title: Fabrication of large perfusable macroporous cell-laden hydrogel scaffolds using microbial transglutaminase.
abstract: In this study, we developed a method to fabricate large, perfusable, macroporous, cell-laden hydrogels. This method is suitable for efficient cell seeding, and can maintain sufficient oxygen delivery and mass transfer. We first loaded three types of testing cells (including NIH 3T3, ADSC and Huh7) into gelatin hydrogel filaments, then cross-linked the cell-laden gelatin hydrogel filaments using microbial transglutaminase (mTGase). In situ cross-linking by mTGase was found to be non-cytotoxic and prevented the scattering of the cells after delivery. The gelatin hydrogel constructs kept the carried cells viable; also, the porosity and permeability were adequate for a perfusion system. Cell proliferation was better under perfusion culture than under static culture. When human umbilical vein endothelial cells were seeded into the constructs, we demonstrated that they stably formed an even coverage on the surface of the hydrogel filaments, serving as a preliminary microvasculature network. We concluded that this method provides a viable solution for cell seeding, oxygen delivery, and mass transfer in large three-dimensional (3-D) tissue engineering. Furthermore, it has the potential for being a workhorse in studies involving 3-D cell cultures and tissue engineering.
authors.affiliation: Institute of Biomedical Engineering, College of Engineering and College of Medicine, National Taiwan University, Taipei City, Taiwan, ROC; Department of Orthopedic Surgery, National Taiwan University Hospital, Taipei City, Taiwan, ROC.
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Chen
authors.affiliation: School of Dental Technology, College of Oral Medicine, Taipei Medical University, Taipei City, Taiwan, ROC.
authors.forename: Kai-Chiang
authors.initials: KC
authors.lastname: Yang
authors.affiliation: Department of Orthopedic Surgery, National Taiwan University Hospital, Taipei City, Taiwan, ROC; Department of Orthopedic Surgery, En Chu Kong Hospital, New Taipei City, Taiwan, ROC.
authors.forename: Chang-Chin
authors.initials: CC
authors.lastname: Wu
authors.affiliation: Department of Orthopedic Surgery, Taoyuan General Hospital, Department of Health, Taoyuan County, Taiwan, ROC.
authors.forename: Jeen-Huei
authors.initials: JH
authors.lastname: Yu
authors.affiliation: Institute of Biomedical Engineering, College of Engineering and College of Medicine, National Taiwan University, Taipei City, Taiwan, ROC.
authors.forename: Feng-Huei
authors.initials: FH
authors.lastname: Lin
authors.affiliation: Department of Orthopedic Surgery, National Taiwan University Hospital, Taipei City, Taiwan, ROC; Department of Orthopedic Surgery, National Taiwan University Hospital, Hsin-Chu Branch, Hsinchu City, Taiwan, ROC. Electronic address: drjssun@gmail.com.
authors.forename: Jui-Sheng
authors.initials: JS
authors.lastname: Sun
mesh_terms: D000818:Animals; D001419:Bacteria / Q000201:enzymology*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005780:Gelatin / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology*; D051379:Mice; D041681:NIH 3T3 Cells; D010539:Permeability / Q000187:drug effects; D016062:Porosity; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D054457:Tissue Scaffolds / Q000737:chemistry*; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D005780:Gelatin; D011503:Transglutaminases
keywords: Cell culture; Hydrogel; Scaffold; Three-dimensional; Tissue engineering