In vitro osteoclastogenesis on textile chitosan scaffold.
Textile chitosan fibre scaffolds were evaluated in terms of interaction with osteoclast-like cells, derived from human primary monocytes. Part of the scaffolds was further modified by coating with fibrillar collagen type I in order to make the surface biocompatible. Monocytes were cultured directly on the scaffolds in the presence of macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor kappaB ligand (RANKL) for up to 18 days. Confocal laser scanning microscopy (CLSM) as well as scanning electron microscopy (SEM) revealed the formation of multinuclear osteoclast-like cells on both the raw chitosan fibres and the collagen-coated scaffolds. The modified surface supported the osteoclastogenesis. Differentiation towards the osteoclastic lineage was confirmed by the microscopic detection of cathepsin K, tartrate resistant acid phosphatase (TRAP), acidic compartments using 3-(2,4-dinitroanillino)-3'-amino-N-methyldipropylamine (DAMP), immunological detection of TRAP isoform 5b, and analysis of gene expression of the osteoclastic markers TRAP, cathepsin K, vitronectin receptor, and calcitonin receptor using reverse transcription-polymerase chain reaction (RT-PCR). The feature of the collagen-coated but also of the raw chitosan fibre scaffolds to support attachment and differentiation of human monocytes facilitates cell-induced material resorption--one main requirement for successful bone tissue engineering.
Heinemann
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C
Max Bergmann Center of Biomaterials and Institute of Materials Science, Dresden University of Technology, Budapester Str. 27, D-01069 Dresden, Germany. christiane.heinemann@tu-dresden.de
Heinemann
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S
Bernhardt
A
A
Lode
A
A
Worch
H
H
Hanke
T
T
D000135:Acid Phosphatase / Q000032:analysis / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D018786:Bone Substitutes / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D056657:Cathepsin K / Q000032:analysis / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D019070:Cell Lineage / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D003094:Collagen / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000235:genetics; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D009000:Monocytes / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D010010:Osteoclasts / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D053245:RANK Ligand / Q000494:pharmacology; D018003:Receptors, Calcitonin / Q000235:genetics; D000071681:Tartrate-Resistant Acid Phosphatase; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000639:trends*
D015415:Biomarkers; D018786:Bone Substitutes; D039302:Integrin alphaVbeta3; D007527:Isoenzymes; D053245:RANK Ligand; D018003:Receptors, Calcitonin; C505688:TNFSF11 protein, human; D016173:Macrophage Colony-Stimulating Factor; D003094:Collagen; D048271:Chitosan; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; C534130:CTSK protein, human; D056657:Cathepsin K