Researcher(s)
- Audrey Orzech, Biochemistry, University of Delaware
Faculty Mentor(s)
- Jason Gleghorn, Biomedical Engineering, University of Delaware
Abstract
The female reproductive tract is lined with mucus, which plays an essential role in protecting epithelial tissues while regulating the local cellular microenvironment. As a dynamic barrier, mucus influences cellular interactions, molecular transport, and host-microbe communication. Therefore, in vitro models that incorporate mucus are needed to investigate how reproductive tract tissues respond to biological and environmental stimuli. We previously developed a synthetic mucus formulation to mimic the biophysical and biochemical properties of cervicovaginal mucus. In the current work, the biocompatibility and flow characteristics of the synthetic mucus were evaluated within a microphysiological (MPS) model of the cervix. Synthetic mucus representing the differing properties of the menstrual and ovulatory phases of the menstrual cycle demonstrated consistent flow through the MPS cassette system, supporting the integration of mucus into the model. Biocompatibility of the mucus was assessed using VK2 vaginal epithelial cells and BJ fibroblasts. Cellular morphology was examined through fluorescent staining of filamentous actin and DNA in nuclei, and inflammatory responses were evaluated by quantifying TNF-α and IL-6 using enzyme-linked immunosorbent assays (ELISAs). TNF-α concentrations remained below detectable limits across experimental conditions, suggesting the mucus did not elicit measurable TNF-α production. MID mucus exposure produced no significant increase in IL-6 production compared with the no mucus control in VK2 epithelial cells or BJ fibroblasts, while LPS treatment induced increased IL-6 production in BJ fibroblasts, confirming cellular responsiveness to an inflammatory stimulus. However, fluorescence imaging of LIVE/DEAD staining revealed not only morphological differences but also changes in cell viability following mucus exposure, indicating changes in cellular phenotype and survival despite minimal cytokine production. The reliable flow of the synthetic mucus formulation through the platform supports its continued optimization into a fully compatible three-dimensional reproductive tract model. This work provides a foundation for future studies investigating reproductive tract biology and host-microbe interactions.