"In a previous study , we developed a novel articular cartilage ECM (ACECM)-derived scaffold using decellularised human joint cartilage. This scaffold could support cell attachment, proliferation, and mesenchymal stem cell differentiation and was used in vivo for cartilage tissue engineering [16,17]. Because cartilage-specific ECM components play an important role in chondrogenesis, as well as in supporting the chondrogenic phenotype [18,19], other biomimetic scaffolds with oriented structures were fabricated using temperature gradient-guided thermal-induced phase separation (TIPS) followed by freeze-drying to mimic the biochemical composition and natural structure of native articular cartilage [20,21]. "
[Show abstract][Hide abstract] ABSTRACT: Background
Osteochondral interface regeneration is challenging for functional and integrated cartilage repair. Various layered scaffolds have been used to reconstruct the complex interface, yet the influence of the permeability of the layered structure on cartilage defect healing remains largely unknown.
We designed and fabricated a novel bilayered scaffold using articular cartilage extracellular matrix (ACECM) and hydroxyapatite (HAp), involving a porous, oriented upper layer and a dense, mineralised lower layer. By optimising the HAp/ACECM ratio, differing pore sizes and porosities were obtained simultaneously in the two layers. To evaluate the effects of permeability on cell behaviour, rabbit chondrocytes were seeded.
Morphological observations demonstrated that a gradual interfacial region was formed with pore sizes varying from 128.2 ± 20.3 to 21.2 ± 3.1 μm. The permeability of the bilayered scaffold decreased with increasing compressive strain and HAp content. Mechanical tests indicated that the interface was stable to bearing compressive and shear loads. Accordingly, the optimum HAp/ACECM ratio (7 w/v%) in the layer to mimic native calcified cartilage was found. Chondrocytes could not penetrate the interface and resided only in the upper layer, where they showed high cellularity and abundant matrix deposition.
Our findings suggest that a bilayered scaffold with low permeability, rather than complete isolation, represents a promising candidate for osteochondral interface tissue engineering.
"Mesenchymal stem cells (MSCs) have been investigated as an alternative to terminally differentiated cells to develop novel treatments for bone and cartilage defects, since they can be easily harvested from several adult tissues and are able to differentiate towards the osteogenic and chondrogenic lineages (Johnstone et al., 1998; Pittenger et al., 1999). Beside bone marrow MSCs (BMSCs), more recently adipose derived mesenchymal stem cells have been successfully used for bone and cartilage applications (Rada et al., 2009; Jung et al., 2010; Rhee et al., 2011; Kang et al., 2012; Choi et al., 2014). In particular, MSCs resident in the infrapatellar fat pad (IFP-MSCs) and knee subcutaneous adipose tissue (ASCs) can be considered appealing alternative cell sources for articular cell-based therapies, thanks to their differentiative potential and ease of harvesting during knee surgery, which causes minimal additional morbidity to patients. "
[Show abstract][Hide abstract] ABSTRACT: Cell-based therapies have recently been proposed for the treatment of degenerative articular pathologies, such as early osteoarthritis, with an emphasis on autologous mesenchymal stem cells (MSCs), as an alternative to terminally differentiated cells. In this study, we performed a donor-matched comparison between infrapatellar fat pad MSCs (IFP-MSCs) and knee subcutaneous adipose tissue stem cells (ASCs), as appealing candidates for cell-based therapies that are easily accessible during surgery. IFP-MSCs and ASCs were obtained from 25 osteoarthritic patients undergoing total knee replacement and compared for their immunophenotype and differentiative potential. Undifferentiated IFP-MSCs and ASCs displayed the same immunophenotype, typical of MSCs (CD13+/CD29+/CD44+/CD73+/CD90+/CD105+/CD166+/CD31-/CD45-). IFP-MSCs and ASCs showed similar adipogenic potential, though undifferentiated ASCs had higher LEP expression compared to IFP-MSCs (p < 0.01). Higher levels of calcified matrix (p < 0.05) and alkaline phosphatase (p < 0.05) in ASCs highlighted their superior osteogenic commitment compared to IFP-MSCs. Conversely, IFP-MSCs pellets showed greater amounts of glycosaminoglycans (p < 0.01) and superior expression of ACAN (p < 0.001), SOX9, COMP (p < 0.001) and COL2A1 (p < 0.05) compared to ASCs pellets, revealing a superior chondrogenic potential. This was also supported by lower COL10A1 (p < 0.05) and COL1A1 (p < 0.01) expression and lower alkaline phosphatase release (p < 0.05) by IFP-MSCs compared to ASCs. The observed dissimilarities between IFP-MSCs and ASCs show that, despite expressing similar surface markers, MSCs deriving from different fat depots in the same surgical site possess specific features. Furthermore, the in vitro peculiar commitment of IFP-MSCs and ASCs from osteoarthritic donors towards the chondrogenic or osteogenic lineage may suggest a preferential use for cartilage and bone cell-based treatments, respectively.
European cells & materials 04/2014; 27:298-311. · 4.89 Impact Factor
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