Rheological Evaluation of a Hydrogel-Based Ovarian Cancer Organoid Model
Discipline: Technology and Engineering
Subcategory: Biomedical Engineering
Session: 1
Room: Farragut North
Astrid Daugherty - FAMU-FSU College of Engineering, National High Magnetic Field Laboratory
Co-Author(s): Tyler Gregory, FAMU-FSU College of Engineering, National High Magnetic Field Laboratory, Tallahassee, Florida; Mary Jean Savitsky, FAMU-FSU College of Engineering, National High Magnetic Field Laboratory, Tallahassee, Florida; Navneet Kaur, FAMU-FSU College of Engineering, National High Magnetic Field Laboratory, Tallahassee, Florida; Jamel Ali, FAMU-FSU College of Engineering, National High Magnetic Field Laboratory, Tallahassee, Florida
Ovarian cancer is a growing global health concern due to an increasing number of diagnoses and poor prognosis associated with the disease. Towards developing a better understanding of ovarian cancer pathophysiology and towards developing more effective therapeutics, three-dimensional (3D) models of ovarian tissue offer significant advantages over traditional 2D models. However, current 3D ovarian models often lack the mechanical attributes of native ovarian cancer tissue and the adjacent extracellular matrix (ECM), which heavily influences cellular behavior. In this investigation we evaluate the hypothesis that modifying the rigidity of an artificially constructed ECM alters the cell viability, proliferation, and structural development of ovarian adenocarcinoma. An alginate-gelatin based hydrogel was used as an artificial EMC, where the stiffness was modulated by altering concentrations of alginate solutions (3%, 4%, 5% w/v) with fixed concentrations of gelatin (4% w/v). After ECM formulation, an ovarian epithelial cell line, SKOV3, was suspended in the ECM and cultured in 3D after which live/dead and proliferation assays were performed over a period of two weeks. Rheological examination of the ECM was carried out to evaluate the mechanical characteristics of the hydrogels, which consisted of three measurement types: time sweeps, frequency sweeps, and flow curves. Finally, SEM imaging was performed on crosslinked hydrogels with and without the incorporation of ovarian cancer cells to observe structural differences in both the ECM and the morphology of the ovarian cancer cells. Results indicate SKOV3 has higher cell viability shortly after seeding in all hydrogels when compared to traditional 2D culture. Rheological analysis confirmed the viscoelastic properties of hydrogels, which are essential for replicating the natural environment of mammalian tissue. Additionally, shear-thinning behavior was observed with increasing shear rate, critical for further applications of a 3D organoid model, such as 3D printing. Generally, a connection between hydrogel stiffness and the ratio of -live-to-dead cells was introduced, exposing how cell growth and survival is altered with various ECM stiffness. Discovery on how model accuracy is affected with varying manufacturing formulations of an organoid model opens avenues for future research, especially in the context of high-throughput screening for ovarian cancer interventions that better mimic real physiological conditions. Ultimately, this research holds the potential to advance early detection and treatment methods for ovarian cancer, addressing a growing healthcare challenge.
Funder Acknowledgement(s): This work was funded by the National Science Foundation (No. EES-2306449, EES-2219558, EES-2000202, CMMI-2000330) and supported by the NSF FAMU CREST Center award (No. EES-1735968). This research work was also supported by The Grainger Foundation Frontiers of Engineering Grant under the National Academy of Sciences Award Number: 200001318. Support was also provided by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number R16GM145595. All the work was performed at the National High Magnetic Field Laboratory, which is supported by National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida. Astrid Daugherty acknowledges generous support from the Dow SURE Program and the Danfoss Undergraduate Engineering Fellowship.
Faculty Advisor: Jamel Ali, jali@eng.famu.fsu.edu
Role: As a primary author of this research project, I directed the planning and execution of all the experiments. I developed the alginate-gelatin based hydrogel to create the artificial extracellular matrix. Consequently, I performed the 2D and 3D cell culture. With the help of the co-authors, I was able to perform all the protocols related to the results, such as the rheological analysis, cell viability and proliferation assays, and SEM imaging. Finally, I designed the presentation and processed all the data collected.