Cell–cell mechanical communications at a large spatial scale (above hundreds of micrometers) have been increasingly recognized in recent decade, which shows importance in tissue-level assembly and morphodynamics. The involved mechanosensing mechanism and resulted physiological functions are still to be fully understood. Recent work showed that traction force sensation in the matrix induces cell communications for self-assembly. Here, based on the experimental model of cell directional migration on Matrigel hydrogel, containing 0.5 mg/ml type I collagen, we studied the mechano-responsive pathways for cell distant communications. Airway smooth muscle (ASM) cells assembled network structure on the hydrogel, whereas stayed isolated individually when cultured on glass without force transmission. Cell directional migration, or network assembly was significantly attenuated by inhibited actomyosin activity, or inhibition of inositol 1,4,5-trisphosphate receptor (IP3R) calcium channel or SERCA pump on endoplasmic reticulum (ER) membrane, or L-type calcium channel on the plasma membrane. Inhibition of integrin β1 with siRNA knockdown reduced cell directional migration and branching assembly, whereas inhibition of cell junctional N-cadherin with siRNA had little effect on distant attractions but blocked branching assembly. Our work demonstrated that the endoplasmic reticulum calcium channels and integrin are mechanosensing signals for cell mechanical communications regulated by actomyosin activity, while N-cadherin is responsible for traction force-induced cell stable connections in the assembly.

Tissues and organs often have specific structures or shapes, which developmental process is coordinated by cells and extracellular matrix modeling. The biomechanical aspect how cells and matrix manage large-spatial constructions for tissue morphogenesis or bioengineering has got increasing attentions, while long-range mechanical communications by cells provide certain insights. Previous work has demonstrated the capability of cells in remodeling matrix structures in distance, however, which biophysical mechanistic studies are still pretty conditional. Here, we investigated the underlying dynamic mechanism of collagen I (COL) fibrillary modeling remotely induced by cell traction force, and the involved cellular mechano-signaling. The research designs were based on large arrays of cell clusters, and with incorporated dynamic tractions, the Molecular Dynamics simulations yielded highly matching outcomes with observed COL fiber clustering in experiments based on large-spatial square, parallelogram, and random-style arrays of cell clusters. The further designed single polygons with variable geometries from triangles to hexagons resulted in predicted structures with assembled COL fibers, which space balance was not maintained when introducing additional contraction at their geometrical centers. The cell cytoskeletal integrity (actin filaments, microtubules), actomyosin contractions, and endoplasmic reticulum calcium channels were essential for the remote fiber inductions, whereas membrane mechanosensitive integrin 1 or Piezo1 alone was less critical in fiber assembly. This work provided new mechanistic insights with dynamic and spatial factors on remote induction of matrix modeling by cells at tissue scale, and the involved cellular mechanism. The assembled biomechanical scaffolds based on pre-designs may lead to applications in tissue engineering.