上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2017, Vol. 37 ›› Issue (7): 885-.doi: 10.3969/j.issn.1674-8115.2017.07.001?

• 论著（基础研究） • 上一篇    下一篇

高压静电法结合冷冻干燥法和离子交联法制备壳聚糖 - 明胶多孔微球

黄芳 1，芮文斌 1，徐丹枫 1，祝宇 1，沈柏用 2，彭承宏 2   

  1. 上海交通大学  医学院附属瑞金医院  1. 泌尿外科， 2. 普外科，上海 200025
  • 出版日期:2017-07-28 发布日期:2017-08-25
  • 通讯作者: 彭承宏，电子信箱：chhpeng@188.com
  • 作者简介:?黄芳（1984—），女，主治医师，博士；电子信箱：fangteng1130@163.com
  • 基金资助:
    国家高新技术研究发展计划（“863”计划）（2008AA02Z417）；国家自然科学基金（30672043，31170938）

Preparation of chitosan-gelatin porous microspheres by high voltage electrostatic method combined with freeze-drying and ionic cross-linking method

HUANG Fang1, RUI Wen-bin1, XU Dan-feng1, ZHU Yu1, SHEN Bai-yong2, PENG Cheng-hong2   

  1. 1. Department of Urology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; 2. Department of General Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2017-07-28 Published:2017-08-25
  • Supported by:
    National High-tech R&D Program of China, “863” Program, 2008AA02Z417; National Natural Science Foundation of China, 30672043, 31170938

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摘要： 目的 · 应用高压静电法结合冷冻干燥（冻干）法和离子交联法制备壳聚糖 – 明胶多孔微球，探讨微球多孔结构形成的影响因 素。方法 · 以三聚磷酸钠（STPP）为交联剂，利用高压静电法结合冻干法和离子交联法制备具有多孔结构的壳聚糖多孔微球和壳聚 糖 – 明胶多孔微球，比较壳聚糖 – 明胶不同体积比、冷冻温度、饱和 STPP 的 85% 乙醇溶液交联固化时间对微球多孔结构的影响，光 镜、扫描电镜、苏木精 - 伊红染色切片观察壳聚糖多孔微球和壳聚糖 – 明胶多孔微球的形态、表面及内部结构、粒径。结果 · 高压静 电结合冻干法和离子交联法制备的微球，球形良好，球体表面及内部均有较多微孔，表面与内部孔径相通。饱和 STPP 的 85% 乙醇溶 液交联固化后的壳聚糖 – 明胶微球结构能在中性溶液中稳定存在，冷冻、冻干与交联反应的先后顺序，交联时间及二次冷冻温度影响 多孔微球的孔径。明胶可以和壳聚糖形成聚电解质复合物，也可以作为致孔剂参与多孔结构的形成。结论 · 高压静电法结合冻干法和 离子交联法制备的壳聚糖 – 明胶多孔微球具有较大的孔径（100 ～ 200 µm），适合细胞的生长及迁移。

关键词: 多孔微球, 壳聚糖, 三维培养, 组织工程

Abstract:

Objective · To prepare chitosan-gelatin porous microspheres by high voltage electrostatic method combined with freeze-drying and ionic cross-linking method and investigate the factors that influence the formation of porous medium.  Methods · Porous chitosan microspheres and chitosangelatin porous microspheres were prepared using high voltage electrostatic method combined with freeze-drying and ionic cross-linking method, with sodium tripolyphosphate (STPP) as crosslinking agent. Factors that affect the porous structure and pore size of porous microspheres were compared, such as different chitosan-gelatin ratio, freezing temperature, curing time with saturated STPP in 85% ethanol solution. The morphology, surface and internal structure, particle size of the porous chitosan microspheres and chitosan-gelatin porous microspheres were observe by using light microscope, scanning electron microscope and hematoxylin-eosin staining.  Results · Microspheres prepared by freeze-drying with an electrostatic and ionic cross-linking method have open, interconnected and highly macroporous, with good spherical surface. Saturated STPP ethanol solution (85% ethanol) was chosen as the crosslinking agent to prevent destruction of the porous structure. The order of freeze and crosslinking, cross-linking time and the second freezing temperature, can influence the pore size of porous microspheres. Gelatin and chitosan can form polyelectrolyte complexes, and can also be used as porogen in porous structure.  Conclusion · The preparation of porous chitosan-gelatin microspheres via this method has a large pore size (diameter 100-200 µm), suitable for cell growth and the migration.

Key words:  porous microspere, chitosan, three-dimensional culture, tissue engineering