中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (8): 1365-1368.doi: 10.3969/j.issn.1673-8225.2010.08.009

• 细胞外基质材料 extracellular matrix materials • 上一篇    下一篇

脱细胞膀胱黏膜下层支架材料的生物学评价

任鹏程1，张旭东1，吕海港1，咎玉玲1，刘  怡1，安丽君2   

  1. 解放军第四军医大学唐都医院，  1全军骨肿瘤研究所，2风湿免疫科，陕西省西安市   710038
  • 出版日期:2010-02-19 发布日期:2010-02-19
  • 通讯作者: 安丽君，主管护师，解放军第四军医大学唐都医院风湿免疫科，陕西省西安市 710038
  • 作者简介:任鹏程，男，1971年生，山西省山阴县人，汉族，1994年解放军第四军医大学毕业，主治医师，主要从事分子生物学研究。 doctorlhg@163.com

Biological evaluation of acellular bladder submucosa materials

Ren Peng-cheng1, Zhang Xu-dong1, Lü Hai-gang1, Jiu Yu-ling1, Liu Yi1, An Li-jun2   

  1. 1 Institute of Orthopedic Oncology of PLA, 2 Department of Rheumatology and Immunology, Tangdu Hospital, Fourth Military Medical University of Chinese PLA, Xi’an  710038, Shaaxi Province, China
  • Online:2010-02-19 Published:2010-02-19
  • Contact: An Li-jun, Nurse-in-charge, Department of Rheumatology and Immunology, Tangdu Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710038, Shaaxi Province, China
  • About author:Ren Peng-cheng, Attending physician, Institute of Orthopedic Oncology of PLA, Tangdu Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710038, Shaaxi Province, China doctorlhg@163.com

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549

被引次数

12

摘要：

背景：脱细胞膀胱黏膜下层是一种天然的细胞外基质生物材料，主要由Ⅰ、Ⅲ型纤维胶原蛋白构成，是一种理想的生物支架材料。
目的：脱细胞猪膀胱组织作为组织工程支架材料的生物学评价。
方法：取适量健康猪膀胱置于PBS 和叠氮钠的混合溶液，浸泡过夜，刮去膀胱黏膜层。用低渗、-80 ℃反复冻融、DNase和RNase混合液消化和NaOH裂解连续方法制备脱细胞膀胱黏膜下层。通过观察其组织学结构特征、DNA残留、细胞毒性、细胞黏附效果及皮下炎症反应等来综合评价脱细胞膀胱黏膜下层的生物相容性。
结果与结论：正常猪的膀胱经改进的脱细胞方法处理后，绝大部分细胞组分被去处，细胞外基质结构保持完好。细胞毒性试验结果显示脱细胞膀胱黏膜下层细胞毒性为1级，DNA提取结果中未见有DNA残留，细胞黏附结果显示猪平滑肌细胞能在脱细胞支架上较好的贴附生长，SD大鼠皮下植入试验显示无明显的炎症反应。结果证实所制备的脱细胞膀胱黏膜下层结构保存完好，有良好的组织相容性，可能成为组织工程修复的替代材料。

关键词: 脱细胞膀胱黏膜下层, 生物相容性, 组织工程膀胱, 猪, 细胞毒性, DNA残留

Abstract:

BACKGROUND: Acellular bladder submucosa is a natural extracellular matrix, which is mainly composed of collagen Ⅰ and Ⅲ. It is regarded as an ideal biological scaffold material.
OBJECTIVE: To evaluate the biocompatibility of acellular bladder submucosa as a tissue engineered scaffold material. 
METHODS: Pig urinary bladder was immersed in the solution of PBS and sodium azide for a night, and the mucosa was removed. Acellular bladder submucosa was prepared using continuous hypotension, freeze-thawed treatment and NaOH spallation. The biocompatibility of acellular bladder submucosa was evaluated through histologic structure, DNA residual, cytotoxicity, cell adhesion, as well as subcutaneous inflammatory reactions. 
RESULTS AND CONCLUSION: The cell components were completely eliminated after decellularization treatment, while the extracellular matrix was remained intact as normal bladder. According to MTT results, cytotoxicity of acellular bladder matrix was assigned to be the first grade. No DNA signal was observed after extraction, and the matrix also supported porcine smooth muscle cell attachment and proliferation. Subcutaneous implantation of the matrix indicated that the acellular bladder submucosa trigger no immunologic rejection reaction obviously. The results demonstrated that: the acellular bladder submucosa prepared here exhibits excellent biocompatibility, which can be used as substitution in tissue-engineering field.

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