Preparation and Mechano-Functional Characterization of PEGylated Fibrin Hydrogels: Impact of Thrombin Concentration.

Three-dimensional (3D) neuronal cultures grown in hydrogels are promising platforms to design brain-like neuronal networks in vitro. However, the optimal properties of such cultures must be tuned to ensure a hydrogel matrix sufficiently porous to promote healthy development but also sufficiently rigid for structural support. Such an optimization is difficult since it implies the exploration of different hydrogel compositions and, at the same time, a functional analysis to validate neuronal culture viability. To advance in this quest, here we present a combination of a rheological protocol and a network-based functional analysis to investigate PEGylated fibrin hydrogel networks with gradually higher stiffness, achieved by increasing the concentration of thrombin. We observed that moderate thrombin concentrations of 10% and 25% in volume shaped healthy networks, although the functional traits depended on the hydrogel stiffness, which was much higher for the latter concentration. Thrombin concentrations of 65% or higher led to networks that did not survive. Our results illustrate the difficulties and limitations in preparing 3D neuronal networks, and stress the importance of combining a mechano-structural characterization of a biomaterial with a functional one.

Rheological Characterization of Three-Dimensional Neuronal Cultures Embedded in PEGylated Fibrin Hydrogels.

Three-dimensional (3D) neuronal cultures are valuable models for studying brain complexity in vitro, and the choice of the bulk material in which the neurons grow is a crucial factor in establishing successful cultures. Indeed, neuronal development and network functionality are influenced by the mechanical properties of the selected material; in turn, these properties may change due to neuron-matrix interactions that alter the microstructure of the material. To advance our understanding of the interplay between neurons and their environment, here we utilized a PEGylated fibrin hydrogel as a scaffold for mouse primary neuronal cultures and carried out a rheological characterization of the scaffold over a three-week period, both with and without cells. We observed that the hydrogels exhibited an elastic response that could be described in terms of the Young's modulus E. The hydrogels without neurons procured a stable E≃420 Pa, while the neuron-laden hydrogels showed a higher E≃590 Pa during the early stages of development that decreased to E≃340 Pa at maturer stages. Our results suggest that neurons and their processes dynamically modify the hydrogel structure during development, potentially compromising both the stability of the material and the functional traits of the developing neuronal network.

Spatiotemporal Organization of Correlated Local Activity within Global Avalanches in Slowly Driven Interfaces.

We study the jerky response of slowly driven fronts in disordered media, just above the depinning transition. We focus on how spatially disconnected clusters of internally correlated activity lead to large-scale velocity fluctuations in the form of global avalanches and identify three different ways in which local activity clusters may organize within a global avalanche, depending on the distance to criticality. Our analysis provides new scaling relations between the power-law exponents of the statistical distributions of sizes and durations of local bursts and global avalanches. Fluid fronts of imbibition in heterogeneous media are taken as a case study to validate these scaling relations.

Capillary rise in Hele-Shaw models of disordered media.

We study the capillary rise of a viscous liquid in large Hele-Shaw models of disordered media, both analytically and experimentally. Compared to the Fries-Dreyer and Lucas-Washburn solutions for capillary rise with and without gravity, our experimental data reveal a systematic deviation at short and intermediate times. The original pressure balance equation leading to Washburn's results is reformulated in order to include an additional resisting term, proportional to the mean velocity of the front h˙, which appears naturally as a result of the geometry of the cell. Analytical solutions h(t) are found for displacements with and without gravity. These new solutions reproduce the experimental results very accurately in Hele-Shaw cells of constant gap thickness, where the capillary pressure can be approximated by a constant. In cells of fluctuating gap thickness, where the capillary pressure fluctuates in space, a small additional pressure contribution is required. This correction that depends on h˙ is also studied.

Roughness and intermittent dynamics of imbibition fronts due to capillary and permeability disorder.

We follow the propagation of an air-liquid interface during forced-flow imbibition of a viscous wetting liquid by a random medium, using a high resolution fast camera. Our model disordered medium mimics an open fracture by a Hele-Shaw cell with a two-valued gap spacing randomly distributed in the fracture (or Hele-Shaw) plane. By systematically varying the imposed flow rate we achieve average imbibition front velocities in the range 0.057

Avalanches and non-Gaussian fluctuations of the global velocity of imbibition fronts.

We present an experimental study of the global velocity V(t) of a viscous fluid interface during forced-flow imbibition in a disordered medium. Our high resolution setup shows that the fronts display an intermittent behavior signature of a burstlike dynamics, with power-law distributed avalanches. When measured at scales comparable to the correlation length, velocity fluctuations follow an asymmetric non-Gaussian distribution, whose skewness increases with decreasing measuring window and/or injection flow rate, offering the effective number of degrees of freedom probed in our experiment.