Oxidized Alginate Dopamine Conjugate: A Study to Gain Insight into Cell/Particle Interactions

Abstract

We had previously synthetized a macromolecular prodrug consisting of oxidized Alginate and dopamine (AlgOx-Da) for a potential application in Parkinson disease (PD). Methods: In the present work, we aimed at gaining an insight into the interactions occurring between AlgOx-Da and SH-SY5Y neuronal cell lines in view of further studies oriented towards PD treatment. With the scope of ascertaining changes in the external and internal structure of the cells, multiple methodologies were adopted. Firstly, fluorescently labeled AlgOx-Da conjugate was synthetized in the presence of fluorescein 5(6)-isothiocyanate (FITC), providing FITC-AlgOx-Da, which did not alter SH-SY5Y cell viability according to the sulforhodamine B test. Furthermore, the uptake of FITC-AlgOx-Da by the SH-SY5Y cells was studied using scanning near-field optical microscopy and assessments of cell morphology over time were carried out using atomic force microscopy. Results: Notably, the AFM methodology confirmed that no relevant damage occurred to the neuronal cells. Regarding the effects of DA on the intracellular reactive oxygen species (ROS) production, AlgOx-Da reduced them in comparison to free DA, while AlgOx did almost not influence ROS production. Conclusions: these findings seem promising for designing in vivo studies aiming at administering Oxidized Alginate Dopamine Conjugate for PD treatment.

Atomic force microscopy in disease‐related studies: Exploring tissue and cell mechanics

“…For instance, coupling AFM with elastic resonant cavity interferometric stress microscopy (ERISM) permits quantification of substrate vertical deformation during indentation (Rheinlaender et al, 2020). Integrating AFM with a high‐resolution fluorescence imaging device enables the detection of morphological changes and fluorescence intensity in neuronal cells during the phagocytosis of fluorescently labeled Parkinson's disease therapeutic drugs (Trapani et al, 2022). The observation of minimal changes in cell height during phagocytosis suggests that the substance does not significantly alter the structural morphology of cells at the cytoskeletal level.…”