Design standards for engineered tissues

Traditional technologies are required to meet specific, quantitative standards of safety and performance. In tissue engineering, similar standards will have to be developed to enable routine clinical use and customized tissue fabrication. In this essay, we discuss a framework of concepts leading towards general design standards for tissue-engineering, focusing in particular on systematic design strategies, control of cell behavior, physiological scaling, fabrication modes and functional evaluation

Myofibrils in Cardiomyocytes Tend to Assemble Along the Maximal Principle Stress Directions

The mechanisms underlying the spatial organization of self-assembled myofibrils in cardiac tissues remain incompletely understood. By modeling cells as elastic solids under active cytoskeletal contraction, we found a good correlation between the predicted maximal principal stress directions and the in vitro myofibril orientations in individual cardiomyocytes. This implies that actomyosin fibers tend to assemble along the maximal tensile stress (MTS) directions. By considering the dynamics of focal adhesion and myofibril formation in the model, we showed that different patterns of myofibril organizations in mature versus immature cardiomyocytes can be explained as the consequence of the different levels of force-dependent remodeling of focal adhesions. Further, we applied the mechanics model to cell pairs and showed that the myofibril organizations can be regulated by a combination of multiple factors including cell shape, cell–substrate adhesions, and cell–cell adhesions. This mechanics model can guide the rational design in cardiac tissue engineering where recapitulating in vivo myofibril organizations is crucial to the contractile function of the heart

Modeling of cardiac muscle thin films: Pre-stretch, passive and active behavior

Recent progress in tissue engineering has made it possible to build contractile bio-hybrid materials that undergo conformational changes by growing a layer of cardiac muscle on elastic polymeric membranes. Further development of such muscular thin films for building actuators and powering devices requires exploring several design parameters, which include the alignment of the cardiac myocytes and the thickness/Young's modulus of elastomeric film. To more efficiently explore these design parameters, we propose a 3-D phenomenological constitutive model, which accounts for both the passive deformation including pre-stretch and the active behavior of the cardiomyocytes. The proposed 3-D constitutive model is implemented within a finite element framework, and can be used to improve the current design of bio-hybrid thin films and help developing bio-hybrid constructs capable of complex conformational changes.Engineering and Applied Science

Traumatic Brain Injury and the Neuronal Microenvironment: A Potential Role for Neuropathological Mechanotransduction

Traumatic brain injury (TBI) is linked to several pathologies for which there is a lack of understanding of disease mechanisms and therapeutic strategies. To elucidate injury mechanisms, it is important to consider how physical forces are transmitted and transduced across all spatial scales of the brain. Although the mechanical response of the brain is typically characterized by its material properties and biological structure, cellular mechanotransduction mechanisms also exist. Such mechanisms can affect physiological processes by responding to exogenous mechanical forces directed through sub-cellular components, such as extracellular matrix and cell adhesion molecules, to mechanosensitive intracellular structures that regulate mechanochemical signaling pathways. We suggest that cellular mechanotransduction may be an important mechanism underlying the initiation of cell and sub-cellular injuries ultimately responsible for the diffuse pathological damage and clinical symptoms observed in TBI, thereby providing potential therapeutic opportunities not previously explored in TBI

Protein-Based Textiles: Bio-Inspired and Bio-Derived Materials for Medical and Non-Medical Applications

The hierarchical structure-dependent function of self-assembling proteins regulates the biochemical and mechanical functions of cells, tissues, and organs. These multi-scale properties make proteins desirable candidates for novel supramolecular materials that require tailored properties and customizable functions. The ability to translate molecular domains of proteins into the bulk production of conformable materials, such as textiles, is restricted by the current limitations in fabrication technologies and the finite abundance of protein starting material. We will review the common features of self-assembling proteins, including their structure-dependent mechanical properties and how these characteristics have inspired techniques for manufacturing protein-based textiles. These technologies coupled with recent advances in recombinant protein synthesis enable the bulk production of fibers and fabrics that emulate the hierarchical function of natural protein networks.Engineering and Applied Science

A Simple Model for Nanofiber Formation by Rotary Jet-Spinning

Nanofibers are microstructured materials that span a broad range of applications from tissue engineering scaffolds to polymer transistors. An efficient method of nanofiber production is rotary jet-spinning (RJS), consisting of a perforated reservoir rotating at high speeds along its axis of symmetry, which propels a liquid, polymeric jet out of the reservoir orifice that stretches, dries, and eventually solidifies to form nanoscale fibers. We report a minimal scaling framework complemented by a semi-analytic and numerical approach to characterize the regimes of nanofiber production, leading to a theoretical model for the fiber radius consistent with experimental observations. In addition to providing a mechanism for the formation of nanofibers, our study yields a phase diagram for the design of continuous nanofibers as a function of process parameters with implications for the morphological quality of fibers.Engineering and Applied Science

Differential Contributions of Conformation Extension and Domain Unfolding to Properties of Fibronectin Nanotextiles

Fibronectin (FN) textiles are built as nanometer-thick fabrics. When uniaxially loaded, these fabrics exhibit a distinct threshold between elastic and plastic deformation with increasing stretch. Fabric mechanics are modeled using an eight-chain network and two-state model, revealing that elastic properties of FN depend on conformational extension of the protein and that plastic deformation depends on domain unfolding. Our results suggest how the molecular architecture of a molecule can be exploited for designer mechanical properties of a bulk material.Engineering and Applied Science

Environ Mol Mutagen

Epigenetic changes such as DNA methylation may be a molecular mechanism through which environmental exposures affect health. Methylation of Alu and long interspersed nucleotide elements (LINE-1) is a well-established measure of DNA methylation often used in epidemiologic studies. Yet, few studies have examined the effects of host factors on LINE-1 and Alu methylation in children. We characterized the relationship of age, sex, and prenatal exposure to persistent organic pollutants (POPs), dichlorodiphenyl trichloroethane (DDT), dichlorodiphenyldichloroethylene (DDE), and polybrominated diphenyl ethers (PBDEs), with DNA methylation in a birth cohort of Mexican-American children participating in the CHAMACOS study. We measured Alu and LINE-1 methylation by pyrosequencing bisulfite-treated DNA isolated from whole blood samples collected from newborns and nine-year old children (n\ue2\u20ac\u2030=\ue2\u20ac\u2030358). POPs were measured in maternal serum during late pregnancy. Levels of DNA methylation were lower in nine-year olds compared to newborns and were higher in boys compared to girls. Higher prenatal DDT/E exposure was associated with lower Alu methylation at birth, particularly after adjusting for cell type composition (P\ue2\u20ac\u2030=\ue2\u20ac\u20300.02 for o,p' -DDT). Associations of POPs with LINE-1 methylation were only identified after examining the co-exposure of DDT/E with PBDEs simultaneously. Our data suggest that repeat element methylation can be an informative marker of epigenetic differences by age and sex and that prenatal exposure to POPs may be linked to hypomethylation in fetal blood. Accounting for co-exposure to different types of chemicals and adjusting for blood cell types may increase sensitivity of epigenetic analyses for epidemiological studies.P01 ES009605/ES/NIEHS NIH HHS/United StatesP01 ES009605/ES/NIEHS NIH HHS/United StatesR01 ES015572/ES/NIEHS NIH HHS/United StatesR01 OH007400/OH/NIOSH CDC HHS/United States2015-04-27T00:00:00

Quality Metrics for Stem Cell-Derived Cardiac Myocytes

Summary Advances in stem cell manufacturing methods have made it possible to produce stem cell-derived cardiac myocytes at industrial scales for in vitro muscle physiology research purposes. Although FDA-mandated quality assurance metrics address safety issues in the manufacture of stem cell-based products, no standardized guidelines currently exist for the evaluation of stem cell-derived myocyte functionality. As a result, it is unclear whether the various stem cell-derived myocyte cell lines on the market perform similarly, or whether any of them accurately recapitulate the characteristics of native cardiac myocytes. We propose a multiparametric quality assessment rubric in which genetic, structural, electrophysiological, and contractile measurements are coupled with comparison against values for these measurements that are representative of the ventricular myocyte phenotype. We demonstrated this procedure using commercially available, mass-produced murine embryonic stem cell- and induced pluripotent stem cell-derived myocytes compared with a neonatal mouse ventricular myocyte target phenotype in coupled in vitro assays

Connexin43 ablation in foetal atrial myocytes decreases electrical coupling, partner connexins, and sodium current

Aims Remodelling and regional gradients in expression of connexins (Cx) are thought to contribute to atrial electrical dysfunction and atrial fibrillation. We assessed the effect of interaction between Cx43, Cx40, and Cx45 on atrial cell-to-cell coupling and inward Na current (INa) in engineered pairs of atrial myocytes derived from wild-type mice (Cx43+/+) and mice with genetic ablation of Cx43 (Cx43−/−). Methods and results Cell pairs were engineered by microcontact printing from atrial Cx43+/+ and Cx43−/− murine myocytes (1 day before birth, 3-5 days in culture). Dual and single voltage clamp were used to measure intercellular electrical conductance, gj, and its dependence on transjunctional voltage, Vj, single gap junction channel conductances, and INa. 3D reconstructions of Cx43, Cx40, and Cx45 immunosignals in gap junctions were made from confocal slices. Full genetic Cx43 ablation produced a decrease in immunosignals of Cx40 to 62 ± 10% (mean ± SE; n= 17) and Cx45 to 66 ± 8% (n= 16). Gj decreased from 80 ± 9 nS (Cx43+/+, n= 17) to 24 ± 2 nS (Cx43−/−, n= 35). Single channel analysis showed a shift in the main peak of the channel histogram from 49 ± 1.7 nS (Cx43+/+) to 67 ± 1.8 nS (Cx43−/−) with a second minor peak appearing at 27 ± 1.5 pS. The dependence of gj on Vj decreased with Cx43 ablation. Importantly, peak INa decreased from −350 ± 44 pA/pF (Cx43+/+) to −154 ± 28 pA/pF (Cx43−/−). Conclusions The dependence of Cx40, Cx45, and INa on Cx43 expression indicates a complex interaction between connexins and INa in the atrial intercalated discs that is likely to be of relevance for arrhythmogenesi