Development of a 3D bioprinted airway smooth muscle model for manipulating structure and measuring contraction

Abstract

The contractile function of airway smooth muscle (ASM) is inextricably linked to its mechanical properties and interaction with the surrounding mechanical environment. As tissue engineering approaches become more commonplace for studying lung biology, the inability to replicate realistic mechanical contexts for ASM will increasingly become a barrier to a fulsome understanding of lung health and disease. To address this knowledge gap, we describe the use of 3D bioprinting technology to generate a novel experimental model of ASM with a wide scope for modulating tissue mechanics. Using a stiffness modifiable alginate-collagen-fibrinogen bioink, we demonstrate that modulating the stiffness of free-floating ASM bare rings is unfeasible; bioink conditions favorable for muscle formation produce structures that rapidly collapse. However, the creation of novel sandwich and spiderweb designs that encapsulate the ASM bundle within stiff acellular load bearing frames successfully created variable elastic loads opposing tissue collapse and contraction. Sandwich and spiderweb constructs demonstrated realistic actin filament organisation, generated significant baseline tone, and responded appropriately to acetylcholine, potassium chloride and cytochalasin D. Importantly, the two designs feasibly simulate different mechanical contexts within the lung. Specifically, the sandwich was relatively compliant and subject to plastic deformation under high contractile loads, whereas the stiffer spiderweb was more robust and only deformed minimally after repeated maximal contractions. Thus, our model represents a new paradigm for studying ASM contractile function in a realistic mechanical context. Moreover, it holds significant capacity to study the effects of ECM composition, multiple cell types and fibrosis on lung health and disease. GRANTSO_LINatural Sciences and Engineering Research Council, Discovery Grant (Adrian West) C_LIO_LIResearch Manitoba, New Investigator Operating Grant (Adrian West) C_LIO_LIChildrens Hospital Research Institute of Manitoba, Operating Grant (Adrian West) C_LIO_LICanadian Foundation for Innovation, John R. Evans Leaders Fund (Adrian West) C_LIO_LIUniversity of Manitoba, Manitoba Graduate Scholarship (Jeffery Osagie) C_LIO_LIResearch Manitoba, Masters Studentship Award (Jeffery Osagie) C_LIO_LIResearch Manitoba, Masters Studentship Award (Sanjana Syeda) C_LIO_LIChildrens Hospital Research Institute of Manitoba, Summer Studentship (Michelle Guimond) C_LIO_LIUniversity of Manitoba, Jack Prior Memorial Undergraduate Student Research Award (Lumiere Parrenas) C_LIO_LIUniversity of Manitoba, Undergraduate Research Award (Ahsen Haroon) C_LIO_LIUniversity of Manitoba, UMSU Undergraduate Research Award (Philip Imasuen) C_LI The grant bodies had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.