山羊骨髓间充质干细胞与纤维蛋白胶修复椎间盘纤维环缺损的组织学比较  

Histological comparison between goat bone marrow mesenchymal stem cells and adhesive fibrin for the repair of annulus fibrosus defect of intervertebral discs

作　　者：徐翔 吴一民[1] 李树文[1] 马丽波[2] 王宇鹏[1] 于英楠[1] 孙涛 张远 任伟 银和平[1] Xu Xiang;Wu Yimin;Li Shuwen;Ma Libo;Wang Yupeng;Yu Yingnan;Sun Tao;Zhang Yuan;Ren Wei;Yin Heping(Department of Minimally Invasive Spinal Surgery,Second Affiliated Hospital of Inner Mongolia Medical College,Huhhot 010030,Inner Mongolia Autonomous Region,China;Department of Nursing,Second Affiliated Hospital of Inner Mongolia Medical College,Huhhot 010030,Inner Mongolia Autonomous Region,China;Department of Emergency,Second Affiliated Hospital of Inner Mongolia Medical College,Huhhot 010030,Inner Mongolia Autonomous Region,China;Department of Surgical Anesthesiology,Inner Mongolia International Hospital,Huhhot 010030,Inner Mongolia Autonomous Region,China;Graduate College,Inner Mongolia Medical University,Huhhot 010059,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古医科大学第二附属医院微创脊柱外科,内蒙古自治区呼和浩特市010030 [2]内蒙古医科大学第二附属医院护理部,内蒙古自治区呼和浩特市010030 [3]内蒙古医科大学第二附属医院急诊科,内蒙古自治区呼和浩特市010030 [4]内蒙古国际蒙医院手术麻醉科,内蒙古自治区呼和浩特市010030 [5]内蒙古医科大学研究生院,内蒙古自治区呼和浩特市010059

出　　处：《中国组织工程研究》2022年第13期1974-1978,共5页Chinese Journal of Tissue Engineering Research

基　　金：内蒙古医科大学科技百万工程课题[YKD2018KJBW(LH)006],项目负责人:徐翔;内蒙古自治区自然科学基金项目(2019BS03034),项目负责人:徐翔。

摘　　要：背景:目前,腰椎间盘摘除术后纤维环破口仍继续存在,容易形成复发型腰椎间盘突出,如何修复纤维环破口是组织工程学的热点问题。目的:比较以明胶海绵为载体的山羊骨髓间充质干细胞移植与纤维蛋白粘合剂修复椎间盘纤维环缺损的能力。方法:从山羊髂骨中提取和分离出骨髓间充质干细胞,并将其进行传代培养。将15只山羊随机分为对照组、粘合组、移植组,每组5只(30个椎间盘)。采用手术器械制备0.75 cm×0.75 cm椎间盘纤维环缺损,粘合组利用纤维蛋白粘合剂对破损的纤维环进行粘合,移植组将负载骨髓间充质干细胞悬液(细胞浓度为5×10^(9) L^(-1),体积为10μL)的明胶海绵(0.75 cm×0.75 cm)放入纤维环缺损中,然后逐层进行缝合。术后6,12周,取出纤维环组织进行苏木精-伊红染色、Masson三色明胶染色、AB-PAS染色和Ⅱ型胶原染色。结果与结论:苏木精-伊红染色观察到移植组中软骨细胞、胶原细胞与骨细胞的数量相比其他两组增多,细胞成熟,修复能力更强;Masson三色明胶染色观察到移植组有更多的成纤维细胞和纤维组织;AB-PAS染色观察到移植组有更多的软骨细胞参与修复;Ⅱ型胶原免疫组化染色发现移植组Ⅱ胶原含量最高;上述结果证实了山羊骨髓间充质干细胞对纤维环缺损有更好的修复能力。BACKGROUND:At present,after lumbar discectomy,annulus fibrosus rupture still exists,easy to form recurrent lumbar disc herniation.How to repair the broken annulus fibrosus is a hot issue in tissue engineering.OBJECTIVE:To compare the ability of goat bone marrow mesenchymal stem cell transplantation with gelatin sponge as carrier and adhesive fibrin repair for annulus fibrosus defects.METHODS:Bone marrow mesenchymal stem cells were extracted and isolated from the goat iliac bone and transferred for culture.Fifteen goats were randomized into control,adhesion,and transplant groups(n=5 per group and 30 discs per group).The 0.75 cm×0.75 cm disc annulus fibrosus defect was made with surgical instruments.In the binding group,the damage was bonded with fibrin adhesive.In the transplant group,the gelatin sponge(0.75 cm×0.75 cm)with bone marrow mesenchymal stem cell suspension(cell concentration:5×10^(9)/L,10μL)was placed into the annulus fibrosus defect and the lesion was stitched layer by layer.At 6 and 12 weeks after surgery,annulus fibrosus tissue was removed for hematoxylin-eosin staining,Masson staining,AB-PAS staining,and type II collagen staining.RESULTS AND CONCLUSION:Hematoxylin-eosin staining results showed that the number of chondrocytes,collagen cells and bone cells in the transplant group increased,the cells were mature and the repair ability was stronger compared with the other two groups.Masson staining demonstrated that the number of fibroblasts and fibrous tissue was more in the transplant group compared with the other two groups.AB-PAS staining exhibited that more chondrocytes in the transplant group were involved in repair compared with the other two groups.Type II collagen staining showed that the content of type II collagen was highest in the transplant group.The above results confirm that goat bone marrow mesenchymal stem cells have a perfect ability to repair annulus fibrosus defects.

关 键 词：骨髓间充质干细胞 纤维蛋白胶 组织学观察 纤维环 Ⅱ型胶原 MASSON染色 AB-PAS染色 苏木精-伊红染色 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]