【摘要】：目的:觀察人體軟骨終板超微結(jié)構(gòu)及頸椎間盤退變的關(guān)系。方法:收集瀘州醫(yī)學(xué)院附屬第一醫(yī)院脊柱外科2013年6月至2015年4月診斷明確并行手術(shù)治療的頸椎病患者軟骨終板及椎間盤85例,同期收集外傷致頸椎骨折(非寰樞椎骨折,術(shù)前無頸椎病癥狀與體征)并行手術(shù)治療的軟骨終板及椎間盤12例作為對照。將頸椎病患者按年齡分組,A組為20~29歲,B組30~39歲,C組40~49歲,D組50~59歲,E組60~70歲,F組為對照組年齡28~46歲,每組包含4個(gè)病例。術(shù)前所有病例均行X線,核磁共振等常規(guī)檢查。術(shù)后觀察各組頸椎軟骨終板及椎間盤HE染色,根據(jù)Thompson椎間盤退變病理分級(jí)標(biāo)準(zhǔn)對其退變程度進(jìn)行分級(jí);Masson染色、透射電鏡照片觀察超微結(jié)構(gòu);Tunel免疫組化,觀察軟骨終板及椎間盤凋亡細(xì)胞。計(jì)量資料采用均數(shù)±標(biāo)準(zhǔn)差(x±s)表示。組間差異比較采用單因素方差分析,P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:HE染色A組,軟骨終板中陷窩細(xì)胞較多,亦可見清晰細(xì)胞核,細(xì)胞周圍纖維結(jié)構(gòu)緊密,鏡下可見少量玻璃樣變性;椎間盤中可見少量細(xì)胞結(jié)構(gòu),纖維組織排列緊密,無斷裂現(xiàn)象。B組,軟骨終板中陷窩細(xì)胞數(shù)量明顯減少,細(xì)胞中仍可見細(xì)胞核等結(jié)構(gòu),細(xì)胞周圍纖維組織排列雜亂;椎間盤中細(xì)胞較少,纖維組織排列較疏松。C組,軟骨終板中陷窩細(xì)胞結(jié)構(gòu)欠清晰,部分細(xì)胞中未見細(xì)胞核,大量細(xì)胞水腫,細(xì)胞周圍可見大量的玻璃樣變性;椎間盤中纖維大量斷裂,膠原纖維變稀疏、雜亂。D組,軟骨終板陷窩細(xì)胞結(jié)構(gòu)欠清晰,大量細(xì)胞核消失,出現(xiàn)空泡現(xiàn)象,細(xì)胞周圍結(jié)締組織大量玻璃樣變性;椎間盤可見大量斷裂的纖維,未見細(xì)胞。e組,軟骨終板中陷窩細(xì)胞明顯減少,細(xì)胞結(jié)構(gòu)不清晰,細(xì)胞周圍大量玻璃樣變性及粘液樣變性;椎間盤可見大量斷裂的膠原纖維,膠原纖維十分稀疏,軟骨終板內(nèi)甚至出現(xiàn)鈣化等一系列退變晚期的表現(xiàn)。而對照組(f組)軟骨終板中細(xì)胞數(shù)量較多,大部分胞核清晰可見,細(xì)胞周圍軟骨纖維排列緊密;椎間盤含大量緊密排列的膠原纖維,其中可見少量細(xì)胞,胞核清晰可見。thompson分級(jí),各組椎間盤及軟骨終板病理變化情況為a組i級(jí)8個(gè),Ⅱ級(jí)2個(gè),Ⅲ級(jí)2個(gè),Ⅳ級(jí)0個(gè),Ⅴ級(jí)0個(gè)。b組i級(jí)2個(gè),Ⅱ級(jí)4個(gè),Ⅲ級(jí)2個(gè),Ⅳ級(jí)4個(gè),Ⅴ級(jí)0個(gè)。c組i級(jí)1個(gè),Ⅱ級(jí)4個(gè),Ⅲ級(jí)4個(gè),Ⅳ級(jí)3個(gè),Ⅴ級(jí)0個(gè)。d組i級(jí)0個(gè),Ⅱ級(jí)1個(gè),Ⅲ級(jí)6個(gè),Ⅳ級(jí)5個(gè),Ⅴ級(jí)0個(gè)。e組i級(jí)0個(gè),Ⅱ級(jí)1個(gè),Ⅲ級(jí)4個(gè),Ⅳ級(jí)7個(gè),Ⅴ級(jí)0個(gè)。f組i級(jí)11個(gè),Ⅱ級(jí)1個(gè),Ⅲ級(jí)0個(gè),Ⅳ級(jí)0個(gè),Ⅴ級(jí)0個(gè)。masson染色見實(shí)驗(yàn)組切片含有大量淡藍(lán)色均一染色結(jié)構(gòu),考慮為退變后的淀粉樣變,部分膠原纖維排列紊亂松散;對照組masson染色見較多深藍(lán)色纖維組織,排列緊密,未見淀粉樣變。掃描電鏡見頸椎病患者軟骨終板細(xì)胞存在不同程度的退變,隨著年齡增加呈現(xiàn)出退變程度遞增的趨勢,部分細(xì)胞可見核變性、溶解,細(xì)胞質(zhì)內(nèi)出現(xiàn)大量的脂質(zhì)、空泡甚至鈣化,細(xì)胞器消失,細(xì)胞功能不完整;細(xì)胞周圍軟骨囊結(jié)構(gòu)紊亂,軟骨囊壁厚度不一;纖維組織亦可見清晰的纖維結(jié)構(gòu)被大量玻璃樣變填充,周期性橫紋減少,結(jié)構(gòu)欠佳。對照組的軟骨終板細(xì)胞,其細(xì)胞核圓滑完整,細(xì)胞質(zhì)均勻,內(nèi)質(zhì)網(wǎng)及線粒體等細(xì)胞器清晰可見,細(xì)胞功能良好;細(xì)胞周圍軟骨囊結(jié)構(gòu)完整,軟骨囊壁厚度均勻;軟骨終板細(xì)胞外基質(zhì)豐富,膠原纖維密集,有明顯的周期性橫紋。tunel免疫組化染色,計(jì)算凋亡率為:a組凋亡率21.5±2.55%,B組凋亡率42.1±2.57%,C組凋亡率47.5±4.68%,D組凋亡率67.4±4.52%,E組凋亡率78.6±4.91%;F組凋亡率15.2±3.12%。實(shí)驗(yàn)組凋亡率均不同程度高于對照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),且隨年齡呈遞增趨勢;其中每組內(nèi)上下終板凋亡率比較,上終板凋亡率雖然在數(shù)值上低于相應(yīng)階段下終板,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。病變節(jié)段椎間盤細(xì)胞凋亡率低于其相鄰上下軟骨終板細(xì)胞凋亡率(P0.05),且差異有統(tǒng)計(jì)學(xué)意義。結(jié)論:(1)頸椎病患者軟骨終板結(jié)構(gòu)的改變與頸椎間盤退變密切相關(guān),且隨年齡增加呈遞增趨勢。(2)頸椎軟骨終板退變的發(fā)生重于頸椎椎間盤退變,頸椎軟骨終板的退變可能是導(dǎo)致頸椎椎間盤退變的啟動(dòng)因素。
[Abstract]:Objective: To observe the relationship between the ultrastructure of cartilage end plate of human body and the degeneration of cervical intervertebral disc. Methods: From June 2013 to April 2015, we collected 85 cases of cervical spondylosis with cervical spondylosis treated by parallel operation from June 2013 to April 2015, and collected trauma-induced cervical fracture (non-atlas vertebral fracture) during the same period. There were 12 cases of cartilage endplates and 12 cases of intervertebral disc treated in parallel operation without symptoms and signs of cervical syndrome before operation. Patients with cervical spondylosis were divided into age group, group A was 20 ~ 29 years, group B was 30 ~ 39 years, group C was 40 ~ 49 years old, group D was 50 ~ 59 years, group E was 60 ~ 70 years old, and F group was 28 ~ 46 years old, including 4 cases in each group. All cases before operation were routine examination such as X-ray and nuclear magnetic resonance. After the operation, the cervical cartilage end plate and intervertebral disc HE staining were observed, and the degree of retrogradation was graded according to the pathological grading standard of Thompson's disc. The ultrastructure was observed by Masson staining and transmission electron microscope. Tunel immunohistochemistry was used to observe the cartilage end plate and intervertebral disc apoptosis cells. The measured data is represented by the standard deviation (x% s). A single factor of variance analysis was used to compare the differences among the groups, and the difference was statistically significant. Results: In the HE staining group A, the cells in the cartilage end plate were more and could also be seen in the clear nucleus, the fiber structure around the cells was compact, a small amount of glass-like degeneration can be seen under the endoscope, a small number of cell structures were seen in the intervertebral disc, and the fibrous tissue was arranged tightly and without fracture. In group B, the number of lacunae cells in the cartilage end plates was significantly reduced, and the cell nucleus and other structures were still visible in the cells, and there was a disordered arrangement of fibrous tissue around the cells, and the cells in the intervertebral disc were less and the fibrous tissue was more loose. In group C, the structure of lacunae cells in the cartilage end plates was not clear, no nuclei were found in some cells, a large number of cell edema was found, a large number of glass-like degeneration was found around the cells, and the fibers in the intervertebral disc were broken in large numbers, and the collagen fibers were sparse and disordered. In group D, the structure of the alveolar cell of the cartilage end plate is clear, a large number of nuclei disappear, a vacuole phenomenon occurs, a large amount of glass-like degeneration around the cell is observed, and a large number of broken fibers can be seen in the intervertebral disc, and the cells are not seen. In e group, the cells in the cartilage end plate were obviously reduced, the cell structure was not clear, a lot of glass-like degeneration and mucus-like degeneration around the cells were found, and a large number of broken collagen fibers were seen in the intervertebral disc, the collagen fibers were very sparse, and there was even calcification in the cartilage end plates. In contrast, the number of cells in the cartilage endplate of the control group (f group) is much larger, most of the nuclei are clearly visible, the pericellular cartilage fibers are arranged tightly, and a large number of closely-arranged collagen fibers are contained in the intervertebral disc, a small amount of cells can be seen, and the nuclei are clearly visible. Thelson classification, the pathological changes of intervertebral disc and cartilage endplate in each group were 8, 鈪，

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