Testing 3D printed biological platform for advancing simulated microgravity and space mechanobiology research

Abstract

The advancement of microgravity simulators is helping many researchers better understanding the impact of the mechanically unloaded space environment on cellular function and disfunction. However, performing microgravity experiments on Earth, using simulators such as the Random Positioning Machine, introduces some unique practical challenges, including air bubble formation and leakage of growth medium from tissue culture flask and plates, all of which limit research progress. Here, we developed an easy-to-use hybrid biological platform designed with the precision of 3D printing technologies combined with PDMS microfluidic fabrication processes to facilitate reliable and reproducible microgravity cellular experiments. The system has been characterized for applications in the contest of brain cancer research by exposing glioblastoma and endothelial cells to 24 h of simulated microgravity condition to investigate the triggered mechanosensing pathways involved in cellular adaptation to the new environment. The platform demonstrated compatibility with different biological assays, i.e., proliferation, viability, morphology, protein expression and imaging of molecular structures, showing advantages over the conventional usage of culture flask. Our results indicated that both cell types are susceptible when the gravitational vector is disrupted, confirming the impact that microgravity has on both cancer and healthy cells functionality. In particular, we observed deactivation of Yap-1 molecule in glioblastoma cells and the remodeling of VE-Cadherin junctional protein in endothelial cells. The study provides support for the application of the proposed biological platform for advancing space mechanobiology research, also highlighting perspectives and strategies for developing next generation of brain cancer molecular therapies, including targeted drug delivery strategies.

Lab-On-a-Chip Technologies for Microgravity Simulation and Space Applications

“…In addition, Silvani et al replicated the in-vivo environment of glioblastoma multiforme (GBM) brain tumors with a microfluidic LOC device [83] and RPM for microgravity simulation [83]. Moreover, in a later work, Silvani utilized 3D printing for the fabrication of the LOC device called microgravity on chip (MOC) (Figure 3b) to cut costs and time for microgravity research for brain cancer cells [68]. This device also eliminated the conventional problems of bubble formation and leakage in RPM [68].…”

“…Moreover, in a later work, Silvani utilized 3D printing for the fabrication of the LOC device called microgravity on chip (MOC) (Figure 3b) to cut costs and time for microgravity research for brain cancer cells [68]. This device also eliminated the conventional problems of bubble formation and leakage in RPM [68]. Diamagnetic levitation is another laboratory-based method to simulate microgravity on earth.…”

Lab-on-a-Chip Technologies for Microgravity Simulation and Space Applications

“…In addition, Silvani et al replicated the in vivo environment of glioblastoma multiforme (GBM) brain tumors with a microfluidic LOC device [ 85 ] and RPM for microgravity simulation [ 85 ]. Moreover, in a later work, Silvani utilized 3D printing for the fabrication of the LOC device called microgravity on chip (MOC) ( Figure 5 b) to cut costs and time for microgravity research for brain cancer cells [ 72 ]. This device also eliminates the conventional problems of bubble formation and leakage in RPM [ 72 ].…”

“…Moreover, in a later work, Silvani utilized 3D printing for the fabrication of the LOC device called microgravity on chip (MOC) ( Figure 5 b) to cut costs and time for microgravity research for brain cancer cells [ 72 ]. This device also eliminates the conventional problems of bubble formation and leakage in RPM [ 72 ].…”

“…Copyright 2020, Authors) ( b ) Top view of microgravity on-chip to research brain cancer cells. (Adapted with permission from [ 72 ], Copyright 2022, Authors).…”