首页|负载单宁酸的双层壳聚糖屏障膜的制备及其生物相容性的研究

负载单宁酸的双层壳聚糖屏障膜的制备及其生物相容性的研究

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目的 制备负载单宁酸的双层壳聚糖膜(CS@TA),测试其活性氧(ROS)清除能力、生物相容性及抗菌性能,研究其作为引导骨再生屏障膜的可行性.方法 采用自蒸发技术先制备出致密的单层壳聚糖(CS)膜,随后采用蒸发干燥、取向冷冻和冷冻干燥技术在单层CS膜上制备出多孔的CS层,从而得到双层CS膜,利用扫描电镜观察其微观结构.将制备好的CS双层膜先接枝 4-羧基苯硼酸形成中间体,按不同比例接枝单宁酸(TA),使用傅里叶红外光谱仪(FITR)分析TA与CS双层膜的相互作用.通过1,1-二苯基-2 苦基肼(DPPH)测试 ROS 清除能力,选取负载 TA 后 ROS 清除效率在 90%以上的双层膜进行后续体外细胞实验,利用CCK-8 及活死细胞染色测试其生物相容性,利用扫描电镜观察 MC3T3-E1 细胞在双层膜多孔面上的黏附情况,通过菌落计数测试其对金黄色葡萄球菌(E.coli)和大肠埃希菌(S.aureus)的抗菌性能.结果 CS@TA 双层膜一面光滑致密,另一面粗糙疏松多孔,截面呈垂直薄膜方向的有序多孔结构,随着TA的加入,双层膜的ROS清除能力先迅速增加后缓慢稳定,CCK-8 及活死细胞染色结果显示加入过多的 TA会明显影响双层膜的生物相容性,细菌稀释涂板计数结果显示加入适量TA的双层膜与未负载TA的双层膜相比,对E.coli和S.aureus具有一定抗菌能力.结论 负载适量TA的双层膜具有较强的 ROS 清除能力,良好的生物学性能,且对E.coli和S.aureus均具有一定的抗菌能力.
Preparation and biocompatibility of a bilayer chitosan barrier membrane loaded with tannic acid
Objective To explore the feasibility of the bilayer chitosan barrier membrane loaded with tannic acid(CS@TA)for guided bone regeneration by exploring the reactive oxygen species(ROS)scavenging ability,bio-compatibility,and antibacterial properties.Methods The single-layer chitosan(CS)film was prepared by self-e-vaporation,and the double-layer CS film was prepared by directional freezing and freeze-drying,and its microstruc-ture was observed by scanning electron microscope.The prepared CS bilayer membrane was grafted with tannic acid(TA)in different proportions,and the interaction between TA and CS bilayer membrane was analyzed by Fourier infrared spectrometer(FITR).The ROS scavenging ability was tested by 1,1-diphenyl-2-picrylhydrazyl(DPPH),and the double-layer membrane of loading TA scavenging efficiency of more than 90%was selected to continue the follow-up experiment.CCK-8 assay and lived dead staining were used to evaluate the survival rate of cell in each groups.MC3T3-E1 cells was adhesion the CS@TA barrier film for studying by SEM.Colony counting was per-formed to test its antibacterial performance against Escherichia coli(E.coli)and Staphylococcus aureus(S.au-reus).Results One side was the smooth,dense while other side was rough,loose and porous,with a longitudinal ordered porous structure in cross-section of the double-layer membrane of CS@TA.With the addition of TA,the ROS scavenging ability of the double-layer membrane first increased rapidly and then stabilized slowly.The results of CCK-8 and lived and dead cells staining showed that excessive TA addition significantly affected the biocompati-bility of the double-layer membrane.The counting results of bacterial dilution coating showed that compared with the double-layer membrane without TA loading,the double-layer membrane had certain antibacterial ability against E.coli and S.aureus when the appropriate amount of TA was added.Conclusion Thus the double-layer with ap-propriate TA loading has strong ROS scavenging ability,good biological performance,and certain antibacterial abil-ity for E.coli and S.aureus.

tannic acidchitosanreactive oxygen speciesantibacterialbarrier membranedirectional freezing

杨肖念、邹多宏

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安徽医科大学口腔医学院,安徽医科大学附属口腔医院,安徽省口腔疾病研究重点实验室,合肥 230032

上海交通大学医学院附属第九人民医院,上海交通大学口腔医学院,上海市口腔医学重点实验室,上海 200011

单宁酸 壳聚糖 活性氧 抗菌 屏障膜 取向冷冻

国家自然科学基金

32171347

2024

安徽医科大学学报
安徽医科大学

安徽医科大学学报

CSTPCD北大核心
影响因子:1.095
ISSN:1000-1492
年,卷(期):2024.59(4)
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