CAJ | 학술논문

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。
배경：재이패점단백창면수복부료적체외세포독성검측중，유우단백분자표면대정전하，1∶9적침제비회사세포취단이도치측정산생오차，영향측정결과。 　　목적：재이유표준적기출상，근거이패점단백특수적성질급사용상태，개진이패점단백창면수복부료적침제비례혹전처리방법。 　　방법：①침제액법：장이패점단백창면수복부료여세포배양액분별이1∶9、1∶131침제비례제비침제액，분별이이패점단백창면수복부료침제액、천연유효침제액、고밀도취을희침제액급세포배양액배양L929소서성섬유세포。②직접접촉법：분별이증류수、이패점단백창면수복부료용액、이갑기아풍급세포배양액배양L929소서성섬유세포。 　　결과여결론：채용침제비1∶9측정양품체외세포독성시，세포산생취단작용，불괄용우양품독성적검측；조정침제비위1∶131후，서응작용화세포취단현상명현강저，제고료검측결과적가신도，현시양품무세포독성。직접접촉법현시양품무세포독성。채용경조정과적침제액법혹직접접촉법균가괄용우이패점단백창면수복부료체외세포독성적검측。
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.