1.Osteogenic potential of human periosteum-derived progenitor cells in PLGA scaffold using allogeneic serum.
Yi-xiong ZHENG ; Jochen RINGE ; Zhong LIANG ; Alexander LOCH ; Li CHEN ; Michael SITTINGER
Journal of Zhejiang University. Science. B 2006;7(10):817-824
The use of periosteum-derived progenitor cells (PCs) combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional (3D) poly-lactic-co-glycolic acid (PLGA) fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic medium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type I collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules of neo-bone tissue formation and the presence of matrix mineralization were confirmed by positive von Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.
Biocompatible Materials
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Bone Development
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Cell Culture Techniques
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methods
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Cell Differentiation
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Cell Survival
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Cells, Cultured
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Collagen
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chemistry
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Humans
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Lactic Acid
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chemistry
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Microscopy, Fluorescence
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Models, Statistical
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Osteogenesis
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Periosteum
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metabolism
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Polyglycolic Acid
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chemistry
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Polymers
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chemistry
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Stem Cells
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cytology
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Tissue Engineering