INEB
INEB
TitleThree-dimensional culture of human osteoblastic cells in chitosan sponges: The effect of the degree of acetylation
Publication TypeJournal Article
2006
AuthorsAmaral, IF, Sampaio, P, Barbosa, MA
JournalJournal of Biomedical Materials Research - Part AJ. Biomed. Mater. Res. Part A
Volume76
Issue2
Pagination335 - 346
Date Published2006///
00219304 (ISSN)
Acetylation, Aggregates, alkaline phosphatase, article, Bone Regeneration, bone tissue, Bone tissue engineering, Cell culture, Cell Culture Techniques, Cell filopodia, cell growth, cell proliferation, Cell Shape, cell structure, cell survival, cell viability, chitosan, Collagen, confocal microscopy, controlled study, Cytoskeletal organization, cytoskeleton, Degree of acetylation (DA), enzyme activity, gel permeation chromatography, human, human cell, Humans, infrared spectroscopy, Microstructure, morphology, osteoblast, Osteoblastic cells, Osteoblasts, Phosphatase activity, porosity, Proteins, Scaffold, Scaffolds, Scanning electron microscopy, Substrates, Tissue engineering
In this investigation, the effect of the degree of acetylation (DA) of chitosan on the behavior of human osteoblastic MG-63 cells cultured in three-dimensional chitosan matrices was assessed. Chitosan sponges with DAs in the range of 4 to 49% were prepared and characterized in terms of microstructure, porosity, and pore size. Collagen sponges were used as 3D control. Cell proliferation was determined using the MTT assay while the retention of the osteoblastic phenotype was monitored by assaying alkaline phosphatase activity. Cell morphology, cytoskeletal organization, and viability were assessed using different microscopy techniques. Chitosan sponges showed a similar microstructure regardless the DA, except for the highest DA used, where a more heterogeneous pore distribution was observed. In terms of cell proliferation, alkaline phosphatase activity and cell viability, cells cultured in chitosan scaffolds performed as well as in the 3D control regardless the DA, except for the highest DA used, where an inhibitory effect on cell proliferation was found. However, while in sponges with DAs ≤ 13% cells attached and spread displaying long cell filopodia and numerous cell-to-cell contacts, in sponges with higher DAs cells tended to remain spherical and grow into spheroid-like cellular aggregates. In the present study, the DA played a key role in determining the affinity of osteoblastic cells towards the substrates, possibly by influencing the nature of the initial adsorbed protein layer. © 2005 Wiley Periodicals, Inc.
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