In vitro osteoclastogenesis on textile chitosan scaffold

Abstract

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.

From the Clinical Problem to the Basic Research—Co-Culture Models of Osteoblasts and Osteoclasts

“…Forty years ago, the first co-culture study was published, which showed that interactions between rat granulosa cells and myocardial cells happen via gap junctions [41]. Since then many other studies, using co-cultures, followed [42,43,44,45]. Traditionally, co-culture models were carried out in 2D.…”

Bridging the gap between traditional cell cultures and bioreactors applied in regenerative medicine: practical experiences with the MINUSHEET perfusion culture system

“…For example, developmental capacity of osteoblasts and osteocytes was investigated with ceramic materials Wang et al 2003;Leukers et al 2005a;Yeatts and Fisher 2011;Bernhardt et al 2011), decellularized spongeous bone (Seitz et al 2007), collagen membranes (Rothamel et al 2004) and mineralized collagen (Gelinsky et al 2004;Bernhardt et al 2008). Further hydroxyapatite scaffolds (Leukers et al 2005b;Detsch et al 2008;da Silva et al 2010a, b), poly-d,l-lactic-coglycolic acid (PLGA) sheets (Shearer et al 2006), iron based metals (Quadbeck et al 2010), bioactive glass (Yue et al 2011), textile chitosan (Heinemann et al 2008(Heinemann et al , 2009(Heinemann et al , 2010, 3D biphasic calcium phosphate scaffolds (Rath et al 2012) and other biocorrodible bone replacement materials (Farack et al 2011) were successfully applied in combination with perfusion culture.…”

Osteoinduction and Antiosteoporotic Performance of Hybrid Biomaterial Chitosan-Bioactive Glass Graft: Effects on Bone Remodeling

“…In fact, an anti-osteoporotic performace was attributed to CH. In a previous study, this molecule was found to inhibit osteoclastic activity, and is thus widely used to treat patients with osteoporosis [37]. One study demonstrated that incorporation of chitosan to the cement prevents the osteoclastic resorption of the composite biomaterial compared to the cement alone [38].…”