A Limb-bud Integrated Microphysiological system to model the effect of Biophysical stimuli in musculoskeletal systems' Organogenesis

Objective

"LIMBO aims to introduce a new class of microphysiological systems (MPSs) that replicate the mechanical microenvironment of the developing limb bud and integrate on-line volumetric spectroscopic analyses. This will provide new insights into biophysical factors in musculoskeletal tissue organogenesis, previously unknown due to technological limitations, and guide regenerative medicine approaches for age-related pathologies. Since Alan Turing's theorization in 1952, developmental biology has relied on the idea that morphogenesis stems from chemical patterns forming from initially uniform substances. Yet, advances in single-cell transcriptomics and lineage tracing have shown that biochemical and genetic instructions alone do not fully explain the self-organizing behavior of developing tissues. A new perspective suggests that mechanical forces are as crucial as gene expression and chemical diffusion, particularly for musculoskeletal tissues development; proper joint formation depends on muscle contractions and foetal movements. A major obstacle to understanding the ""sculpting forces"" driving tissue growth, shape, and differentiation patterns is the lack of technologies to measure the biophysical parameters of developing tissues, replicate the cell scale mechanical milieu, and monitor its temporospatial evolution. MPSs are gaining traction as tools to dissect developmental processes and observe molecular-scale dynamics. However, replicating the mechanics of developing musculoskeletal tissues - affected by cyclical strains, growth-generated internal stresses, and topography/localized curvature induced cues - presents peculiar engineering challenges. I propose a multidisciplinary approach that combines mouse embryonal limb bud cells with an MPS technology for cyclical mechanical stimulation, micro fibrous scaffolds, and acoustic cell patterning to replicate different mechanical cues, and with confocal Raman Spectroscopy Imaging to assess tissues biomolecular time evolution."