Abstract
Innervation plays a key role in the development and homeostasis of organs and tissues of the orofacial complex. Among these structures, teeth are peculiar organs as they are not innervated until later stages of development. Furthermore, the implication of neurons in tooth initiation, morphogenesis and differentiation is still controversial. Co-cultures constitute a valuable method to investigate and manipulate the interactions of nerve fibers with their target organs in a controlled and isolated environment. Conventional co-cultures between neurons and their target tissues have already been performed, but these cultures do not offer optimal conditions that are closely mimicking the in vivo situation. Indeed, specific cell populations require different culture media in order to preserve their physiological properties. In this study we evaluate the usefulness of a microfluidics system for co-culturing mouse trigeminal ganglia and developing teeth. This device allows the application of specific media for the appropriate development of both neuronal and dental tissues. The results show that mouse trigeminal ganglia and teeth survive for long culture periods in this microfluidics system, and that teeth maintain the attractive or repulsive effect on trigeminal neurites that has been observed in vivo. Neurites are repealed when co-cultured with embryonic tooth germs, while postnatal teeth exert an attractive effect to trigeminal ganglia-derived neurons. In conclusion, microfluidics system devices provide a valuable tool for studying the behavior of neurons during the development of orofacial tissues and organs, faithfully imitating the in vivo situation.
Highlights
Innervation plays a key role in both development and homeostasis of organs and tissues (Kumar and Brockes, 2012; Pagella et al, 2014)
CONVENTIONAL CO-CULTURES OF EMBRYONIC TRIGEMINAL GANGLIA AND TOOTH GERMS During tooth development, axons projecting from trigeminal nerves approach and innervate the dental follicle mesenchyme, but they do not enter the dental papilla mesenchyme until later stages of development (Figure 1)
Recent studies suggest that innervation is pivotal during embryonic development and regeneration of many tissues and organs (Pagella et al, 2014)
Summary
Innervation plays a key role in both development and homeostasis of organs and tissues (Kumar and Brockes, 2012; Pagella et al, 2014). It has been demonstrated that innervation is necessary for the development and maintenance of taste buds (Oakley et al, 1998; Mistretta et al, 1999; Sun and Oakley, 2002; Oakley and Witt, 2004) In this context, it is important to better understand the role of sensory innervation during odontogenesis. Teeth develop as a result of sequential and reciprocal interactions between the oral ectoderm and cranial neural crest-derived mesenchyme These interactions give rise to epithelial-derived ameloblasts and mesenchyme-derived odontoblasts that are responsible for the formation of enamel and dentin, respectively (Thesleff et al, 1995; Mitsiadis and Graf, 2009).
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