Maziar Aghvami

Nonlinear strain stiffening is not sufficient to explain how far cells can feel on fibrous protein gels.

Recent observations suggest that cells on fibrous extracellular matrix materials sense mechanical signals over much larger distances than they do on linearly elastic synthetic materials. In this work, we systematically investigate the distance fibroblasts can sense a rigid boundary through fibrous gels by quantifying the spread areas of human lung fibroblasts and 3T3 fibroblasts cultured on sloped collagen and fibrin gels. The cell areas gradually decrease as gel thickness increases from 0 to 150 μm, with characteristic sensing distances of >65 μm below fibrin and collagen gels, and spreading affected on gels as thick as 150 μm. These results demonstrate that fibroblasts sense deeper into collagen and fibrin gels than they do into polyacrylamide gels, with the latter exhibiting characteristic sensing distances of <5 μm. We apply finite-element analysis to explore the role of strain stiffening, a characteristic mechanical property of collagen and fibrin that is not observed in polyacrylamide, in facilitating mechanosensing over long distances. Our analysis shows that the effective stiffness of both linear and nonlinear materials sharply increases once the thickness is reduced below 5 μm, with only a slight enhancement in sensitivity to depth for the nonlinear material at very low thickness and high applied traction. Multiscale simulations with a simplified geometry predict changes in fiber alignment deep into the gel and a large increase in effective stiffness with a decrease in substrate thickness that is not predicted by nonlinear elasticity. These results suggest that the observed cell-spreading response to gel thickness is not explained by the nonlinear strain-stiffening behavior of the material alone and is likely due to the fibrous nature of the proteins.

Multiscale Mechanical Simulations of Cell Compacted Collagen Gels

Engineered tissues are commonly stretched or compressed (i.e., conditioned) during culture to stimulate extracellular matrix (ECM) production and to improve the mechanical properties of the growing construct. The relationships between mechanical stimulation and ECM remodeling, however, are complex, interdependent, and dynamic. Thus, theoretical models are required for understanding the underlying phenomena so that the conditioning process can be optimized to produce functional engineered tissues. Here, we continue our development of multiscale mechanical models by simulating the effect of cell tractions on developing isometric tension and redistributing forces in the surrounding fibers of a collagen gel embedded with explants. The model predicted patterns of fiber reorganization that were similar to those observed experimentally. Furthermore, the inclusion of cell compaction also changed the distribution of fiber strains in the gel compared to the acellular case, particularly in the regions around the cells where the highest strains were found.

A Combined In Vitro Imaging and Multi-Scale Modeling System for Studying the Role of Cell Matrix Interactions in Cutaneous Wound Healing.

Many cell types remodel the extracellular matrix of the tissues they inhabit in response to a wide range of environmental stimuli, including mechanical cues. Such is the case in dermal wound healing, where fibroblast migrate into and remodel the provisional fibrin matrix in a complex manner that depends in part on the local mechanical environment and the evolving multi-scale mechanical interactions of the system. In this study, we report on the development of an image-based multi-scale mechanical model that predicts the short-term (24 hours), structural reorganization of a fibrin gel by fibroblasts. These predictive models are based on an in vitro experimental system where clusters of fibroblasts (i.e., explants) were spatially arranged into a triangular geometry onto the surface of fibrin gels that were subjected to either Fixed or Free in-plane mechanical constraints. Experimentally, regional differences in short-term structural remodeling and cell migration were observed for the two gel boundary conditions. A pilot experiment indicated that these small differences in the short-term remodeling of the fibrin gel translate into substantial differences in long-term (4 weeks) remodeling, particularly in terms of collagen production. The multi-scale models were able to predict some regional differences in remodeling and qualitatively similar reorganization patterns for the two boundary conditions. However, other aspects of the model, such as the magnitudes and rates of deformation of gel, did not match the experiments. These discrepancies between model and experiment provide fertile ground for challenging model assumptions and devising new experiments to enhance our understanding of how this multi-scale system functions. These efforts will ultimately improve the predictions of the remodeling process, particularly as it relates to dermal wound healing and the reduction of patient scarring. Such models could be used to recommend patient-specific mechanical-based treatment dependent on parameters such as wound geometry, location, age, and health.

Pressure Drop and Heat Transfer of Fully Developed, Laminar Flow in Rough, Rectangular Microchannels

In this paper, a Gaussian, isotropic distribution has been applied to model the wall roughness in microchannels of rectangular cross-section in order to investigate the friction factor and convection coefficient of fully developed, laminar and incompressible flows. As the hydraulic diameter of (micro-) channels decreases, the surface to volume ratio increases rapidly. As a result, the surface phenomena and the effect of roughness become more significant. There is a need for a better understanding of the effect of wall roughness on fluid flow characteristics in microchannels which make them well suited for a wide variety of unique cooling applications. The results of this research indicate that the friction factor and convection heat transfer coefficient increase by increasing the relative roughness of channel. The effect of changing the aspect ratio has also been investigated in this research. In addition, results are compared with experimental data obtained by different researchers.Copyright

Fiber Network Models Predict Enhanced Cell Mechanosensing on Fibrous Gels

The propagation of mechanical signals through nonlinear fibrous tissues is much more extensive than through continuous synthetic hydrogels. Results from recent studies indicate that increased mechanical propagation arises from the fibrous nature of the material rather than the strain-stiffening property. The relative importance of different parameters of the fibrous network structure to this propagation, however, remains unclear. In this work, we directly compared the mechanical response of substrates of varying thickness subjected to a constant cell traction force using either a nonfibrous strain-stiffening continuum-based model or a volume-averaged fiber network model consisting of two different types of fiber network structures: one with low fiber connectivity (growth networks) and one with high fiber connectivity (Delaunay networks). The growth network fiber models predicted a greater propagation of substrate displacements through the model and a greater sensitivity to gel thickness compared to the more connected Delaunay networks and the nonlinear continuum model. Detailed analysis of the results indicates that rotational freedom of the fibers in a network with low fiber connectivity is critically important for enhanced, long-range mechanosensing. Our findings demonstrate the utility of multiscale models in predicting cells mechanosensing on fibrous gels, and they provide a more complete understanding of how cell traction forces propagate through fibrous tissues, which has implications for the design of engineered tissues and the stem cell niche.

A Comparative Assessment of Implant Site Viability in Humans and Rats

Our long-term objective is to devise methods to improve osteotomy site preparation and, in doing so, facilitate implant osseointegration. As a first step in this process, we developed a standardized oral osteotomy model in ovariectomized rats. There were 2 unique features to this model: first, the rats exhibited an osteopenic phenotype, reminiscent of the bone health that has been reported for the average dental implant patient population. Second, osteotomies were produced in healed tooth extraction sites and therefore represented the placement of most implants in patients. Commercially available drills were then used to produce osteotomies in a patient cohort and in the rat model. Molecular, cellular, and histologic analyses demonstrated a close alignment between the responses of human and rodent alveolar bone to osteotomy site preparation. Most notably in both patients and rats, all drilling tools created a zone of dead and dying osteocytes around the osteotomy. In rat tissues, which could be collected at multiple time points after osteotomy, the fate of the dead alveolar bone was followed. Over the course of a week, osteoclast activity was responsible for resorbing the necrotic bone, which in turn stimulated the deposition of a new bone matrix by osteoblasts. Collectively, these analyses support the use of an ovariectomy surgery rat model to gain insights into the response of human bone to osteotomy site preparation. The data also suggest that reducing the zone of osteocyte death will improve osteotomy site viability, leading to faster new bone formation around implants.

Biophysical regulation of osteotomy healing: An animal study

BACKGROUND Osteotomies have been performed for centuries yet there remains a remarkable lack of consensus on optimal methods for cutting bone. There is universal agreement, however, that preserving cell viability is critical. PURPOSE To identify mechanobiological parameters influencing bone formation after osteotomy site preparation. MATERIALS AND METHODS A murine maxillary osteotomy model was used to evaluate healing. Computational modeling characterized stress and strain distributions in the osteotomy, as well as the magnitude and distribution of heat generated by drilling. The impact of osteocyte death and bone composition were assessed using molecular and cellular assays. RESULTS The phases of osteotomy healing in mice align closely with results in large animals; in addition, molecular analyses extended our understanding of osteoprogenitor cell proliferation, differentiation, and mineralization. Computational analyses provided insights into temperature changes caused by drilling and the mechanobiological state in the healing osteotomies, while concomitant cellular assays correlate drill speed with osteocyte apoptosis and bone resorption. Even when drilling was controlled, trabeculated, spongy (Type III) bone healed faster than densely lamellar (Type I) bone because of the abundance of Wnt responsive osteoprogenitor cells in the former. CONCLUSIONS These data provide a mechanobiological framework for evaluating tools and technologies designed to improve osteotomy site preparation.

Fibroblast-Mediated Fiber Realignment in Fibrin Gels

When fibroblasts are added to a fibrin gel, the cells rapidly compact the gel and produce a fiber alignment pattern that depends in part on the cell traction forces, gel geometry, and gel mechanical constraints [1]. Over time the fibrin is digested and replaced with cell synthesized collagen and other extracellular matrix (ECM) proteins that follow the initial alignment pattern of the gel [2]. This remodeling process proceeds in a complex and integrated manner that is influenced by the mechanical environment [3]. In order to better understand fibroblast-fibrin interactions and the remodeling process, we obtained time-lapse images of the development of fiber alignment between clusters of dermal fibroblasts (i.e., explants) in a fibrin gel. The experimental results were then compared to a model that incorporated the effects of traction forces on ECM reorganization.© 2013 ASME

A thermal and biological analysis of bone drilling

With the introduction of high-speed cutting tools, clinicians have recognized the potential for thermal damage to the material being cut. Here, we developed a mathematical model of heat transfer caused by drilling bones of different densities and validated it with respect to experimentally measured temperatures in bone. We then coupled these computational results with a biological assessment of cell death following osteotomy site preparation. Parameters under clinical control, e.g., drill diameter, rotational speed, and irrigation, along with patient-specific variables such as bone density were evaluated in order to understand their contributions to thermal damage. Predictions from our models provide insights into temperatures and thresholds that cause osteocyte death and that can ultimately compromise stability of an implant.

Roughness Effect on Pressure Drop for Electroosmotic (EO) Flow in Microtubes

This paper presents a theoretical model of the roughness effect on friction factor and pressure drop of fully developed, laminar flow in microtubes by considering the effect of the electrical double layer. The EDL potential distribution is calculated using the Poisson–Boltzmann equation and then the velocity profile is obtained by solving the fluid momentum equation with a body force term. The wall roughness in microtubes is modeled by utilizing a Gaussian, isotropic distribution. It is found that the effect of roughness is to increase the friction factor and pressure drop of the electroosmotic flow in microtubes.Copyright