Substrate softness promotes terminal differentiation of human keratinocytes without altering their ability to proliferate back into a rigid environment

Archives of Dermatological Research
Choua YaRomain Debret

Abstract

Substrate stiffness is a key regulator of cell behavior. To investigate how mechanical properties of cell microenvironment affect the human keratinocyte, primary cells were seeded on polyacrylamide hydrogels of different compliances (soft: 4 kPa, medium: 14 kPa, rigid: 45 kPa) in comparison with glass coverslip (> GPa). Keratinocyte spreading and proliferation were strongly decreased on the softest hydrogel, while no significant difference was observed between medium, rigid hydrogels and glass coverslip, and cells' viability was comparable in all conditions after 72 h. We then performed a RNA-seq to compare the transcriptomes from keratinocytes cultured for 72 h on the softest hydrogel or on coverslips. The cells on the soft hydrogel showed a strong increase in the expression of late differentiation marker genes from the epidermal differentiation complex (1q21) and the antioxidant machinery. In parallel, these cells displayed a significant loss of expression of the matrix receptors (integrin α6 and β1) and the EGF receptor. However, when these cells were replated on a plastic culture plate (> GPa), they were able to re-engage the proliferation machinery with a strong colony-formation efficiency. Overall, using low-calcium diffe...Continue Reading

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Jun 24, 2021·Journal of Dental Research·Y XieL Chen

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