Ndatenda nekushanyira Nature.com. Shanduro yebrowser yauri kushandisa ine tsigiro yeCSS shoma. Kuti uwane mibairo yakanaka, tinokurudzira kushandisa bhurawuza nyowani (kana kudzima inoenderana modhi muInternet Explorer). Zvichakadaro, kuti tive nechokwadi chekuenderera mberi nerutsigiro, isu ticharatidza saiti isina masitaera uye JavaScript.
Kugadzwa kwemhando dzemhuka dzemodic shanduko (MC) hwaro hwakakosha hwekudzidza MC. Makumi mashanu neina eNew Zealand White tsuro akakamurwa kuita sham-operation boka, tsandanyama kudyara boka (ME boka) uye nucleus pulposus implantation boka (NPE boka). Muboka reNPE, intervertebral disc yakaratidzwa neanterolateral lumbar surgical approach uye tsono yakashandiswa kubaya L5 vertebral body pedyo neplate yekupedzisira. NP yakatorwa kubva kuL1 / 2 intervertebral disc nesirinji uye yakapinza mairi. Kuchera gomba mu subchondral bone. Nzira dzekuvhiya uye nzira dzekuchera muboka rekuisa tsandanyama uye sham-operation boka raive rakafanana neaya ari muNP implantation group. Muboka reME, chidimbu chemhasuru chakaiswa mugomba, nepo muboka resham-operation, hapana chakaiswa mugomba. Mushure mekuvhiyiwa, MRI scanning uye molecular biological test yakaitwa. Chiratidzo muboka reNPE chakachinja, asi pakanga pasina shanduko iri pachena yechiratidzo muboka rekushanda-sham neboka reME. Histological observation yakaratidza kuti kuwedzera kwenyama kusina kujairika kwakaonekwa panzvimbo yekusimwa, uye kutaura kweIL-4, IL-17 uye IFN-γ kwakawedzerwa muboka reNPE. Kuiswa kweNP mu subchondral bone kunogona kuumba mhando yemhuka yeMC.
Modic shanduko (MC) zvironda zve vertebral endplates uye padyo nemwongo wepfupa unoonekwa pamagnetic resonance imaging (MRI). Izvo zvinowanzoitika muvanhu vane zviratidzo zvakabatana1. Zvidzidzo zvakawanda zvakasimbisa kukosha kweMC nekuda kwekushamwaridzana kwayo nemarwadzo ekudzokera shure (LBP) 2,3. de Roos et al.4 uye Modic et al.5 vakazvimiririra vakatanga vakatsanangura zvitatu zvakasiyana-siyana zve subchondral zviratidzo zvisiri izvo mu vertebral bone marrow. Modic type I shanduko ndeye hypointense paT1-weighted (T1W) sequences uye hyperintense paT2-weighted (T2W) sequences. Ichi chironda chinoratidza fissure endplates uye padyo nevascular granulation tissue mumwongo. Modic type II shanduko dzinoratidza chiratidzo chepamusoro pane ese T1W uye T2W kutevedzana. Mune rudzi urwu rwekukuvadza, kupera kweplate kunogona kuwanikwa, pamwe chete ne histological fatty replacement yepedyo yemapfupa emapfupa. Modic type III shanduko inoratidza yakaderera chiratidzo muT1W uye T2W kutevedzana. Sclerotic maronda anoenderana nemaendplates akaonekwa6. MC inoonekwa sechirwere chechirwere chemuzongoza uye yakanyatsobatanidzwa nezvirwere zvakawanda zvinopera zvemusana7,8,9.
Tichifunga nezve data iripo, zvidzidzo zvakati wandei zvakapa ruzivo rwakadzama mune etiology uye pathological nzira dzeMC. Albert et al. akataura kuti MC inogona kukonzerwa ne disc herniation8. Hu et al. inonzi MC kune yakanyanya disc degeneration10. Kroc akaronga pfungwa ye "mukati disc rupture," iyo inotaura kuti repetitive disc trauma inogona kutungamirira ku microtears mu endplate. Mushure mekuumbwa kwakatsemuka, kupera kweplate neiyo nucleus pulposus (NP) inogona kukonzeresa mhinduro ye autoimmune, izvo zvinotungamira mukukudziridzwa kweMC11. Ma et al. vakagovana maonero akafanana uye vakashuma kuti NP-induced autoimmunity inobata basa rakakosha pathogenesis yeMC12.
Immune system masero, kunyanya maCD4 + T mubatsiri lymphocytes, anoita basa rakakosha pathogenesis ye autoimmunity13. Iyo ichangobva kuwanikwa Th17 subset inogadzira proinflammatory cytokine IL-17, inosimudzira chemokine kutaura, uye inokurudzira T masero munhengo dzakakuvadzwa kuti abudise IFN-γ14. Masero eTh2 anoitawo basa rakasiyana mune pathogenesis yemhinduro dzekudzivirira. Kutaura kwe IL-4 semumiririri Th2 sero kunogona kutungamirira kune zvakakomba immunopathological effects15.
Kunyangwe zvidzidzo zvekiriniki zvakawanda zvakaitwa paMC16,17,18,19,20,21,22,23,24, pachine kushomeka kwemhando dzekuyedza dzemhuka dzinogona kutevedzera maitiro eMC anowanzoitika muvanhu uye anogona inoshandiswa kuongorora etiology kana marapirwo matsva akadai seanonangwa kurapa. Parizvino, mhando shoma dzemhuka dzeMC dzakashumwa kuti dzidzidze nzira dzepathological.
Zvichienderana nedzidziso ye autoimmune yakatsanangurwa naAlbert naMa, chidzidzo ichi chakagadzira yakapfava uye inoberekana tsuro MC modhi ne autotransplanting NP padhuze nedhiri vertebral end plate. Zvimwe zvinangwa ndezvekutarisa histological maitiro emhando dzemhuka uye kuongorora maitiro chaiwo eNP mukuvandudza kweMC. Kuti izvi zviitike, tinoshandisa matekiniki akadai semolecular biology, MRI, uye histological zvidzidzo kudzidza mafambiro eMC.
Tsuro mbiri dzakafa nekubuda ropa munguva yekuvhiyiwa, uye tsuro ina dzakafa panguva yekuvhiyiwa panguva yeMRI. Tsuro makumi mana nesere dzakasara dzakararama uye dzakaratidza kusazvibata kana zviratidzo zvetsinga mushure mekuvhiyiwa.
MRI inoratidza kuti kusimba kwechiratidzo chemukati wakamisikidzwa mumakomba akasiyana kwakasiyana. Chiratidzo chekusimba kweL5 vertebral body muboka reNPE zvishoma nezvishoma zvakashanduka pa12, 16 uye mavhiki e20 mushure mekuiswa (T1W sequence yakaratidza chiratidzo chakaderera, uye kutevedzana kweT2W kwakaratidza chiratidzo chakavhenganiswa pamwe nechiratidzo chakaderera) (Fig. 1C), nepo MRI inooneka. yemamwe mapoka maviri ezvikamu zvakabatanidzwa zvakaramba zvakagadzikana panguva imwe chete (Fig. 1A, B).
(A) Mumiririri akatevedzana maMRIs etsuro lumbar spine pa3 nguva. Hapana kusarudzika kwechiratidzo kwakawanikwa mumifananidzo yeboka resham-operation. (B) Zviratidzo zvechiratidzo chemuviri we vertebral muboka re ME zvakafanana neavo vari muboka rekushanda-sham, uye hapana shanduko yakakosha yechiratidzo inoonekwa panzvimbo yekubatanidza nekufamba kwenguva. (C) Muboka reNPE, chiratidzo chepasi chinonyatsooneka mumutsara weT1W, uye chiratidzo chakasanganiswa uye chiratidzo chepasi chinonyatsooneka mumutsara weT2W. Kubva panguva ye12-vhiki kusvika kunguva ye20-vhiki, zvikwangwani zvepamusoro zvakakomberedza zviratidzo zvakaderera muT2W kutevedzana zvinoderera.
Zviri pachena pfupa hyperplasia inogona kuonekwa panzvimbo yekusimwa kwemuviri we vertebral muboka reNPE, uye pfupa hyperplasia inoitika nokukurumidza kubva ku12 kusvika kumavhiki e20 (Fig. 2C) kana ichienzaniswa neboka reNPE, hapana kuchinja kukuru kunoonekwa mumuenzaniso we vertebral. miviri; Sham boka uye ME boka (Fig. 2C) 2A, B).
(A) Pamusoro pemuviri we vertebral pane chikamu chakasimwa chakanyanya kutsetseka, gomba rinoporesa zvakanaka, uye hapana hyperplasia mumutumbi wevertebral. (B) Chimiro chenzvimbo yakasimwa muboka reME yakafanana neiyo muboka rekushanda kwesham, uye hapana shanduko iri pachena mukuonekwa kwenzvimbo yakasimwa nekufamba kwenguva. (C) Bone hyperplasia yakaitika panzvimbo yakasimwa muboka reNPE. Iyo pfupa hyperplasia yakawedzera nekukurumidza uye yakatowedzera kuburikidza ne intervertebral disc kusvika kune contralateral vertebral body.
Histological analysis inopa ruzivo rwakadzama pamusoro pekuumbwa kwemapfupa. Mufananidzo 3 unoratidza mafoto ezvikamu zvekuvhiyiwa zvakasvibiswa neH&E. Muchikwata che sham-operation, chondrocytes yakanyatsorongeka uye hapana kuwedzera kwesero kwakawanikwa (Fig. 3A). Mamiriro ezvinhu muboka reME akanga akafanana neaya ari muboka resham-operation (Fig. 3B). Zvisinei, muboka reNPE, nhamba yakawanda yechondrocytes uye kupararira kweNP-sero semasero akaonekwa panzvimbo yekuisa (Fig. 3C);
(A) Trabeculae inogona kuonekwa pedyo neplate yekupedzisira, iyo chondrocytes yakanyatsorongedzwa neuniform cell size uye chimiro uye hapana kuwedzera (40 times). (B) Mamiriro enzvimbo yekusimwa muboka reME yakafanana neyeboka re sham. Trabeculae uye chondrocytes zvinogona kuonekwa, asi hapana kupararira kuri pachena panzvimbo yekusimwa (40 times). (B) Zvinogona kuonekwa kuti chondrocytes uye NP-sero masero anowedzera zvakanyanya, uye chimiro uye ukuru hwechondrocytes hazvina kuenzana (40 times).
Kutaura kwe interleukin 4 (IL-4) mRNA, interleukin 17 (IL-17) mRNA, uye interferon γ (IFN-γ) mRNA yakaonekwa mumapoka maviri eNPE neME. Apo mazinga ekutaura kwemajini anotarirwa akaenzaniswa, mazwi ejeni eIL-4, IL-17, uye IFN-γ akawedzera zvakanyanya muboka reNPE zvichienzaniswa neboka reME uye sham operation boka (Fig. 4) (P <0.05). Kuenzaniswa neboka rekushanda kwesham, mazinga ekutaura eIL-4, IL-17, uye IFN-γ muboka reME akawedzera zvishoma uye haana kusvika pakuchinja kwenhamba (P> 0.05).
Iyo mRNA kutaura kweIL-4, IL-17 uye IFN-γ muboka reNPE yakaratidza chimiro chepamusoro kupfuura avo vari muboka rekuita sham uye ME boka (P <0.05).
Kusiyana neizvi, mazinga ekutaura muboka reME haana kuratidza musiyano wakakosha (P> 0.05).
Ongororo yeWestern blot yakaitwa pachishandiswa masoja ekudzivirira chirwere anotengeswa kurwisa IL-4 uye IL-17 kusimbisa iyo yakashandurwa mRNA kutaura pateni. Sezvinoratidzwa muFigure 5A, B, zvichienzaniswa neboka reME uye sham operation boka, mapuroteni mazinga eIL-4 uye IL-17 muboka reNPE akawedzera zvakanyanya (P <0.05). Kuenzaniswa neboka rekushanda kwesham, mazinga eprotein eIL-4 uye IL-17 muboka reME akatadzawo kusvika pakuchinja kwakakosha (P> 0.05).
(A) Mapuroteni mazinga eIL-4 uye IL-17 muboka reNPE akanga akakwirira zvikuru kupfuura avo vari muboka reME neboka re placebo (P <0.05). (B) Western blot histogram.
Nekuda kwehuwandu hushoma hwemasamples evanhu akawanikwa panguva yekuvhiyiwa, zvakajeka uye zvakadzama zvidzidzo zvepathogenesis yeMC zvakati kuoma. Takaedza kumisikidza mhando yemhuka yeMC kuti tidzidze maitiro ayo anogona kuitika. Panguva imwecheteyo, kuongororwa kweradiological, histological evaluation uye molecular biological evaluation yakashandiswa kutevera nzira yeMC yakakonzerwa neNP autograft. Nekuda kweizvozvo, iyo NP yekumisikidza modhi yakakonzera shanduko zvishoma nezvishoma mukusimba kwechiratidzo kubva ku12-vhiki kusvika ku20-vhiki-yenguva mapoinzi (yakasanganiswa yakaderera chiratidzo muT1W kutevedzana uye yakaderera chiratidzo muT2W sequences), zvichiratidza shanduko yetishu, uye histological uye molecular. ongororo yebhayoloji yakasimbisa mhedzisiro yeongororo yeradiological.
Mhedzisiro yekuyedza iyi inoratidza kuti shanduko yekuona uye histological yakaitika panzvimbo yekukanganisa kwemuviri wevertebral muboka reNPE. Panguva imwecheteyo, kutaura kweIL-4, IL-17 uye IFN-γ majini, pamwe neIL-4, IL-17 uye IFN-γ zvakaonekwa, zvichiratidza kuti kukanganisa kwe autologous nucleus pulposus tissue mu vertebral. muviri unogona kukonzera nhevedzano yechiratidzo uye morphological shanduko. Zviri nyore kuwana kuti zviratidzo zvezviratidzo zvemuviri we vertebral wemuenzaniso wemhuka (chiratidzo chakaderera mumutsara weT1W, chiratidzo chakavhenganiswa uye chiratidzo chakaderera mumutsara weT2W) zvakafanana zvikuru neaya emasero emusana wemunhu, uye maitiro eMRI zvakare. simbisa kucherechedzwa kwehistology uye gross anatomy, ndiko kuti, shanduko mumasero emuviri wevertebral inofambira mberi. Kunyange zvazvo mhinduro yekuputika kunokonzerwa nekushungurudzika kwakanyanya kunogona kuonekwa nokukurumidza mushure mekuputika, migumisiro yeMRI yakaratidza kuti kuwedzera zvishoma nezvishoma kuchinja kwechiratidzo kwakaonekwa mavhiki e12 mushure mekuputika uye kwakaramba kusvika kumavhiki e20 pasina zviratidzo zvekupora kana kuchinjwa kweMRI kuchinja. Mhedzisiro iyi inoratidza kuti autologous vertebral NP inzira yakavimbika yekumisikidza inofambira mberi MV mutsuro.
Iyi yekubaya modhi inoda hunyanzvi hwakakwana, nguva, uye kushanda nesimba. Mukuedza kwekutanga, kuparadzanisa kana kunyanya kukurudzira kweparavertebral ligamentous structures kunogona kukonzera kuumbwa kwe vertebral osteophytes. Kuchengetedza kunofanira kutorwa kuti kusakuvadza kana kutsamwisa madhisiki ari pedyo. Sezvo kudzika kwekupinda kunofanirwa kudzorwa kuti tiwane mibairo inowirirana uye inoberekana, isu nemaoko takagadzira plug nekucheka sheath ye3 mm kureba tsono. Kushandisa iyi plug kunovimbisa yunifomu yekuchera kudzika mukati me vertebral body. Mukuedza kwekutanga, vanachiremba vatatu vemapfupa vakabatanidzwa mukuvhiya vakawana 16-gauge tsono dziri nyore kushanda nadzo pane 18-gauge tsono kana dzimwe nzira. Kuti udzivise kubuda ropa kwakanyanya panguva yekuchera, kubata tsono ichiri kwechinguva kunopa gomba rekupinza rakakodzera, zvichiratidza kuti imwe dhigirii yeMC inogona kudzorwa nenzira iyi.
Kunyangwe zvidzidzo zvakawanda zvakanangana neMC, zvishoma zvinozivikanwa nezve etiology uye pathogenesis yeMC25,26,27. Zvichienderana nezvidzidzo zvedu zvakapfuura, takaona kuti autoimmunity ine basa rakakosha mukuitika nekuvandudzwa kweMC12. Ichi chidzidzo chakaongorora huwandu hwekutaura kweIL-4, IL-17, uye IFN-γ, iyo ndiyo nzira huru yekusiyanisa yeCD4 + masero mushure mekukurudzira antigen. Muchidzidzo chedu, tichienzaniswa neboka rakashata, boka reNPE raive nepamusoro kutaura kweIL-4, IL-17, uye IFN-γ, uye mazinga eprotein e IL-4 uye IL-17 akanga akakwirirawo.
Clinically, IL-17 mRNA kutaura inowedzerwa muNP masero kubva kuvarwere vane disc herniation28. Kuwedzera IL-4 uye IFN-γ mazinga ekutaura akawanikwawo mune acute non-compressive disc herniation modhi kana ichienzaniswa neane hutano kutonga29. IL-17 inobata basa rinokosha mukuputika, kukuvadzwa kwetishu mu autoimmune zvirwere30 uye inowedzera simba rekudzivirira zvirwere kune IFN-γ31. Kuwedzeredzwa IL-17-mediated tishu kukuvara kwave kuchishumwa muMRL/lpr mice32 uye autoimmunity-inobatwa mice33. IL-4 inogona kudzivisa kutaura kweproinflammatory cytokines (yakadai seIL-1β uye TNFα) uye macrophage activation34. Yakanzi mRNA kutaura kweIL-4 kwakanga kwakasiyana muboka reNPE kana ichienzaniswa neIL-17 uye IFN-γ panguva imwe chete; Iyo mRNA kutaura kweIFN-γ muboka reNPE yaive yakanyanya kukwirira kupfuura mune mamwe mapoka. Nokudaro, IFN-γ kugadzirwa kunogona kuva murevereri wemhinduro yekuputika inokonzerwa neNP intercalation. Zvidzidzo zvakaratidza kuti IFN-γ inobudiswa nemhando dzakasiyana-siyana dzemasero, kusanganisira activated type 1 mubatsiri T masero, masero emuurayi echisikigo, uye macrophages35,36, uye inokosha pro-inflammatory cytokine inokurudzira mhinduro dzemuviri37.
Ichi chidzidzo chinoratidza kuti autoimmune mhinduro inogona kuve nechekuita mukuitika uye kukura kweMC. Luoma et al. akawana kuti zviratidzo zve MC uye NP yakakurumbira zvakafanana paMRI, uye zvose zvinoratidza chiratidzo chepamusoro muT2W sequence38. Mamwe ma cytokines akasimbiswa kuve akabatana zvakanyanya nekuitika kweMC, senge IL-139. Ma et al. yakaratidza kuti kukwira kumusoro kana kudzika kweNP kunogona kuva nesimba guru pakuitika nekuvandudzwa kweMC12. Bobechko40 naHerzbein et al.41 vakashuma kuti NP is immunotolerant tissue isingagoni kupinda muvascular circulation kubva pakuberekwa. NP protrusions inopinza miviri yekunze muropa, nekudaro kuyananidza maitiro emuno autoimmune42. Autoimmune reactions inogona kukonzera huwandu hukuru hwezvidziviriro zvemuviri, uye kana zvinhu izvi zvichiramba zvichiratidzwa kune tishu, zvinogona kukonzera shanduko mukusaina43. Muchidzidzo ichi, kuwandisa kweIL-4, IL-17 uye IFN-γ ndiwo maitiro ekudzivirira zvirwere, zvichiwedzera kuratidza hukama hwepedyo pakati peNP neMCs44. Iyi modhi yemhuka inotevedzera zvakanaka NP budiriro uye kupinda mundiro yekupedzisira. Iyi nzira yakawedzera kuratidza kukanganisa kwe autoimmunity paMC.
Sezvinotarisirwa, iyi modhi yemhuka inotipa chikuva chinobvira chekudzidza MC. Nekudaro, iyi modhi ichine zvimwe zvinogumira: chekutanga, panguva yekutarisa mhuka, dzimwe tsuro dzepakati-kati dzinoda kusimbiswa pakuongorora histological uye molecular biology, saka dzimwe mhuka "dzinobva dzarega kushandiswa" nekufamba kwenguva. Chechipiri, kunyange zvazvo mapoinzi matatu akaiswa muchidzidzo ichi, zvinosuwisa, takangotevedzera rudzi rumwe chete rweMC (Modic type I change), saka hazvina kukwana kumiririra chirongwa chekukura kwechirwere chevanhu, uye nguva yakawanda inoda kuiswa zviri nani cherechedza shanduko dzese dzechiratidzo. Chechitatu, shanduko muchimiro chetishu inogona kunyatso kuratidzwa nehistological staining, asi mamwe hunyanzvi matekiniki anogona kuratidza zvirinani shanduko ye microstructural mune iyi modhi. Semuenzaniso, polarized light microscopy yakashandiswa kuongorora kuumbwa kwefibrocartilage mune rabbit intervertebral discs45. Iyo yakareba-yakareba mhedzisiro yeNP paMC uye endplate inoda kumwe kudzidza.
Makumi mashanu mana mana eNew Zealand tsuro chena (huremu hunenge 2.5-3 kg, makore 3-3.5 mwedzi) dzakakamurwa zvisina tsarukano kuita sham oparesheni boka, tsandanyama kuisirwa boka (ME boka) uye tsinga midzi yekudyara boka (NPE boka). Maitiro ese ekuedzwa akatenderwa neEthics Committee yeTianjin Hospital, uye nzira dzekuyedza dzakaitwa zvakanyatsoenderana neyakabvumidzwa nhungamiro.
Kumwe kuvandudzwa kwakaitwa kune nzira yekuvhiya yeS. Sobajima 46. Tsuro imwe neimwe yakaiswa mu lateral recumbency position uye anterior pamusoro pezvishanu zvakatevedzana lumbar intervertebral discs (IVDs) yakaratidzwa kushandisa posterolateral retroperitoneal approach. Tsuro imwe neimwe yakapihwa general anesthesia (20% urethane, 5 ml/kg kuburikidza netsinga yenzeve). A longitudinal skin incision yakagadzirwa kubva kumucheto kwezasi kwembabvu kusvika kumucheto wepelvic, 2 cm ventral kune paravertebral tsandanyama. Iyo yakarurama anterolateral spine kubva kuL1 kusvika kuL6 yakaratidzwa nekupinza uye kusina kuchena kwechidimbu chepamusoro-soro chemukati, retroperitoneal tissue, uye mhasuru (Fig. 6A). Iyo disc level yakatemwa kushandisa pelvic brim seye anatomical landmark yeL5-L6 disc level. Shandisa 16-gauge puncture tsono kuti uboora gomba pedyo neplate yekupedzisira yeL5 vertebra kusvika pakadzika 3 mm (Fig. 6B). Shandisa 5-ml sirinji kuti ushuve autologous nucleus pulposus muL1-L2 intervertebral disc (Fig. 6C). Bvisa nucleus pulposus kana tsandanyama zvichienderana nezvinodiwa neboka rega rega. Mushure mekunge gomba rekubooresa radzika, sutures inonyura inoiswa pane yakadzika fascia, superficial fascia uye ganda, uchichenjerera kuti usakuvadza periosteal tissue ye vertebral body panguva yekuvhiyiwa.
(A) Iyo L5-L6 disc inoratidzwa kuburikidza ne posterolateral retroperitoneal approach. (B) Shandisa 16-gauge tsono kuchera gomba pedyo neL5 endplate. (C) Autologous MFs inokohwa.
General anesthesia yaishandiswa ne20% urethane (5 ml / kg) inoshandiswa kuburikidza nenzeve yenzeve, uye lumbar spine radiographs yakadzokororwa pa12, 16, uye mavhiki e20 mushure mekushanda.
Tsuro dzakabayirwa nejekiseni intramuscular ye ketamine (25.0 mg / kg) uye intravenous sodium pentobarbital (1.2 g / kg) pa12, 16 uye mavhiki makumi maviri mushure mekuvhiyiwa. Muzongoza wose wakabviswa kuti histological analysis uye kunyatsoongorora kwakaitwa. Quantitative reverse transcription (RT-qPCR) uye Western blotting yakashandiswa kuona shanduko mune immune factor.
Kuongororwa kweMRI kwakaitwa mutsuro vachishandisa 3.0 T yekiriniki magineti (GE Medical Systems, Florence, SC) yakashongedzerwa ne orthogonal lemb coil receiver. Tsuro dzakaitwa anesthetized ne 20% urethane (5 mL/kg) kuburikidza netsinga yenzeve ndokubva yaiswa supine mukati memagineti nenharaunda yelumbar iri pakati pe5-inch diameter circular surface coil (GE Medical Systems). Coronal T2-inorema localizer mifananidzo (TR, 1445 ms; TE, 37 ms) yakawanikwa kutsanangura nzvimbo ye lumbar disc kubva kuL3-L4 kusvika kuL5-L6. Sagittal plane T2-weighted slices yakawanikwa nemaitiro anotevera: fast spin-echo sequence nekudzokorora nguva (TR) ye2200 ms uye nguva echo (TE) ye70 ms, matrix; nzvimbo yekuona ye 260 uye zvisere zvinokurudzira; Ukobvu hwekucheka hwaive 2 mm, gap raive 0.2 mm.
Mushure mokunge mufananidzo wekupedzisira watorwa uye tsuro yekupedzisira yakaurayiwa, sham, mhasuru-yakaiswa mukati, uye NP discs zvakabviswa kuti histological kuongororwa. Mishonga yakagadziriswa mu 10% neutral buffered formalin kwevhiki ye1, yakasvibiswa ne ethylenediaminetetraacetic acid, uye parafini yakakamurwa. Matishu akaiswa muparafini uye akachekwa kuita sagittal zvikamu (5 μm gobvu) uchishandisa microtome. Zvikamu zvakasvibiswa ne hematoxylin uye eosin (H&E).
Mushure mekuunganidza intervertebral discs kubva kutsuro muboka rega rega, RNA yakazara yakatorwa uchishandisa UNIQ-10 column (Shanghai Sangon Biotechnology Co., Ltd., China) maererano nemirairo yemugadziri uye ImProm II reverse transcription system (Promega Inc. , Madison, WI, USA). Reverse transcription yakaitwa.
RT-qPCR yakaitwa uchishandisa Prism 7300 (Applied Biosystems Inc., USA) uye SYBR Green Jump Start Taq ReadyMix (Sigma-Aldrich, St. Louis, MO, USA) maererano nemirairo yemugadziri. Iyo PCR reaction vhoriyamu yaive 20 μl uye yaive ne1.5 μl yecDNA yakanyungudutswa uye 0.2 μM yega yega yekutanga. Zvinyorwa zvakagadzirirwa naOligoPerfect Designer (Invitrogen, Valencia, CA) uye yakagadzirwa naNanjing Golden Stewart Biotechnology Co., Ltd. (China) (Tafura 1). Mamiriro ezvinhu anotevera ekutenderera kwemabhasikoro akashandiswa: yekutanga polymerase activation step pa 94 ° C ye 2 min, ipapo 40 cycles ye 15 s imwe neimwe pa 94 ° C ye template denaturation, annealing ye 1 min pa 60 ° C, kuwedzera, uye fluorescence. kuyerwa kwakaitwa kweminiti 1 pa72°C. Ese masampuli akakwidziridzwa katatu uye avhareji kukosha kwakashandiswa RT-qPCR kuongororwa. Data yeAmplification yakaongororwa uchishandisa FlexStation 3 (Molecular Devices, Sunnyvale, CA, USA). IL-4, IL-17, uye IFN-γ gene expression yakajairwa kune endogenous control (ACTB). Relative expression levels of target mRNA akaverengwa achishandisa 2-ΔΔCT nzira.
Yese mapuroteni akatorwa kubva mumatishu achishandisa tishu homogenizer muRIPA lysis buffer (ine protease uye phosphatase inhibitor cocktail) uye ipapo centrifuged pa13,000 rpm kwemaminetsi makumi maviri pa4 ° C kuti abvise tsvina yematishu. Makumi mashanu micrograms eprotein akatakurwa munzira, akaparadzaniswa ne10% SDS-PAGE, uye ndokuzoendeswa kune PVDF membrane. Kuvharira kwakaitwa mu5% nonfat yakaoma mukaka muTris-buffered saline (TBS) ine 0.1% Pakati pe20 ye1 h pakamuri tembiricha. Iyo membrane yakavharwa netsuro anti-decorin primary antibody (diluted 1:200; Boster, Wuhan, China) (diluted 1:200; Bioss, Beijing, China) usiku hwose pa 4 ° C uye yakaita pamazuva echipiri; ine masoja ekudzivirira chirwere chechipiri (mbudzi inorwisa tsuro immunoglobulin G pa1:40,000 dilution) yakasanganiswa nehorseradish peroxidase (Boster, Wuhan, China) kweawa imwe patembiricha yemumba. Western blot zviratidzo zvakaonekwa nekuwedzera chemiluminescence pane chemiluminescent membrane mushure meX-ray irradiation. Pakuongorora densitometric, mablots akaongororwa uye akaverengerwa pachishandiswa BandScan software uye mhedzisiro yacho yakaratidzirwa sereshiyo yechinangwa chegene immunoreactivity kune tubulin immunoreactivity.
Masvomhu akaitwa pachishandiswa SPSS16.0 software package (SPSS, USA). Dhiyabhorosi yakaunganidzwa panguva yekudzidza yakaratidzwa seyanoreva ± chiyero chekutsauka (kureva ± SD) uye yakaongororwa uchishandisa nzira imwe chete yakadzokororwa matanho ekuongororwa kwekusiyana (ANOVA) kuona kusiyana pakati pemapoka maviri. P <0.05 yainzi statistically yakakosha.
Nokudaro, kugadzwa kwemuenzaniso wemhuka weMC nekuisa autologous NPs mumuviri we vertebral uye kuita macroanatomical observation, MRI kuongorora, histological evaluation uye molecular biological analysis inogona kuva chinhu chinokosha chekuongorora nekunzwisisa nzira dzeMC dzevanhu nekugadzira nzira itsva yekurapa. kupindira.
Matauriro echinyorwa ichi: Han, C. et al. Mhuka yemuenzaniso weModic shanduko yakagadzwa nekuisa autologous nucleus pulposus mu subchondral bone ye lumbar spine. Sci. Rep. 6, 35102: 10.1038/srep35102 (2016).
Weishaupt, D., Zanetti, M., Hodler, J., uye Boos, N. Magnetic resonance imaging ye lumbar spine: kuwanda kwe disc herniation uye kuchengetwa, nerve root compression, end plate abnormalities, uye facet joint osteoarthritis mune vanozvipira vasina zviratidzo. . rate. Radiology 209, 661-666, doi: 10.1148/radiology.209.3.9844656 (1998).
Kjaer, P., Korsholm, L., Bendix, T., Sorensen, JS, uye Leboeuf-Eed, K. Modic shanduko uye hukama hwavo kune zvakawanikwa mumakiriniki. European Spine Journal: kubudiswa kwepamutemo kweEuropean Spine Society, European Society of Spinal Deformity, uye European Society yeCervical Spine Research 15, 1312-1319, doi: 10.1007 / s00586-006-0185-x (2006).
Kuisma, M., nevamwe. Modic inoshanduka mu lumbar vertebral endplates: kuwanda uye kushamwaridzana nekurwadziwa kwepashure uye sciatica muvashandi vechirume vezera repakati. Spine 32, 1116–1122, doi:10.1097/01.brs.0000261561.12944.ff (2007).
de Roos, A., Kressel, H., Spritzer, K., uye Dalinka, M. MRI yemapfupa emapfupa anoshanduka pedyo neplate yekupedzisira muchirwere chinopera che lumbar spine. AJR. American Journal yeRadiology 149, 531-534, doi: 10.2214 / ajr.149.3.531 (1987).
Modic, MT, Steinberg, PM, Ross, JS, Masaryk, TJ, uye Carter, JR Degenerative disc chirwere: kuongororwa kwe vertebral marrow kuchinja neMRI. Radiology 166, 193-199, doi: 10.1148/radiology.166.1.3336678 (1988).
Modic, MT, Masaryk, TJ, Ross, JS, uye Carter, JR Imaging ye degenerative disc chirwere. Radiology 168, 177-186, doi: 10.1148/radiology.168.1.3289089 (1988).
Jensen, TS, nevamwe. Predictors of neovertebral endplate (Modic) chiratidzo chekuchinja muhuwandu hwevanhu. European Spine Journal: Official Publication yeEuropean Spine Society, European Society of Spinal Deformity, uye European Society yeCervical Spine Research, Division 19, 129-135, doi: 10.1007 / s00586-009-1184-5 (2010).
Albert, HB naMannisch, K. Modic anochinja mushure me lumbar disc herniation. European Spine Journal : Official Publication yeEuropean Spine Society, European Society of Spinal Deformity uye European Society yeCervical Spine Research 16, 977-982, doi: 10.1007 / s00586-007-0336-8 (2007).
Kerttula, L., Luoma, K., Vehmas, T., Gronblad, M., uye Kaapa, E. Modic type I shanduko inogona kufanotaura nekukurumidza kufambira mberi deformational disc degeneration: 1-gore rinotarisira kudzidza. European Spine Journal 21, 1135-1142, doi: 10.1007/s00586-012-2147-9 (2012).
Hu, ZJ, Zhao, FD, Fang, XQ uye Fan, SW Modic shanduko: zvinogona zvikonzero uye mupiro kune lumbar disc degeneration. Medical Hypotheses 73, 930–932, doi: 10.1016/j.mehy.2009.06.038 (2009).
Krok, HV Internal disc rupture. Disc prolapse matambudziko pamusoro pemakore makumi mashanu. Muzongoza (Phila Pa 1976) 11, 650–653 (1986).
Nguva yekutumira: Zvita-13-2024