中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (6): 906-910.doi: 10.3969/j.issn.2095-4344.2017.06.015

• 药物控释材料 drug delivery materials • 上一篇    下一篇

胍基壳聚糖的止血性能和细胞毒性

姚心培1，2，荆妙蕾1，关  静2
  

  1. 1天津工业大学纺织学部，天津市  300387；2军事医学科学院卫生装备研究所，天津市  300161
  • 收稿日期:2017-01-12 出版日期:2017-02-28 发布日期:2017-03-16
  • 通讯作者: 荆妙蕾，硕士，副教授，天津工业大学纺织学部，天津市 300387 关静，博士，副研究员，军事医学科学院卫生装备研究所，天津市 300161
  • 作者简介:姚心培，女，1992年生，湖北省宜昌市人，汉族，2017年天津工业大学毕业，工程硕士，主要从事生物材料的止血性能研究。
  • 基金资助:

    国家自然科学基金(31570956)

Hemostatic property and cytotoxicity of the guanidine modified chitosan

Yao Xin-pei1, 2, Jing Miao-lei1, Guan Jing2 
  

  1. 1Department of Textile, Tianjin Polytechnic University, Tianjin 300387, China; 2Institute of Medical Equipment, Academy of Military Medical Sciences, Tianjin 300161, China
  • Received:2017-01-12 Online:2017-02-28 Published:2017-03-16
  • Contact: Jing Miao-lei, Master, Associate professor, Department of Textile, Tianjin Polytechnic University, Tianjin 300387, China Guan Jing, M.D., Associate researcher, Institute of Medical Equipment, Academy of Military Medical Sciences, Tianjin 300161, China
  • About author:Yao Xin-pei, Master, Department of Textile, Tianjin Polytechnic University, Tianjin 300387, China; Institute of Medical Equipment, Academy of Military Medical Sciences, Tianjin 300161, China
  • Supported by:

    the National Natural Science Foundation of China, No. 31570956

PDF

279

被引次数

2

摘要：

文章快速阅读：

 

文题释义：
体外凝血指标：活化部分凝血活酶时间、凝血酶原时间、凝血酶时间是常规的凝血检查项目，用于检验凝血系统出止血能力。通过对此3项指标测试，从而判断材料对内源性凝血途径、外源性凝血途径和共同途径的作用情况。
细胞毒性评价指标：MTT比色法是细胞毒性的常用检测方法，能反映活细胞的生长状况。采用MTT比色法计算出细胞增殖率，依据标准鉴定材料的毒性等级。为了更直观的反映材料作用后活死细胞数量，又采用荧光染色方法将活死细胞用绿色及红色荧光加以区分，充分说明了材料的细胞毒性水平。

背景：壳聚糖是一类具有良好止血、抗菌和吸附能力的生物材料，在其氨基处引入胍基基团能使其抗菌性和吸附性能提高，是化工分离的常用材料。然而，接枝胍基后对壳聚糖的凝血性能是否有影响，却鲜有研究。
目的：测试胍基壳聚糖的体外凝血性能及细胞毒性，探讨其作为止血材料的可能性。
方法：以壳聚糖和精氨酸为原料，NHS和EDC为偶联剂制备胍基壳聚糖粉末，通过全血凝固时间和3项体外凝血指标(活化部分凝血活酶时间、凝血酶原时间、凝血酶时间)对胍基壳聚糖的凝血能力进行初步评价。将胍基壳聚糖的浸提液与小鼠成纤维细胞共同培养，通过MTT比色法和荧光显微镜观察其对细胞生长状况的影响。
结果与结论：①胍基化壳聚糖缩短了全血凝固时间，具有一定的止血能力；②体外凝血指标结果表明，胍基化壳聚糖不通过传统的内外源性凝血途径发生作用，目前无确切研究证明胍基基团能促进凝血，提示该作用可能依赖于壳聚糖；③MTT检测结果显示，胍基壳聚糖细胞增殖率为87.8%，毒性反应为1级；④荧光染色照片显示活细胞数量很多，死细胞很少，说明胍基壳聚糖对细胞的毒性作用低。

关键词: 生物材料, 材料相容性, 胍基, 壳聚糖, 体外凝血, 细胞毒性, 国家自然科学基金

Abstract:

BACKGROUND: Chitosan is a kind of biomaterial with good hemostatic, antibacterial and absorption properties. The guanidine modified chitosan holding better antibacterial and absorption abilities is a commonly used chemical separation material. However, its coagulation property is still unclear.
OBJECTIVE: To evaluate the in vitro hemostatic property and cytotoxicity of the guanidine modified chitosan, and to explore its availability as a hemostatic material.
METHODS: The guanidine modified chitosan was synthesized with the raw materials of chitosan and arginine, and the coupling agents of EDC and NHS. The coagulation ability was evaluated preliminarily through the whole blood coagulation time, and three in vitro coagulation indexes (activated partial thromboplastin time, prothrombin time and thrombin time). The guanidine modified chitosan extracts were co-cultured with the mouse fibroblasts, and then the cell growth was observed by MTT assay and fluorescence microscope.
RESULTS AND CONCLUSION: The guanidine modified chitosan shortened the whole blood coagulation time, suggesting its hemostatic property. In vitro coagulation results indicated that the guanidine modified chitosan exerted the effect not through the traditional endogenous and extrinsic coagulation pathways. Guanidino group has not been proved to be hemostatic, so the chitosan may be the effective component. MTT assay showed that the cell proliferation rate was 87.8% and the toxicity level was grade 1. Fluorescent staining observed abundant live cells, and few dead cells, indicating that the guanidine modified chitosan exhibits low cytotoxicity. 

Key words: Guanidine, Chitosan, Hemostasis, Cell Proliferation, Tissue Engineering

中图分类号: