Development of three-dimensional semi-solid hydrogel matrices for ratiometric fluorescence sensing of Amyloid β peptide and imaging in SH-SY5 cells: Improvement of point of care diagnosis of Alzheimer's disease biomarker

Abstract

Alzheimer's is a neurodegenerative disease with high morbidity and mortality in the elderly, so, detection of its biomarker for definite diagnosis of Alzheimer's in the early stage of disease is a challenge. Amyloid beta peptide (Aβ) chosen as an Alzheimer's biomarker. Here, we developed novel, semi-solid, three-dimensional hydrogel matrices for ratiometric fluorescence detection of Aβ. This assay's great performance stems from the employment of a hybrid conjugate composed of Rhodamine B (RB), Carbon dots (CDs), and an Aβ probe entrapped in Polyvinyl alcohol (PVA), and then detection of fluorescence resonance energy transfer (FRET) that occurs in the presence of AuNP/target-Aβ, as a result of hybridization. The RB-CDs' fluorescence (at 582 nm and 675 nm under 430 nm excitation) is quenched in the presence of AuNPs, while the ratio of fluorescence (I₅₈₂/I₆₇₅) is increased by the addition of Aβ target, and shows a linear relationship in the range of 75 pM-250 nM, with a detection limit of 0.5 pM. Furthermore, the assay possesses strong selectivity for Aβ compared to other proteins, and different quantities of a human serum sample successfully analyzed with excellent sensitivity, satisfactory precision, and reliability. Due to distribution of Aβ in SH-SY5 human neuroblastoma cells, extending this UV-Vis-NIR full-range responsive CDs bio-probe to imaging of Aβ in cells. In both fixed and living SH-SY5 cells, the nanoprobe delivers a clear signal to the Aβ target. Because of its high sensitivity, selectivity, biocompatibility and affordability, this nanoprobe is a good option for early Alzheimer's disease diagnosis.

Keywords: Alzheimer's; Amyloid beta peptide (Aβ) biomarker; Hydrogel film; Multicolor bioimaging; Ratiometric fluorescence assay; SH-SY5 human neuroblastoma cells.