Adenosina-loaded chitosan nanoparticles improve mitochondrial membrane potential in chronic -adrenergic stimulated cardiac myoblasts

DESIDERIO JC; GODOY COTO J; CIANCIO C; BOGGAN SIMAL G; ENNIS IL

Congreso; Reunión anual de la Sociedad Argentina de Fisiología (SAFIS); 2023

ADENOSINE-LOADED CHITOSAN NANOPARTICLES IMPROVE MITOCHONDRIAL MEMBRANE POTENTIAL IN CHRONIC ßADRENERGIC STIMULATED CARDIAC MYOBLASTS (R90) Desiderio J1, Godoy Coto J1, Ciancio MC1, Boggan Simal G2, Ennis IL1 1 Centro de Investigaciones Cardiovasculares ?Dr. Horacio E. Cingolani?, Facultad de Ciencias Médicas, UNLP ? CONICET. 2 Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas (INIFTA), UNLP ? CONICET. Introduction: Sustained ß-adrenergic overstimulation impairs both function and structure in cardiomyocytes, ultimately leading to cardiac failure. Chitosan is obtained from crustaceans and is useful for biomedical uses due to its capability to form nanoparticles that can transport drugs within. Adenosine is a widely used cardioprotective drug, but its short half-life is a limiting factor for long-term treatments. Objective: To develop an efficient adenosine drug delivery system and explore if adenosine-loaded nanoparticles can prevent mitochondrial damage produced by ß-adrenergic chronic overstimulation in H9c2 cells. Methods: Blank and adenosine-loaded chitosan nanoparticles (B-CNP and A-CNP) were synthesized by ionic gelation. Their size and entrapment efficiency was assessed. Spectrophotometry was used to obtain chitosan (CS), adenosine and CNP absorbance spectra. Mitochondrial membrane potential was assessed with TMRE probe. H9c2 cells were cultured for 48 h with 10 mM Isoproterenol (ISO). 24 h prior experiments cells were treated with adenosine 10 μM (Ade), B-CNP or A-CNP. Results are shown as mean ± SEM (n) and considered statistically different, otherwise p-value is stated. Normality was assessed and t-tests or two-way ANOVA were carried out. Results: CS absorbance spectra showed a linear behavior at 330nm (r2=0.972), wavelength used to assess synthesis CS yield. CNP showed a similar spectra and behavior at 330 nm. Adenosine absorbance spectra showed a peak ~260nm and linear behavior in that wavelength (r2=0.974). Hydrodynamic radius of B-CNP and A-CNP were similar (d.nm, B-CNP: 302.3±77.8 (5), A-CNP: 342.6±57.8 (4)). Entrapment efficiency (%) of A-CNP was in 42.91±5.80 (8). FCCP and ISO treatment depolarized H9c2 mitochondria (%C, FCCP: 76.6±4.7 (4), ISO: 85.4±4.6 (12)). A-CNP treatment prevented mitochondrial depolarization both in FCCP and ISO treatments (%C, FCCP: 84.2±5.7 (2), ISO: 106.0±10.2 (3)). Conclusion: We developed a system to quantify both CS synthesis yield and adenosine loading in the CNP. Adenosine loading did not change CNP size. A-CNP may protect the mitochondria against chemically depolarization and ß-adrenergic chronic overstimulation. Further experiments are needed to elucidate the exact mechanisms involved and other potential beneficial effects. TOPIC AREA: CARDIOVASCULAR PHYSIOLOGY-HYPERTENSION