实用医学杂志
實用醫學雜誌
실용의학잡지
THE JOURNAL OF PRACTICAL MEDICINE
2014年
7期
1041-1044
,共4页
支架%髓核%脱细胞%形态学%细胞毒性
支架%髓覈%脫細胞%形態學%細胞毒性
지가%수핵%탈세포%형태학%세포독성
Scaffold%Nucleus pulposus%Decellularization%Morphology%Cytotoxicity
目的：运用去污剂及核酸酶法制备兔脱细胞髓核支架，体外评价该支架的形态学及细胞毒性改变，探讨其作为组织工程髓核支架的可行性。方法：取健康成年新西兰大白兔髓核组织50枚，随机分为新鲜对照组（n＝25）和脱细胞组（n＝25），后者用0．3％TritonX-100＋0．5％脱氧胆酸钠联合核酸酶进行脱细胞处理24 h ，通过肉眼观、病理染色及扫描电镜观察髓核形态改变、细胞脱除和细胞外基质变化情况，并取兔骨髓间充质干细胞为对象，运用CCK-8法及死活细胞染色法在体外评价该支架的细胞毒性。结果：髓核脱细胞支架形态保持良好，细胞清除彻底，细胞外胶原基质结构破坏较小，与正常髓核相比，支架组织及其浸提液对兔骨髓间充质干细胞无明显毒副作用。结论：TritonX-100与脱氧胆酸钠联合核酸酶可有效脱除髓核细胞成分，较好地保留细胞外基质结构，且该支架对骨髓间充质干细胞无明显毒副作用，可以作为髓核组织工程研究的理想材料。
目的：運用去汙劑及覈痠酶法製備兔脫細胞髓覈支架，體外評價該支架的形態學及細胞毒性改變，探討其作為組織工程髓覈支架的可行性。方法：取健康成年新西蘭大白兔髓覈組織50枚，隨機分為新鮮對照組（n＝25）和脫細胞組（n＝25），後者用0．3％TritonX-100＋0．5％脫氧膽痠鈉聯閤覈痠酶進行脫細胞處理24 h ，通過肉眼觀、病理染色及掃描電鏡觀察髓覈形態改變、細胞脫除和細胞外基質變化情況，併取兔骨髓間充質榦細胞為對象，運用CCK-8法及死活細胞染色法在體外評價該支架的細胞毒性。結果：髓覈脫細胞支架形態保持良好，細胞清除徹底，細胞外膠原基質結構破壞較小，與正常髓覈相比，支架組織及其浸提液對兔骨髓間充質榦細胞無明顯毒副作用。結論：TritonX-100與脫氧膽痠鈉聯閤覈痠酶可有效脫除髓覈細胞成分，較好地保留細胞外基質結構，且該支架對骨髓間充質榦細胞無明顯毒副作用，可以作為髓覈組織工程研究的理想材料。
목적：운용거오제급핵산매법제비토탈세포수핵지가，체외평개해지가적형태학급세포독성개변，탐토기작위조직공정수핵지가적가행성。방법：취건강성년신서란대백토수핵조직50매，수궤분위신선대조조（n＝25）화탈세포조（n＝25），후자용0．3％TritonX-100＋0．5％탈양담산납연합핵산매진행탈세포처리24 h ，통과육안관、병리염색급소묘전경관찰수핵형태개변、세포탈제화세포외기질변화정황，병취토골수간충질간세포위대상，운용CCK-8법급사활세포염색법재체외평개해지가적세포독성。결과：수핵탈세포지가형태보지량호，세포청제철저，세포외효원기질결구파배교소，여정상수핵상비，지가조직급기침제액대토골수간충질간세포무명현독부작용。결론：TritonX-100여탈양담산납연합핵산매가유효탈제수핵세포성분，교호지보류세포외기질결구，차해지가대골수간충질간세포무명현독부작용，가이작위수핵조직공정연구적이상재료。
Objective To use detergents and nucleic acid enzyme to prepare scaffold of extracellular ma-trix , then assess the morphological and cytotoxic changes in vitro , and explore the feasibility of this type of scaffold as an ideal tissue-engineering scaffold. Methods Fifty pieces of fresh nucleus pulposus were randomly divided into a fresh control group and a decellularized group. The specimens in decellularized group were treated with 0.3%Tri-ton X-100, 0.5%sodium deoxycholate, and nuclease for 24 h. Morphological changes were studied by macroscopy, pathological staining and scanning electron microscopy. Cytotoxicity was determined by CCK-8 and LIVE/DEAD Viability/Cytotoxicity Assay Kit in vitro. Results The shape of scaffold was maintained,and the extracellular ma-trix was presented while the cells disappeared after decellularization. As compared with the fresh tissue , the scaffold and its extracts had no cytotoxicity to rabbit bone marrow stem cells. Conclusions Almost all the cells have been removed while the extracellular matrix is reserved , and the scaffold has no cytotoxicity to the seed cells. The decel-lularized scaffold can be used as an ideal substance to fabricate tissue-engineering nucleus pulposus.
