组织工程化同种异体脱细胞脊髓治疗脊髓损伤及其机制的实验研究

项目名称： 组织工程化同种异体脱细胞脊髓治疗脊髓损伤及其机制的实验研究

项目编号： No.30872600

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 任先军

作者单位： 中国人民解放军第三军医大学

项目金额： 29万元

中文摘要： 脊髓损伤是一种常见的中枢神经系统损伤，可造成患者严重的感觉、运动功能障碍，但由于中枢神经系统中轴突再生能力弱，并有髓鞘、胶质瘢痕等抑制因素的存在，致使患者神经功能恢复效果极差。组织工程技术的出现为脊髓损伤的治疗带来了希望。虽然在组织工程研究中，已有多种天然的与合成的高分子材料用于脊髓损伤修复，但仍需进一步改进以符合组织工程脊髓修复的需要。我们尝试将脱细胞脊髓基质材料用以构建组织工程脊髓支架，以满足组织工程脊髓修复的解剖和生理学需要。本研究通过冻融结合化学萃取法制备脱细胞脊髓支架，并进一步优化该制备方法，应用HE染色、髓鞘染色及扫描电镜等方法评价制备脱细胞脊髓支架的可行性，然后通过对支架的成分分析、免疫原性分析、细胞毒性、组织相容性，血液相容性、全身毒性反应等检测评价脱细胞脊髓支架的生物安全性。结果显示，脊髓的细胞外基质成份在支架中得以保留，该支架具有高度仿生的三维结构，生物相容性好，为组织工程脊髓支架的研究提供了理论依据。

中文关键词： 脊髓损伤；脱细胞脊髓支架；组织工程；细胞毒性；生物安全性

英文摘要： Spinal cord injury (SCI) is a trauma with high incidence in the central nervous system (CNS), which usually results in severe impairment of sensary, motor and autonomic function below the level of injury.Because the CNS axons lack the ability for spontaneous regeneration- - further compounded by chemical (myelin inhibitors) and physical (e.g. glial scar) barriers to regeneration after spinal cord injury, patients normally experience poor functional recovery.This has presented tissue engineering technology as an alternative strategy for the treatment of spinal cord injury.Various natural and synthetic polymeric materials have been used to promote the functional recovery after spinal cord injury. Nevertheless, further improvements are needed for all previously-reported approaches, which mimic a native extracellular matrix. So we tried to use the acellular spinal cord matrix material to construct a spinal cord tissue engineering scaffold that meet the anatomical and physiological needs. In the research, The acellular spinal cord scaffold was prepared by freeze thawing and chemical extraction medthod. The program of decellulated was made and optimized. The acellular spinal cord scaffold was estimated by HE stain and Immunostaining, Myelin Staining, Scanning Electron Microscopy Analysis. The biological safety of acellular spinal cord scaffold was detected by component analysis, immunogenicity analysis, cytotoxicity, histocompatibility, blood compatibility, general toxicity reaction. the results showed that extracellular matrix components within the spinal cord retained in the scaffold, which possessed highly biomimetic three-dimensional structure, biocompatibility and cell affinity. Thus we provided rationale for constructing the ideal tissue engineering scaffold of spinal cord.

英文关键词： spinal cord injury; acellular spinal cord scaffold; tissue engineering; cytotoxicity; histocompatibility