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Controlling the mechanical environment in bioreactors represents a key element in the reactors' optimization. Positive effects of fluid flow in three-dimensional bioreactors have been observed, but local stresses at cell scale remain unknown. These effects led to the development of numerical tools to assess the micromechanical environment of cells in bioreactors. Recently, new possible scaffold geometry has emerged: granular packings. In the present study, the primary goal was to compare the efficiency of such a scaffold to the other ones from literature in terms of wall shear stress levels and distributions. To that aim, three different types of granular packings were generated through discrete element method, and computational fluid dynamics was used to simulate the flow within these packings. Shear stress levels and distributions were determined. A linear relationship between shear stress and inlet velocity was observed, and its slope was similar to published data. The distributions of normalized stress were independent of the inlet velocity and were highly comparable to those of widely used porous scaffolds. Granular packings present similar features to more classical porous scaffolds and have the advantage of being easy to manipulate and seed. The methods of this work are generalizable to the study of other granular packing configurations.

An engineered three dimensional (3D) in vitro cell culture system was designed with the goal of inducing and controlling in vitro osteogenesis in a reproducible manner under conditions more similar to the in vivo bone microenvironment than traditional two-dimensional (2D) models. This bioreactor allows efficient mechanical loading and perfusion of an original cubic calcium phosphate bioceramic of highly controlled composition and structure. This bioceramic comprises an internal portion containing homogeneously interconnected macropores surrounded by a dense layer, which minimises fluid flow bypass around the scaffold. This dense and flat layer permits the application of a homogeneous loading on the bioceramic while also enhancing its mechanical strength. Numerical modelling of constraints shows that the system provides direct mechanical stimulation of cells within the scaffold. Experimental results establish that under perfusion at a steady flow of 2 µL/min, corresponding to 3 ≤ Medium velocity ≤ 23 µm/s, mouse calvarial cells grow and differentiate as osteoblasts in a reproducible manner, and lay down a mineralised matrix. Moreover, cells respond to mechanical loading by increasing C-fos expression, which demonstrates the effective mechanical stimulation of the culture within the scaffold. In summary, we provide a "proof-of-concept" for osteoblastic cell culture in a controlled 3D culture system under perfusion and mechanical loading. This model will be a tool to analyse bone cell functions in vivo, and will provide a bench testing system for the clinical assessment of bioactive bone-targeting molecules under load.