A standalone bioreactor system to deliver compressive load under perfusion flow to hBMSC-seeded 3D chitosan-graphene templates

Abstract

The availability of engineered biological tissues holds great potential for both clinical applications and basic research in a life science laboratory. A prototype standalone perfusion/compression bioreactor system was proposed to address the osteogenic commitment of stem cells seeded onboard of 3D chitosan-graphene (CHT/G) templates. Testing involved the coordinated administration of a 1 mL/min medium flow rate together with dynamic compression (1% strain at 1 Hz; applied twice daily for 30 min) for one week. When compared to traditional static culture conditions, the application of perfusion and compression stimuli to human bone marrow stem cells using the 3D CHT/G template scaffold induced a sizable effect. After using the dynamic culture protocol, there was evidence of a larger number of viable cells within the inner core of the scaffold and of enhanced extracellular matrix mineralization. These observations show that our novel device would be suitable for addressing and investigating the osteogenic phenotype commitment of stem cells, for both potential clinical applications and basic research.

Small-Scale Fluidized Bed Bioreactor for Long-Term Dynamic Culture of 3D Cell Constructs and in vitro Testing

“…Of note, differences in fluidization level among bioreactors, specially until day 4, are a result of setup inherent variability and beads adjusting to the dynamic environment. These characteristics are similar to other parallelised small-scale technologies ( Tai et al, 2015 ; Qian et al, 2016 ; Lovecchio et al, 2019 ; Valls-Margarit et al, 2019 ), and their validation demonstrates that the prototype could enable simultaneous multiple condition observations and/or establishment of co-culture protocols. It improves the throughput character of the sFBB, as well as, the diverse range of applications from cellular expansion to investigation of cellular responses to external agents and stresses (e.g., hydrodynamic forces, differentiation factors), and paracrine effects of different cells types under co-culture, for example.…”

Micromechanical Behavior of TPMS Scaffolds for Bone Tissue Engineering

“…The FE simulations were performed under different compression magnitudes (6%, 8%, and 10% ramp compression over the height of the construct, i.e., progressive reduction of height during the compression period) and progressive compression periods (10, 20, and 30 s), following numerical and bioreactor studies available in the literature aiming to mimic the adaptation of the scaffold–hydrogel construct to the biological environment. [ 16,23,33,34 ] This resulted in a total of 81 simulation cases, allowing for the evaluation of the effect of the macromechanical compression magnitude and strain rate on the micromechanical environment of each cell substrate provided by the specific TPMS scaffold.…”

Analysis of Intracellular Magnesium and Mineral Depositions during Osteogenic Commitment of 3D Cultured Saos2 Cells

“…To this aim, images acquired with optical microscope were processed, using ImageJ software (National Institute of Health, USA), in order to determine the size of the red (Alizarina Red staining) area. This was calculated using the maximum entropy threshold-based image segmentation method [47]. Four separate fields out of each single microphograph were analysed.…”