Comparative functional aspects of 3T3 fibroblasts in 2D and 3D cell- culture environment

Abstract

Abstract Tissue engineering is a multipurpose and valuable tool extensively employed for tissue architecture, and the evaluation of drug efficacy, with other applications. Its particular focus on techniques aimed at restoring or replacing part or entire tissues of replaceable organs. The development of in-vitro bioengineered skin models holds significant potential for clinical utilization. Most existing skin bioengineering methods primarily rely on two-dimensional (2D) cell cultures, which, however, possess notable limitations, particularly in the context of assessing the safety and effectiveness of topical pharmaceutical agents’ due absence of dermal-epidermal interaction. In contrast, three-dimensional (3D) skin bioengineering model enabling the epidermal keratinocytes culture in combination with fibroblasts embedded within 3D matrices. In this study, we have designed a comprehensive experimental approach to investigate the role of Swiss 3T3 cells and their variants pre-exposed to a single pulse of varying doses of Mitomycin C collagen-embedded in supporting the growth of keratinocytes. We examined the proliferation and viability of 3T3 cells to assess their ability to provide growth of epidermal cells in an embedded niche. Additionally, we analysed the levels of Keratinocyte Growth Factor, paracrine secretion by fibroblasts in both 2D and 3D cell culture condition. Furthermore, we conducted a histological comparison of the epidermal layers constructed under 2D and 3D cell culture conditions, evaluating the expression of various epidermal markers to gain insights into their structural and functional differences. This study sheds light on the pivotal role of 2D and 3D skin bioengineering models and their potential applications in dermatological research and pharmaceutical development.