Multi-factor optimization and interaction analysis for enhanced nanobubble formulation and performance of Dasatinib

The objective of this study was to develop and optimize PLGA nanobubbles as a delivery system for dasatinib. In this study, we developed nanobubbles with a perfluoropentane core and a PLGA outer shell, optimizing their properties using a three-level Box-Behnken design. We thoroughly examined the physical and chemical attributes of dasatinib-loaded nanobubbles, including size, surface charge, and structure, and assessed drug encapsulation and release, as well as investigating cell uptake and antitumor effectiveness. The encapsulation efficiency, particle size, and zeta potential of the nanobubbles varied from 15.12 % to 69.23 %, 96.78 nm-767.72 nm, and-38.56 mV to-12.34 mV, respectively, in 29 trials using a Box-Behnken design. Employing Response Surface Methodology, we optimized the formulation, resulting in an ideal combination: 160.35 mg dasatinib, 500 mg PLGA, 2.0 % w/v PVA, and stirring at 6000 rpm, achieving a high global desirability value of 0.971. These nanobubbles demonstrated an encapsulation efficiency of 80.12 % and a loading capacity of 24.34 %, with a low viscosity of 6.1 cP. Nanobubbles showed notable differences in drug release with and without ultrasound. Under sonication, 59.42 % of dasatinib was released in 6 h compared to 35.34 % without sonication, and after 24 h, 76.89 % was released without ultrasound, while nearly 99.58 % was released with ultrasound. Stability tests at different temperatures showed consistent encapsulation efficiency and particle size, especially at lower temperatures. Safety evaluations indicated no hemolysis, and cellular uptake studies revealed around a 2.5-fold increase in fluorescence intensity in cells exposed to Dasatinib Nanobubbles with ultrasound. Cytotoxicity tests demonstrated enhanced sensitivity, with IC50 values of 86.82 mu M for free dasatinib, 63.58 mu M for nano- bubbles without ultrasound, and 44.58 mu M for nanobubbles with ultrasound. The PLGA nanobubbles efficiently encapsulated dasatinib, boosting cellular uptake and cytotoxicity, underscoring their potential for enhanced cancer therapy, particularly with ultrasound.