da ka'idar autoimmune da Albert da Ma suka gabatar, wannan binciken ya kafa samfurin MC zomo mai sauƙi kuma mai iya sakewa ta hanyar sarrafa kansa ta NP kusa da farantin ƙarshen vertebral. Sauran maƙasudai shine kiyaye halayen tarihi na samfuran dabbobi da kimanta takamaiman hanyoyin NP a cikin haɓakar MC. Don wannan karshen, muna amfani da fasaha irin su ilmin kwayoyin halitta, MRI, da nazarin tarihi don nazarin ci gaban MC.
Zomaye biyu sun mutu sakamakon zubar jini a lokacin tiyata, kuma zomaye hudu sun mutu yayin maganin safiya yayin MRI. Ragowar zomaye 48 sun tsira kuma ba su nuna alamun halayya ko jijiya ba bayan tiyata.
MRI yana nuna cewa ƙarfin siginar nama da aka haɗa a cikin ramuka daban-daban ya bambanta. Siginar siginar L5 na jiki a cikin ƙungiyar NPE a hankali ya canza a 12, 16 da 20 makonni bayan shigarwa (T1W jerin ya nuna ƙananan sigina, kuma jerin T2W ya nuna alamar gauraye da ƙananan sigina) (Fig. 1C), yayin da MRI ya bayyana. na sauran rukunoni biyu na sassan da aka haɗa sun kasance da kwanciyar hankali a daidai wannan lokacin (Fig. 1A, B).
(A) Wakilin jerin MRI na zomo lumbar kashin baya a lokacin 3. Ba a sami rashin daidaituwa na sigina ba a cikin hotunan ƙungiyar ta sham-operation. (B) Abubuwan siginar siginar vertebral a cikin ƙungiyar ME suna kama da waɗanda ke cikin ƙungiyar sham-aiki, kuma babu wani canji mai mahimmanci da aka lura a wurin sakawa a tsawon lokaci. (C) A cikin ƙungiyar NPE, ƙananan sigina yana bayyane a fili a cikin jerin T1W, kuma siginar da aka haɗa da ƙananan sigina suna bayyane a cikin jerin T2W. Daga lokacin makwanni 12 zuwa lokacin makwanni 20, manyan sigina masu yawa da ke kewaye da ƙananan sigina a cikin jerin T2W suna raguwa.
Za'a iya ganin hyperplasia na kasusuwa a fili a wurin da aka sanyawa na jiki na vertebral a cikin kungiyar NPE, kuma hyperplasia na kashi yana faruwa da sauri daga 12 zuwa 20 makonni (Fig. 2C) idan aka kwatanta da kungiyar NPE, babu wani canji mai mahimmanci da aka samu a cikin samfurin vertebral. jiki; Ƙungiyar Sham da ƙungiyar ME (Fig. 2C) 2A, B).
(A) saman jikin kashin baya a sashin da aka dasa yana da santsi sosai, ramin ya warke sosai, kuma babu hyperplasia a jikin kashin baya. (B) Siffar wurin da aka dasa a cikin rukunin ME yana kama da wanda ke cikin rukunin aiki na sham, kuma babu wani canji na zahiri a bayyanar da aka dasa a cikin lokaci. (C) Ciwon kai na kashi ya faru a wurin da aka dasa a cikin ƙungiyar NPE. Ƙunƙarar hyperplasia na kashi ya ƙaru da sauri kuma har ma ya kara ta hanyar intervertebral diski zuwa jikin kashin baya.
Binciken tarihi yana ba da ƙarin cikakkun bayanai game da samuwar kashi. Hoto na 3 yana nuna hotunan sassan da aka yi bayan tiyata da aka lalata da H&E. A cikin ƙungiyar sham-aiki, chondrocytes an tsara su da kyau kuma ba a gano yaduwar kwayar halitta ba (Fig. 3A). Halin da ke cikin ƙungiyar ME ya kasance daidai da wanda ke cikin ƙungiyar sham-aiki (Fig. 3B). Duk da haka, a cikin ƙungiyar NPE, an lura da yawan ƙwayar chondrocytes da haɓakar ƙwayoyin NP-kamar a wurin da aka dasa (Fig. 3C);
(A) Ana iya ganin trabeculae kusa da farantin ƙarshen, chondrocytes an tsara su da kyau tare da girman tantanin halitta da siffa kuma babu yaduwa (sau 40). (B) Yanayin wurin dasawa a cikin rukunin ME yayi kama da na rukunin sham. Ana iya ganin trabeculae da chondrocytes, amma babu wani yaduwa a fili a wurin dasawa (sau 40). (B) Ana iya ganin cewa chondrocytes da NP-kamar sel suna karuwa sosai, kuma siffar da girman chondrocytes ba daidai ba ne (sau 40).
Maganar interleukin 4 (IL-4) mRNA, interleukin 17 (IL-17) mRNA, da interferon γ (IFN-γ) mRNA an lura dasu a duka kungiyoyin NPE da ME. Lokacin da aka kwatanta matakan maganganu na kwayoyin da aka yi niyya, maganganun kwayoyin halitta na IL-4, IL-17, da IFN-γ sun karu sosai a cikin ƙungiyar NPE idan aka kwatanta da na kungiyar ME da ƙungiyar aikin sham (Fig. 4). (P <0.05). Idan aka kwatanta da ƙungiyar aiki na sham, matakan magana na IL-4, IL-17, da IFN-γ a cikin ƙungiyar ME sun karu kaɗan kawai kuma ba su kai ga canjin ƙididdiga ba (P> 0.05).
Maganar mRNA na IL-4, IL-17 da IFN-γ a cikin ƙungiyar NPE sun nuna haɓaka mafi girma fiye da waɗanda ke cikin rukunin aiki na sham da ƙungiyar ME (P <0.05).
Ya bambanta, matakan magana a cikin ƙungiyar ME ba su nuna wani bambanci mai mahimmanci (P> 0.05).
Anyi nazarin ɓangarorin Yamma ta amfani da samfuran rigakafi na kasuwanci akan IL-4 da IL-17 don tabbatar da canjin yanayin bayyanar mRNA. Kamar yadda aka nuna a cikin Figures 5A, B, idan aka kwatanta da ƙungiyar ME da ƙungiyar aikin sham, matakan furotin na IL-4 da IL-17 a cikin ƙungiyar NPE sun karu sosai (P <0.05). Idan aka kwatanta da ƙungiyar aiki na sham, matakan furotin na IL-4 da IL-17 a cikin ƙungiyar ME kuma sun kasa isa ga canje-canje masu mahimmanci (P> 0.05).
(A) Matakan furotin na IL-4 da IL-17 a cikin ƙungiyar NPE sun fi girma fiye da waɗanda ke cikin ƙungiyar ME da ƙungiyar placebo (P <0.05). (B) Histogram na Yamma.
Saboda ƙayyadaddun adadin samfuran ɗan adam da aka samu yayin tiyata, cikakkun bayanai dalla-dalla game da cututtukan MC suna da ɗan wahala. Mun yi ƙoƙarin kafa samfurin dabba na MC don yin nazarin yuwuwar hanyoyin sa. A lokaci guda, an yi amfani da kimantawa na rediyo, ƙididdigar tarihin tarihi da kuma nazarin kwayoyin halitta don bin tsarin MC wanda NP autograft ya haifar. A sakamakon haka, ƙirar ƙirar NP ta haifar da canji a hankali a cikin ƙarfin sigina daga 12-mako zuwa 20-mako-mako (makonni na 20-mako) (haɗin ƙananan sigina a cikin jerin T1W da ƙananan sigina a cikin jerin T2W), yana nuna canje-canje na nama, da tarihin tarihi da kwayoyin halitta. kimantawar nazarin halittu sun tabbatar da sakamakon binciken na rediyo.
Sakamakon wannan gwaji ya nuna cewa canje-canje na gani da tarihin tarihi sun faru a wurin da aka yi amfani da vertebral a cikin ƙungiyar NPE. A lokaci guda, bayyanar il-4, IL-17 da IFN-γ-17 da Ilyn-γ, IF-17 da Ilyn-γ, IF-17 da IFN-γ, IF-17 da IFN-γ 4 and jiki na iya haifar da jerin sigina da sauye-sauyen yanayi. Yana da sauƙi a gano cewa siginar siginar jikin vertebral na samfurin dabba (ƙananan sigina a cikin jerin T1W, sigina mai gauraya da ƙananan sigina a cikin jerin T2W) suna da kama da na ƙwayoyin vertebral na mutum, da kuma halayen MRI ma. tabbatar da abubuwan lura na histology da gross anatomy, wato, canje-canje a cikin ƙwayoyin jikin vertebral suna ci gaba. Kodayake amsawar ƙwayar cuta ta haifar da mummunan rauni na iya bayyana ba da daɗewa ba bayan huda, sakamakon MRI ya nuna cewa ci gaba da karuwa da canje-canjen sigina ya bayyana 12 makonni bayan huda kuma ya ci gaba har zuwa makonni 20 ba tare da wani alamun farfadowa ba ko kuma canza canjin MRI. Waɗannan sakamakon sun nuna cewa autologous vertebral NP hanya ce ta dogara don kafa MV mai ci gaba a cikin zomaye.
Wannan samfurin huda yana buƙatar isasshen ƙwarewa, lokaci, da ƙoƙarin tiyata. A cikin gwaje-gwaje na farko, rarrabawa ko haɓakar haɓakar sifofin ligamentous na paravertebral na iya haifar da samuwar osteophytes na vertebral. Ya kamata a kula don kada a lalata ko harzuka fayafai da ke kusa. Tunda zurfin shigar dole ne a sarrafa shi don samun daidaito da sakamako mai iya sakewa, da hannu muka yi filogi ta hanyar yanke kullin allura mai tsayin mm 3. Amfani da wannan filogi yana tabbatar da zurfin hakowa iri ɗaya a jikin kashin baya. A gwaje-gwaje na farko, likitocin kashi uku da ke cikin aikin sun sami alluran ma'auni 16 mafi sauƙin aiki tare da alluran ma'auni 18 ko wasu hanyoyin. Don guje wa zubar da jini mai yawa a lokacin hakowa, riƙe da allura har yanzu na ɗan lokaci zai ba da ramin shigar da ya fi dacewa, yana nuna cewa ana iya sarrafa wani matakin MC ta wannan hanyar.
Kodayake yawancin karatu sun yi niyya ga MC, an san kadan game da ilimin etiology da pathogenesis na MC25,26,27. Dangane da karatun da muka yi a baya, mun gano cewa ciwon kai yana taka muhimmiyar rawa wajen faruwa da ci gaban MC12. Wannan binciken yayi nazarin ƙididdigar ƙididdiga na IL-4, IL-17, da IFN-γ, waɗanda sune manyan hanyoyin bambance-bambancen ƙwayoyin CD4 + bayan haɓakar antigen. A cikin bincikenmu, idan aka kwatanta da ƙungiyar mara kyau, ƙungiyar NPE tana da mafi girman magana na IL-4, IL-17, da IFN-γ, kuma matakan furotin na IL-4 da IL-17 sun kasance mafi girma.
A asibiti, IL-17 mRNA magana yana ƙaruwa a cikin sel NP daga marasa lafiya da diski herniation28. An ƙara yawan matakan IL-4 da IFN-γ a cikin wani babban samfurin diski mara ƙarfi idan aka kwatanta da kulawar lafiya29. IL-17 yana taka muhimmiyar rawa a kumburi, raunin nama a cikin cututtuka na autoimmune30 kuma yana haɓaka amsawar rigakafi ga IFN-γ31. An sami rahoton raunin nama mai haɓakawa na IL-17 a cikin MRL / lpr mice32 da mice33 mai saurin kamuwa da cuta. IL-4 na iya hana maganganun cytokines na proinflammatory (irin su IL-1β da TNFa) da kuma kunnawa macrophage34. An ba da rahoton cewa bayanin mRNA na IL-4 ya bambanta a cikin ƙungiyar NPE idan aka kwatanta da IL-17 da IFN-γ a lokaci guda; Maganar mRNA na IFN-γ a cikin ƙungiyar NPE ya fi girma fiye da na sauran ƙungiyoyi. Sabili da haka, samar da IFN-γ na iya zama mai shiga tsakani na mayar da martani ta hanyar haɗin gwiwar NP. Nazarin ya nuna cewa IFN-γ an samar da shi ta nau'in sel masu yawa, ciki har da nau'in 1 mai taimakawa T Kwayoyin da aka kunna, kwayoyin kisa na halitta, da macrophages35,36, kuma shine cytokine mai mahimmanci na pro-inflammatory wanda ke inganta martani na rigakafi37.
Wannan binciken yana nuna cewa amsawar autoimmune na iya shiga cikin abin da ya faru da ci gaban MC. Luoma et al. gano cewa siginar siginar MC da fitattun NP suna kama da MRI, kuma duka biyu suna nuna babban sigina a cikin jerin T2W38. An tabbatar da cewa wasu cytokines suna da alaƙa da abin da ya faru na MC, kamar IL-139. Ma et al. ya ba da shawarar cewa haɓakar sama ko ƙasa na NP na iya samun babban tasiri akan abin da ya faru da ci gaban MC12. Bobechko40 da Herzbein et al.41 sun ruwaito cewa NP wani nau'i ne na rigakafi wanda ba zai iya shiga cikin jijiyoyin jini daga haihuwa ba. Fitowar NP suna gabatar da gawarwakin waje a cikin wadatar jini, ta haka ne ke yin sulhu tsakanin halayen autoimmune na gida42. Haɗin kai na autoimmune zai iya haifar da adadi mai yawa na abubuwan rigakafi, kuma lokacin da waɗannan abubuwan ke ci gaba da fallasa su zuwa kyallen takarda, suna iya haifar da canje-canje a cikin sigina43. A cikin wannan binciken, overexpression na IL-4, IL-17 da IFN-γ sune abubuwan da suka dace na rigakafi, suna kara tabbatar da kusanci tsakanin NP da MCs44. Wannan samfurin dabba yana kwaikwayi nasarar NP da shigarwa cikin farantin ƙarshen. Wannan tsari ya kara bayyana tasirin rashin lafiyar jiki ga MC.
Kamar yadda aka zata, wannan samfurin dabba yana ba mu damar yin amfani da dandamali don nazarin MC. Duk da haka, wannan samfurin har yanzu yana da wasu iyakoki: na farko, a lokacin lokacin lura da dabba, wasu zomaye masu tsaka-tsaki suna buƙatar a ba da su don gwajin ilimin tarihi da kwayoyin halitta, don haka wasu dabbobin "sun kasa amfani" a kan lokaci. Abu na biyu, duk da cewa an saita maki uku a cikin wannan binciken, amma abin takaici, mun tsara nau'in MC guda ɗaya kawai (Nau'in Modic I Change), don haka bai isa ya wakilci tsarin ci gaban cututtukan ɗan adam ba, kuma ana buƙatar saita ƙarin lokacin don haka. mafi kyawun lura da duk canje-canjen sigina. Na uku, sauye-sauyen tsarin nama za a iya nunawa a fili ta hanyar tabo na tarihi, amma wasu fasahohin na musamman na iya bayyana sauye-sauyen ƙananan ƙwayoyin cuta a cikin wannan ƙirar. Misali, an yi amfani da microscopy mai haske na polarized don nazarin samuwar fibrocartilage a cikin fayafai intervertebral zomo45. Tasirin dogon lokaci na NP akan MC da farantin karfe yana buƙatar ƙarin nazari.
Maza 54 na New Zealand fararen zomaye (nauyin kimanin 2.5-3 kg, shekaru 3-3.5 watanni) an raba su cikin dazuzzuka zuwa rukunin aikin sham, ƙungiyar shigar da tsoka (ME group) da ƙungiyar jijiyoyi (NPE group). Kwamitin da'a na asibitin Tianjin ya amince da dukkan hanyoyin gwaji, kuma an aiwatar da hanyoyin gwaji bisa ka'idojin da aka amince da su.
An yi wasu gyare-gyare ga fasahar tiyatar S. Sobajima 46. Kowane zomo an sanya shi a cikin matsayi na gefe kuma an fallasa saman gaba na fayafai guda biyar na lumbar intervertebral diski (IVDs) ta hanyar amfani da tsarin retroperitoneal na baya. An ba kowane zomo maganin sa barci na gabaɗaya (20% urethane, 5 ml/kg ta jijiyar kunne). An yi tsinkayar fata mai tsayi daga ƙananan gefen haƙarƙari zuwa ƙwanƙwasa, 2 cm na ventral zuwa tsokoki na paravertebral. An fallasa kashin baya na gefen dama daga L1 zuwa L6 ta hanyar kaifi da rarrabuwar kawuna na nama mai zurfi, nama na retroperitoneal, da tsokoki (Fig. 6A). An ƙaddara matakin diski ta amfani da ƙwanƙolin ƙashin ƙugu a matsayin alama ta jiki don matakin diski na L5-L6. Yi amfani da allurar huɗa mai ma'auni 16 don haƙa rami kusa da ƙarshen farantin vertebra na L5 zuwa zurfin 3 mm (Fig. 6B). Yi amfani da sirinji na 5-ml don yin amfani da ƙwayar ƙwayar cuta ta autologous a cikin L1-L2 intervertebral diski (Fig. 6C). Cire pulposus na tsakiya ko tsoka bisa ga buƙatun kowace ƙungiya. Bayan da rami mai zurfi ya zurfafa, ana sanya suturar da za a iya ɗauka a kan zurfin fascia, fascia na sama da fata, kula da kada ya lalata nama na periosteal na jikin kashin baya yayin tiyata.
(A) Ana fallasa diski na L5-L6 ta hanyar hanyar retroperitoneal na baya. (B) Yi amfani da allura mai ma'auni 16 don haƙa rami kusa da ƙarshen L5. (C) Ana girbe MFs masu sarrafa kansa.
An gudanar da maganin sa barci na yau da kullum tare da 20% urethane (5 ml / kg) wanda aka gudanar ta hanyar jijiya na kunne, kuma an sake maimaita radiyo na lumbar a 12, 16, da 20 makonni bayan aiki.
An sadaukar da zomaye ta hanyar allurar ketamine (25.0 mg / kg) da sodium pentobarbital (1.2 g / kg) a cikin 12, 16 da 20 makonni bayan tiyata. An cire dukkanin kashin baya don nazarin tarihi kuma an yi bincike na gaske. An yi amfani da rubutun juzu'i mai ƙima (RT-qPCR) da ɓarkewar Yamma don gano canje-canjen abubuwan rigakafi.
An yi gwaje-gwajen MRI a cikin zomaye ta amfani da 3.0 T na asibiti magnet (GE Medical Systems, Florence, SC) sanye take da mai karɓan gaɓar hannu na orthogonal. An kwantar da zomaye tare da 20% urethane (5 mL/kg) ta hanyar jijiya na kunne sannan kuma a sanya supine a cikin maganadisu tare da yankin lumbar da ke tsakiya akan madaidaicin madauwari na 5-inch (GE Medical Systems). An samo hotunan Coronal T2 masu nauyi (TR, 1445 ms; TE, 37 ms) don ayyana wurin diski na lumbar daga L3-L4 zuwa L5-L6. Sagittal jirgin sama T2-nauyi yanka aka samu tare da wadannan saituna: sauri spin-echo jerin tare da maimaita lokaci (TR) na 2200 ms da echo lokaci (TE) na 70 ms, matrix; filin gani na 260 da takwas masu motsa jiki; Tsawon kauri shine 2 mm, rata shine 0.2 mm.
Bayan da aka ɗauki hoto na ƙarshe kuma an kashe zomo na ƙarshe, an cire sham ɗin, da ke cikin tsoka, da fayafai na NP don bincikar tarihi. An gyara nama a cikin 10% tsaka-tsaki buffered formalin na mako 1, an lalata shi da ethylenediaminetetraacetic acid, kuma an raba paraffin. An saka tubalan nama a cikin paraffin kuma an yanke su cikin sassan sagittal (kauri 5 μm) ta amfani da microtome. An lalata sassan da hematoxylin da eosin (H&E).
Bayan tattara fayafai na intervertebral daga zomaye a cikin kowane rukuni, an fitar da jimlar RNA ta amfani da ginshiƙi na UNIQ-10 (Shanghai Sangon Biotechnology Co., Ltd., China) bisa ga umarnin masana'anta da tsarin jujjuyawar ImProm II (Promega Inc. , Madison, WI, Amurka). An yi juyar da rubutun.
An yi RT-qPCR ta amfani da Prism 7300 (Applied Biosystems Inc., USA) da SYBR Green Jump Start Taq ReadyMix (Sigma-Aldrich, St. Louis, MO, Amurka) bisa ga umarnin masana'anta. Girman amsawar PCR shine 20 μl kuma ya ƙunshi 1.5 μl na diluted cDNA da 0.2 μM na kowane firamare. OligoPerfect Designer (Invitrogen, Valencia, CA) ne ya tsara abubuwan da aka tsara da Nanjing Golden Stewart Biotechnology Co., Ltd. (China) (Table 1). An yi amfani da yanayin hawan keke na thermal mai zuwa: matakin kunnawa na farko na polymerase a 94 ° C na 2 min, sannan 40 hawan keke na 15 s kowane a 94 ° C don ƙididdige samfuri, annealing na 1 min a 60 ° C, tsawo, da haske. An yi ma'auni na 1 min a 72 ° C. An haɓaka duk samfuran sau uku kuma an yi amfani da matsakaicin ƙimar don nazarin RT-qPCR. An bincika bayanan haɓakawa ta amfani da FlexStation 3 (Na'urorin Molecular, Sunnyvale, CA, Amurka). IL-4, IL-17, da IFN-γ maganganun kwayoyin halitta an daidaita su zuwa kulawar endogenous (ACTB). An ƙididdige matakan maganganun dangi na mRNA mai niyya ta amfani da hanyar 2-ΔΔCT.
An fitar da jimlar furotin daga kyallen takarda ta amfani da nama homogenizer a cikin RIPA lysis buffer (wanda ya ƙunshi protease da phosphatase inhibitor hadaddiyar giyar) sa'an nan kuma a tsakiya a 13,000 rpm na 20 min a 4 ° C don cire tarkacen nama. An ɗora nauyin microgram 50 na furotin a kowane layi, an raba shi da 10% SDS-PAGE, sannan a tura shi zuwa membrane na PVDF. An yi toshewa a cikin 5% busasshiyar madara mara kitse a cikin Tris-buffered saline (TBS) mai ƙunshe da 0.1% Tween 20 na 1 h a zafin daki. An yi amfani da membrane na zomo anti-decorin primary antibody (diluted 1:200; Boster, Wuhan, China) (diluted 1:200; Bioss, Beijing, China) na dare a 4 ° C kuma ya amsa a rana ta biyu; tare da na biyu antibody (akuya anti-zomo immunoglobulin G a 1:40,000 dilution) hade da horseradish peroxidase (Boster, Wuhan, China) na 1 hour a dakin zafin jiki. An gano sigina na toshewa ta Yamma ta hanyar haɓakar kemiluminescence akan membran chemiluminescent bayan haskakawar X-ray. Don bincike na densitometric, an duba abubuwan da aka lalata kuma an ƙididdige su ta amfani da software na BandScan kuma an bayyana sakamakon a matsayin rabon immunoreactivity na ƙwayoyin cuta zuwa tubulin immunoreactivity.
An yi lissafin ƙididdiga ta amfani da kunshin software na SPSS16.0 (SPSS, Amurka). Bayanan da aka tattara a lokacin binciken an bayyana su azaman ma'anar ± daidaitattun daidaituwa (ma'anar ± SD) kuma an yi nazari ta hanyar amfani da matakan maimaita maimaitawa na bambance-bambance (ANOVA) don ƙayyade bambance-bambance tsakanin ƙungiyoyin biyu. P <0.05 an yi la'akari da mahimmancin ƙididdiga.
Don haka, kafa samfurin dabba na MC ta hanyar shigar da NPs na autologous a cikin jikin vertebral da kuma yin macroanatomical lura, MRI bincike, histological kimantawa da nazarin halittu nazarin halittu na iya zama wani muhimmin kayan aiki don tantancewa da fahimtar hanyoyin MC na ɗan adam da haɓaka sabbin hanyoyin warkewa. shisshigi.
Yadda za a buga wannan labarin: Han, C. et al. An kafa samfurin dabba na Modic canje-canje ta hanyar dasa ƙwayar ƙwayar cuta ta autologous a cikin ƙashin ƙasa na kashin baya na lumbar. Sci. Wakili 6, 35102: 10.1038/srep35102 (2016).
Weishaupt, D., Zanetti, M., Hodler, J., da Boos, N. Magnetic resonance imaging na lumbar kashin baya: yaduwa na diski herniation da kuma riƙewa, jijiya tushen matsawa, karshen farantin abnormalities, da facet hadin gwiwa osteoarthritis a asymptomatic sa kai. . ƙimar. Radiology 209, 661-666, doi:10.1148/radiology.209.3.9844656 (1998).
Kjaer, P., Korsholm, L., Bendix, T., Sorensen, JS, da Leboeuf-Eed, K. Modic canje-canje da dangantakar su da binciken asibiti. Jaridar Turai ta Turai: Jama'ar Jami'in Societo na Turai, da jama'ar Turai ta Turai na Cervical Pine Bincike na 15, 1312-1319, doi: 10.1007 / s00586-006-018.
Kuisma, M., et al. Canje-canje na Modic a cikin ƙananan ƙwayoyin vertebral na lumbar: yaduwa da haɗin gwiwa tare da ƙananan ciwon baya da sciatica a cikin ma'aikatan maza na tsakiya. Kashin baya 32, 1116–1122, doi:10.1097/01.brs.0000261561.12944.ff (2007).
de Roos, A., Kressel, H., Spritzer, K., Da Dalinka, M. MRI na kasusuwan kasusuwa ya canza kusa da ƙarshen farantin a cikin cututtuka na degenerative na kashin baya na lumbar. AJR. Jaridar Amurka ta Radiology 149, 531-534, doi: 10.2214/ajr.149.3.531 (1987).
Modic, MT, Steinberg, PM, Ross, JS, Masaryk, TJ, da Carter, JR Degenerative disc cuta: kimantawa na vertebral marrow canje-canje tare da MRI. Radiology 166, 193-199, doi:10.1148/radiology.166.1.3336678 (1988).
Modic, MT, Masaryk, TJ, Ross, JS, da Carter, JR Imaging na degenerative diski cuta. Radiology 168, 177-186, doi: 10.1148/radiology.168.1.3289089 (1988).
Jensen, TS, et al. Masu tsinkayar siginar ƙarshen ƙarshen neovertebral (Modic) suna canzawa a cikin yawan jama'a. Jaridar Spine na Turai: Official Publication of the European Spine Society, European Society of Spinal Deformity, da European Society for Cervical Spine Research, Division 19, 129-135, doi: 10.1007/s00586-009-1184-5 (2010).
Albert, HB da Mannisch, K. Modic canje-canje bayan lumbar disc herniation. Jaridar Spine na Turai : Official Publication of the European Spine Society, European Society of Spinal Deformity and the European Society for Cervical Spine Research 16, 977-982, doi: 10.1007/s00586-007-0336-8 (2007).
Kerttula, L., Luoma, K., Vehmas, T., Gronblad, M., da Kaapa, E. Modic nau'in I canje-canje na iya hango ko hasashen ci gaba da ci gaba da ɓarnawar ɓarna na ɓarna da sauri: nazarin mai yiwuwa na 1 na shekara. Jaridar Spine na Turai 21, 1135-1142, doi: 10.1007/s00586-012-2147-9 (2012).
Hu, ZJ, Zhao, FD, Fang, XQ da Fan, SW Modic canje-canje: abubuwan da za a iya haifar da su da gudummawa ga lalata diski na lumbar. Maganganun Magunguna 73, 930-932, doi: 10.1016/j.mehy.2009.06.038 (2009).
Krok, HV Tsagewar diski na ciki. Matsalolin da ke faruwa a cikin diski sama da shekaru 50. Spine (Phila Pa 1976) 11, 650-653 (1986).
Lokacin aikawa: Dec-13-2024