【摘要】：[目的]觀察以新型絲素蛋白多孔支架復(fù)合兔髓核細(xì)胞體外構(gòu)建組織工程化髓核的可行性。[方法]分離培養(yǎng)兔髓核細(xì)胞,與絲素蛋白多孔支架在體外復(fù)合培養(yǎng),建立組織工程化髓核模型,通過(guò)掃描電鏡、HE染色、甲苯胺藍(lán)染色、Ⅱ型膠原免疫組化以及酶聯(lián)免疫吸附測(cè)定觀察細(xì)胞在支架上1周和3周的生長(zhǎng)及增殖情況。[結(jié)果]培養(yǎng)1周后,掃描電鏡顯示細(xì)胞呈球狀均勻地貼附在支架內(nèi)部。細(xì)胞-支架復(fù)合體HE染色可見(jiàn)支架內(nèi)部有大量髓核細(xì)胞,甲苯胺蘭染色陽(yáng)性,Ⅱ型膠原免疫組化染色陽(yáng)性。培養(yǎng)3周后,掃描電鏡顯示細(xì)胞成層黏附于支架表面,細(xì)胞重疊生長(zhǎng),分泌大量細(xì)胞外基質(zhì),HE染色可見(jiàn)支架內(nèi)部有大量髓核細(xì)胞填滿(mǎn)支架孔隙并分泌大量細(xì)胞外基質(zhì),甲苯胺蘭染色陽(yáng)性,Ⅱ型膠原免疫組化染色陽(yáng)性。酶聯(lián)免疫吸附測(cè)定:3周組蛋白多糖含量(52.4±4.5)ng/ml明顯高于1周組(29.3±3.6)ng/ml,P0.05,3周組的II型膠原含量(24.3±1.8)ng/ml明顯高于1周組(15.16±1.5)ng/ml,P0.05。[結(jié)論]新型絲素蛋白多孔支架復(fù)合兔髓核細(xì)胞體外培養(yǎng)生長(zhǎng)良好,分泌大量類(lèi)似髓核樣細(xì)胞外基質(zhì),可以用于體外構(gòu)建組織工程化髓核。
[Abstract]:[objective] to observe the feasibility of constructing tissue engineered nucleus pulposus with a new fibroin porous scaffold combined with rabbit nucleus pulposus cells in vitro. [methods] the rabbit nucleus pulposus cells were isolated and cultured in vitro. The model of tissue engineered nucleus pulposus was established by co-culture with silk fibroin porous scaffold. Scanning electron microscopy, HE staining and toluidine blue staining were used. The growth and proliferation of type 鈪，

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