CAJ | 학술논문

目的以骨基质明胶和软骨基质构建一体化纤维环-髓核双相支架,并检测其理化性能及细胞相容性.方法制备中空骨基质明胶环,并注入脱细胞软骨匀浆,经冷冻干燥、交联后制备成一体化纤维环-髓核双相支架.行Hoechst 33258、天狼星红、HE染色,扫描电镜观察支架内部结构,检测支架孔隙率和吸水率,检测双相支架复水后的力学性能.分离山羊纤维环和髓核细胞,接种至双相支架的相应部位,体外培养48 h,扫描电镜、活/死细胞染色评价支架与细胞的生物相容性.结果镜下Hoechst33258染色未见细胞残留,天狼星红染色阳性,HE染色示两部分结合紧密.扫描电镜可见支架呈多孔结构,孔隙相连通,纤维环相孔径为(401.4±13.1) μm,髓核相孔径为(112.4±21.8) μm.支架孔隙率为73.37％±2.56％,支架吸水率为655.7％±78.6％.支架压缩弹性模量为(49.06±15.57)kPa,小于正常椎间盘的(135.9±28.9)kPa,但在同一数量级.扫描电镜观察细胞黏附于支架表面,细胞周围有基质分泌,live/dead细胞染色示细胞在支架上活性良好.结论以天然骨基质明胶和软骨基质构建的一体化纤维环-髓核双相支架,无免疫原性,具有良好的孔径和孔隙率,支架两部分连接处结合紧密,在结构、生化成分及生物力学性能上与椎间盘组织相似,且具有良好的生物相容性.
목적 이골기질명효화연골기질구건일체화섬유배-수핵쌍상지가,병검측기이화성능급세포상용성.방법 제비중공골기질명효배,병주입탈세포연골균장,경냉동간조、교련후제비성일체화섬유배-수핵쌍상지가.행Hoechst 33258、천랑성홍、HE염색,소묘전경관찰지가내부결구,검측지가공극솔화흡수솔,검측쌍상지가복수후적역학성능.분리산양섬유배화수핵세포,접충지쌍상지가적상응부위,체외배양48 h,소묘전경、활/사세포염색평개지가여세포적생물상용성.결과 경하Hoechst33258염색미견세포잔류,천랑성홍염색양성,HE염색시량부분결합긴밀.소묘전경가견지가정다공결구,공극상련통,섬유배상공경위(401.4±13.1) μm,수핵상공경위(112.4±21.8) μm.지가공극솔위73.37％±2.56％,지가흡수솔위655.7％±78.6％.지가압축탄성모량위(49.06±15.57)kPa,소우정상추간반적(135.9±28.9)kPa,단재동일수량급.소묘전경관찰세포점부우지가표면,세포주위유기질분비,live/dead세포염색시세포재지가상활성량호.결론 이천연골기질명효화연골기질구건적일체화섬유배-수핵쌍상지가,무면역원성,구유량호적공경화공극솔,지가량부분련접처결합긴밀,재결구、생화성분급생물역학성능상여추간반조직상사,차구유량호적생물상용성.
Objective To fabricate an integrated annulus fibrosus-nucleus pulposus biphasic scaffolds based on bone matrix gelatin and acellular cartilage matrix,and to detect its property and cell compatibility.Methods An integrated annulus fibrosus-nucleus pulposus biphasic scaffold was fabricated by the following steps: preparing the hollow bone matrix gelatin ring,injecting the acellular cartilage homogenate into the center of the bone matrix gelatin ring,and freeze drying.Sample slices were stained with Hoechst 33258,picrosirius and HE.The internal structure of the scaffold was observed under a scanning electron microscope.The porosity and water absorption of the scaffold were also evaluated.Compressive mechanical property under wet situation was tested.The annulus fibrosus and nucleus pulposus cells were isolated from sheep disc and separately implanted into the corresponding sites of the scaffold,and biocompatibility of the scaffold was evaluated by scanning electron microscope and live/dead cell staining.Results Hoechst 33258 staining showed no residual cells,picrosirius staining was positive,and HE staining showed two parts linked closely.Under scanning electron microscope,the scaffold had porous structure,and the average pore size was 401.4±13.1 μm for annulus fibrosus,and 112.4±21.8 μm for nucleus pulposus.The porosity and water absorption of the scaffold was 73.37％±2.56％ and 655.7％±78.6％,respectively.The average compressive elastic modulus of the scaffold (49.06 ±15.57) kpa was smaller than that of the native disc (135.9±28.9) kPa.Scanning electron microscope showed cells adhered on the scaffold surface,with secreted matrix around them,and live/dead cells staining showed cells with good activity on scaffolds.Conclusion The integrated annulus fibrosus-nucleus pulposus scaffold based on bone matrix gelatin and cartilage matrix is an ideal artificial disc material,in view of well pore size,closely linked boundary,and good biocompatibility.