CAJ | 학술논문

背景:在应用组织工程修复周围神经的研究中,载体的选择至关重要.理想的载体应与细胞外基质类似,与活体细胞有良好的生物相容性.目的:评价豚鼠海马神经干细胞与胶原蛋白海绵和明胶海绵的生物相容性,探讨它们作为周围神经组织工程载体材料的可行性.设计:随机对照实验.单位:郑州大学第一附属医院耳鼻咽喉科,郑州大学基础医学院解剖教研室.材料:实验于2005-07/2005-12在郑州大学基础医学院解剖教研室神经生物学研究室完成.健康新生(＜24 h)杂色豚鼠12只,清洁级,雌雄不限,体质量50-70 g,由郑州大学医学院实验动物中心提供.方法:新生(＜24 h)豚鼠,10 g/L水合氯醛腹腔麻醉,体积分数为0.75乙醇消毒,无菌操作分离出海马组织.体外无血清培养海马神经干细胞,传至第2代,将浓度调整为1×1010 L-1后分别与胶原蛋白海绵、明胶海绵联合体外培养,一周后取材,进行细胞计数,倒置相差显微镜及扫描电镜观察,并测定两种材料与细胞的吸附率. 主要观察指标:①观察神经干细胞在胶原蛋白海绵和明胶海绵上的生长、黏附情况,并测定其细胞总数与载体材料的吸附率.②免疫细胞化学染色分化细胞形态观察.结果:①神经干细胞可以在胶原蛋白海绵和明胶海绵上生长,逐渐黏附,胶原蛋白海绵细胞吸附率高于明胶海绵(37.17%和14.87%,x2=4.819,P＜0.05).②对照组、胶原蛋白海绵组、明胶海绵组细胞总数差异无显著性[(53.17±3.5)×104,(53.25±2.6)×104,(52.04±4.05)×104,F=0.233,P＞0.05].结论:胶原蛋白及明胶两种材料,尤其是胶原蛋白,具有良好的神经干细胞相容性,可以作为周围神经组织工程的支架材料应用于临床.
배경:재응용조직공정수복주위신경적연구중,재체적선택지관중요.이상적재체응여세포외기질유사,여활체세포유량호적생물상용성.목적:평개돈서해마신경간세포여효원단백해면화명효해면적생물상용성,탐토타문작위주위신경조직공정재체재료적가행성.설계:수궤대조실험.단위:정주대학제일부속의원이비인후과,정주대학기출의학원해부교연실.재료:실험우2005-07/2005-12재정주대학기출의학원해부교연실신경생물학연구실완성.건강신생(＜24 h)잡색돈서12지,청길급,자웅불한,체질량50-70 g,유정주대학의학원실험동물중심제공.방법:신생(＜24 h)돈서,10 g/L수합록철복강마취,체적분수위0.75을순소독,무균조작분리출해마조직.체외무혈청배양해마신경간세포,전지제2대,장농도조정위1×1010 L-1후분별여효원단백해면、명효해면연합체외배양,일주후취재,진행세포계수,도치상차현미경급소묘전경관찰,병측정량충재료여세포적흡부솔. 주요관찰지표:①관찰신경간세포재효원단백해면화명효해면상적생장、점부정황,병측정기세포총수여재체재료적흡부솔.②면역세포화학염색분화세포형태관찰.결과:①신경간세포가이재효원단백해면화명효해면상생장,축점점부,효원단백해면세포흡부솔고우명효해면(37.17%화14.87%,x2=4.819,P＜0.05).②대조조、효원단백해면조、명효해면조세포총수차이무현저성[(53.17±3.5)×104,(53.25±2.6)×104,(52.04±4.05)×104,F=0.233,P＞0.05].결론:효원단백급명효량충재료,우기시효원단백,구유량호적신경간세포상용성,가이작위주위신경조직공정적지가재료응용우림상.
BACKGROUND: The selection of carrier plays an essential role in the research of applying tissue-engineering to fix the peripheral nerves. An ideal carrier would be one that is similar to extracellular matrix and that it has biocompatibility with in vivo cells.OBJECTIVE: To evaluate the biocompatibility of hippocampal neural stem cells with collagen and gelatin sponge in vitro and to probe into the feasibility of using the materials as biomaterial scaffold in peripheral nerve tissue engineering.DESIGN: A randomized and controlled trial.SETTING: Department of Otorhinolaryngology of the First Affiliated Hospital of Zhengzhou University; Department of Anatomy of the School of Basic Medical Sciences of Zhengzhou University.MATERIALS: The experiment was carried out from July to December 2005 at the Laboratory of Neurobiology of the Department of Anatomy of the School College of Basic Medical Sciences of Zhengzhou University.Twelve New born (＜ 24 hours) clean grade guinea pigs of either gender with a body mass of 50-70 g, were provided by the Experimental Animals Center of Zhengzhou University School of Medicine.METHODS: The new born (＜ 24 hours) guinea pigs were anesthetized intraperitoneally with 10 g/L chloral hydrate and sterilized in 0.75 volume fraction of alcohol. Hippocampal tissue was resected from the brain under a surgical microscope. Hippocampus neural stem cells were cultured in vitro. The cultured cells of two generations were suspended at a density of 1 ×1010L-1 and respectively combined with collagen and gelatin sponge.The number of cells was counted and histological changes were observed under an inverted phase microscope and scanning electron microscope after 7 days, and the adhesion rate of the two materials to the cells were measured.MAIN OUTCOME MEASURES: ①Growth of the neural stem cells and their adhesion to the collagen and gelatin sponge were observed and the total number of the cells and adhesion rate with carrier were measured. ②The morphological changes of differentiation cells by immunocytochemical staining.RESULTS: ①Hippocampus neural stem cells could grew on the collagen and gelatin sponge and attached to them gradually. The adsorption rate of collagen was higher than thai of gelatin sponge (37.17 % and 14.87 %,x2=4.819,P ＜ 0.05). ② There was no significant difference in the total number of the cells in the control group, collagen group and gelatin sponge group [(53.17±3.5)×104,(53.25±2.6)×104, (52.04±4.05)×104,F=0.233,P ＞ 0.05].CONCLUSION: Two biomaterials (collagen and glutin sponge), especially collagen, have good biocompatibility with hippocampus neural stem cells from the guinea pigs and can be used safely as scaffold materials in peripheral nerve tissue engineering.