中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (14): 1371-1378.doi: 10.4103/1673-5374.137590

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

动态培养温敏胶原为细胞外基质构建组织工程周围神经的优势

  

  • 收稿日期:2014-04-25 出版日期:2014-07-25 发布日期:2014-07-25
  • 基金资助:

    中国国家自然科学基金(31071222),吉林省科技发展计划项目(20080738),吉林大学白求恩医学科研支持计划——前沿交叉学科创新项目(2013106023)

Dynamic culture of a thermosensitive collagen hydrogel as an extracellular matrix improves the construction of tissue-engineered peripheral nerve

Lanfeng Huang 1, Rui Li 2, Wanguo Liu 3, Jin Dai 1, Zhenwu Du 4, Xiaonan Wang 1, Jianchao Ma 1, Jinsong Zhao 5   

  1. 1 Department of Joint Surgery, Orthopedics Hospital of the Second Hospital of Jilin University, Changchun, Jilin Province, China
    2 Centre of Hand & Foot Surgery and Reparative & Reconstructive Surgery, Orthopedics Hospital of the Second Hospital of Jilin University, Changchun, Jilin Province, China
    3 Department of Orthopedics Surgery, the Third Hospital of Jilin University, Changchun, Jilin Province, China
    4 Institute of Orthopedics, the Second Hospital of Jilin University, Changchun, Jilin Province, China
    5 Department of Ophthalmology, the Second Hospital of Jilin University, Changchun, Jilin Province, China
  • Received:2014-04-25 Online:2014-07-25 Published:2014-07-25
  • Contact: Jinsong Zhao, M.D., Ph.D., Department of Ophthalmology, the Second Hospital of Jilin University, Changchun 130041, Jilin Province, China, jinsong@jlu.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 31071222; Jilin Province Science and Technology Development Project in China, No. 20080738; and the Frontier Interdiscipline Program of Norman Bethune Health Science Center of Jilin University in China, No. 2013106023.

摘要:

构建组织工程复合物修复周围神经损伤是一种新的治疗策略,但在种子细胞黏附到支架的过程中常发生大量细胞丢失。实验利用温敏胶原与聚乳酸联合作为细胞外基质,与骨髓间充质细胞联合在体外构建组织工程周围神经复合物,并使培养皿以0.5Hz频率在水平面上下35°摆动的方式进行动态培养。结果发现骨髓间充质细胞在聚乳酸支架周围形成的膜结构,骨髓间充质细胞排列整齐,胶原填充在空隙中,骨髓间充质细胞黏附于聚乳酸纤维上。总DNA定量分析表明,与单纯I型胶原构建的支架相比,温敏胶原构建的支架中骨髓间充质细胞DNA含量较高;且经动态培养的温敏胶原构建的支架中骨髓间充质细胞的DNA含量高于静态培养。提示动态培养的组织工程复合物可以保持充足的种子细胞数量,有效避免了原始构建阶段的细胞流失,而且种子细胞在材料中的分布良好。

关键词: 神经再生, 周围神经, 生物材料, 细胞外基质, 组织工程, 神经支架, 骨髓间充质细胞, 温敏胶原, 聚乳酸, 动态培养, NSFC grant

Abstract:

Tissue engineering technologies offer new treatment strategies for the repair of peripheral nerve injury, but cell loss between seeding and adhesion to the scaffold remains inevitable. A thermosensitive collagen hydrogel was used as an extracellular matrix in this study and combined with bone marrow mesenchymal stem cells to construct tissue-engineered peripheral nerve composites in vitro. Dynamic culture was performed at an oscillating frequency of 0.5 Hz and 35° swing angle above and below the horizontal plane. The results demonstrated that bone marrow mesenchymal stem cells formed membrane-like structures around the poly-L-lactic acid scaffolds and exhibited regular alignment on the composite surface. Collagen was used to fill in the pores, and seeded cells adhered onto the poly-L-lactic acid fibers. The DNA content of the bone marrow mesenchymal stem cells was higher in the composites constructed with a thermosensitive collagen hydrogel compared with that in collagen I scaffold controls. The cellular DNA content was also higher in the thermosensitive collagen hydrogel composites constructed with the thermosensitive collagen hydrogel in dynamic culture than that in static culture. These results indicate that tissue-engineered composites formed with thermosensitive collagen hydrogel in dynamic culture can maintain larger numbers of seeded cells by avoiding cell loss during the initial adhesion stage. Moreover, seeded cells were distributed throughout the material.

Key words: nerve regeneration, peripheral nerve, biomaterials, extracellular matrix, tissue engineering, nerve scaffold, bone marrow mesenchymal stem cells, thermosensitive collagen hydrogel, poly-L-lactic acid, dynamic culture, NSFC grant, neural regeneration