20.500.12556/RUL-182020 Characterization of diabetes-related biomechanical and structural alterations in human deep fasciae using atomic force microscopy and multimodal multiphoton imaging Background: Type 2 diabetes mellitus (T2DM) is associated with connective-tissue complications, but effects on human deep fascia remain poorly understood. This study employed atomic force microscopy (AFM) nanoindentation and multiphoton imaging to quantify region-specific biomechanical and structural changes of deep fasciae in T2DM. Methods: Fascia lata, thoracolumbar fascia, and plantar fascia from 17 T2DM and 17 control male deceased (<24 h post-mortem) donors were analyzed. AFM was used to quantify nanoscale stiffness (Young’s modulus) under hydrated conditions. Label-free multiphoton microscopy comprising Second Harmonic Generation (SHG), Two-Photon Excited Fluorescence (TPEF), and Fluorescence Lifetime Imaging Microscopy (FLIM) was used to assess collagen organization, autofluorescence, and fluorescence decay kinetics. Results: AFM revealed significantly increased stiffness in diabetic plantar fascia (87 ± 3.1 vs 40 ± 1.2 kPa, p = 0.004) and fascia lata (28 ± 1.2 vs 16 ± 0.5 kPa, p < 0.001) compared to controls, while thoracolumbar fascia stiffness did not differ significantly between groups. Stiffness changes showed no significant association with age or body mass index. SHG forward-to-backward ratios did not differ significantly between groups, suggesting preserved collagen supramolecular organization. However, TPEF ratios were significantly elevated in diabetic plantar fascia and fascia lata, suggesting increased accumulation of autofluorescent species. FLIM revealed a significant decrease in mean fluorescence lifetime (τₘ) in diabetic plantar fascia, indicative of altered collagen microenvironment, possibly due to glycation or oxidative modifications. Conclusion: T2DM induces site-specific biomechanical stiffening and biochemical remodeling of human deep fascia, particularly in load-bearing regions. These changes suggest mechanisms involving glycation and oxidative stress, providing mechanistic insights into musculoskeletal complications associated with diabetic fasciopathy. advanced glycation end-products atomic force microscopy deep fascia extracellular matrix remodeling multiphoton imaging type 2 diabetes mellitus napredni končni produkti glikacije mikroskopija atomskih sil globoka fascija preoblikovanje zunajceličnega matriksa večfotonsko slikanje sladkorna bolezen tipa 2 true false true Angleški jezik Slovenski jezik Članek v reviji 2026-04-22 13:19:15 2026-04-22 13:19:26 2026-04-24 15:19:42 0000-00-00 00:00:00 2026 0 0 11 str. Vol. 98, art. 103209 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616 1532-3072 10.1016/j.tice.2025.103209 258042627 RAZ_Ugwoke_Chiedozie_Kenneth_2025.pdf RAZ_Ugwoke_Chiedozie_Kenneth_2025.pdf 1 38E4054D51713179EDD312AFA25A7C9C d15db775cf10964cb0ed26cf2d45f8d2e027191c4ba41c052b271588d5e93a4c accea5f9-3e3d-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=232910 https://www.sciencedirect.com/science/article/pii/S0040816625004914 0 e22b0366-3e3c-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=232909 Medicinska fakulteta Biotehniška fakulteta 0 0 0