摘要
目的:为验证温敏型可注射性水凝胶壳聚糖-甘油磷酸钠(chitosan/glycerophosphate,C/GP)作为神经支架的可行性,将体外诱导的神经元样细胞与温敏型C/GP凝胶共培养,观察其生物相容性。方法:实验于2006-03/12在白求恩国际和平医院实验室完成。①实验材料:壳聚糖由浙江金壳生物化学有限公司提供,β-甘油磷酸钠由FlukaAG公司提供。②实验方法:脱乙酰度91%的壳聚糖200mg溶于9mLHCl溶液中,甘油磷酸钠560mg溶于1mL去离子水中,消毒后室温下按2∶体积比将甘油磷酸钠溶液加入壳聚糖溶液中制备温敏型水凝胶3C/GP。实验分为3组:由骨髓基质细胞诱导分化的神经元样细胞与C/GP复合物共培养者为实验组;神经元样细胞与完全培养基共培养者为细胞对照组;C/GP复合物加完全培养基共培养者为C/GP对照组。③观察指标:光镜下观察各组细胞形态变化,各组在培养24h后MTT法酶标仪测吸光度(A值),连续观察7d绘制生长曲线。结果:神经元样细胞在C/GP凝胶呈三维分布,胞体变大,逐渐伸出突起,呈单极、双极或多极,部分相邻细胞的突起连接。神经元样细胞的生长曲线为"S"型,第1,2天为潜伏期,第3～5天为对数生长期,6d后细胞数量开始减少,实验组的A值较细胞对照组略低,两组在7d后A值均有所降低,但各个时间点差异均无显著性意义(P>0.05)。结论:神经元样细胞与C/GP凝胶有良好的生物相容性,温敏型可注射性C/GP凝胶能作为神经工程的载体材料。
AIM： To explore the feasibility of applying injective thermosensitive chitosan/glycerophosphate （C/GP） gelatin as nerve scaffolds, and observe the biocompatibility of in vitro induced neuron-like cells co-cultured with C/GP gelatin. METHODS, The experiment was performed at Bethun International Peace Hospital frgm March to December in 2006.（1） Chitosan was offered by Golden-Shell Biochemical Co., Ltd. And β-sodium glycerophosphate was provided by Fluka AG Company.（2）200 mg chitosan （with a degree of deacetylation 91%） was dissolved in 9 mL HCI solution. Then 560 mg sodium glycarophosphate solution dissolved in 1 mL deionized water was added to chitosan solution according to the ratio of 2：3 to mix C/GP solution at room temperature. The experiment was divided into 3 groups： experimental group： the neuron-like cells induced by marrow stromal cells were co-cultured with C/GP composite; cell comparison group： the neuron-like cells were co-cultured with complete medium; C/GP comparison group： the C/GP composite was co-cultured with complete medium. （3）Morphological changes of cells was detected under light microscope, and MTT method was used to determine the absorbance （A） value after 24 hours of culture. Seven days later, the growth curve was drawn. RESULTS： Neuron-like cells on C/GP gelatin exhibited larger cell bodies and sent out neurites in three-dimensional culture conditions. They were unipolar, bipolar or multi-polar. Classical sigmoidal growth curves of the two groups were quite similar： the first two days was latent period, 3-5 days was logarithm trophophase, and then the number of the cells decreased from the sixth day. The A value of experimental group was slightly lower than that in cell comparison group, and reduced in the two groups after the seventh day. There were no significant differences at each time point （P 〉 0.05）. CONCLUSION： Biocompatibility between C/GP and neuron-like cells is very good. Ther thermosensitive C/GP gelatin is a po
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第48期9692-9695,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
