Array-Based Innervated salivary gland microphysiologic system

About This Grant

Abstract The goal of this R21 project is to develop a modular microphysiologic system (MPS) capable of static and optional perfused culture of three dimensional (3D) organoids/tissues in array format. This MPS will be demonstrated by culturing innervated salivary gland tissue mimetics (iSGm). We will test the hypothesize that basal perfusion (Aim 1) and/or innervation (Aim 2) will preserve the acinar cell phenotype and functional response to neurotransmitter stimulation in cultures >14 days. Recent advances in tissue-chip technology have catalyzed the development of MPS as in vitro alternatives to use of animals for tissue/organ modeling and preclinical drug development. While many advances in biomaterials, tissue engineering and microfluidics have enabled the culture of complex tissue mimetics, most MPS fail to fully recapitulate native organs that are innervated. The autonomic nervous system (ANS) regulates the function of most organs in the body and is critical for in vivo salivary gland organogenesis, function and regeneration following injury. This project will exploit established expertise in single-side open microbubble (MB) array technology and ultra-thin (400 nm thick) microporous nanomembrane technology developed independently. We will couple these technologies to demonstrate the proposed MPS by leveraging several technical innovations to create and bond a two-side open MB array to the nanomembrane. Salivary gland tissue mimetics will be cultured in the MB array on the apical side of the nanomembrane juxtaposed to neuronal tissue in the basal compartment. The pore size of the nanomembrane will be chosen to prevent nerve cell transmigration. The gap between the MB array and the nanomembrane, created by a pressure sensitive adhesive (PSA), will be filled with a hydrogel engineered to provide a based to the MB for salivary gland cell seeding and to provide a matrix for 3D neurite extension replicating native tissue physiology. Diffusion, fluid flows and shear stresses through the device will be simulated using COMSOL modeling to test design parameters to inform the device design. The growth and function of SGm and iSGm in the MPS will be compared to SGm culture in standard single-side opened MB arrays with and without basal compartment perfusion. Successful completion of this project will advance a MPS for co-culturing similar-sized 3D tissue mimetics in an array format coupled to a second organ system separated by a nanomembrane with or without a gap. The latter can be filled with hydrogel to support a third organ system or as in this project, optimized to support 3D neurite extension and target tissue innervation. The proposed MPS is truly capable of high-content testing and can be used to rapidly and cost effectively test drugs for off-target effects on SG function in the drug development process.