【摘要】：目的探討新型絲素蛋白多孔支架復(fù)合經(jīng)PKH26標(biāo)記的兔髓核細(xì)胞初步構(gòu)建組織工程髓核的可行性。方法消化分離兔髓核細(xì)胞,培養(yǎng)獲取P1代細(xì)胞,對P1代髓核細(xì)胞行番紅O以及Ⅱ型膠原免疫組織化學(xué)染色;PKH26標(biāo)記兔髓核細(xì)胞,MTT法檢測標(biāo)記前后髓核細(xì)胞增殖情況,將標(biāo)記后的細(xì)胞接種支架,體外培養(yǎng)4 d后,將細(xì)胞-支架復(fù)合物植入裸鼠皮下,體內(nèi)培養(yǎng)12周后,進(jìn)行活體熒光成像技術(shù)檢測、HE染色、甲苯胺藍(lán)染色、番紅O染色和Ⅱ型膠原免疫組化染色。結(jié)果 P1代髓核細(xì)胞番紅O染色陽性,Ⅱ型膠原免疫組織化學(xué)染色陽性;標(biāo)記后細(xì)胞熒光強(qiáng)度分布均勻,標(biāo)記前后髓核細(xì)胞光密度(OD)值比較差異無統(tǒng)計(jì)學(xué)意義(P0.05);活體成像技術(shù)顯示裸鼠皮下植入的細(xì)胞-支架復(fù)合體呈現(xiàn)強(qiáng)烈熒光;HE染色見多孔支架內(nèi)壁有大量髓核細(xì)胞填滿并分泌大量細(xì)胞外基質(zhì);甲苯胺藍(lán)染色、番紅O染色及Ⅱ型膠原免疫組化染色均呈陽性,細(xì)胞周圍大量細(xì)胞外基質(zhì)分泌。結(jié)論以新型絲素蛋白多孔支架復(fù)合兔髓核細(xì)胞經(jīng)體內(nèi)培養(yǎng)形成的類髓核樣組織,可用于組織工程化髓核的構(gòu)建。
[Abstract]:Objective to investigate the feasibility of constructing tissue engineered nucleus pulposus with fibroin porous scaffold combined with PKH26 labeled rabbit nucleus pulposus cells. Methods the rabbit nucleus pulposus cells were digested and isolated, and P1 passage cells were cultured and stained with phenanthin O and type 鈪，

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