Biology subjects

Fougnier, D.

Publications and source records attributed to Fougnier, D..

3 recordsLinked to original sources

bioRxiv2025

3D osteocyte networks under Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS)

Although osteocytes are known to play a key role in skeletal mechano-adaptation, few in vitro models have investigated how pulsatile mechanical stimuli influence the properties of 3D osteocyte networks. Here we design and develop a microfluidic based in vitro model to study 3D osteocyte networks cultured under Pulsatile Unidirectional Fluid Flow Stimuli (PUFFS). Digital light projection stereolithography was used to design and fabricate a three-chambered PDMS microfluidic chip. Model osteocytes (murine MLO-Y4) were encapsulated in collagen matrix within the chip to form self-assembled three-dimensional (3D) cell networks. Daily stimulus in the form of PUFFS was then applied for upto 21 days. A combination of experiments, computational simulation and analytical modeling was used to characterize the mechanical environment experienced by embedded cells during PUFFS. Viability, morphology, cell-connectivity, expression of key proteins, and gene expression, and real-time calcium signaling within 3D osteocyte networks were characterized at select time-points and compared to static conditions. Results show that PUFFS stimulation at 0.33 and 1.66 Hz can initiate mechanotransduction via calcium signals that are propagated across the network of collagen encapsulated osteocytes via Cx43 junctions. Furthermore, osteocytes cultured in these devices maintain expression of several key osteocyte genes for up to 21d. Taken together, this model can potentially serve as a testbed to study how 3D osteocyte networks respond to dynamic mechanical stimulation relevant to skeletal tissues.

bioengineering↗

bioRxiv2023

Droplet bioprinting of acellular and cell-laden structures at high-resolutions

Advances in Digital Light Processing (DLP) based (bio) printers have made printing of intricate structures at high resolution possible using a wide range of photosensitive bioinks. A typical setup of a DLP bioprinter includes a vat or reservoir filled with liquid bioink, which presents challenges in terms of cost associated with bioink synthesis, high waste, and gravity-induced cell settling, contaminations, or variation in bioink viscosity during the printing process. Here, we report a vat-free, low-volume, waste-free droplet bioprinting method capable of rapidly printing 3D soft structures at high resolution using model bioinks. A multiphase many-body dissipative particle dynamics (mDPD) model was developed to simulate the dynamic process of droplet-based DLP printing and elucidate the roles of surface wettability and bioink viscosity. Process variables such as light intensity, photo-initiator concentration, and bioink formulations were optimized to print 3D soft structures ([~]0.4 to 3 kPa) with an XY resolution of 38 {+/-} 1.5 m and Z resolution of 237{+/-}5.4 m. To demonstrate its versatility, droplet bioprinting was used to print a range of acellular 3D structures such as a lattice cube, a Mayan pyramid, a heart-shaped structure, and a microfluidic chip with endothelialized channels. Droplet bioprinting, performed using model C3H/10T1/2 cells, exhibited high viability (90%) and cell spreading. Additionally, microfluidic devices with internal channel network lined with endothelial cells showed robust monolayer formation while osteoblast-laden constructs showed mineral deposition upon osteogenic induction. Overall, droplet bioprinting could be a low-cost, no-waste, easy-to-use, method to make customized bioprinted constructs for a range of biomedical applications.

bioengineering↗

bioRxiv2023

Printing double network tough hydrogels using Temperature-Controlled Projection Stereolithography (TOPS)

We report a new method to shape double-network (DN) hydrogels into customized microscale 3D structures that exhibit superior mechanical properties in both tension and compression. A one-pot prepolymer formulation containing photo-cross-linkable acrylamide and thermo-reversible sol-gel {kappa}-carrageenan with a suitable crosslinker, and photo-initiator/absorbers are optimized. A new TOPS system is utilized to photo-polymerize the primary acrylamide network into a 3D structure above the sol-gel transition of {kappa}-carrageenan (80{degrees}C), while cooling down generates the secondary physical {kappa}-carrageenan network to realize tough DN hydrogel structures. 3D structures, printed with high lateral (37m) and vertical (180m) resolutions and superior 3D design freedoms (internal voids), exhibit ultimate stress and strain of 200 kPa and 2400% respectively under tension, and simultaneously exhibit high compression stress of 15 MPa with a strain of 95%, both with high recovery rates. The roles of swelling, necking, self-healing, cyclic loading, dehydration, and rehydration on the mechanical properties of printed structures are also investigated. To demonstrate the potential of this technology to make mechanically reconfigurable flexible devices, we print an axicon lens and show that a Bessel beam can be dynamically tuned via user-defined tensile stretching of the device. This technique can be broadly applied to other hydrogels to make novel smart multifunctional devices for a range of applications.

bioengineering↗