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Biology subjects

Day, J. H.

Publications and source records attributed to Day, J. H..

2 recordsLinked to original sources

Layer-by-Layer Fabrication of 3D Hydrogel Structures Using Open Microfluidics

Patterning and 3D fabrication techniques have enabled the use of hydrogels for a number of applications including microfluidics, sensors, separations, and tissue engineering in which form fits function. Devices such as reconfigurable microvalves or implantable tissues have been created using lithography or casting techniques. Here, we present a novel open microfluidic patterning method that utilizes surface tension forces to pattern hydrogel layers on top of each other, producing 3D hydrogel structures. We use a patterning device to form a temporary open microfluidic channel on an existing gel layer, allowing the controlled flow of unpolymerized gel in regions defined by the device. Once the gel is polymerized, the patterning device can then be removed, and subsequent layers added to create a multi-layered 3D structure. The use of open-microfluidic and surface tension-based methods to define the shape of each layer enables patterning to be performed with a simple pipette, minimizing dead-volume and shear stress applied on the fluid. Our method is compatible with unmodified (native) biological hydrogels, or other non-biological materials with fluid properties compatible with capillary flow. With our open-microfluidic layer-by-layer fabrication method, we demonstrate the capability to build agarose and type I collagen structures featuring asymmetric designs, multiple components, overhanging features, and cell laden regions.

bioengineering

Investigating Fibroblast-Induced Collagen Gel Contraction Using a Dynamic Microscale Platform

Mechanical forces have long been recognized as fundamental drivers in biological processes, such as embryogenesis, tissue formation and disease regulation. The collagen gel contraction (CGC) assay has served as a classic tool in the field of mechanobiology to study cell-induced contraction of extracellular matrix (ECM), which plays an important role in inflammation and wound healing. In a conventional CGC assay, cell-laden collagen is loaded into a cell culture vessel (typically a well plate) and forms a disk-shaped gel adhering to the bottom of the vessel. The decrement in diameter or surface area of the gel is used as a parameter to quantify the degree of cell contractility. In this study, we developed a microscale CGC assay with an engineered well plate insert that uses surface tension forces to load and manipulate small volumes (14 {micro}L) of cell-laden collagen. The system is easily operated with two pipetting steps and the microscale device moves dynamically as a result of cellular forces. We used a straightforward one-dimensional measurement as the gel contraction readout. We adapted a conventional lung fibroblast CGC assay to demonstrate the functionality of the device, observing significantly more gel contraction when human lung fibroblasts were cultured in serum-containing media versus serum-free media (p[≤]0.01). We further cocultured eosinophils and fibroblasts in the system, two important cellular components that lead to fibrosis in asthma, and observed that soluble factors from eosinophils significantly increase fibroblast-mediated gel contraction (p[≤]0.01). Our microscale CGC device provides a new method for studying downstream ECM effects of intercellular cross talk using 7-35 fold less cell-laden gel than traditional CGC assays.

cell biology