Coupling curvature-dependent and shear stress-stimulated neotissue growth in dynamic bioreactor cultures: a 3D computational model of a complete scaffold.

The main challenge in tissue engineering consists in understanding and controlling the growth process of in vitro cultured neotissues toward obtaining functional tissues. Computational models can provide crucial information on appropriate bioreactor and scaffold design but also on the bioprocess environment and culture conditions. In this study, the development of a 3D model using the level set method to capture the growth of a microporous neotissue domain in a dynamic culture environment (perfusion bioreactor) was pursued. In our model, neotissue growth velocity was influenced by scaffold geometry as well as by flow- induced shear stresses. The neotissue was modeled as a homogenous porous medium with a given permeability, and the Brinkman equation was used to calculate the flow profile in both neotissue and void space. Neotissue growth was modeled until the scaffold void volume was filled, thus capturing already established experimental observations, in particular the differences between scaffold filling under different flow regimes. This tool is envisaged as a scaffold shape and bioprocess optimization tool with predictive capacities. It will allow controlling fluid flow during long-term culture, whereby neotissue growth alters flow patterns, in order to provide shear stress profiles and magnitudes across the whole scaffold volume influencing, in turn, the neotissue growth

Influence of flow rate and scaffold pore size on cell behavior during mechanical stimulation in a flow perfusion bioreactor.

Mechanically stimulating cell-seeded scaffolds by flow-perfusion is one approach utilized for developing clinically applicable bone graft substitutes. A key challenge is determining the magnitude of stimuli to apply that enhances cell differentiation but minimizes cell detachment from the scaffold. In this study, we employed a combined computational modeling and experimental approach to examine how the scaffold mean pore size influences cell attachment morphology and subsequently impacts upon cell deformation and detachment when subjected to fluid-flow. Cell detachment from osteoblast-seeded collagen-GAG scaffolds was evaluated experimentally across a range of scaffold pore sizes subjected to different flow rates and exposure times in a perfusion bioreactor. Cell detachment was found to be proportional to flow rate and inversely proportional to pore size. Using this data, a theoretical model was derived that accurately predicted cell detachment as a function of mean shear stress, mean pore size, and time. Computational modeling of cell deformation in response to fluid flow showed the percentage of cells exceeding a critical threshold of deformation correlated with cell detachment experimentally and the majority of these cells were of a bridging morphology (cells stretched across pores). These findings will help researchers optimize the mean pore size of scaffolds and perfusion bioreactor operating conditions to manage cell detachment when mechanically simulating cells via flow perfusion. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc

Design and Topology Optimisation of Tissue Scaffolds

Tissue restoration by tissue scaffolding is an emerging technique with many potential applications. While it is well-known that the structural properties of tissue scaffolds play a critical role in cell regrowth, it is usually unclear how optimal tissue regeneration can be achieved. This thesis hereby presents a computational investigation of tissue scaffold design and optimisation. This study proposes an isosurface-based characterisation and optimisation technique for the design of microscopic architecture, and a porosity-based approach for the design of macroscopic structure. The goal of this study is to physically define the optimal tissue scaffold construct, and to establish any link between cell viability and scaffold architecture. Single-objective and multi-objective topology optimisation was conducted at both microscopic and macroscopic scales to determine the ideal scaffold design. A high quality isosurface modelling technique was formulated and automated to define the microstructure in stereolithography format. Periodic structures with maximised permeability, and theoretically maximum diffusivity and bulk modulus were found using a modified level set method. Microstructures with specific effective diffusivity were also created by means of inverse homogenisation. Cell viability simulation was subsequently conducted to show that the optimised microstructures offered a more viable environment than those with random microstructure. The cell proliferation outcome in terms of cell number and survival rate was also improved through the optimisation of the macroscopic porosity profile. Additionally artificial vascular systems were created and optimised to enhance diffusive nutrient transport. The formation of vasculature in the optimisation process suggests that natural vascular systems acquire their fractal shapes through self-optimisation

Design and Topology Optimisation of Tissue Scaffolds

Tissue restoration by tissue scaffolding is an emerging technique with many potential applications. While it is well-known that the structural properties of tissue scaffolds play a critical role in cell regrowth, it is usually unclear how optimal tissue regeneration can be achieved. This thesis hereby presents a computational investigation of tissue scaffold design and optimisation. This study proposes an isosurface-based characterisation and optimisation technique for the design of microscopic architecture, and a porosity-based approach for the design of macroscopic structure. The goal of this study is to physically define the optimal tissue scaffold construct, and to establish any link between cell viability and scaffold architecture. Single-objective and multi-objective topology optimisation was conducted at both microscopic and macroscopic scales to determine the ideal scaffold design. A high quality isosurface modelling technique was formulated and automated to define the microstructure in stereolithography format. Periodic structures with maximised permeability, and theoretically maximum diffusivity and bulk modulus were found using a modified level set method. Microstructures with specific effective diffusivity were also created by means of inverse homogenisation. Cell viability simulation was subsequently conducted to show that the optimised microstructures offered a more viable environment than those with random microstructure. The cell proliferation outcome in terms of cell number and survival rate was also improved through the optimisation of the macroscopic porosity profile. Additionally artificial vascular systems were created and optimised to enhance diffusive nutrient transport. The formation of vasculature in the optimisation process suggests that natural vascular systems acquire their fractal shapes through self-optimisation

Overcoming conventional modeling limitations using image- driven lattice-boltzmann method simulations for biophysical applications

The challenges involved in modeling biological systems are significant and push the boundaries of conventional modeling. This is because biological systems are distinctly complex, and their emergent properties are results of the interplay of numerous components/processes. Unfortunately, conventional modeling approaches are often limited by their inability to capture all these complexities. By using in vivo data derived from biomedical imaging, image-based modeling is able to overcome this limitation. In this work, a combination of imaging data with the Lattice-Boltzmann Method for computational fluid dynamics (CFD) is applied to tissue engineering and thrombogenesis. Using this approach, some of the unanswered questions in both application areas are resolved. In the first application, numerical differences between two types of boundary conditions: “wall boundary condition” (WBC) and “periodic boundary condition” (PBC), which are commonly utilized for approximating shear stresses in tissue engineering scaffold simulations is investigated. Surface stresses in 3D scaffold reconstructions, obtained from high resolution microcomputed tomography images are calculated for both boundary condition types and compared with the actual whole scaffold values via image-based CFD simulations. It is found that, both boundary conditions follow the same spatial surface stress patterns as the whole scaffold simulations. However, they under-predict the absolute stress values approximately by a factor of two. Moreover, it is found that the error grows with higher scaffold porosity. Additionally, it is found that the PBC always resulted in a lower error than the WBC. In a second tissue engineering study, the dependence of culture time on the distribution and magnitude of fluid shear in tissue scaffolds cultured under flow perfusion is investigated. In the study, constructs are destructively evaluated with assays for cellularity and calcium deposition, imaged using µCT and reconstructed for CFD simulations. It is found that both the shear stress distributions within scaffolds consistently increase with culture time and correlate with increasing levels of mineralized tissues within the scaffold constructs as seen in calcium deposition data and µCT reconstructions. In the thrombogenesis application, detailed analysis of time lapse microscopy images showing yielding of thrombi in live mouse microvasculature is performed. Using these images, image-based CFD modeling is performed to calculate the fluid-induced shear stresses imposed on the thrombi’s surfaces by the surrounding blood flow. From the results, estimates of the yield stress (A critical parameter for quantifying the extent to which thrombi material can resist deformation and breakage) are obtained for different blood vessels. Further, it is shown that the yielding observed in thrombi occurs mostly in the outer shell region while the inner core remains intact. This suggests that the core material is different from the shell. To that end, we propose an alternative mechanism of thrombogenesis which could help explain this difference. Overall, the findings from this work reveal that image-based modeling is a versatile approach which can be applied to different biomedical application areas while overcoming the difficulties associated with conventional modeling

Simulation of cell seeding and retention in a disordered polymeric scaffold

Historically, bone repair has been performed using materials like metals, ceramics, cements and bioactive glass. The major problem with all these materials is that they do not perform the necessary non-structural functions of bone. Engineered tissue, created by growing bone cells on porous biodegradable material (scaffold), will address this issue with current bone repair techniques. Improving engineered tissue treatments requires a thorough understanding of factors affecting cell seeding and proliferation inside a disordered porous material which is not feasible using current experimental techniques. A model for particle transport in a disordered porous material that can predict the particle deposition pattern will be useful to understand the factors influencing particle transport and retention. Such a model has applications ranging from biomedical, microfluidics, environmental and water treatment. Currently available models for filtration or contaminant transport in a porous media either consider the porous media to be uniform or do not predict the particle deposition pattern. We develop an image-based computational model, which incorporates the structure of a disordered porous material, to study the effect of flow and material internal structure on particle transport and deposition, which was then applied to cell seeding. Particle motion and attachment inside the porous material is controlled by a deterministic convection component, obtained by numerically solving the Stokes Equation using FEM; a stochastic diffusion component, modeled using a random walk process, and an electrostatic component estimated using analytical expressions for the interaction of a colloid particle with a surface. Our simulations show that the Peclet number has a significant effect on the cell attachment pattern in a scaffold. At low Peclet numbers, cell attachment is concentrated at the inlet region of the scaffold, while cells penetrate deeper into the scaffold with increasing Peclet number. Additionally, the seeding pattern is found to vary considerably with internal pore structure. Visualization of the data indicates that attachment clusters at low velocity diffusion dominated zones

Pluripotency state affects the mechanical phenotype of the embryonic stem cell nucleus

PhDThe thesis aims at investigating the connection between nucleus mechanical characteristics with pluripotency state and differentiation associated with altered cell gene expression levels. The project investigates the deformation characteristics of the cell nucleus during unconfined compression in a 3D cell-seeded agarose constructs. The studies report modification in the mechanical behaviour of the nucleus in different embryonic stem cell phenotypes based on various pluripotent states (naïve or primed states) or following triggering of early differentiation. A multi-scale model is also presented to simulate dynamic details of mechanical perturbation to cells during compression. The first chapter presents a review of the relevant literature to introduce current progress in the related research field and the second chapter describes the general methods used in the thesis including cell culture, agarose construct preparation, construct compression and microscopy recording. The third chapter presents findings of studies involving the application of compression to embryonic stem cells in naïve and primed sate within agarose scaffolds. A range of parameters relating to the relative cell/nucleus morphological modifications are recorded with analysis and discussion. Chapter four presents studies that investigate the early differentiation of embryonic stem cells from either the naïve and primed pluripotency, achieved by altering cell culture condition, and further reveals the nuclear mechanical characteristic changes. The fifth chapter describes a multi-scale model developed to simulating the 3D cell-seeded agarose compression reported in previous chapters. This model is also used to estimate cell mechanical parameters and show accurate deformation detail in different locations within the construct. A final discussion of the thesis is provided in chapter 6 with a plan for future work.China Scholarship Counci

Continuum Modeling and Simulation in Bone Tissue Engineering

Bone tissue engineering is currently a mature methodology from a research perspective. Moreover, modeling and simulation of involved processes and phenomena in BTE have been proved in a number of papers to be an excellent assessment tool in the stages of design and proof of concept through in-vivo or in-vitro experimentation. In this paper, a review of the most relevant contributions in modeling and simulation, in silico, in BTE applications is conducted. The most popular in silico simulations in BTE are classified into: (i) Mechanics modeling and sca old design, (ii) transport and flow modeling, and (iii) modeling of physical phenomena. The paper is restricted to the review of the numerical implementation and simulation of continuum theories applied to di erent processes in BTE, such that molecular dynamics or discrete approaches are out of the scope of the paper. Two main conclusions are drawn at the end of the paper: First, the great potential and advantages that in silico simulation o ers in BTE, and second, the need for interdisciplinary collaboration to further validate numerical models developed in BTE.Ministerio de Economía y Competitividad del Gobierno España DPI2017-82501-