【摘要】： 目的 采用手術(shù)方法建立大鼠膝關(guān)節(jié)骨軟骨缺損及表面軟骨缺損動(dòng)物模型,通過檢測(cè)膠原表達(dá)變化探討不同關(guān)節(jié)軟骨缺損自我修復(fù)的能力,通過檢測(cè)基質(zhì)金屬蛋白酶(matrix metalloproteinase,MMP)在損后傷關(guān)節(jié)軟骨組織中的表達(dá)變化,探討損傷導(dǎo)致關(guān)節(jié)軟骨退變的病理學(xué)機(jī)制。 方法 取雌性SD大鼠48只,隨機(jī)分成A、B、C三組,A組18只,在雙膝關(guān)節(jié)股骨髕面制作骨軟骨缺損直徑3.0mm,深度3.0mm,以缺損底部出現(xiàn)滲血為宜;B組18只,在雙膝關(guān)節(jié)股骨髁間制作表面軟骨缺損寬度0.3mm,深度0.3mm,缺損底部無滲血,不穿透軟骨下骨板;C組12只,雙膝關(guān)節(jié)只做關(guān)節(jié)囊切開,不做任何處置直接縫合,為假手術(shù)對(duì)照組。分別于術(shù)后4、8、12W三個(gè)時(shí)間點(diǎn)取材,實(shí)驗(yàn)組每個(gè)時(shí)間點(diǎn)處死6只大鼠,對(duì)照組每個(gè)時(shí)間點(diǎn)處死4只大鼠。取雙膝關(guān)節(jié)原切口,充分顯露股骨遠(yuǎn)端關(guān)節(jié)面,于股骨髁上垂直于骨干截下股骨遠(yuǎn)端。將標(biāo)本放置于固定脫鈣后制作石蠟切片,行HE、甲苯胺蘭染色、Ⅰ型膠原、Ⅱ型膠原、MMP-2、MMP-3、MMP-9免疫組化染色。 結(jié)果 A組術(shù)后4W缺損中有少量新生組織生成,8W及12W可見到纖維組織填充,三個(gè)時(shí)間點(diǎn)修復(fù)組織細(xì)胞外基質(zhì)Ⅰ型膠原免疫組化染色陽(yáng)性,Ⅱ型膠原免疫組化染色陰性,關(guān)節(jié)軟骨組織中MMP-2、MMP-3、MMP-9表達(dá)增高(與C組比較有顯著性差異,P＜0.05)。B組表面軟骨缺損4W及8W未見修復(fù)跡象,12W可見微量纖維組織填充,其細(xì)胞外基質(zhì)Ⅰ型膠原免疫組化染色陽(yáng)性,Ⅱ型膠原免疫組化染色陰性,B組關(guān)節(jié)軟骨組織術(shù)后三個(gè)時(shí)間點(diǎn)MMP-2、MMP-3、MMP-9表達(dá)均增高(與C組比較有顯著性差異,P＜0.05)。C組三個(gè)時(shí)間點(diǎn)均為正常關(guān)節(jié)軟骨組織,Ⅰ型膠原免疫組化染色陰性,Ⅱ型膠原免疫組化染色陽(yáng)性,MMP-2、MMP-3、MMP-9低度表達(dá),無形態(tài)學(xué)異常改變。 結(jié)論 大鼠骨軟骨缺損在間充質(zhì)細(xì)胞的參與下以纖維形式進(jìn)行自我修復(fù),修復(fù)組織無法恢復(fù)正常關(guān)節(jié)面的平整,是非透明軟骨組織。表面關(guān)節(jié)軟骨缺損由于未穿透軟骨下骨板,無間充質(zhì)細(xì)胞參與,很難實(shí)現(xiàn)自我修復(fù),隨著時(shí)間推移缺損周圍的軟骨變薄,出現(xiàn)不同程度的退變。通過手術(shù)方法可以在大鼠膝關(guān)節(jié)建立合理的骨軟骨缺損及表面軟骨缺損動(dòng)物模型,并可以用來考察移植方法修復(fù)軟骨缺損的療效。機(jī)械性損傷可以導(dǎo)致關(guān)節(jié)軟骨細(xì)胞外基質(zhì)成分發(fā)生改變,喪失其原有的生物學(xué)特性而退變,基質(zhì)降解素(MMP-3)和明膠酶(MMP-2、9)在損傷后的軟骨組織中表達(dá)增高,使細(xì)胞外基質(zhì)的降解增加,是導(dǎo)致關(guān)節(jié)軟骨退變的重要因素。
[Abstract]:Objective to establish an animal model of osteochondral defect and surface cartilage defect of knee joint in rats by surgical method, and to explore the ability of self-repair of different articular cartilage defects by detecting the changes of collagen expression. By detecting the expression of matrix metalloproteinase (matrix metalloproteinase,MMP) in articular cartilage after injury, the pathological mechanism of articular cartilage degeneration caused by injury was investigated. Methods Forty eight female SD rats were randomly divided into three groups: group A (n = 18), group B (n = 18). The surface cartilage defects were made in the femoral condyle of both knee joints with a width of 0.3 mm and a depth of 0.3 mm. There was no bleeding at the bottom of the defect and no penetration of the subchondral bone plate, while in group C, 12 knees were treated with joint capsule incision without any disposal and direct suture, which was used as the sham operation control group. 6 rats were killed at each time point in the experimental group and 4 rats in the control group were killed at each time point. The joint surface of the distal femur was fully exposed through the original incision of the knee joint, and was perpendicular to the shaft of the femoral condyle to amputate the distal femur. The specimens were fixed and decalcified to make paraffin sections. The specimens were stained with HE, toluidine blue, type 鈪，

本文編號(hào)：2278176