Biophysical Regulations of Epigenetic State and Notch Signaling in Neural Development Using Microgroove Substrates

TY - JOUR

T1 - Biophysical Regulations of Epigenetic State and Notch Signaling in Neural Development Using Microgroove Substrates

AU - Hsu, Chia Chen

AU - Serio, Andrea

AU - Gopal, Sahana

AU - Gelmi, Amy

AU - Chiappini, Ciro

AU - Desai, Ravi A.

AU - Stevens, Molly M.

N1 - Publisher Copyright: © 2022 The Authors. Published by American Chemical Society.

PY - 2022/7/27

Y1 - 2022/7/27

N2 - A number of studies have recently shown how surface topography can alter the behavior and differentiation patterns of different types of stem cells. Although the exact mechanisms and molecular pathways involved remain unclear, a consistent portion of the literature points to epigenetic changes induced by nuclear remodeling. In this study, we investigate the behavior of clinically relevant neural populations derived from human pluripotent stem cells when cultured on polydimethylsiloxane microgrooves (3 and 10 μm depth grooves) to investigate what mechanisms are responsible for their differentiation capacity and functional behavior. Our results show that microgrooves enhance cell alignment, modify nuclear geometry, and significantly increase cellular stiffness, which we were able to measure at high resolution with a combination of light and electron microscopy, scanning ion conductance microscopy (SICM), and atomic force microscopy (AFM) coupled with quantitative image analysis. The microgrooves promoted significant changes in the epigenetic landscape, as revealed by the expression of key histone modification markers. The main behavioral change of neural stem cells on microgrooves was an increase of neuronal differentiation under basal conditions on the microgrooves. Through measurements of cleaved Notch1 levels, we found that microgrooves downregulate Notch signaling. We in fact propose that microgroove topography affects the differentiation potential of neural stem cells by indirectly altering Notch signaling through geometric segregation and that this mechanism in parallel with topography-dependent epigenetic modulations acts in concert to enhance stem cell neuronal differentiation.

AB - A number of studies have recently shown how surface topography can alter the behavior and differentiation patterns of different types of stem cells. Although the exact mechanisms and molecular pathways involved remain unclear, a consistent portion of the literature points to epigenetic changes induced by nuclear remodeling. In this study, we investigate the behavior of clinically relevant neural populations derived from human pluripotent stem cells when cultured on polydimethylsiloxane microgrooves (3 and 10 μm depth grooves) to investigate what mechanisms are responsible for their differentiation capacity and functional behavior. Our results show that microgrooves enhance cell alignment, modify nuclear geometry, and significantly increase cellular stiffness, which we were able to measure at high resolution with a combination of light and electron microscopy, scanning ion conductance microscopy (SICM), and atomic force microscopy (AFM) coupled with quantitative image analysis. The microgrooves promoted significant changes in the epigenetic landscape, as revealed by the expression of key histone modification markers. The main behavioral change of neural stem cells on microgrooves was an increase of neuronal differentiation under basal conditions on the microgrooves. Through measurements of cleaved Notch1 levels, we found that microgrooves downregulate Notch signaling. We in fact propose that microgroove topography affects the differentiation potential of neural stem cells by indirectly altering Notch signaling through geometric segregation and that this mechanism in parallel with topography-dependent epigenetic modulations acts in concert to enhance stem cell neuronal differentiation.

KW - epigenetics

KW - micropatterning

KW - neural stem cell

KW - neural tissue engineering

KW - neuron

KW - Notch signaling

KW - topography

UR - http://www.scopus.com/inward/record.url?scp=85135216432&partnerID=8YFLogxK

U2 - 10.1021/acsami.2c01996

DO - 10.1021/acsami.2c01996

M3 - Article

AN - SCOPUS:85135216432

SN - 1944-8244

VL - 14

SP - 32773

EP - 32787

JO - ACS Applied Materials and Interfaces

JF - ACS Applied Materials and Interfaces

IS - 29

ER -