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ʻO ke kūkulu ʻana i nā hiʻohiʻona holoholona o ka hoʻololi modic (MC) he kumu koʻikoʻi no ke aʻo ʻana iā MC. Ua māhele ʻia he kanalimakumamāhā mau lāpaki keʻokeʻo o Aotearoa i ka hui sham-operation, ka hui hoʻokomo ʻiʻo (ME group) a me ka hui hoʻokomo nucleus pulposus (NPE group). I loko o ka hui NPE, ua hōʻike ʻia ka disc intervertebral ma o ke ʻano ʻokiʻoki lumbar anterolateral a ua hoʻohana ʻia kahi nila e hou i ke kino vertebral L5 kokoke i ka papa hope. Ua unuhi ʻia ʻo NP mai ka disc intervertebral L1/2 e kahi syringe a ua hoʻokomo ʻia i loko. Ke ʻeli ʻana i kahi lua i loko o ka iwi subchondral. ʻO nā kaʻina hana ʻokiʻoki a me nā ʻano ʻeli ʻana i ka hui hoʻokomo ʻiʻo a me ka hui sham-operation ua like ia me nā mea i loko o ka hui hoʻokomo NP. I loko o ka hui ME, ua kau ʻia kahi ʻāpana ʻiʻo i loko o ka lua, ʻoiai i loko o ka hui sham-operation, ʻaʻohe mea i kau ʻia i loko o ka lua. Ma hope o ke ʻoki ʻana, ua hana ʻia ka scan MRI a me ka hoʻāʻo ʻana i ka biology molecular. Ua loli ka hōʻailona ma ka hui NPE, akā ʻaʻohe loli hōʻailona maopopo i ka hui sham-operation a me ka hui ME. Ua hōʻike ka nānā ʻana o Histological ua ʻike ʻia ka ulu ʻana o nā ʻiʻo maʻamau ʻole ma kahi i hoʻokomo ʻia ai, a ua hoʻonui ʻia ka hōʻike ʻana o IL-4, IL-17 a me IFN-γ i ka hui NPE. ʻO ka hoʻokomo ʻana o NP i loko o ka iwi subchondral hiki ke hana i kahi kumu hoʻohālike holoholona o MC.
ʻO nā loli Modic (MC) he mau ʻeha o nā papa hope vertebral a me ka iwi iwi e pili ana i ʻike ʻia ma ke kiʻi magnetic resonance imaging (MRI). He mea maʻamau lākou i nā poʻe me nā hōʻailona pili1. Ua hoʻokūpaʻa nā haʻawina he nui i ke koʻikoʻi o MC ma muli o kona pilina me ka ʻeha hope haʻahaʻa (LBP)2,3. Ua wehewehe kūʻokoʻa ʻo de Roos et al.4 lāua ʻo Modic et al.5 i ʻekolu mau ʻano like ʻole o nā ʻano hōʻailona subchondral i loko o ka iwi iwi vertebral. ʻO nā loli ʻano Modic I he hypointense ma nā kaʻina T1-weighted (T1W) a me hyperintense ma nā kaʻina T2-weighted (T2W). Hōʻike kēia ʻeha i nā papa hope fissure a me nā ʻiʻo granulation vascular e pili ana i loko o ka iwi iwi. Hōʻike nā loli ʻano Modic II i ka hōʻailona kiʻekiʻe ma nā kaʻina T1W a me T2W. Ma kēia ʻano lesion, hiki ke loaʻa ka luku ʻana o ka endplate, a me ka hoʻololi momona histological o ka iwi iwi e pili ana. Hōʻike nā loli ʻano Modic III i ka hōʻailona haʻahaʻa ma nā kaʻina T1W a me T2W. Ua ʻike ʻia nā ʻeha sclerotic e pili ana i nā endplates6. Ua manaʻo ʻia ʻo MC he maʻi pathological o ka iwi kuamoʻo a pili loa ia me nā maʻi degenerative he nui o ka iwi kuamoʻo7,8,9.
Ke noʻonoʻo nei i ka ʻikepili i loaʻa, ua hāʻawi kekahi mau haʻawina i nā ʻike kikoʻī i ke kumu a me nā ʻano pathological o MC. Ua hōʻike ʻo Albert et al. ua hiki ke hoʻokumu ʻia ʻo MC e ka herniation disc8. Ua hoʻopili ʻo Hu et al. iā MC i ka degeneration disc koʻikoʻi10. Ua hāpai ʻo Kroc i ke kumumanaʻo o ka "rupture disc kūloko," e ʻōlelo ana e hiki i ka trauma disc repetitive ke alakaʻi i nā microtears ma ka endplate. Ma hope o ka hoʻokumu ʻia ʻana o ka cleft, ʻo ka luku ʻia ʻana o ka endplate e ka nucleus pulposus (NP) ke hoʻoulu i kahi pane autoimmune, kahi e alakaʻi hou aku ai i ka hoʻomohala ʻana o MC11. Ua kaʻana like ʻo Ma et al. i kahi manaʻo like a ua hōʻike ʻo NP-induced autoimmunity i kahi kuleana koʻikoʻi i ka pathogenesis o MC12.
ʻO nā pūnaewele ʻōnaehana pale, ʻoiai nā lymphocytes kōkua CD4+ T, he kuleana koʻikoʻi ko lākou i ka pathogenesis o ka autoimmunity13. ʻO ka subset Th17 i ʻike hou ʻia e hana i ka proinflammatory cytokine IL-17, paipai i ka hōʻike ʻana o ka chemokine, a hoʻoulu i nā pūnaewele T i loko o nā ʻōpū i hōʻino ʻia e hana i ka IFN-γ14. He kuleana kūikawā ko nā pūnaewele Th2 i ka pathogenesis o nā pane pale. ʻO ka hōʻike ʻana o IL-4 ma ke ʻano he cell Th2 koho e hiki ke alakaʻi i nā hopena immunopathological koʻikoʻi15.
ʻOiai ua nui nā haʻawina lapaʻau i hana ʻia ma MC16,17,18,19,20,21,22,23,24, aia nō ka nele o nā hiʻohiʻona hoʻokolohua holoholona kūpono e hiki ke hoʻohālike i ke kaʻina hana MC e hana pinepine ʻia i loko o ke kanaka a hiki ke hoʻohana ʻia e noiʻi i ke kumu a i ʻole nā lāʻau lapaʻau hou e like me ka lāʻau lapaʻau i kuhikuhi ʻia. A hiki i kēia lā, ua hōʻike ʻia kekahi mau hiʻohiʻona holoholona o MC e aʻo ana i nā ʻano pathological ma lalo.
Ma muli o ke kumumanaʻo autoimmune i hāpai ʻia e Albert lāua ʻo Ma, ua hoʻokumu kēia haʻawina i kahi kumu hoʻohālike MC rabbit maʻalahi a hiki ke hana hou ʻia ma o ka hoʻoili pono ʻana iā NP kokoke i ka papa hope vertebral i wili ʻia. ʻO nā pahuhopu ʻē aʻe e nānā i nā ʻano histological o nā hiʻohiʻona holoholona a loiloi i nā ʻano hana kikoʻī o NP i ka hoʻomohala ʻana o MC. No kēia hopena, hoʻohana mākou i nā ʻano hana e like me ka biology molecular, MRI, a me nā haʻawina histological e aʻo ai i ka holomua o MC.
Ua make ʻelua mau lāpaki i ke kahe ʻana o ke koko i ka wā o ke ʻoki ʻana, a ua make ʻehā mau lāpaki i ka wā o ka anesthesia i ka wā o ka MRI. Ua ola nā lāpaki he 48 i koe a ʻaʻole i hōʻike i nā hōʻailona o ke ʻano a i ʻole ka neurological ma hope o ke ʻoki ʻana.
Hōʻike ka MRI he ʻokoʻa ka ikaika o ka hōʻailona o ke kino i hoʻokomo ʻia i loko o nā lua like ʻole. Ua loli mālie ka ikaika o ka hōʻailona o ke kino vertebral L5 i ka hui NPE ma 12, 16 a me 20 mau pule ma hope o ke kau ʻana (ua hōʻike ka moʻo T1W i ka hōʻailona haʻahaʻa, a ua hōʻike ka moʻo T2W i ka hōʻailona hui pū ʻia me ka hōʻailona haʻahaʻa) (Kiʻi 1C), ʻoiai ua noho paʻa nā hiʻohiʻona MRI o nā hui ʻelua ʻē aʻe o nā ʻāpana i hoʻokomo ʻia i ka manawa like (Kiʻi 1A, B).
(A) Nā MRI kaʻina hana o ka iwi kuamoʻo lumbar rabbit ma 3 mau manawa. ʻAʻohe ʻano ʻē o nā hōʻailona i loaʻa i nā kiʻi o ka hui sham-operation. (B) Ua like nā ʻano hōʻailona o ke kino vertebral i ka hui ME me nā mea i loko o ka hui sham-operation, a ʻaʻohe loli hōʻailona koʻikoʻi i ʻike ʻia ma kahi hoʻokomo i ka hala ʻana o ka manawa. (C) I loko o ka hui NPE, ʻike maopopo ʻia ka hōʻailona haʻahaʻa ma ke kaʻina T1W, a ʻike maopopo ʻia ka hōʻailona hui a me ka hōʻailona haʻahaʻa ma ke kaʻina T2W. Mai ka wā 12-pule a hiki i ka wā 20-pule, emi nā hōʻailona kiʻekiʻe sporadic e hoʻopuni ana i nā hōʻailona haʻahaʻa ma ke kaʻina T2W.
Hiki ke ʻike ʻia ka hyperplasia iwi maopopo ma kahi i hoʻokomo ʻia ai ke kino vertebral i loko o ka hui NPE, a ʻoi aku ka wikiwiki o ka hyperplasia iwi mai 12 a 20 mau pule (Kiʻi 2C) i hoʻohālikelike ʻia me ka hui NPE, ʻaʻohe loli koʻikoʻi i ʻike ʻia i nā kino vertebral i hoʻohālike ʻia; hui Sham a me ka hui ME (Kiʻi 2C) 2A,B).
(A) He laumania loa ka ʻili o ke kino vertebral ma ka ʻāpana i hoʻokomo ʻia, hoʻōla maikaʻi ka lua, a ʻaʻohe hyperplasia i loko o ke kino vertebral. (B) Ua like ke ʻano o ka wahi i hoʻokomo ʻia i loko o ka hui ME me ko ka hui hana hoʻopunipuni, a ʻaʻohe loli maopopo i ke ʻano o ka wahi i hoʻokomo ʻia i ka hala ʻana o ka manawa. (C) Ua hana ʻia ka hyperplasia iwi ma kahi i hoʻokomo ʻia i loko o ka hui NPE. Ua piʻi koke ka hyperplasia iwi a ua hoʻolōʻihi ʻia ma o ka disc intervertebral i ke kino vertebral contralateral.
Hāʻawi ka loiloi histological i ka ʻike kikoʻī e pili ana i ka hoʻokumu ʻana o ka iwi. Hōʻike ka Kiʻi 3 i nā kiʻi o nā ʻāpana postoperative i hoʻoluʻu ʻia me H&E. I loko o ka hui sham-operation, ua hoʻonohonoho maikaʻi ʻia nā chondrocytes a ʻaʻohe ʻike ʻia o ka hoʻonui ʻana o nā cell (Kiʻi 3A). Ua like ke kūlana ma ka hui ME me ko ka hui sham-operation (Kiʻi 3B). Eia naʻe, i loko o ka hui NPE, ua ʻike ʻia kahi nui o nā chondrocytes a me ka hoʻonui ʻana o nā cell NP-like ma kahi i hoʻokomo ʻia ai (Kiʻi 3C);
(A) Hiki ke ʻike ʻia nā trabeculae kokoke i ka papa hope, ua hoʻonohonoho pono ʻia nā chondrocytes me ka nui like a me ke ʻano o nā cell a ʻaʻohe hoʻonui ʻana (40 mau manawa). (B) Ua like ke kūlana o ke kahua hoʻokomo i loko o ka hui ME me ko ka hui hoʻopunipuni. Hiki ke ʻike ʻia nā trabeculae a me nā chondrocytes, akā ʻaʻohe hoʻonui maopopo ma ke kahua hoʻokomo (40 mau manawa). (B) Hiki ke ʻike ʻia e hoʻonui nui ʻia nā chondrocytes a me nā cell NP-like, a ʻaʻole like ke ʻano a me ka nui o nā chondrocytes (40 mau manawa).
Ua ʻike ʻia ka hōʻike ʻana o ka interleukin 4 (IL-4) mRNA, interleukin 17 (IL-17) mRNA, a me ka interferon γ (IFN-γ) mRNA ma nā hui NPE a me ME. I ka hoʻohālikelike ʻia ʻana o nā pae hōʻike o nā genes i manaʻo ʻia, ua hoʻonui nui ʻia nā hōʻike gene o IL-4, IL-17, a me IFN-γ ma ka hui NPE i hoʻohālikelike ʻia me nā mea o ka hui ME a me ka hui hana hoʻopunipuni (Kiʻi 4) (P < 0.05). I ka hoʻohālikelike ʻia me ka hui hana hoʻopunipuni, ua piʻi iki wale nā pae hōʻike o IL-4, IL-17, a me IFN-γ ma ka hui ME a ʻaʻole i hiki i ka loli helu (P > 0.05).
Ua hōʻike ka hōʻike ʻana o ka mRNA o IL-4, IL-17 a me IFN-γ i ka hui NPE i kahi ʻano kiʻekiʻe loa ma mua o nā mea i loko o ka hui hana hoʻopunipuni a me ka hui ME (P < 0.05).
I ka hoʻohālikelike ʻana, ʻaʻole i hōʻike ʻia nā pae hōʻike ma ka hui ME i kahi ʻokoʻa koʻikoʻi (P>0.05).
Ua hana ʻia ka loiloi Western blot me ka hoʻohana ʻana i nā antibodies i loaʻa ma ke kālepa e kūʻē iā IL-4 a me IL-17 e hōʻoia i ke ʻano hōʻike mRNA i hoʻololi ʻia. E like me ka mea i hōʻike ʻia ma nā Kiʻi 5A, B, i hoʻohālikelike ʻia me ka hui ME a me ka hui hana hoʻopunipuni, ua hoʻonui nui ʻia nā pae protein o IL-4 a me IL-17 i ka hui NPE (P < 0.05). I ka hoʻohālikelike ʻia me ka hui hana hoʻopunipuni, ʻaʻole i hiki i nā pae protein o IL-4 a me IL-17 i ka hui ME ke hoʻololi i nā loli koʻikoʻi (P> 0.05).
(A) ʻOi aku ke kiʻekiʻe o nā pae protein o IL-4 a me IL-17 i ka hui NPE ma mua o nā mea i loko o ka hui ME a me ka hui placebo (P < 0.05). (B) Histogram blot Komohana.
Ma muli o ka palena o nā hāpana kanaka i loaʻa i ka wā o ke ʻoki kino, he mea paʻakikī iki nā haʻawina maopopo a kikoʻī e pili ana i ka pathogenesis o MC. Ua hoʻāʻo mākou e hoʻokumu i kahi kumu hoʻohālike holoholona o MC e aʻo i kāna mau ʻano pathological hiki. I ka manawa like, ua hoʻohana ʻia ka loiloi radiological, ka loiloi histological a me ka loiloi biological molecular e hahai i ke kaʻina o MC i hoʻokumu ʻia e ka NP autograft. ʻO ka hopena, ua hopena ke kumu hoʻohālike NP implantation i kahi loli mālie i ka ikaika hōʻailona mai nā manawa 12-pule a i 20-pule (hōʻailona haʻahaʻa i hui pū ʻia ma nā kaʻina T1W a me ka hōʻailona haʻahaʻa ma nā kaʻina T2W), e hōʻike ana i nā loli o nā ʻiʻo, a ua hōʻoia nā loiloi histological a me nā molecular biological i nā hopena o ke aʻo radiological.
Hōʻike nā hopena o kēia hoʻokolohua ua loaʻa nā loli ʻike maka a me nā histological ma kahi o ka hoʻopiʻi kino vertebral i ka hui NPE. I ka manawa like, ua ʻike ʻia ka hōʻike ʻana o nā genes IL-4, IL-17 a me IFN-γ, a me IL-4, IL-17 a me IFN-γ, e hōʻike ana i ka hoʻopiʻi ʻana o ka ʻiʻo autologous nucleus pulposus i loko o ke kino vertebral e hiki ke hana i kahi moʻo o nā loli hōʻailona a me ke ʻano. He mea maʻalahi ke ʻike ʻana he like loa nā ʻano hōʻailona o nā kino vertebral o ke kumu hoʻohālike holoholona (hōʻailona haʻahaʻa ma ke kaʻina T1W, hōʻailona hui ʻia a me ka hōʻailona haʻahaʻa ma ke kaʻina T2W) me nā cell vertebral kanaka, a hōʻoia pū nā ʻano MRI i nā nānā ʻana o ka histology a me ka anatomy gross, ʻo ia hoʻi, he holomua nā loli i nā cell kino vertebral. ʻOiai ʻo ka pane mumū i hoʻokumu ʻia e ka trauma acute e ʻike koke ʻia ma hope o ka puncture, ua hōʻike nā hopena MRI ua ʻike ʻia nā loli hōʻailona e piʻi mau ana i 12 mau pule ma hope o ka puncture a hoʻomau a hiki i 20 mau pule me ka ʻole o nā hōʻailona o ka hoʻōla ʻana a i ʻole ka hoʻohuli ʻana o nā loli MRI. Hōʻike kēia mau hopena he ala hilinaʻi ka autologous vertebral NP no ka hoʻokumu ʻana i ka MV holomua i nā lāpaki.
Pono kēia kumu hoʻohālike puncture i ke akamai kūpono, ka manawa, a me ka hoʻoikaika ʻoki kino. I nā hoʻokolohua mua, ʻo ka dissection a i ʻole ka hoʻoulu nui ʻana o nā ʻano ligamentous paravertebral e hopena i ka hoʻokumu ʻia ʻana o nā osteophytes vertebral. Pono e mālama pono ʻaʻole e hōʻino a hoʻonāukiuki paha i nā disc e pili ana. ʻOiai pono e hoʻomalu ʻia ka hohonu o ke komo ʻana e loaʻa ai nā hopena kūlike a hiki ke hana hou ʻia, ua hana lima mākou i kahi plug ma ka ʻoki ʻana i ka ʻūhā o kahi nila 3 mm ka lōʻihi. ʻO ka hoʻohana ʻana i kēia plug e hōʻoiaʻiʻo i ka hohonu o ka wili like ʻana i loko o ke kino vertebral. I nā hoʻokolohua mua, ʻekolu mau kauka orthopedic i komo i ka hana i ʻike he maʻalahi ka hana ʻana me nā nila 16-gauge ma mua o nā nila 18-gauge a i ʻole nā ʻano hana ʻē aʻe. I mea e pale aku ai i ke kahe nui ʻana o ke koko i ka wā wili, ʻo ka paʻa ʻana i ka nila no kekahi manawa e hāʻawi i kahi lua hoʻokomo kūpono, e hōʻike ana e hiki ke hoʻomalu ʻia kekahi kekelē o MC ma kēia ʻano.
ʻOiai ua kālele ʻia ka MC e nā haʻawina he nui, ʻaʻole i ʻike nui ʻia e pili ana i ke kumu a me ka pathogenesis o MC25,26,27. Ma muli o kā mākou mau haʻawina ma mua, ua ʻike mākou he kuleana koʻikoʻi ka autoimmunity i ka hanana a me ka hoʻomohala ʻana o MC12. Ua nānā kēia haʻawina i ka hōʻike quantitative o IL-4, IL-17, a me IFN-γ, ʻo ia nā ala hoʻokaʻawale nui o nā cell CD4+ ma hope o ka hoʻoulu ʻana o ka antigen. I kā mākou haʻawina, i hoʻohālikelike ʻia me ka hui maikaʻi ʻole, ʻoi aku ka kiʻekiʻe o ka hōʻike ʻana o ka hui NPE o IL-4, IL-17, a me IFN-γ, a ʻoi aku hoʻi ke kiʻekiʻe o nā pae protein o IL-4 a me IL-17.
Ma ke ʻano lapaʻau, ua hoʻonui ʻia ka hōʻike ʻana o ka IL-17 mRNA i loko o nā pūnaewele NP mai nā maʻi me ka herniation disc28. Ua ʻike ʻia hoʻi ka piʻi ʻana o nā pae hōʻike IL-4 a me IFN-γ i loko o kahi kumu hoʻohālike herniation disc non-compressive acute i hoʻohālikelike ʻia me nā kaohi olakino29. He kuleana koʻikoʻi ko IL-17 i ka mumū, ka ʻeha o nā ʻiʻo i nā maʻi autoimmune30 a hoʻonui i ka pane pale i ka IFN-γ31. Ua hōʻike ʻia ka ʻeha o nā ʻiʻo i hoʻonui ʻia e IL-17 i nā ʻiole MRL/lpr32 a me nā ʻiole autoimmunity-susceptible33. Hiki i ka IL-4 ke kāohi i ka hōʻike ʻana o nā cytokines proinflammatory (e like me IL-1β a me TNFα) a me ka hoʻoulu ʻana o ka macrophage34. Ua hōʻike ʻia ua ʻokoʻa ka hōʻike ʻana o ka mRNA o IL-4 i ka hui NPE i hoʻohālikelike ʻia me IL-17 a me IFN-γ i ka manawa like; ʻOi aku ka kiʻekiʻe o ka hōʻike ʻana o ka mRNA o IFN-γ i ka hui NPE ma mua o nā hui ʻē aʻe. No laila, ʻo ka hana ʻana o IFN-γ ke lilo i mea waena o ka pane mumū i hoʻokumu ʻia e ka intercalation NP. Ua hōʻike ʻia nā haʻawina ua hana ʻia ʻo IFN-γ e nā ʻano cell helper like ʻole, me nā cell T helper type 1 i hoʻāla ʻia, nā cell pepehi kanaka kūlohelohe, a me nā macrophages35,36, a he cytokine pro-inflammatory koʻikoʻi ia e paipai ana i nā pane pale37.
Hōʻike kēia haʻawina e pili ana paha ka pane autoimmune i ka hanana a me ka hoʻomohala ʻana o MC. Ua ʻike ʻo Luoma et al. ua like nā ʻano hōʻailona o MC a me NP koʻikoʻi ma MRI, a hōʻike lāua ʻelua i ka hōʻailona kiʻekiʻe ma ka moʻo T2W38. Ua hōʻoia ʻia kekahi mau cytokines e pili pono ana me ka hanana o MC, e like me IL-139. Ua hōʻike ʻo Ma et al. ʻo ka puka ʻana i luna a i lalo paha o NP he mana nui paha i ka hanana a me ka hoʻomohala ʻana o MC12. Ua hōʻike ʻo Bobechko40 lāua ʻo Herzbein et al.41 he ʻiʻo immunotolerant ʻo NP ʻaʻole hiki ke komo i ke kahe koko mai ka hānau ʻana. Hoʻokomo nā puka NP i nā kino haole i loko o ke koko, no laila e hoʻoponopono ana i nā hopena autoimmune kūloko42. Hiki i nā hopena Autoimmune ke hoʻoulu i ka nui o nā mea pale, a i ka wā e hōʻike mau ʻia ai kēia mau mea i nā ʻiʻo, hiki iā lākou ke hana i nā loli i ka hōʻailona43. Ma kēia haʻawina, ʻo ka overexpression o IL-4, IL-17 a me IFN-γ nā mea pale maʻamau, e hōʻoia hou ana i ka pilina pili ma waena o NP a me MCs44. Hoʻohālike maikaʻi kēia kumu hoʻohālike holoholona i ka holomua o NP a me ke komo ʻana i ka papa hope. Ua hōʻike hou kēia kaʻina hana i ka hopena o ka autoimmunity ma MC.
E like me ka mea i manaʻo ʻia, hāʻawi kēia kumu hoʻohālike holoholona iā mākou i kahi kahua hiki ke aʻo i ka MC. Eia nō naʻe, aia nō kekahi mau palena o kēia kumu hoʻohālike: ʻo ka mea mua, i ka wā o ka nānā ʻana i nā holoholona, pono e pepehi ʻia kekahi mau lāpaki waena no ka hoʻāʻo ʻana i ka biology histological a me ka molecular, no laila "hāʻule kekahi mau holoholona mai ka hoʻohana ʻana" i ka hala ʻana o ka manawa. ʻO ka lua, ʻoiai ua hoʻonohonoho ʻia ʻekolu mau manawa i loko o kēia haʻawina, ʻo ka mea pōʻino, ua hoʻohālike wale mākou i hoʻokahi ʻano MC (Modic type I change), no laila ʻaʻole lawa ia e hōʻike i ke kaʻina hana hoʻomohala maʻi kanaka, a pono e hoʻonohonoho ʻia nā manawa hou aʻe e nānā pono i nā loli hōʻailona āpau. ʻO ke kolu, hiki ke hōʻike maopopo ʻia nā loli i ke ʻano o nā ʻiʻo e ka hoʻoluʻu histological, akā hiki i kekahi mau ʻano hana kūikawā ke hōʻike maikaʻi i nā loli microstructural i loko o kēia kumu hoʻohālike. No ka laʻana, ua hoʻohana ʻia ka microscopy māmā polarized e kālailai i ka hoʻokumu ʻia ʻana o ka fibrocartilage i loko o nā disc intervertebral rabbit45. Pono nā hopena lōʻihi o NP ma MC a me ka endplate i ka noiʻi hou aku.
Ua hoʻokaʻawale ʻia he kanalimakumamāhā mau lāpaki keʻokeʻo kāne o Aotearoa (ma kahi o 2.5-3 kg ke kaumaha, makahiki 3-3.5 mahina) i loko o ka hui hana hoʻopunipuni, ka hui hoʻokomo ʻiʻo (hui ME) a me ka hui hoʻokomo aʻa nerve (hui NPE). Ua ʻāpono ʻia nā kaʻina hana hoʻokolohua a pau e ke Kōmike Ethics o ka Halemai ʻo Tianjin, a ua hoʻokō ʻia nā ʻano hana hoʻokolohua e like me nā alakaʻi i ʻāpono ʻia.
Ua hana ʻia kekahi mau hoʻomaikaʻi ʻana i ke ʻano hana ʻoki kino o S. Sobajima 46. Ua kau ʻia kēlā me kēia lāpaki ma ke kūlana moe ʻaoʻao a ua hōʻike ʻia ka ʻili mua o ʻelima mau disc intervertebral lumbar (IVD) me ka hoʻohana ʻana i kahi ala posterolateral retroperitoneal. Ua hāʻawi ʻia kēlā me kēia lāpaki i ka anesthesia laulā (20% urethane, 5 ml/kg ma o ke aʻa pepeiao). Ua hana ʻia kahi ʻoki ʻili longitudinal mai ka lihi haʻahaʻa o nā iwi ʻaoʻao a i ka pelvic brim, 2 cm ventral a i nā ʻiʻo paravertebral. Ua hōʻike ʻia ka iwi kuamoʻo anterolateral ʻākau mai L1 a i L6 ma o ka ʻoki ʻoi a me ka blunt o ka ʻiʻo subcutaneous ma luna, ka ʻiʻo retroperitoneal, a me nā ʻiʻo (Kiʻi 6A). Ua hoʻoholo ʻia ka pae disc me ka hoʻohana ʻana i ka pelvic brim ma ke ʻano he hōʻailona anatomical no ka pae disc L5-L6. E hoʻohana i kahi nila puncture 16-gauge e wili i kahi lua kokoke i ka papa hopena o ka vertebra L5 a hiki i ka hohonu o 3 mm (Kiʻi 6B). E hoʻohana i kahi syringe 5-ml e omo i ka autologous nucleus pulposus i loko o ka L1-L2 intervertebral disc (Kiʻi 6C). Wehe i ka nucleus pulposus a i ʻole ka ʻiʻo e like me nā koi o kēlā me kēia hui. Ma hope o ka hohonu ʻana o ka lua wili, kau ʻia nā sutures absorbable ma ka fascia hohonu, ka fascia superficial a me ka ʻili, e mālama pono ʻaʻole e hōʻino i ka ʻiʻo periosteal o ke kino vertebral i ka wā o ke ʻoki ʻana.
(A) Hōʻike ʻia ka disc L5–L6 ma o kahi ala posterolateral retroperitoneal. (B) E hoʻohana i kahi nila 16-gauge e wili i kahi lua kokoke i ka L5 endplate. (C) ʻOhi ʻia nā Autologous MFs.
Ua hāʻawi ʻia ka anesthesia laulā me 20% urethane (5 ml/kg) i hāʻawi ʻia ma o ke aʻa pepeiao, a ua hana hou ʻia nā radiographs lumbar spine ma 12, 16, a me 20 mau pule ma hope o ke ʻoki ʻana.
Ua pepehi ʻia nā lāpaki ma o ka hoʻokomo ʻana i ke ketamine intramuscular (25.0 mg/kg) a me ka sodium pentobarbital intravenous (1.2 g/kg) i 12, 16 a me 20 mau pule ma hope o ke ʻoki ʻana. Ua wehe ʻia ka iwi kuamoʻo holoʻokoʻa no ka nānā ʻana i ka histological a ua hana ʻia ka nānā maoli. Ua hoʻohana ʻia ka quantitative reverse transcription (RT-qPCR) a me Western blotting e ʻike i nā loli i nā mea pale.
Ua hana ʻia nā hoʻokolohua MRI i nā lāpaki me ka hoʻohana ʻana i kahi magnet lapaʻau 3.0 T (GE Medical Systems, Florence, SC) i lako me kahi mea hoʻokipa coil lālā orthogonal. Ua hoʻomaʻemaʻe ʻia nā lāpaki me 20% urethane (5 mL/kg) ma o ke aʻa pepeiao a laila kau ʻia ma luna o ka magnet me ka ʻāpana lumbar i kau ʻia ma luna o kahi coil ʻili poepoe 5-ʻīniha ke anawaena (GE Medical Systems). Ua loaʻa nā kiʻi localizer Coronal T2-weighted (TR, 1445 ms; TE, 37 ms) e wehewehe i kahi o ka disc lumbar mai L3-L4 a i L5-L6. Ua loaʻa nā ʻāpana Sagittal plane T2-weighted me nā hoʻonohonoho aʻe: kaʻina spin-echo wikiwiki me ka manawa hana hou (TR) o 2200 ms a me ka manawa echo (TE) o 70 ms, matrix; kahua ʻike o 260 a me ʻewalu stimuli; ʻO ka mānoanoa o ka ʻoki ʻana he 2 mm, ʻo ka hakahaka he 0.2 mm.
Ma hope o ka lawe ʻia ʻana o ke kiʻi hope loa a me ka pepehi ʻia ʻana o ka lāpaki hope loa, ua wehe ʻia ka sham, nā ʻiʻo i hoʻokomo ʻia, a me nā disc NP no ka hoʻokolokolo histological. Ua hoʻopaʻa ʻia nā ʻiʻo i loko o ka formalin buffered 10% no 1 pule, decalcified me ka ethylenediaminetetraacetic acid, a ua ʻoki ʻia ka paraffin. Ua hoʻokomo ʻia nā poloka ʻiʻo i loko o ka paraffin a ʻoki ʻia i nā ʻāpana sagittal (5 μm ka mānoanoa) me ka hoʻohana ʻana i ka microtome. Ua hoʻoluʻu ʻia nā ʻāpana me ka hematoxylin a me ka eosin (H&E).
Ma hope o ka hōʻiliʻili ʻana i nā disc intervertebral mai nā lāpaki i kēlā me kēia hui, ua unuhi ʻia ka RNA holoʻokoʻa me ka hoʻohana ʻana i kahi kolamu UNIQ-10 (Shanghai Sangon Biotechnology Co., Ltd., Kina) e like me nā kuhikuhi a ka mea hana a me kahi ʻōnaehana hoʻololi hope ImProm II (Promega Inc., Madison, WI, USA). Ua hana ʻia ke hoʻololi hope.
Ua hana ʻia ʻo RT-qPCR me ka hoʻohana ʻana i kahi Prism 7300 (Applied Biosystems Inc., USA) a me SYBR Green Jump Start Taq ReadyMix (Sigma-Aldrich, St. Louis, MO, USA) e like me nā kuhikuhi a ka mea hana. ʻO ka nui o ka hopena PCR he 20 μl a loaʻa iā ia he 1.5 μl o ka cDNA i hoʻoheheʻe ʻia a me 0.2 μM o kēlā me kēia primer. Ua hoʻolālā ʻia nā Primers e OligoPerfect Designer (Invitrogen, Valencia, CA) a hana ʻia e Nanjing Golden Stewart Biotechnology Co., Ltd. (Kina) (Papa 1). Ua hoʻohana ʻia nā kūlana paikikala wela ma lalo nei: ka hana hoʻāla polymerase mua ma 94°C no 2 min, a laila 40 mau pōʻaiapuni o 15 s kēlā me kēia ma 94°C no ka denaturation template, annealing no 1 min ma 60°C, extension, a me fluorescence. ua hana ʻia nā ana no 1 min ma 72°C. Ua hoʻonui ʻia nā laʻana a pau i ʻekolu manawa a ua hoʻohana ʻia ka waiwai awelika no ka loiloi RT-qPCR. Ua kālailai ʻia nā ʻikepili hoʻonui me ka hoʻohana ʻana iā FlexStation 3 (Molecular Devices, Sunnyvale, CA, USA). Ua hoʻonohonoho ʻia ka hōʻike ʻana o IL-4, IL-17, a me IFN-γ i ka mana endogenous (ACTB). Ua helu ʻia nā pae hōʻike pili o ka mRNA target me ka hoʻohana ʻana i ke ʻano 2-ΔΔCT.
Ua unuhi ʻia ka protein holoʻokoʻa mai nā ʻiʻo me ka hoʻohana ʻana i kahi homogenizer ʻiʻo i loko o ka RIPA lysis buffer (loaʻa kahi cocktail inhibitor protease a me phosphatase) a laila centrifuged ma 13,000 rpm no 20 min ma 4°C e wehe i nā ʻōpala ʻiʻo. Ua hoʻouka ʻia he kanalima micrograms o ka protein no kēlā me kēia ala, i hoʻokaʻawale ʻia e 10% SDS-PAGE, a laila hoʻoili ʻia i kahi membrane PVDF. Ua hana ʻia ka pale ʻana i loko o ka waiū maloʻo nonfat 5% i loko o ka saline Tris-buffered (TBS) e loaʻa ana he 0.1% Tween 20 no 1 h ma ka mahana o ka lumi. Ua hoʻoulu ʻia ka membrane me ka rabbit anti-decorin primary antibody (diluted 1:200; Boster, Wuhan, Kina) (diluted 1:200; Bioss, Beijing, Kina) i ka pō ma 4°C a ua pane i nā lā ʻelua; me ka antibody lua (kao anti-rabbit immunoglobulin G ma 1:40,000 dilution) i hui pū ʻia me ka horseradish peroxidase (Boster, Wuhan, Kina) no 1 hola ma ka mahana o ka lumi. Ua ʻike ʻia nā hōʻailona Western blot e ka hoʻonui ʻia ʻana o ka chemiluminescence ma ka membrane chemiluminescent ma hope o ka irradiation X-ray. No ka nānā ʻana o ka densitometric, ua scan ʻia a helu ʻia nā blots me ka hoʻohana ʻana i ka polokalamu BandScan a ua hōʻike ʻia nā hopena ma ke ʻano he ratio o ka immunoreactivity gene target i ka immunoreactivity tubulin.
Ua hana ʻia nā helu helu me ka hoʻohana ʻana i ka pūʻolo polokalamu SPSS16.0 (SPSS, USA). Ua hōʻike ʻia nā ʻikepili i hōʻiliʻili ʻia i ka wā o ke aʻo ʻana ma ke ʻano he awelika ± ʻokoʻa maʻamau (awelika ± SD) a ua kālailai ʻia me ka hoʻohana ʻana i ka loiloi anakahi i hana hou ʻia o hoʻokahi ala o ka variance (ANOVA) e hoʻoholo i nā ʻokoʻa ma waena o nā hui ʻelua. Ua manaʻo ʻia ʻo P < 0.05 he koʻikoʻi ka helu.
No laila, ʻo ka hoʻokumu ʻana i kahi kumu hoʻohālike holoholona o MC ma o ke kau ʻana i nā NP autologous i loko o ke kino vertebral a me ka hana ʻana i ka nānā ʻana macroanatomical, ka nānā ʻana o MRI, ka loiloi histological a me ka nānā ʻana i ka biology molecular e lilo i mea hana koʻikoʻi no ka loiloi ʻana a me ka hoʻomaopopo ʻana i nā ʻano hana o ka MC kanaka a me ka hoʻomohala ʻana i nā hana therapeutic hou.
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