Department of Biology, Hacettepe University, Ankara, Turkey.
Department of Chemistry, Hacettepe University, Ankara, Turkey.
J Biomater Sci Polym Ed. 2021 Jun;32(8):1024-1039. doi: 10.1080/09205063.2021.1892472. Epub 2021 Mar 11.
The development of novel biocompatible and cost effective cryogel membrane which shows enhanced antimicrobial properties in order to use for several approaches such as wound dressing, scaffold or food packaging was aimed in this study. A super macro porous lysozyme imprinted cryogel membranes showing antibacterial effect against both Gram-positive and Gram-negative bacteria were prepared by using molecular imprinting technique. N-methacryloyl-(L)-histidine methyl ester (MAH) was used as the pseudo specific ligand and complexed with Cu in order to provide metal ion coordination between MAH and template molecule (lysozyme). Comparing the antibacterial activity of different lysozyme concentrations, cryogel membranes were prepared in three different concentrations. To synthesize Poly (hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methylester) P(HEMA-MAH) cryogel membrane, free radical polymerization initiated by N, N, N', N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) was carried out at -12 °C. The characterization of the lysozyme imprinted cryogel membrane was accomplished by using scanning electron microscopy (SEM), swelling degree measurements and Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) spectroscopy. The cytotoxicity test of produced membrane was performed by using mouse fibroblast cell line L929. The antibacterial activity of P(HEMA-MAH) lysozyme molecular imprinted [P(HEMA-MAH) Lyz-MIP] cryogel membranes against () and () were determined by Kirby-Bauer membranes diffusion and viable cell counting methods. When the antibacterial effect of P(HEMA-MAH) Lyz-MIP cryogel membranes were evaluated, it was found that P(HEMA-MAH) Lyz-MIP cryogel membranes had stronger antibacterial effects against Gram-negative bacteria even in low lysozyme concentrations. In addition, 100% bacterial inhibition was detected for both of two bacteria at increasing lysozyme concentrations.
本研究旨在开发新型生物相容性、成本效益高的冷冻凝胶膜,使其具有增强的抗菌性能,可用于多种方法,如伤口敷料、支架或食品包装。通过使用分子印迹技术,制备了具有超微孔结构的溶菌酶印迹冷冻凝胶膜,对革兰氏阳性菌和革兰氏阴性菌均具有抗菌作用。N-甲基丙烯酰基-(L)-组氨酸甲酯(MAH)用作伪配体,并与 Cu 络合,以提供 MAH 与模板分子(溶菌酶)之间的金属离子配位。比较不同溶菌酶浓度的抗菌活性,制备了三种不同浓度的冷冻凝胶膜。为了合成聚(羟乙基甲基丙烯酸酯-N-甲基丙烯酰基-(L)-组氨酸甲酯)P(HEMA-MAH)冷冻凝胶膜,在-12°C 下通过 N, N, N', N'-四甲基乙二胺(TEMED)和过硫酸铵(APS)引发自由基聚合。通过扫描电子显微镜(SEM)、溶胀度测量和傅里叶变换红外光谱-衰减全反射(FTIR-ATR)光谱对溶菌酶印迹冷冻凝胶膜进行了表征。通过使用小鼠成纤维细胞系 L929 进行了膜的细胞毒性测试。通过 Kirby-Bauer 膜扩散法和活菌计数法测定了 P(HEMA-MAH)溶菌酶分子印迹[P(HEMA-MAH)Lyz-MIP]冷冻凝胶膜对()和()的抗菌活性。当评估 P(HEMA-MAH)Lyz-MIP 冷冻凝胶膜的抗菌效果时,发现即使在低溶菌酶浓度下,P(HEMA-MAH)Lyz-MIP 冷冻凝胶膜对革兰氏阴性菌也具有更强的抗菌作用。此外,在增加溶菌酶浓度时,两种细菌的抑菌率均达到 100%。