(W1030-08-48) Drug Loaded Spanlastics in 3D Bioprinted Hydrogels for Chemotherapy

Purpose: The purpose of this study is to fabricate Dox-loaded Spanlastics (Span-Dox), bioprint nanocarrier-loaded hydrogels, and assess their anticancer activity in breast cancer cells (MCF-7 cells).

Methods: The Spanlastics (Span) was made in two different ratios, 70:30 and 80:20 using Span 60 and Tween 80, respectively (Table 1b). The particle size, zeta potential, and polydispersity index were observed using the Malvern Zetasizer Nano-ZS (Table 1a). Doxorubicin Hydrochloride (Dox) was loaded into both formulations and the encapsulation efficiency was observed by UV absorbance of the supernatant obtained from centrifugation using the Eppendorf Centrifuge 5804R 15 amp version. Span-B was loaded with Doxorubicin (Span-Dox) at different concentrations, 0.5%w/v and 1%w/v. The morphology of Span and Span-Dox were viewed with Scanning Electron Microscope (JEOL JSM-5600). Varying concentrations of Sodium Alginate was tested for printability using the FRESH 3D printing method. 5% Sodium Alginate was mixed with Span-Dox and printed into 6-well plates using Allevi 1 3DBio printer. MCF-7 cells were grown to 80% confluency and then treated with Span-B and Span-Dox 0.5 and Span-Dox 1. The cell viability at different timepoints were observed and analyzed. Again, the cellular uptake of Doxorubicin was demonstrated by comparing the fluorescence of treated cell groups by applying Alamar blue dye and viewing under Olympus CBH IX83 fluorescence microscope.

Results: Span-B was selected as lead formulation due to higher entrapment efficiency, 40%, as compared to the 80:20 which was 30% (Table 1C). The morphology of Span and Span-Dox were viewed under SEM at different scales showing the spherical nano vesicular nature of the Spanlastics (Figure 1b).  After bioprinting with 2% and 5% Sodium Alginate, 5% proved to be best in terms of viscosity and shape which was physically observed (Figure1a). The treatment of MCF-7 cells with Span and Span-Dox  showed that Span-Dox had significant effect on cell viability and the higher the concentration of Doxorubicin in Spanlastics, the lower the percentage cell viability (Figure 2b). Also, the Span-Dox delivered Dox into the nucleus of the MCF7 cells which showed a red color as seen in Figure 2a.

Conclusion: The 70:30 Span60 to Tween 80 ratio gave the best Span-Dox formulation evidenced by particle size, zeta potential and PDI, as well as encapsulation efficiency. Sodium Alginate 5% loaded Span-Dox was ‘FRESH’ 3D-printed. Span-Dox reduced MCF-7 cell viability dose-dependently.

References: Alaaeldin E, et. al. Spanlastics as an efficient delivery system for the enhancement of thymoquinone anticancer efficacy: Fabrication and cytotoxic studies against breast cancer cell lines. J Drug Deliv Sci Technol. 2021;65:102725. doi:10.1016/j.jddst.2021.102725
Bizuayehu HM, et. al. Burden of 30 cancers among men: Global statistics in 2022 and projections for 2050 using population-based estimates. Cancer. 2024;130(21):3708-3723. doi:10.1002/cncr.35458
Abdulkafi, Ali & Abdal-Hammid, Shaimaa & Alhammid, Abd. (2023). A Modern Method For The Delivery Of Nanovesicular Drugs, Spanlastics. Journal of Pharmacy. 4. 1-11. 10.47310/iarjp.2023.v04i06.001.

Acknowledgements: This was supported by NSF REU Site: Ole Miss Nanoengineering Summer REU Program ENG/EEC/EPSCoR 2148764 and the PharmE3D lab of the Department of Pharmaceutics and Drug Delivery, University of Mississippi.