中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (26): 4199-4204.doi: 10.3969/j.issn.2095-4344.2017.26.017

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壳聚糖藻酸盐支架与脂肪间充质干细胞复合构建的组织工程脊髓修复急性脊髓损伤

郑华斌1，罗  琳1，陈  路2
  

  1. 1西南医科大学附属中医医院骨科，四川省泸州市  646000；2川北医学院附属医院骨科，四川省南充市  637000
  • 收稿日期:2017-04-10 出版日期:2017-09-18 发布日期:2017-09-28
  • 通讯作者: 罗琳，主治医师，西南医科大学附属中医医院，四川省泸州市646000
  • 作者简介:郑华斌，男，1970年生，四川省泸州市人，汉族，副主任医师，主要从事脊髓损伤方面的研究。
  • 基金资助:
    四川省教育厅项目(16ZB0229)

Tissue-engineered spinal cord construction by chitosan alginate scaffold and adipose-derived mesenchymal stem cells in the treatment of acute spinal cord injury

Zheng Hua-bin1, Luo Lin1, Chen Lu2
  

  1. 1Department of Orthopedics, Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Department of Orthopedics, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
  • Received:2017-04-10 Online:2017-09-18 Published:2017-09-28
  • Contact: Luo Lin, Attending physician, Department of Orthopedics, Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Zheng Hua-bin, Associate chief physician, Department of Orthopedics, Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:

    the Project of Sichuan Provincial Education Department, No. 16ZB0229

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摘要：

文章快速阅读：

 

 

文题释义：
理想的脊髓组织工程支架材料：材料表面有利于细胞的黏附和增殖；生物相容性较好；具有生物降解吸收性，待组织内新组织长成后，支架材料能够被降解为没有毒性的小分子，降解的小分子通过代谢排出；支架材料能够按照预定的形态诱导细胞生长；具有可塑性；具有三维立体结构；具有适宜的力学性能；材料容易加工、来源广泛。
 
背景：组织工程化脊髓为脊髓损伤的修复带来了新的治疗思路。
目的：探讨壳聚糖藻酸盐支架与脂肪间充质干细胞复合的组织工程脊髓修复大鼠急性脊髓损伤的效果。
方法：取48只SD大鼠，建立脊髓半横断模型，随机分4组干预，模型组直接缝合硬膜，细胞组在脊髓断端周边移植脂肪间充质干细胞，支架组在脊髓损伤区植入壳聚糖藻酸盐支架，实验组在脊髓损伤区植入组织工程脊髓(壳聚糖藻酸盐支架与CM-Dil标记脂肪间充质干细胞复合物)。移植后1，3，7周，评估大鼠后肢运动功能；移植后7周取脊髓标本，进行免疫荧光染色和苏木精-伊红染色。
结果与结论：①后肢运动功能：移植后3，7周，实验组后肢运动功能强于模型组、细胞组及支架组(P < 0.05)，细胞组、支架组后肢运动功能强于模型组(P < 0.05)；②免疫荧光染色：模型组无神经丝蛋白200、神经元特异性烯醇化酶、胶原纤维酸性蛋白阳性细胞；支架组可见神经元特异性烯醇化酶阳性细胞、胶原纤维酸性蛋白阳性细胞及少量神经丝蛋白200阳性细胞；细胞组未见神经丝蛋白200、神经元特异性烯醇化酶阳性细胞，可见大量的胶原纤维酸性蛋白阳性细胞；实验组见大量神经元特异性烯醇化酶阳性细胞，少量CM-Dil标记的脂肪间充质干细胞、较多的胶原纤维酸性蛋白阳性细胞及神经丝蛋白200阳性细胞；③苏木精-伊红染色：模型组有大量成纤维细胞和炎性细胞浸润，有较大的脊髓空洞；支架组无脊髓空洞，脊髓和支架材料之间连接紧密；细胞组有脊髓空洞；实验组无脊髓空洞，脊髓损伤区有各种形态细胞形成；④结果表明：壳聚糖藻酸盐支架与脂肪间充质干细胞复合的组织工程脊髓，可促进急性脊髓损伤的修复。

关键词: 生物材料, 材料相容性, 壳聚糖, 藻酸盐, 脂肪间充质干细胞, 组织工程, 大鼠, 脊髓损伤

Abstract:

BACKGROUND: Tissue-engineered spinal cord has brought new treatment ideas for spinal cord repair.
OBJECTIVE: To construct the tissue-engineered spinal cord by chitosan alginate scaffold and adipose-derived mesenchymal stem cells (ADMSCs), and to investigate its repairing effects on acute spinal cord injury in rats.
METHODS: The spinal cord hemisection model was established in 48 Sprague-Dawley rats and then, rat models were randomly divided into four groups: model group, scaffold group, ADMSCs group and tissue-engineered spinal cord group, followed by direct suturing of the dura mater, implantation of chitosan alginate scaffold, implantation of ADMSCs, and implantation of tissue-engineered spinal cord, respectively. The limb motor function of rats was evaluated based on the Basso-Beattie-Bresnahan score at 1, 3, 7 weeks after transplantation. Immunofluorescence staining and hematoxylin-eosin staining of the spinal cord specimens were performed at 7 weeks after transplantation.
RESULTS AND CONCLUSION: (1) Motor function of the hind limb: At 3 and 7 weeks after transplantation, the Basso-Beattie-Bresnahan score was highest in the tissue-engineered spinal cord followed, followed by the ADMSCs and scaffold groups, and lowest in the model group (P < 0.05). (2) Immunofluorescence and hematoxylin-eosin staining: in the model group, the spinal cord injury area was infiltrated with a large number of fibroblasts and inflammatory cells. In the scaffold group, there was no scar formation; neuron-specific enolase-positive cells, glial fibrillary acidic protein-positive cells and a few neurofilament protein 200-positive cells were found in the junctional area. In the ADMSCs group, the spinal cord injury area was filled with scar tissue, and a large number of glial fibrillary acidic protein-positive cells were found. In the tissue-engineered spinal cord group, there was no scar tissue, and there were a large number of neuron-specific enolase-positive cells, a small amount of CM-Dil-labeled ADMSCs and more glial fibrillary acidic protein-positive cells. Neurofilament protein 200 positive cells were connected to each other at the junction. These findings indicate that the tissue-engineered spinal cord constructed by chitosan alginate scaffold and ADMSCs can promote spinal cord repair after acute spinal cord injury. 

Key words: Chitosan, Alginates, Stem Cells, Tissue Enigneering

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