摘要
背景:细胞支架是细胞生长的载体,其孔径是影响组织工程脊髓疗效的重要因素之一。目的:通过将神经干细胞与不同孔径的聚乳酸-羟基乙酸(poly lactic-co-glycolic acid,PLGA)支架体外复合培养,筛选确立组织工程脊髓支架材料的最佳孔径。方法:取传第1代的神经干细胞悬液50μL(细胞数1010L-1),分别种植在孔径200～300μm、400～500μm的PLGA支架上复合培养7d,得到两种组织工程脊髓。30只大鼠均建立脊髓损伤模型,造模后分为3组,分别在脊髓缺损处立即填塞上述两种组织工程脊髓,空白对照组在缺损处不进行材料移植。倒置相差显微镜及电镜下观察神经干细胞在PLGA支架中的生长增殖与分布,MTT检测两种组织工程脊髓所含神经干细胞的相对数量,采用BBB运动功能评分比较不同孔径的组织工程脊髓的移植疗效。结果与结论:镜下神经干细胞在各孔径材料上均紧密贴附并增殖分化,组织相容性良好。共培养7d后,孔径200～300μmPLGA支架组、孔径400～500μmPLGA支架组的吸光度值基本相似(P>0.05),说明PLGA支架的孔径大小对培养的神经干细胞增殖数量无明显影响。移植第4周与空白对照组比较,孔径200～300μmPLGA支架组、孔径400～500μmPLGA支架组大鼠的神经功能均有不同程度恢复,BBB运动功能评分均明显升高(P<0.05),且孔径200～300μm的PLGA支架其移植效果更好。
BACKGROUND：Cytoskeleton is a carrier of cell growth,and its pore caliber is one of the most important factors to affect the curative effect of tissue engineered spinal cord.OBJECTIVE：To explore the optimal pore size of poly lactic-co-glycolic acid（PLGA）scaffolds for tissue engineered spinal cord by in vitro culture of neural stem cells（NSCs）and various pore sizes of PLGA scaffolds.METHODS：50μL（cell number 1010/L）NSCs suspension at passage 1 was separately seeded on 200-300μm,400-500μm PLGA stent for 7 days.Two sorts of tissue engineered spinal cord were constructed in vitro.Thirty rat models of spinal cord injury were established,and then assigned to 3 groups.The detect sites of these models were filled with above-mentioned spinal cord immediately,but the blank control was not treated with any material.The cells growth and proliferation implanted on PLGA were observed by phase contrast microscope and scanning electron microscope.Relative number of NSCs in two tissue engineered spinal cords was measured by MTT assay.The effects of transplantation with tissue engineered spinal cord were evaluated by the BBB scale.RESULTS AND CONCLUSION：Neural stem cells implanted on different pore size scaffolds were seen growing by phase contrast microscope and scanning electron microscope,with good histocompatibility.After 7-day coculture,absorbance was similar between 200-300μm PLGA and 400-500μm PLGA groups（P0.05）.These indicated that the pore size had no effects on NSC number.At week 4 following transplantation,in the blank control group,neural function was recovered to different degrees in the 200-300μm PLGA and 400-500μm PLGA groups.BBB motor functional score was significantly increased（P0.05）.The pore size of 200-300μm utilized in fabricating tissue engineered spinal cord has the best transplantation effect as compared to others.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第3期393-396,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
宁波市自然科学基金资助项目(2008A610093)
课题名称"静脉移植骨髓间充质干细胞治疗大鼠脑梗塞的MRI检测和疗效观察研究"~~
