Photo-crosslinkable hydrogel and 3D rapid prototyping technology application for primary hepatocyte culture
Date Issued
2015
Date
2015
Author(s)
Liao, Chih-Hsun
Abstract
3D printing moved from being theoretical to a reality, and in recent years 3D printers have become cheaper to produce. Nowadays, 3D bio-printing has been widely used in tissue engineering and regenerative medicine. This rapid prototyping technology provide a great opportunity to fabricate 3D scaffolds without problems such as restricted geometric shapes in traditional methods. In this research, we modified an open-source 3D printer project, originally printed thermoplastic polymer which was not support soft tissue and cell encapsulation. Thus, we designed a bio-printer including photo-crosslinking module and redesign syringe pump extruder of 3D printer. In the experiments, we used gelatin hydrogel, instead of thermoplastics, as main material of scaffold. Gelatin is commonly used in pharmaceutics, food, cosmetics, as well as cell culture. However, gelatin has poor mechanical properties at 37˚C to be the main structure of a tissue scaffold. Therefore, further processing is required to strengthen for gelatin. In this study, we demonstrated optimized methacrylation of gelatin forming a highly hydrophilic photo-crosslinkable hydrogel. The hydrogel is analyzed by NMR spectra to monitor degree of functionalization. Because of complexity of liver, it is hard to create in vitro liver model. Moreover, isolated primary hepatocytes rapidly lose its mature hepatic functions after culturing in 2D environment due to loss of ECM and intercellular interaction. We applied 3D bio-printing technology in creating hepatocyte laden hydrogel scaffold. Porous hydrogel scaffold was design by CAD software and printed by our well-calibrated 3D bio-printer reaching 300μm precision. Also scanning electron microscopy analysis was involved to confirm porous structure of the hydrogel scaffold offering cell attachment and nutrient dispersion well. In our research, primary hepatocytes expressed mature hepatic markers after 5 days cultured in our 3D hydrogel scaffolds. We believed this technology opens new possibilities for the application of 3D bio-printing in liver tissue engineering.
Subjects
tissue engineering
hepatocyte
bio-printing
hydrogel
regenerative medicine
Type
thesis
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