The Janus Role of Adhesion in Chondrogenesis

Abstract

Tackling the first stages of the chondrogenic commitment is essential to drive chondrogenic differentiation to healthy hyaline cartilage and minimize hypertrophy. During chondrogenesis, the extracellular matrix continuously evolves, adapting to the tissue adhesive requirements at each stage. Here, we take advantage of previously developed nanopatterns, in which local surface adhesiveness can be precisely tuned, to investigate its effects on prechondrogenic condensation. Fluorescence live cell imaging, immunostaining, confocal microscopy and PCR analysis are used to follow the condensation process on the nanopatterns. Cell tracking parameters, condensate morphology, cell–cell interactions, mechanotransduction and chondrogenic commitment are evaluated in response to local surface adhesiveness. Results show that only condensates on the nanopatterns of high local surface adhesiveness are stable in culture and able to enter the chondrogenic pathway, thus highlighting the importance of controlling cell–substrate adhesion in the tissue engineering strategies for cartilage repair.

Mesenchymal Stem Cells Isolated from Paediatric Paravertebral Adipose Tissue Show Strong Osteogenic Potential

“…Specific staining revealed that the cells were positive for CD29, CD44, CD73, CD90, CD105 and CD166 and negative for CD14, CD31, CD34 and CD45 (adipose-derived mesenchymal stem cells; normal, human (ATCC PCS-500-011)). The cell line has been used to date for various research applications, including adipose tissue regeneration [ 21 ], osteointegration in bioactive coatings [ 22 ], chondrogenesis [ 23 ] and neurogenesis [ 24 ].…”

Nanoscale ligand density modulates gap junction intercellular communication of cell condensates during chondrogenesis

“…We have previously developed a technique to produce RGD-dendrimer-based nanopatterns with liquid-like order and controlled spacing on large surfaces areas, thus being fully compatible with normal cell culture protocols [31]. These nanopatterns allowed us to control cell adhesion and differentiation of hASCs towards different lineages [31][32][33][34]36]. We previously found that S90 nanopatterns produce larger and mechanically more stable prechondrogenic cell condensates [32,33,36].…”

“…These nanopatterns allowed us to control cell adhesion and differentiation of hASCs towards different lineages [31][32][33][34]36]. We previously found that S90 nanopatterns produce larger and mechanically more stable prechondrogenic cell condensates [32,33,36]. Cells on pristine and nanopatterned substrates immediately start aggregating, while cells on FN coatings first proliferate and adopt a monolayer configuration, from which a few small cell condensates develop after 3 to 5 days in culture [32].…”

“…qPCR and data analysis qPCR was performed as previously described [36], with the Sso Advanced Universal SYBR Green Supermix kit (Bio-Rad, 1725271) in an Applied Biosystems StepOnePlus Real-Time PCR Machine (Thermo Fisher Scientific). Commercial primer pairs were used for VCL (Bio-Rad, qHsaCID0020885) and GJA1 (Bio-Rad, qHsaCID0012977), as well as B2M (Bio-Rad, qHsaCID0015347) and RPL24 (Bio-Rad, qHsaCID0038677) as housekeeping genes.…”

Dual Function Molecules and Processes in Cell Fate Decision: A Preface to the Special Issue

“…The first contribution provided by Casanellas et al [ 5 ] regards the Janus role of cell adhesion in differentiation. Specifically, the authors focus on chondrogenesis, which implies a continuous extracellular matrix remodeling to maintain healthy hyaline cartilage and minimize hypertrophy.…”