Abstract
Salivary gland bioengineering requires understanding the interaction between salivary epithelium and surrounding tissues. An important component of salivary glands is the presence of neurons. No previous studies have investigated how neurons and salivary epithelial cells interact in an in vitro co-culture model. In this study, we describe the self-organization of neurons around salivary epithelial cells in co-culture, in a similar fashion to what occurs in native tissue. We cultured primary mouse cortical neurons (m-CN) with a salivary epithelial cell line (Par-C10) on growth factor-reduced Matrigel (GFR-MG) for 4 days. After this time, co-cultures were compared with native salivary glands using confocal microscopy. Our findings indicate that m-CN were able to self-organize basolaterally to salivary epithelial cell clusters in a similar manner to what occurs in native tissue. These results indicate that this model can be developed as a potential platform for studying neuron-salivary epithelial cell interactions for bioengineering purposes.
Highlights
Hyposalivation results in poor oral health and negatively affects patients’ quality of life[1]
Neurons have been shown to thrive in co-culture with endothelial cells[7] and skeletal muscle cells[8], and while the role of parasympathetic innervation of salivary glands has been investigated in embryonic submandibular organ explant cultures[9], no studies to date have investigated co-cultures of neurons and salivary epithelial single cells
The main goal of this study was to verify the existence of precursor conditions under which co-cultures of neuron-salivary epithelial cells physically organize in a similar fashion to native tissue
Summary
Hyposalivation results in poor oral health and negatively affects patients’ quality of life[1]. The most prominent cause of this condition is primary Sjögren’s Syndrome (SS), an autoimmune disease that affects salivary and lacrimal glands, but it can result from a wide variety of oral and systemic diseases, such as diabetes, depression, fibromyalgia, and HIV/AIDS, in addition to iatrogenic causes such as radiation therapy, chemotherapy, and drugs[2]. Understanding neuron-salivary epithelial cell interactions is an important step for the development of a viable artificial salivary gland. In order to develop an in vitro model to study functional interactions between neurons and salivary epithelial cells, the following conditions have to be demonstrated: 1) ability of cells to thrive when cultured together, 2) that neurons do not cause salivary epithelial disruption, or vice versa, and 3) ability of neurons to localize on the basolateral surface of salivary epithelial cell clusters
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