Extracellular matrix-enriched polymeric scaffolds as a substrate for hepatocyte cultures: in vitro and in vivo studies.
Tissue engineering is a promising approach to developing hepatic tissue suitable for the functional replacement of a failing liver. The aim of the present study was to investigate whether an extracellular cell matrix obtained from fibroblasts-cultured within scaffolds of hyaluronic acid (HYAFF) could influence the proliferation rate and survival of rat hepatocytes both during long-term culture and after in vivo transplantation. Cultures were evaluated by histological and morphological analysis, a proliferation assay and metabolic activity (albumin secretion). Hepatocytes cultured in extracellular matrix-enriched scaffolds exhibited a round cellular morphology and re-established cell-cell contacts, growing into aggregates of several cells along and/or among fibers in the fabric. Hepatocytes were able to secrete albumin up to 14 days in culture. In vivo results demonstrated the biocompatibility of HYAFF-11 implanted in nude mice, in which hepatocytes maintained small well-organised aggregates until the 35th day. In conclusion, the presence of a fibroblast-secreted extracellular matrix improved the biological properties of the hyaluronan scaffold, favoring the survival and morphological integrity of hepatocytes in vitro and in vivo.
Zavan
B
B
Department of Histology, Microbiology and Medical Biotecnology, University of Padova, Italy. barbara.zavan@unipd.it
Brun
P
P
Vindigni
V
V
Amadori
A
A
Habeler
W
W
Pontisso
P
P
Montemurro
D
D
Abatangelo
G
G
Cortivo
R
R
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology / Q000637:transplantation*; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D019164:Liver, Artificial; D008422:Materials Testing; D051379:Mice; D016513:Mice, SCID; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods*
D001672:Biocompatible Materials; D006820:Hyaluronic Acid