柠檬酸-EDC/NHS胶原凝胶修复破损纤维环  

Annular repair using citric acid-EDC/NHS collagen gel in rats

作　　者：王言[1] 孙超[1] 黄博[1] 刘欢[1] 袁懿[2] 郭宇[3] 周跃[1] 

机构地区：[1]第三军医大学新桥医院骨科,重庆400037 [2]第三军医大学药学院,重庆400038 [3]第三军医大学大坪医院野战外科研究所影像科,重庆400042

出　　处：《第三军医大学学报》2016年第10期1078-1084,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81271982;81472076);国家杰出青年科学基金(81401801)~~

摘　　要：目的探讨利用柠檬酸-EDC/NHS一型胶原凝胶修复纤维环损伤,减缓椎间盘退变的效果。方法使用鼠尾肌腱一型胶原凝胶,柠檬酸(CA)、1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺(EDC)和N-羟基琥珀酰亚胺(NHS)作为交联剂,将一型胶原凝胶与交联剂按不同质量比混合制成一型胶原混合凝胶。建立鼠尾椎间盘纤维环穿刺模型。清洁级SD大鼠48只,分为假手术组(n=8)、CA-EDC/NHS凝胶(CA6)治疗组(n=10)、CA-EDC/NHS凝胶(CA3)治疗组(n=10)、EDC/NHS凝胶(CA0)治疗组(n=10)、穿刺未治疗组(n=10)。分别于1、2、4周行小动物X摄片、小动物MRI检查,统计分析不同时间点椎间隙高度系数、髓核像素值及Pfirrmann分级,于第4周时将大鼠处死行组织学切片观察。结果 4周后,CA-EDC/NHS凝胶(CA6)治疗组的鼠尾椎间盘对比假手术组仅发生了轻微的退变,基本保持了髓核组织的完整性,在椎间盘高度系数(均数79%)、髓核像素值(均数126.9)和Pfirrmann分级结果(平均1.3分)均优于其余两个治疗组和穿刺未治疗组(P<0.05)。CA-EDC/NHS凝胶(CA6)治疗组的组织学观察显示,针道的周围没有发现炎性细胞浸润和瘢痕组织,针道尽头可见一团凝胶状物质在断裂的纤维环组织之间形成了桥接使其闭合。结论柠檬酸-EDC/NHS胶原凝胶具有一定的修复破损纤维环,减缓椎间盘退变的能力,并且与柠檬酸剂量存在相关性。Objective To explore the effect of citric acid（ CA）-1-ethyl-3-（ 3-dimethylaminopropyl）carbodiimide（ EDC） / N-hydroxysuccinimide（ NHS） collagen gel on repairing annular defects. Methods Type I collagen was extracted from the rat tail tendon and crosslinked with CA at different mass ratios using EDC and NHS as crosslinking reagents to prepare 4 kinds of collagen gels. Forty-eight adult SD rats were divided into sham-operation group（ n = 8）,CA6 group（ n = 10） which was punctured and injected with CAEDC / NHS collagen gel（ CA6）,CA3 group（ n = 10） which was punctured and injected with CA-EDC / NHS collagen gel（ CA3）,CA0 group（ n = 10） which was punctured and injected with EDC / NHS collagen gel（ CA0）,and model group（ n = 10） which was punctured and untreated. X-ray filming and magnetic resonance imaging（ MRI） were performed before only puncture and at the 1st,2nd,and 4th week after puncture. At each time point,disc height index（ % DHI）,voxel count and modified MRI Pfirrmann grading were collected and analyzed. All animals were killed at the 4th week for the morphological study. Results The discs in the second group showed only slight degeneration compared with the healthy discs,and the results of % DHI（ average 79%）,voxel count（ average 126. 9）,Pfirrmann grading（ average grade 1. 3） and morphology in the CA6 group indicated less degeneration tendency compared with the other 3 puncture groups at the 4th week（ P〈0. 05）. The annular fibrosus was partially repaired by the collagen gels that bridged the defects. Conclusions CA-EDC / NHS collagen gel is capable of repairing annular defects induced by needle puncture,which may be closely related with the dose of CA.

关 键 词：纤维环修复 柠檬酸 1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺 N-羟基琥珀酰亚胺 鼠尾穿刺模型 

分 类 号：R332[医药卫生—人体生理学] R341.26[医药卫生—基础医学]