中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (11): 793-795.

• 论著 • 上一篇    下一篇

羧甲基壳聚糖膜对人皮肤黑素细胞的生物相容性研究

孔玉龙1,王克玉2,张秀文3,马伟元1   

  1. 1. 山东大学齐鲁医院皮肤科
    2. 济南市山东大学齐鲁医院皮肤科
    3. 山东大学齐鲁医院
  • 收稿日期:2014-02-14 修回日期:2014-07-07 出版日期:2014-11-15 发布日期:2014-11-01
  • 通讯作者: 王克玉 E-mail:wangkeyu@medmail.com.cn
  • 基金资助:
    山东省自然科学基金

Biocompatibility of carboxymethyl chitosan membranes with human skin melanocytes

  • Received:2014-02-14 Revised:2014-07-07 Online:2014-11-15 Published:2014-11-01

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摘要： 目的 探讨羧甲基壳聚糖膜对健康人黑素细胞的生物相容性，探索羧甲基壳聚糖膜搭载黑素细胞的可行性。 方法 使用流延法联合戊二醛交联法制备羧甲基壳聚糖膜，黑素细胞分离培养技术分离培养健康人黑素细胞，使用HMB45染色、MTT法、NaOH裂解法、酪氨酸酶活性检测等方法检测羧甲基壳聚糖膜对黑素细胞功能的影响。 结果 倒置显微镜下观察，黑素细胞在羧甲基壳聚糖膜上分布均匀，形态正常。免疫组化染色显示，贴附于羧甲基壳聚糖膜的黑素细胞抗HMB45单克隆抗体染色阳性。利用MTT法测定的黑素细胞增殖曲线显示，羧甲基壳聚糖可以支持黑素细胞的正常生长，黑素合成量（A值为0.083 ± 0.015）与培养皿所培养的黑素细胞（A值0.066 ± 0.008）比较，差异无统计学意义（t = 2.38，P > 0.01）；黑素细胞酪氨酸酶活性（A值0.234 ± 0.083）与培养皿培养的黑素细胞（A值0.241 ± 0.061）比较，差异无统计学意义（t = 0.23，P > 0.05）。 结论 羧甲基壳聚糖膜可以维持黑素细胞的正常功能，具有较好的生物相容性。

关键词: 黑素细胞, 壳聚糖, 膜，人工, 生物相容性材料

Abstract: Kong Yulong, Wang Keyu, Zhang Xiuwen, Ma Weiyuan. Department of Dermatology, Qilu Hospital of Shandong University, Jinan 250012, China Corresponding author: Wang Keyu, Email: wangkeyu@medmail.com.cn 【Abstract】 Objective To study the biocompatibility of carboxymethyl chitosan （CMCS） membrane with melanocytes from healthy human skin, and to investigate the feasibility to transport and carry melanocytes by using CMCS membrane. Methods CMCS membrane was prepared by a casting method combined with a glutaraldehyde-based cross-linking method. Melanocytes were isolated from the foreskin of healthy men, and subjected to primary culture and subculture. The third-passage melanocytes were classified into two groups to be cultured on the CMCS membrane （test group） or traditional culture plates （control group）. Methyl thiazolyl tetrazolium （MTT） assay was performed to evaluate the proliferative activity of melanocytes, and a sodium hydroxide-based lysis method to determine melanin content. HMB45 staining was conducted, and tyrosinase activity was estimated for melanocytes. Results Inverted microscopy showed that melanocytes were evenly distributed on the CMCS membrane with a normal shape. The melanocytes adherent to the CMCS membrane stained positive for anti-HMB45 monoclonal antibody. The growth curve of the melanocytes on the CMCS membrane, which was obtained from MTT assay, demonstrated that CMCS membrane could support the normal growth of melanocytes. No significant difference was observed between the test group and control group in melanin content （0.083 ± 0.015 vs. 0.066 ± 0.008, t = 2.38, P > 0.01） or tyrosinase activity （0.234 ± 0.083 vs. 0.241 ± 0.061, t = 0.23, P > 0.05）. Conclusion CMCS membrane can maintain the normal biological activity of melanocytes and have good biocompatibility with skin melanocytes.

Key words: Melanocytes, Chitosan, Membranes, artificial, Biocompatible materials

中图分类号: 

  • R751