Abstract
Guiding cell culture via engineering extracellular microenvironment has attracted tremendous attention due to its appealing potentials in the repair, maintenance, and development of tissues or even whole organs. However, conventional biofabrication technologies are usually less productive in fabricating microscale three-dimensional (3D) constructs because of the strident requirements in processing precision and complexity. Here we present an optical µ-printing technology to rapidly fabricate 3D microscaffold arrays for 3D cell culture and cell-scaffold interaction studies on a single chip. Arrays of 3D cubic microscaffolds with cubical sizes matching the single-cell size were fabricated to facilitate cell spreading on suspended microbeams so as to expose both apical and basal cell membranes. We further showed that the increasing of the cubical size of the microscaffolds led to enhanced spreading of the seeded human mesenchymal stem cells and activation of mechanosensing signaling, thereby promoting osteogenesis. Moreover, we demonstrated that the spatially selective modification of the surfaces of suspended beams with a bioactive coating (gelatin methacrylate) via an in-situ printing process allowed tailorable cell adhesion and spreading on the 3D microscaffolds.
Highlights
Microscale or nanoscale topographies have revealed their essential influence on regulating cell behavior and fate[1,2,3]
We demonstrated that a dynamic optical projection stereolithography (DOPsL) technology can quickly pattern various kinds of hydrogels with or without seeding of cells into user-defined 3D extracellular microenvironments for cell study[34, 35]
A polymer material (i.e. SU-8 photoresist) with good mechanical property as well as excellent chemical resistance and nonirritant characteristics is used to fabricate a variety of 3D cubic microscaffolds, whose cubical sizes are tens of micrometers, to mimic the natural structure of bone lacunae[38]
Summary
Microscale or nanoscale topographies have revealed their essential influence on regulating cell behavior and fate[1,2,3]. The minimum feature size of droplet-based bioprinters is around 60 μm[27], which hinders the application of these technologies in the fabrication of micrometer-scale microscaffolds for cell studies. Range of 3D microscaffolds with ultrafine features for 3D cell culture studies, e.g. cell migration[29], cell differentiation[30], and cell morphology[31] Such a single-spot scanning technology is limited by its low yield in the fabrication of large volume or large quantities of ultrafine microstructures[28]. An in-situ printing technique is developed to selectively deposit gelatin methacrylate (GelMA) on the suspended beam as bioactive coating material to guide cell adhesion and spreading Such a two-material based printing protocol offers a more biomimetic 3D microplatform with tunable biochemical and structural properties for controlled cell culture and migration studies, e.g. guided bone regeneration
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