RESUMO
A detailed understanding of the cell adhesion on polymeric surfaces is required to improve the performance of biomaterials. Quartz crystal microbalance with dissipation (QCM-D) as a surface-sensitive technique has the advantage of label-free and real-time monitoring of the cell-polymer interface, providing distinct signal patterns for cell-polymer interactions. In this study, QCM-D was used to monitor human fetal osteoblastic (hFOB) cell adhesion onto polycaprolactone (PCL) and chitosan (CH) homopolymer films as well as their blend films (75:25 and 25:75). Complementary cell culture assays were performed to verify the findings of QCM-D. The thin polymer films were successfully prepared by spin-coating, and relevant properties, i.e., surface morphology, ζ-potential, wettability, film swelling, and fibrinogen adsorption, were characterized. The adsorbed amount of fibrinogen decreased with an increasing percentage of chitosan in the films, which predominantly showed an inverse correlation with surface hydrophilicity. Similarly, the initial cell sedimentation after 1 h resulted in lesser cell deposition as the chitosan ratio increased in the film. Furthermore, the QCM-D signal patterns, which were measured on the homopolymer and blend films during the first 18 h of cell adhesion, also showed an influence of the different interfacial properties. Cells fully spread on pure PCL films and had elongated morphologies as monitored by fluorescence microscopy and scanning electron microscopy (SEM). Corresponding QCM-D signals showed the highest frequency drop and the highest dissipation. Blend films supported cell adhesion but with lower dissipation values than for the PCL film. This could be the result of a higher rigidity of the cell-blend interface because the cells do not pass to the next stages of spreading after secretion of their extracellular matrix (ECM) proteins. Variations in the QCM-D data, which were obtained at the blend films, could be attributed to differences in the morphology of the films. Pure chitosan films showed limited cell adhesion accompanied by low frequency drop and low dissipation.
RESUMO
The objective of the study is, for the first time, to construct a new near infrared (NIR) fluorophore, spectrophotometric, colorimetric, ratiometric, and turn-on probe (CSME) based on chromenylium cyanine platform decorated with methionine biomolecule to provide an efficient solution for critical shortcoming to be encountered for analysis of hazardous Hg2+ in environment and living cell. The CSME structure and its interaction with Hg2+ ion were evaluated by NMR, FTIR, MS, UV-Vis and fluorescence methods as well as Density Functional Theory (DFT) calculations. The none fluorescence CSME having spirolactam ring only interacted with Hg2+ in aqueous solution including competing ions. This interaction caused the fluorescence CSME with opened spirolactam form which exhibited spectral and colorimetric changes in the NIR region. The probe based on UV-Vis and fluorescence techniques respond in 90 s, has wide linear ranges (for UV-Vis: 6.29 × 10-8 - 1.86 × 10-4 M; for fluorescence: 9.49 × 10-9 - 1.13 × 10-5 M), and has a lower Limit of Detection (LOD) value (for fluorescence: 4.93 × 10-9 M, 0.99 ng/mL) than the value predicted by the US Environmental Protection Agency (EPA) organization. Hg2+ analysis was performed in drinking and tap water with low Relative Standard Deviation (RSD) values and high recovery. Smartphone and living cell applications were successfully performed for colorimetric sensing Hg2+ in real samples and 3T3 cells, respectively.
