Cycloastragenol as an Exogenous Enhancer of Chondrogenic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells. A Morphological Study.
Marta Anna SzychlinskaGiovanna CalabreseSilvia RavalliNunziatina Laura ParrinelloStefano FortePaola CastrogiovanniElisabetta PricocoRosa ImbesiSergio CastorinaRosalia LeonardiMichelino Di RosaGiuseppe MusumeciPublished in: Cells (2020)
Stem cell therapy and tissue engineering represent a promising approach for cartilage regeneration. However, they present limits in terms of mechanical properties and premature de-differentiation of engineered cartilage. Cycloastragenol (CAG), a triterpenoid saponin compound and a hydrolysis product of the main ingredient in Astragalus membranaceous, has been explored for cartilage regeneration. The aim of this study was to investigate CAG's ability to promote cell proliferation, maintain cells in their stable active phenotype, and support the production of cartilaginous extracellular matrix (ECM) in human adipose-derived mesenchymal stem cells (hAMSCs) in up to 28 days of three-dimensional (3D) chondrogenic culture. The hAMSC pellets were cultured in chondrogenic medium (CM) and in CM supplemented with CAG (CAG-CM) for 7, 14, 21, and 28 days. At each time-point, the pellets were harvested for histological (hematoxylin and eosin (H&E)), histochemical (Alcian-Blue) and immunohistochemical analysis (Type I, II, and X collagen, aggrecan, SOX9, lubricin). After excluding CAG's cytotoxicity (MTT Assay), improved cell condensation, higher glycosaminoglycans (sGAG) content, and increased cell proliferation have been detected in CAG-CM pellets until 28 days of culture. Overall, CAG improved the chondrogenic differentiation of hAMSCs, maintaining stable the active chondrocyte phenotype in up to 28 days of 3D in vitro chondrogenic culture. It is proposed that CAG might have a beneficial impact on cartilage regeneration approaches.
Keyphrases
- extracellular matrix
- mesenchymal stem cells
- cell therapy
- stem cells
- cell proliferation
- endothelial cells
- tissue engineering
- transcription factor
- cell cycle
- induced apoptosis
- wound healing
- type diabetes
- high throughput
- binding protein
- metabolic syndrome
- bone marrow
- cell cycle arrest
- single cell
- insulin resistance
- cell death
- oxidative stress
- signaling pathway