Transcriptomic Changes Towards Osteogenic Differentiation of Mesenchymal Stem Cells on 3D Printed GelMA/CNC Hydrogel Under Pulsatile Pressure Environment.

Biomimetic soft hydrogels used in bone tissue engineering frequently produce unsatisfactory outcomes. Here, we investigated how human bone-marrow-derived mesenchymal stem cells (hBMSCs) differentiated into early osteoblasts on remarkably soft three-dimensional (3D) hydrogel (70 ± 0.00049 Pa). Specifically, hBMSCs seeded onto cellulose nanocrystals (CNC) incorporated methacrylate gelatin (GelMA) hydrogels were subjected to pulsatile pressure stimulation (PPS) of 5-20 kPa for seven days. The PPS stimulates cellular processes such as mechanotransduction, cytoskeletal distribution, prohibition of oxidative stress, calcium homeostasis, osteogenic marker gene expression, and osteo-specific cytokine secretions in hBMSCs on soft substrates. The involvement of Piezo 1 was the main ion channel involved in mechanotransduction. Additionally, RNA-sequencing results reveal differential gene expression concerning osteogenic differentiation, bone mineralization, ion channel activity, and focal adhesion. Our findings suggest a practical and highly scalable method for promoting stem cell commitment to osteogenesis on soft matrices for clinical reconstruction. This article is protected by copyright. All rights reserved.