藻酸盐凝胶支架在生物学修复椎间盘退行性变中的应用  

Alginate scaffold in the repair of lumbar intervertebral degenerative disc by biological method

作　　者：赵鑫[1] 黄师[1] 严宁[1] 刘超乾[1] 侯铁胜[1] 

机构地区：[1]解放军第二军医大学长海医院骨科,上海市200433

出　　处：《中国组织工程研究与临床康复》2008年第1期73-76,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:藻酸盐凝胶不仅在体外培养细胞使用,还可应用于体内组织工程修复。实验以藻酸盐凝胶为支架复合骨髓间充质干细胞,观察修复兔腰椎间盘退行性变的能力。方法:实验于2006-03/2007-03在解放军第二军医大学长海医院骨科实验室完成。①密度梯度分离法分离培养兔骨髓间充质干细胞,配制1.2%藻酸钠水溶液,进行体外骨髓间充质干细胞/藻酸盐复合物的制备,观察细胞生长。②18只新西兰大白兔手术建立腰椎间盘退行性变模型,随机分为3组:对照组、空白组和治疗组。在手术建立椎间盘退行性变模型结束时,在治疗组椎间隙,注射干细胞复合藻酸盐凝胶支架,对照组注射骨髓间充质干细胞,空白组不予任何干预。③应用核磁共振观察椎间盘手术前后高度和T2信号变化;术后12周取材大体观察组织结构;免疫组织化学检测Ⅱ型胶原含量,分光光度法测量蛋白多糖的含量,观察椎间盘退行性变修复效果。结果:18只新西兰大白兔均进入结果分析。①镜下观察支架陷窝内为干细胞,生长良好;将支架行冰冻切片,观察细胞位于网络内,形态良好。②术后12周治疗组MRI测量椎间隙高度和T2信号与术前相比变化最小,与空白组和对照组比较差异有显著性意义(P<0.05)。③术后12周取材大体观察,空白组和对照组的椎间盘,髓核与纤维环不清,中央区域的髓核为纤维组织所替代。而治疗组的椎间盘轻度退行性变,组织结构尚可分清,周围未发现炎症反应,藻酸盐完全降解。④术后12周治疗组蛋白多糖含量高于空白组和对照组(P<0.05),Ⅱ型胶原含量低于空白组和对照组(P<0.05)。结论:动物实验证实藻酸盐支架复合骨髓间充质干细胞具有修复退行性变椎间盘能力,藻酸盐可以做为修复椎间盘退行性变研究的生物学材料。AIM： Alginate gel can be used in cell culture in vitro and tissue repair in vivo. This study observed the effect of bone marrow mesenchymal stem cells （BMSCs） co-cultured with alginate gel on repairing the degeneration of lumbar intervertebral disc. METHODS： The experiment was performed at the laboratory of Department of Orthopedics, Changhai Hospital of Second Military Medical University of Chinese PLA from March 2006 to March 2007. （1）BMSCs were isolated by density gradient centrifugation and cultured with 1.2% alginate solution in vitro. Cell growth was observed. （2）Models of degenerative intervertebral disc were established in eighteen New Zealand rabbits and the animals were randomly divided into three groups： in treatment group, BMSCs encapsulated in alginate beads were transplanted into the degenerative disc; in control group, only BMSCs were transplanted; in blank group, no cells were transplanted. （3）The changes in intervertebral disc height and T2 signal were detected by MRI. Gross histological observation was performed at 12 weeks postoperatively. Type Ⅱ collagen content was detected by immunohistochemistry, and proteoglycan was quantitated by spectrophotometer to observe the repair effect. RESULTS： All 18 rabbits were included in the final analysis. （1）Microscopic observation showed that BMSCs were located at scaffold lacune and grew well. By frozen section, the cells were situated in the scaffold. （2）Measured by MRI, the changes in disc height and T2 value in postoperative 12 weeks were not obvious compared with preoperative ones, but significantly different compared with the blank group and control group （P 〈 0.05）. （3）In gross observation, the nucleus pulposus and annulus fibrosus of intervertebral disc in blank and control groups were not clear, and the nucleus pulposus in central region were replaced by fibrous tissues. While in treatment group, the degeneration was relieved, and the tissue structure was still clear; no inflammatory reaction was found a

关 键 词：藻酸盐 骨髓间充质干细胞 椎间盘 治疗 

分 类 号：R318.08[医药卫生—生物医学工程]