Fabrication of single gel with different mechanical stiffness using three-dimensional mold

The hydrogel is considering as functional substrates for three-dimensional (3D) environment mimicking the native tissue in vitro. To get the cell or tissue culture result in different stiffness, researchers used separate gel at different times. Sometimes these results are manipulated by surrounding environment. To overcome this, we prepared a single hydrogel with different young modulus using a 3 D printed mold and cell culture and tissue culture was performed to check the functional capacity. In this proposed device we successfully produced a multiproperties agarose hydrogel on a single platform. We designed different mold pattern to confirm that this gel formation technique can be used for any types of design and many different concentrated hydrogels can be combined together. The cell and tissue culture results showed that even in same planer surface, each gel solely maintains their own physical properties and control the cell and tissue adherence and proliferation behavior. The protocol is fairly simple and reproducible. The design helps to produce consistent gel thickness, shape, and size. This 3 D mold has provided a new way tostudy the fundamental cellular responses to engineered microenvironments that may have a high implementation in both biological and healthcare-related applications. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 6-11, 2019.