CAJ | 학술논문

背景:骨组织工程的支架材料主要作用为模拟细胞体内生长的空间环境,为细胞形成骨组织提供三维支架载体,但目前尚缺乏理想载体材料.目的:应用纤维蛋白胶作为细胞支架进行兔骨髓基质细胞立体培养,探讨其作为骨组织工程支架材料的可行性.设计、时间及地点:单一样本观察,于2007 09/2008 01在吉林大学中日联谊医院及天津科技大学机械工程学院完成.材料:将纤维蛋白原和凝血酶按不同比例混合,制备不同强度的纤维蛋白胶.1月龄雄性新两兰大耳白兔1只,体质量0.25 kg.方法:以兔骨髓基质细胞作为种子细胞在CO2孵箱中进行传代培养后,收集扩增的骨髓基质细胞与支架材料纤维蛋白胶复合后再进行培养4周.主要观察指标:采用相筹显微镜观察纤维蛋白胶中培养的骨髓基质细胞的生长状况,以苏木精伊红染色观察不同时期纤维蛋白胶中骨髓基质细胞的活性,以电子显微镜观察培养4周后骨髓皋质细胞超微结构变化.结果:骨髓基质细胞在不同强度的纤维蛋白胶中的牛长状态不同,在低强度纤维蛋白胶中活性好,扩增迅速,而在高强度纤维蚩白胶中细胞生长缓慢,数量少或逐渐死亡.电镜观察纤维蛋白原和凝血酶比例为4:1的纤维蛋白胶培养4周的基质细胞,各细胞器清晰可见,细胞具有良好活性.结论:骨髓基质细胞在低强度形成固体时的纤维蛋白胶中能够良好扩增生长,以纤维蛋白原和凝血酶比例为4:1的效果最佳.
배경:골조직공정적지가재료주요작용위모의세포체내생장적공간배경,위세포형성골조직제공삼유지가재체,단목전상결핍이상재체재료.목적:응용섬유단백효작위세포지가진행토골수기질세포입체배양,탐토기작위골조직공정지가재료적가행성.설계、시간급지점:단일양본관찰,우2007 09/2008 01재길림대학중일련의의원급천진과기대학궤계공정학원완성.재료:장섬유단백원화응혈매안불동비례혼합,제비불동강도적섬유단백효.1월령웅성신량란대이백토1지,체질량0.25 kg.방법:이토골수기질세포작위충자세포재CO2부상중진행전대배양후,수집확증적골수기질세포여지가재료섬유단백효복합후재진행배양4주.주요관찰지표:채용상주현미경관찰섬유단백효중배양적골수기질세포적생장상황,이소목정이홍염색관찰불동시기섬유단백효중골수기질세포적활성,이전자현미경관찰배양4주후골수고질세포초미결구변화.결과:골수기질세포재불동강도적섬유단백효중적우장상태불동,재저강도섬유단백효중활성호,확증신속,이재고강도섬유치백효중세포생장완만,수량소혹축점사망.전경관찰섬유단백원화응혈매비례위4:1적섬유단백효배양4주적기질세포,각세포기청석가견,세포구유량호활성.결론:골수기질세포재저강도형성고체시적섬유단백효중능구량호확증생장,이섬유단백원화응혈매비례위4:1적효과최가.
BACKGROUND: In tissue engineering, three-dimensional biodegradable scaffolds are generally used as a basic structure for cell anchorage, proliferation. Currently, no perfect scaffold is available. OBJECTIVE: To observe the growth of rabbit bone marrow stromal cells (BMSCs) cultured in different-intensity three-dimensional fibrin glue in vitro, and to discuss the feasibility of fibrin glue used as a scaffold material of bone tissue engineering. DESIGN, TIME AND SETTING: The single sample observational study was performed at the China-Japan Union Hospital of Jilin University and School of Mechanical Engineering of Tianjin University of Technology from September 2007 to January 2008. MATERIALS: Fibrinogen and thrombin were mixed at various proportions, and prepared into different intensity fibrin glue. A month-old male New Zealand white rabbits, weighing 0.25 kg was used in this study. METHODS: Rabbit BMSCs were cultured and serial subcultivation in a CO2 incubator. And then the amplified BMSCs were collected and continue to be cultured in different intensity fibrin glue for 4 weeks. MAIN OUTCOME MEASURES：Observation of growing BMSCs is performed using the phase contrast microscope. The activity of BMSCs in fibrin glue at different stages was observed using hematoxylin-eosin staining. The ultrastructural changes of BMSCs were observed which had been cultivated in fibrin glue for 4 weeks. RESULTS: After growing in fibrin glue for 4 weeks, BMSCs showed strongly active status in low intensity fibrin glue and growing slowly or dying in high intensity fibrin glue. Under the electron microscope, BMSCs following 4 weeks culture in fibrin glue (proportation of fibrinogen and thrombin was 4:1) were found, with visible cellular organs, and BMSCs had good activities. CONCLUSION: BMSCs can spread and proliferate quickly in low intensity fibrin glue. The optimal proportion of fibrinogen and thrombin is 4: 1.