Endogenous Bioelectrical Modulation of Redox, Vascular Support, and Extracellular Matrix-Related Markers in Human Fibroblasts Following the REAC CO-IBZ Protocol
Sara Cruciani, Vania Fontani, Arianna Rinaldi, Giuseppe Garroni, Diletta Serra, Salvatore Rinaldi, Margherita MaioliBackground: Fibroblasts are active regulators of tissue architecture, extracellular matrix organization, vascular support, oxidative balance, and stress-related cellular responses. The present study aimed to characterize the molecular response of human foreskin fibroblasts (HFF1) following exposure to the standardized Radio Electric Asymmetric Conveyer (REAC) Circulatory Optimization—Inside Blue Zone (CO-IBZ) protocol. Methods: Cells were exposed to nine standardized CO-IBZ sessions. RT-qPCR showed significant upregulation of SIRT1, NOX4, and p16, whereas VEGF, p21, and HAS2 showed directional increases that did not reach statistical significance. Total antioxidant capacity, nitrite levels, and intracellular ROS showed small non-significant changes. Quantitative immunofluorescence analysis showed significant increases in NOX4, PARP1, and vWF fluorescence intensity, whereas SIRT1 and collagen type III exhibited directional, non-significant increases. Results: Taken together, the transcriptional and biochemical findings, together with quantitative immunofluorescence measurements of protein markers, define a CO-IBZ-associated molecular profile involving redox-regulatory, vascular-support-related, cell-cycle- and stress-response-related, and extracellular matrix-related markers. Conclusions: These results complement previous in vitro characterizations of the REAC ACT-IBZ and MO-IBZ protocols and provide a basis for further investigation in primary cells and more complex biological models.