大鼠脱细胞脊髓的形态学观察

Morphology study of decellularized rat spinal cord

中文关键词:  脊髓损伤  组织工程  脱细胞脊髓支架  大鼠

英文关键词:Spinal cord injury  Tissue engineering  Decellularized spinal cord  Rat

中文摘要:

  【摘要】 目的：观察大鼠脱细胞脊髓的形态特点，探讨其作为脊髓组织工程支架的可行性。方法：取11条SD大鼠脊髓，采用冻融+化学萃取法进行脱细胞处理，其中2条采用HE染色观察脱细胞情况，立春红2R-亮绿SF染色观察脱髓鞘情况；3条取颈、胸、腰段横切片行Mallory三色胶原染色和透射电镜检查，3条取颈、胸、腰段纵切片行Mallory三色胶原染色，3条取颈、胸、腰段贴片行扫描电镜检查，观察其形态构成等参数，比较不同部位材料内部结构的差异。结果：经脱细胞处理的脊髓呈乳白色扁长条状，干燥，体积较处理前略变小，韧性及强度增加，表面脊膜包裹完好，无异味。HE染色显示脊髓内未见细胞残留；丽春红2R-亮绿SF染色证实神经细胞轴突脱髓鞘完全。颈、胸及腰段脊髓横断面均呈网格状结构，纵断面均呈平行管状结构。颈、胸及腰段横断面网格大小及纵断面平行管状结构间距行统计学两两比较，颈段网格长径明显大于胸段（P＜0.05），胸段网格短径明显小于腰段（P=0.05），其余均无统计学差异（P＞0.05）。结论：SD大鼠脊髓经脱细胞处理后颈、胸和腰段脱细胞彻底，保留了原脊髓的主体结构，可作为脊髓组织工程的支架。

英文摘要:

  【Abstract】 Objective：To observate the morphology of rat decellularized spinal cord and to investigate the its feasibility of working as spinal cord tissue engineering scaffolds.Method：Eleven SD rat spinal cords adopt were processed by the method of freeze-thaw and chemically extracted for the preparation of decellularized and demyelinating spinal cord scaffold material.Two scaffold materials were processed for HE staining to observe acellular and Ponceau Red 2R-Brilliant Green SF staining to observe demyelinating.3 cross-sectional specimens from cervical，thoracic and lumbar sections were stained by Mallory trichrome collagen and observed under transmission electron microscope.3 longitudinal specimens from cervical，thoracic and lumbar sections were stained by Mallory trichrome collagen，and 3 specimens from cervical，thoracic and lumbar sections were observed under transmission electron microscope.Their morphology and inner structure of varied site were observed and compared.Result：The decellularized spinal cord was cream color and displayed flat long strip，with the features of dry，slightly decreased volume and increased flexibility and strength，it had complete superficial duramater and no unusual smell.HE staining indicated no cell in the spinal cord；Ponceau Red 2R-Brilliant Green SF staining showed complete demyelination.Cross-section specimen presented with grid structure in cervical，thoracic and lumbar spinal cord，and longitudinal specimen presented with parallel-tube structure.The grid size and longitudinal distance between the parallel tube of cervical，thoracic and lumbar specimen were compared mutually，the results showed longer diameter of cervical grid than thoracic ones（P＜0.05），and the short diameter of lumbar grid larger than thoracic ones（P=0.05），while no significance remained for the rest.Conclusion：After relative management，spinal cord of cervical，thoracic and lumbar present with complete decellularization，the main structure of the original spinal cords are retained，which is apt to be used as spinal cord tissue engineering scaffold.