title: Open-Porous Hydroxyapatite Scaffolds for Three-Dimensional Culture of Human Adult Liver Cells.
abstract: Liver cell culture within three-dimensional structures provides an improved culture system for various applications in basic research, pharmacological screening, and implantable or extracorporeal liver support. Biodegradable calcium-based scaffolds in such systems could enhance liver cell functionality by providing endothelial and hepatic cell support through locally elevated calcium levels, increased surface area for cell attachment, and allowing three-dimensional tissue restructuring. Open-porous hydroxyapatite scaffolds were fabricated and seeded with primary adult human liver cells, which were embedded within or without gels of extracellular matrix protein collagen-1 or hyaluronan. Metabolic functions were assessed after 5, 15, and 28 days. Longer-term cultures exhibited highest cell numbers and liver specific gene expression when cultured on hydroxyapatite scaffolds in collagen-1. Endothelial gene expression was induced in cells cultured on scaffolds without extracellular matrix proteins. Hydroxyapatite induced gene expression for cytokeratin-19 when cells were cultured in collagen-1 gel while culture in hyaluronan increased cytokeratin-19 gene expression independent of the use of scaffold in long-term culture. The implementation of hydroxyapatite composites with extracellular matrices affected liver cell cultures and cell differentiation depending on the type of matrix protein and the presence of a scaffold. The hydroxyapatite scaffolds enable scale-up of hepatic three-dimensional culture models for regenerative medicine applications.
authors.affiliation: Department of Mechanical Engineering and Materials Science, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Anthony
authors.identifier: 0000-0001-9188-1234
authors.initials: A
authors.lastname: Finoli
authors.affiliation: Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15203, USA.
authors.forename: Eva
authors.initials: E
authors.lastname: Schmelzer
authors.affiliation: Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15203, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Over
authors.affiliation: Department of Mechanical Engineering and Materials Science, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors.forename: Ian
authors.initials: I
authors.lastname: Nettleship
authors.affiliation: Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15203, USA; Department of Bioengineering, McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15203, USA.
authors.forename: Joerg C
authors.initials: JC
authors.lastname: Gerlach
mesh_terms: D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D017886:Durapatite / Q000737:chemistry*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D019164:Liver, Artificial; D008297:Male; D009924:Organ Culture Techniques / Q000295:instrumentation*; D016062:Porosity; D066330:Printing, Three-Dimensional; D023822:Tissue Engineering / Q000295:instrumentation; D054457:Tissue Scaffolds
chemical_list: D016326:Extracellular Matrix Proteins; D017886:Durapatite