The monitoring of some parameters, such as pressure loads, temperature, and glucose level in sweat on the plantar surface, is one of the most promising approaches for evaluating the health state of the diabetic foot and for preventing the onset of inflammatory events later degenerating in ulcerative lesions. This work presents the results of sensors microfabrication, experimental characterization and FEA-based thermal analysis of a 3D foot-insole model, aimed to advance in the development of a fully custom smart multisensory hardware–software monitoring platform for the diabetic foot. In this system, the simultaneous detection of temperature-, pressure- and sweat-based glucose level by means of full custom microfabricated sensors distributed on eight reading points of a smart insole will be possible, and the unit for data acquisition and wireless transmission will be fully integrated into the platform. Finite element analysis simulations, based on an accurate bioheat transfer model of the metabolic response of the foot tissue, demonstrated that subcutaneous inflamed lesions located up to the muscle layer, and ischemic damage located not below the reticular/fat layer, can be successfully detected. The microfabrication processes and preliminary results of functional characterization of flexible piezoelectric pressure sensors and glucose sensors are presented. Full custom pressure sensors generate an electric charge in the range 0–20 pC, proportional to the applied load in the range 0–4 N, with a figure of merit of 4.7 ± 1 GPa. The disposable glucose sensors exhibit a 0–6 mM (0–108 mg/dL) glucose concentration optimized linear response (for sweat-sensing), with a LOD of 3.27 µM (0.058 mg/dL) and a sensitivity of 21 µA/mM cm² in the PBS solution. The technical prerequisites and experimental sensing performances were assessed, as preliminary step before future integration into a second prototype, based on a full custom smart insole with enhanced sensing functionalities.

The dis(re)organization of the cytoskeletal actin in enterocytes mediates epithelial barrier dys(re)function, playing a key role in modulating epithelial monolayer’s integrity and remodeling under transition from physiological to pathological states. Here, by fluorescence-based morphological and morphometric analyses, we detected differential responses of cytoskeletal actin in intestinal epithelial Caco-2 cell monolayers at two different stages of their spontaneous differentiation, i.e., undifferentiated cells at 7 days post-seeding (dps) and differentiated enterocyte-like cells at 21 dps, upon challenge in vitro with the inflammation-mimicking stimulus of phorbol-12-myristate-13-acetate (PMA). In addition, specific responses were found in the presence of the natural dipeptide carnosine detecting its potential counteraction against PMA-induced cytoskeletal alterations and remodeling in differentiated Caco-2 monolayers. In such an experimental context, by both immunocytochemistry and Western blot assays in Caco-2 monolayers, we identified the expression of the allograft inflammatory factor 1 (AIF-1) as protein functionally related to both inflammatory and cytoskeletal pathways. In 21 dps monolayers, particularly, we detected variations of its intracellular localization associated with the inflammatory stimulus and its mRNA/protein increase associated with the differentiated 21 dps enterocyte-like monolayer compared to the undifferentiated cells.