Biomimetic hydrogels have garnered increased interest due to their considerable potential for use in various fields, such as tissue engineering, 3D cell cultivation, and drug delivery. The primary challenge for applying hydrogels in tissue engineering is accurately evaluating their mechanical characteristics. In this context, we propose a method using scanning ion conductance microscopy (SICM) to determine the rigidity of living human breast cancer cells MCF-7 cells grown on a soft, self-assembled Fmoc-FF peptide hydrogel. Moreover, it is demonstrated that the map of Young’s modulus distribution obtained by the SICM method allows for determining the core location. The Young’s modules for MCF-7 cells decrease with the substrate stiffening, with values of 1050 Pa, 835 Pa, and 600 Pa measured on a Petri dish, Fmoc-FF hydrogel, and Fmoc-FF/chitosan hydrogel, respectively. A comparative analysis of the SICM results and the data obtained by atomic force microscopy was in good agreement, allowing for the use of a composite cell–substrate model (CoCS) to evaluate the ‘soft substrate effect’. Using the CoCS model allowed us to conclude that the MCF-7 softening was due to the cells’ mechanical properties variations due to cytoskeletal changes. This research provides immediate insights into changes in cell mechanical properties resulting from different soft scaffold substrates.

Calcium is a key macroelement involved in a range of physiological processes in the body, and its concentration in blood is an important diagnostic indicator in various diseases. This work presents a novel rapid method for the point-of-care determination of total calcium content in patient blood, by applying a drop of capillary blood from a finger onto a hydrogel. Gelatin hydrogel, modified with an optical sensor for calcium, Arsenazo III, was used as a platform for the separation of blood into plasma and erythrocytes. A comparative analysis of various types of hydrogel materials (polyacrylamide, PVA, Fmoc-FF, carbomer, carbopol, gelatin) was performed, demonstrating that among the studied systems, only gelatin hydrogel is suitable as a platform for the determination of calcium in blood plasma. The binding of calcium ions from blood plasma with the calcium sensor embedded in the hydrogel leads to a change in the absorption spectrum of the system, enabling photometric determination of calcium concentrations below and above the normal range in blood plasma. Therefore, this rapid assay allows monitoring of calcium metabolism disorders in the human organism. The method is characterized by its speed, simplicity of sample preparation, and potential for integration into clinical practice.