中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2008年
10期
1943-1949
,共7页
黎晖%陈东风%刘金元%周健洪%杜少辉%李伊为%邓汝东%张赛霞%曾和平
黎暉%陳東風%劉金元%週健洪%杜少輝%李伊為%鄧汝東%張賽霞%曾和平
려휘%진동풍%류금원%주건홍%두소휘%리이위%산여동%장새하%증화평
壳聚糖-明胶-碱性纤维生长因子支架%骨髓间充质干细胞%增殖%体外%干细胞组织工程
殼聚糖-明膠-堿性纖維生長因子支架%骨髓間充質榦細胞%增殖%體外%榦細胞組織工程
각취당-명효-감성섬유생장인자지가%골수간충질간세포%증식%체외%간세포조직공정
背景:干细胞的分化潜能与培养条件有密切关系,改变支架材料的表面特性,三维结构,增加生长因子均可实现对干细胞增殖分化的控制.目的:制备适合骨髓间充质干细胞附着生长的、具有最佳孔隙率和孔隙结构的药物缓释组织工程支架--三维大孔支架,提供能促进多能干细胞生长的微环境.设计:重复测量设计.单位:广州中医药大学基础医学院解剖教研室.材料:实验所用健康成年 SD 大鼠由广州中医药大学实验动物中心提供.壳聚糖、碱性成纤维细胞生长因子购自Sigma公司.方法:实验于2003-03/2006-12主要在广州中医药大学基础医学院解剖教研室完成.采用冷冻干燥的方法,用不同比例的壳聚糖.碱性成纤维细胞生长因子-明胶依次混匀,通过控制冷冻、复温和干燥时间处理使其具有最佳孔隙率和孔结构,制备具缓释碱性成纤维细胞生长因子功能的三维大孔支架.取SD大鼠股骨和胫骨骨髓,分离、培养骨髓间充质干细胞并移植于缓释碱性成纤维细胞生长因子的三维大孔支架上进行三维培养,与无碱性成纤维细胞生长因子的支架对照.实验过程中对动物的处置符合动物伦理学标准.主要观察指标:用ELISA和扫描电镜观察支架的三维结构和缓释性能,用苏木精-伊红染色、MTT、细胞计数及扫描电镜方法观察缓释碱性成纤维细胞生长因子的三维大孔支架对骨髓间充质干细胞生长状态和细胞活力的影响.结果:含有碱性成纤维细胞生长因子的三维大孔支架有碱性成纤维细胞生长因子缓释性能,孔隙尺寸与不含碱性成纤维细胞生长因子的支架三维结构相比,差异无显著性意义(P>0.05).含有碱性成纤维细胞生长因子的三维大孔支架能提高在支架上立体培养的骨髓间充质干细胞存活率,促进骨髓间充质干细胞黏附、增殖和活力,与不含碱性成纤维细胞生长因子的支架相比,差异有显著性意义(P<0.05).结论:含有碱性成纤维细胞生长因子的三维大孔支架能缓释碱性成纤维细胞生长因子,有利于在支架上立体培养的骨髓间充质干细胞存活,为其在组织工程中的应用打下基础.
揹景:榦細胞的分化潛能與培養條件有密切關繫,改變支架材料的錶麵特性,三維結構,增加生長因子均可實現對榦細胞增殖分化的控製.目的:製備適閤骨髓間充質榦細胞附著生長的、具有最佳孔隙率和孔隙結構的藥物緩釋組織工程支架--三維大孔支架,提供能促進多能榦細胞生長的微環境.設計:重複測量設計.單位:廣州中醫藥大學基礎醫學院解剖教研室.材料:實驗所用健康成年 SD 大鼠由廣州中醫藥大學實驗動物中心提供.殼聚糖、堿性成纖維細胞生長因子購自Sigma公司.方法:實驗于2003-03/2006-12主要在廣州中醫藥大學基礎醫學院解剖教研室完成.採用冷凍榦燥的方法,用不同比例的殼聚糖.堿性成纖維細胞生長因子-明膠依次混勻,通過控製冷凍、複溫和榦燥時間處理使其具有最佳孔隙率和孔結構,製備具緩釋堿性成纖維細胞生長因子功能的三維大孔支架.取SD大鼠股骨和脛骨骨髓,分離、培養骨髓間充質榦細胞併移植于緩釋堿性成纖維細胞生長因子的三維大孔支架上進行三維培養,與無堿性成纖維細胞生長因子的支架對照.實驗過程中對動物的處置符閤動物倫理學標準.主要觀察指標:用ELISA和掃描電鏡觀察支架的三維結構和緩釋性能,用囌木精-伊紅染色、MTT、細胞計數及掃描電鏡方法觀察緩釋堿性成纖維細胞生長因子的三維大孔支架對骨髓間充質榦細胞生長狀態和細胞活力的影響.結果:含有堿性成纖維細胞生長因子的三維大孔支架有堿性成纖維細胞生長因子緩釋性能,孔隙呎吋與不含堿性成纖維細胞生長因子的支架三維結構相比,差異無顯著性意義(P>0.05).含有堿性成纖維細胞生長因子的三維大孔支架能提高在支架上立體培養的骨髓間充質榦細胞存活率,促進骨髓間充質榦細胞黏附、增殖和活力,與不含堿性成纖維細胞生長因子的支架相比,差異有顯著性意義(P<0.05).結論:含有堿性成纖維細胞生長因子的三維大孔支架能緩釋堿性成纖維細胞生長因子,有利于在支架上立體培養的骨髓間充質榦細胞存活,為其在組織工程中的應用打下基礎.
배경:간세포적분화잠능여배양조건유밀절관계,개변지가재료적표면특성,삼유결구,증가생장인자균가실현대간세포증식분화적공제.목적:제비괄합골수간충질간세포부착생장적、구유최가공극솔화공극결구적약물완석조직공정지가--삼유대공지가,제공능촉진다능간세포생장적미배경.설계:중복측량설계.단위:엄주중의약대학기출의학원해부교연실.재료:실험소용건강성년 SD 대서유엄주중의약대학실험동물중심제공.각취당、감성성섬유세포생장인자구자Sigma공사.방법:실험우2003-03/2006-12주요재엄주중의약대학기출의학원해부교연실완성.채용냉동간조적방법,용불동비례적각취당.감성성섬유세포생장인자-명효의차혼균,통과공제냉동、복온화간조시간처리사기구유최가공극솔화공결구,제비구완석감성성섬유세포생장인자공능적삼유대공지가.취SD대서고골화경골골수,분리、배양골수간충질간세포병이식우완석감성성섬유세포생장인자적삼유대공지가상진행삼유배양,여무감성성섬유세포생장인자적지가대조.실험과정중대동물적처치부합동물윤리학표준.주요관찰지표:용ELISA화소묘전경관찰지가적삼유결구화완석성능,용소목정-이홍염색、MTT、세포계수급소묘전경방법관찰완석감성성섬유세포생장인자적삼유대공지가대골수간충질간세포생장상태화세포활력적영향.결과:함유감성성섬유세포생장인자적삼유대공지가유감성성섬유세포생장인자완석성능,공극척촌여불함감성성섬유세포생장인자적지가삼유결구상비,차이무현저성의의(P>0.05).함유감성성섬유세포생장인자적삼유대공지가능제고재지가상입체배양적골수간충질간세포존활솔,촉진골수간충질간세포점부、증식화활력,여불함감성성섬유세포생장인자적지가상비,차이유현저성의의(P<0.05).결론:함유감성성섬유세포생장인자적삼유대공지가능완석감성성섬유세포생장인자,유리우재지가상입체배양적골수간충질간세포존활,위기재조직공정중적응용타하기출.
BACKGROUND: Stem cell differentiation potential is strongly correlated with culture condition. The alteration in scaffold material surface function, three dimensional (3D) structure, and addition of growth factors can control stem cell proliferation and differentiation.OBJECTIVE: To develop 3D macroporous scaffolds with optimal porosity and porous structure to provide a microenvironment that promotes the growth of multi-potent stem cells.DESIGN: Repetitive measurement.SETTING: Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine.MATERIALS: Healthy adult SD rats were provided by the Experimental Animal Center in Guangzhou University of Chinese Medicine. Chitosan and basic fibroblast growth factor (bFGF) were purchased from Sigma Corporation (St. Louis,MO).METHODS: The experiment was performed at the Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine from March 2003 to December 2006. Using a freeze-drying method, 3D macroporous scaffolds made of different ratios of chitosan-gelatin with bFGF were fabricated that could release bFGF with controlled porosity and porous structure. Bone marrow was obtained from the femur and tibia of SD rats, and mesenchymal stem cells (MSCs) were isolated, cultured and seeded on the scaffolds with bFGF. MSCs seeded on scaffolds with no bFGF served as control. The procedure during experiment was accorded with animal ethical requirements.MAIN OUTCOME MEASURES: 3D structure and release performance of the scaffolds were observed by ELISA and scanning electron microscope; the effect of 3D macroporous scaffolds that released bFGF on MSC growth and viability were observed by HE staining, MTT, cell counting and SEM.RESULTS: There was no significant difference in pore size between scaffolds with and without bFGF (P > 0.05). Scaffolds with bFGF significantly improved MSC survival rate, promoted cell adhesion, proliferation, and viability compared with scaffolds without bFGF (P < 0.05).CONCLUSION: The results suggest that 3D macroporous scaffolds with bFGF release improve MSC survival on scaffolds,and lay a foundation for its application in tissue engineering.