羧甲基壳聚糖膜对人皮肤黑素细胞的生物相容性研究  

Biocompatibility of carboxymethyl chitosan membranes with human skin melanocytes

作　　者：孔玉龙[1] 王克玉[1] 张秀文[1] 马伟元[1] 

机构地区：[1]山东大学齐鲁医院皮肤科,济南250012

出　　处：《中华皮肤科杂志》2014年第11期793-795,共3页Chinese Journal of Dermatology

基　　金：山东省自然科学基金（Y2008C78）

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（4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.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 glutaraldehydebased 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.

关 键 词：黑素细胞 壳聚糖 膜 人工 生物相容性材料 

分 类 号：R318.08[医药卫生—生物医学工程]