Development and Characterization of Biomimetic Structures for use in 3D in Vitro Models

Abstract

In vitro studies have a significant impact on the progression of novel therapies, cancer and stem cell research, and the drug discovery process. However, the current standard for in vitro models primarily relies on two-dimensional (2D) cell cultures, which fail to fully replicate the intricate cellular architecture and response to microenvironmental cues. To address this limitation, three-dimensional (3D) cell cultures have emerged as more accurate representations of the in vivo cellular environment. Tissue-engineered scaffolds are particularly promising in this regard. However, existing technologies for assessing 3D cell cultures are predominantly limited to endpoint assays, offering only a snapshot of cellular behavior. In this study, our goal is to overcome this challenge by employing continuous electrical assessment of 3D cell cultures cultivated on electroactive composite scaffolds composed of poly(3,4- ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) and MXenes. The scaffold architectures are created using the freeze-drying technique, and we investigate the influence of different scaffold compositions on morphology, swelling, and biocompatibility. By combining the electroactive properties of PEDOT:PSS with the distinctive characteristics of MXenes, we aim to develop scaffolds that not only support cell growth but also enable real-time, noninvasive monitoring of cellular behavior. Our study surpasses conventional optical-based assessments by introducing continuous electrical measurements as a means of tracking the dynamics of 3D cell cultures. The electroactive composite scaffolds facilitate the detection of electrical signals associated with crucial cellular activities like attachment, proliferation, and differentiation. This approach provides real-time insights into the behavior and response of cells to their microenvironment, fostering a deeper understanding of cellular processes and interactions within the 3D scaffold. To evaluate the performance of the electroactive composite scaffolds, we culture them with diverse cell types and assess essential parameters such as cell attachment, viability, growth, and marker expression. By integrating the benefits of 3D cell culture and continuous electrical assessment, our study aims to comprehensively characterize scaffold-cell interactions and their potential applications in tissue engineering and regenerative medicine. This research has the potential to advance the field of in vitro models by offering a more accurate representation of cellular responses in a 3D microenvironment, facilitating the development of enhanced therapies, and enriching our comprehension of complex biological processes.

Date of Award	Aug 2023
Original language	American English
Supervisor	Anna-Maria Pappa (Supervisor)