Department of Operative Dentistry and Periodontology, University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany.
Clin Oral Investig. 2011 Feb;15(1):65-73. doi: 10.1007/s00784-009-0357-2. Epub 2009 Dec 5.
Mouthwashes containing protective enzymes are required especially for patients suffering from xerostomia. The present study aimed to investigate the possibilities of modulating the immobilisation of lysozyme in the in situ pellicle layer. In situ formed pellicles were incubated in vitro for 10 min with various enzymatic buffer solutions containing lysozyme and additive enzymes such as transglutaminase or trypsin as well as polyphenolic compounds (cistus tea). After the rinses, the pellicle samples were incubated in collected whole saliva or in desorption solutions for 0, 20 and 40 min and the enzyme activities were measured. Furthermore, accumulation of lysozyme in the pellicle was visualised in ultrathin sections of the pellicle using the gold immunolabelling technique and transmission electron microscopy. Hen egg white lysozyme was accumulated in the in situ pellicle tenaciously. Up to 2.8-fold higher activities than in controls were observed. The addition of transglutaminase did not enhance the immobilisation of lysozyme activity, whereas the polyphenolic compound had no adverse effect. Accumulation of lysozyme in the acquired pellicle was confirmed by gold immunolabelling. Targeted and tenacious immobilisation of lysozyme in the acquired pellicle is possible. Poylphenolic compounds might be a relevant additive for mouthwashes containing lysozyme.
含有保护酶的漱口水特别适用于口干症患者。本研究旨在探讨调节溶菌酶在原位黏膜层固定化的可能性。原位形成的黏膜在体外用含有溶菌酶和添加酶(如转谷氨酰胺酶或胰蛋白酶)以及多酚化合物(乳香茶)的各种酶缓冲液孵育 10 分钟。漱口后,将黏膜样本在收集的全唾液或解吸溶液中孵育 0、20 和 40 分钟,并测量酶活性。此外,使用金免疫标记技术和透射电子显微镜在黏膜的超薄切片中可视化溶菌酶在黏膜中的积累。鸡蛋白溶菌酶在原位黏膜中牢固地积累。与对照组相比,观察到高达 2.8 倍的活性增加。转谷氨酰胺酶的添加并没有增强溶菌酶活性的固定化,而多酚化合物没有不良影响。金免疫标记证实了溶菌酶在获得性黏膜中的积累。溶菌酶在获得性黏膜中的靶向和牢固固定化是可能的。多酚化合物可能是含溶菌酶漱口水的相关添加剂。