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https://hdl.handle.net/1822/3941
Título: | Osteogenic induction of human bone marrow-derived mesenchymal progenitor cells in novel synthetic polymer-hydrogel matrices |
Autor(es): | Endres, M. Hutmacher, D. W. Salgado, A. J. Kaps, C. Ringe, J. Reis, R. L. Sittinger, M. Brandwood, A. Schantz, J. T. |
Data: | 2003 |
Editora: | Mary Ann Liebert |
Revista: | Tissue Engineering |
Citação: | "Tissue Engineering". ISSN 1076-3279. 9:4 (Aug. 2003) 689-702. |
Resumo(s): | The aim of this project was to investigate the in vitro osteogenic potential of human mesenchymal progenitor cells in novel matrix architectures built by means of a three-dimensional bioresorbable synthetic framework in combination with a hydrogel. Human mesenchymal progenitor cells (hMPCs) were isolated from a human bone marrow aspirate by gradient centrifugation. Before in vitro engineering of scaffold-hMPC constructs, the adipogenic and osteogenic differentiation potential was demonstrated by staining of neutral lipids and induction of bone-specific proteins, respectively. After expansion in monolayer cultures, the cells were enzymatically detached and then seeded in combination with a hydrogel into polycaprolactone (PCL) and polycaprolactone-hydroxyapatite (PCL-HA) frameworks. This scaffold design concept is characterized by novel matrix architecture, good mechanical properties, and slow degradation kinetics of the framework and a biomimetic milieu for cell delivery and proliferation. To induce osteogenic differentiation, the specimens were cultured in an osteogenic cell culture medium and were maintained in vitro for 6 weeks. Cellular distribution and viability within three-dimensional hMPC bone grafts were documented by scanning electron microscopy, cell metabolism assays, and confocal laser microscopy. Secretion of the osteogenic marker molecules type I procollagen and osteocalcin was analyzed by semiquantitative immunocytochemistry assays. Alkaline phosphatase activity was visualized by p-nitrophenyl phosphate substrate reaction. During osteogenic stimulation, hMPCs proliferated toward and onto the PCL and PCL-HA scaffold surfaces and metabolic activity increased, reaching a plateau by day 15. The temporal pattern of bone-related marker molecules produced by in vitro tissue-engineered scaffold-cell constructs revealed that hMPCs differentiated better within the biomimetic matrix architecture along the osteogenic lineage. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/3941 |
DOI: | 10.1089/107632703768247386 |
ISSN: | 1076-3279 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: | 3B’s - Artigos em revistas/Papers in scientific journals |