贻贝粘蛋白创面修复敷料体外细胞毒性的检测方法  

Cytotoxicity tests for the mussel adhesive protein dressing for wound healing

作　　者：刘茜[1,2] 兰华林 顾铭[4] 高敏 王召旭[1] 

机构地区：[1]中国食品药品检定研究院,生物材料与组织工程室,北京市100050 [2]江苏省医疗器械检验所,生物安全评价中心,江苏省南京市210022 [3]重庆大学生物工程学院,教育部生物流变科学与技术重点实验室,重庆市400044 [4]中国科学院过程工程研究所,生化工程国家重点实验室,北京市100190

出　　处：《中国组织工程研究》2013年第38期6785-6790,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家科技部项目(2011BAZ03243)~~

摘　　要：背景:在贻贝粘蛋白创面修复敷料的体外细胞毒性检测中,由于蛋白分子表面带正电荷,1∶9的浸提比会使细胞聚团而导致测定产生误差,影响测定结果。目的:在已有标准的基础上,根据贻贝粘蛋白特殊的性质及使用状态,改进贻贝粘蛋白创面修复敷料的浸提比例或前处理方法。方法:①浸提液法:将贻贝粘蛋白创面修复敷料与细胞培养液分别以1∶9、1∶131浸提比例制备浸提液,分别以贻贝粘蛋白创面修复敷料浸提液、天然乳胶浸提液、高密度聚乙烯浸提液及细胞培养液培养L929小鼠成纤维细胞。②直接接触法:分别以蒸馏水、贻贝粘蛋白创面修复敷料溶液、二甲基亚砜及细胞培养液培养L929小鼠成纤维细胞。结果与结论:采用浸提比1∶9测定样品体外细胞毒性时,细胞产生聚团作用,不适用于样品毒性的检测;调整浸提比为1∶131后,絮凝作用和细胞聚团现象明显降低,提高了检测结果的可信度,显示样品无细胞毒性。直接接触法显示样品无细胞毒性。采用经调整过的浸提液法或直接接触法均可适用于贻贝粘蛋白创面修复敷料体外细胞毒性的检测。BACKGROUND：In the cytotoxicity test of the mussel adhesive protein dressing for wound healing, because of positive charge properties of the protein, when extracting ratio is 1：9, the cel s exhibit poly-group phenomenon that results in errors in the cytotoxicity test of mussel adhesive protein samples. OBJECTIVE：According to the existing standards, to improve the leaching proportion and pretreatment of mussel adhesive protein dressing for wound healing based on the special properties and working condition of mussel adhesive protein. METHODS：（1） Extract method：Extract solution of mussel adhesive protein dressing was prepared with mussel adhesive protein dressing and cel medium at extracting ratios of 1：9 and 1：131. Then, L929 cel s were cultured in extract solutions of mussel adhesive protein dressing, natural latex and high-density polyethylene, respectively. （2） Method of direct contact：Distil ed water, solutions of mussel adhesive protein dressing, dimethyl sulfoxide and cel medium were used to culture L929 cel s. RESULTS AND CONCLUSION：When the extracting ratio was 1：9, the cel s agglomerated which is not suitable for cytotoxicity test. When the extracting ratio was 1：131, flocculated sediment and cel aggregation disappeared, the cytotoxicity test results showed no cytotoxicity with higher reliability. Direct contact method showed the samples had no cytotoxicity. The extract method with adjusted extracting radio or direct contact method can be applied to test the in vitro cytotoxicity of mussel adhesive protein dressing for wound healing.

关 键 词：粘蛋白类 生物相容性材料 生物敷料 细胞毒性 免疫 

分 类 号：R318[医药卫生—生物医学工程]