Complex Tissue Geometries Research Articles

3D printable phantom for mimicking electrical properties of dermal tissue.

Skin cancer is one of the most ubiquitous forms of cancer that is often overdiagnosed or missed by traditional diagnostic techniques. Bioimpedance spectroscopy (BIS) is a technology that aims to take advantage of the variations in electrical properties of tissue to identify ectopic formations. It is difficult to develop BIS technologies without obtaining tumor tissue samples. One solution is to use a "tissue phantom," a synthetic structure that mimics the properties of tissue. Current solutions using natural biomaterials, such as gelatin, have not been able to create complex tissue geometries that are vital to honing BIS diagnostics. However, semi-synthetic polymers, such has gelatin methacrylate (GelMA), offer the benefits of possessing similar electrical properties to their respective source biomaterial while being 3D printable. In this work, we first measured the impedance of porcine dermal tissue. We then applied these impedance measurements to create an electrically accurate tissue phantom using a photocurable hydrogel, GelMA, and varying concentrations of NaCl, aluminum powder, and titanium dioxide powder.

A model for mechanosensitive cell migration in dynamically morphing soft tissues

Processes such as embryonic development and wound healing involve a complex coupled interplay between cellular contractility and migration, as well as extracellular matrix (ECM) mechanics and remodeling. Despite intense efforts to describe biomechanical interactions between cells and the surrounding extracellular matrix (ECM), the regulatory principles of the coupled cell migration and contractility within dynamically morphing tissues remain poorly understood. Existing theories dealing with cell/ECM contractility do not account for the evolution of cell concentration that is a result of cell migration, which in turn inadvertently influences contractility and vice-versa. Here, we propose a mechanosensitive transient nonlinear theory to connect cell migration and contractility inside 3D ECM, while accounting for ”active” surface effects. Moreover, we introduce a finite element implementation to account for coupling cell migration and contractility capturing the transient large deformation response in complex tissue geometries. Motivated by microtissue experimental setups, simulation results presented in this work, capture changes in the bulk shape of morphing tissues as well as the evolution of the cell concentration for wounded and intact microtissues. The presented theoretical model and computational framework enable us to interpret experimental data, isolate factors that dominate the mechanical behavior, and conduct hypothesis driven research.

Extracellular Matrix Microparticles Improve GelMA Bioink Resolution for 3D Bioprinting at Ambient Temperature.

Current bioinks for 3D bioprinting, such as gelatin-methacryloyl, are generally low viscosity fluids at room temperature, requiring specialized systems to create complex geometries. Adding decellularized extracellular matrix microparticles derived from porcine tracheal cartilage to gelatin-methacryloyl creates a yield stress fluid capable of forming self-supporting structures. This bioink blend performs similarly at 25°C to gelatin-methacryloyl alone at 15°C in linear resolution, print fidelity, and tensile mechanics. This method lowers barriers to manufacturing complex tissue geometries and removes the need for cooling systems.

Comparison of fluorescence lifetime and multispectral imaging for quantitative multiplexing in biological tissue.

Fluorescence lifetime (FLT) multiplexing and multispectral imaging (MSI) are both frequently employed for in vitro and ex vivo biological studies. In vivo applications of MSI for deep seated fluorophores require consideration of diffusive light propagation in biological tissue. We have previously shown that a well-known redshift of fluorescence spectra in diffusive medium induces a fluorophore cross-talk, which cannot be accounted for even with known optical properties of the medium. In contrast, FLT measurements remain largely unaffected by light propagation in tissue, enabling zero cross-talk and accurate relative quantification. While a fully quantitative estimation of fluorophore concentrations requires depth resolved tomographic imaging, this is often not possible due to the difficulty of estimating tissue optical properties and modelling light propagation in complex tissue geometries. Here, we experimentally investigate the performance of planar (non-tomographic) MSI and FLT multiplexing for the quantitative recovery of multiple near-infrared fluorophores embedded in 4-8 mm thick tissue. We show that FLT multiplexing provides a superior quantification accuracy (error < 10%) compared to MSI (error = 20-107%) in tissue. The error rates for MSI increased with tissue thickness and can be directly attributed to the spectral redshift induced cross-talk between emission spectra. Our data indicate that planar FLT multiplexing can provide high quantification accuracy in thick biological tissue without a need for optical property estimation, thereby offering an important validation tool for rapid quantification of fluorophore concentrations in bulk tissue.

A new prospect in magnetic nanoparticle-based cancer therapy: Taking credit from mathematical tissue-mimicking phantom brain models

Distribution patterns/performance of magnetic nanoparticles (MNPs) was visualized by computer simulation and experimental validation on agarose gel tissue-mimicking phantom (AGTMP) models. The geometry of a complex three-dimensional mathematical phantom model of a cancer tumor was examined by tomography imaging. The capability of mathematical model to predict distribution patterns/performance in AGTMP model was captured. The temperature profile vs. hyperthermia duration was obtained by solving bio-heat equations for four different MNPs distribution patterns and correlated with cell death rate. The outcomes indicated that bio-heat model was able to predict temperature profile throughout the tissue model with a reasonable precision, to be applied for complex tissue geometries. The simulation results on the cancer tumor model shed light on the effectiveness of the studied parameters.

Mechanical Properties, Cytocompatibility and Manufacturability of Chitosan:PEGDA Hybrid-Gel Scaffolds by Stereolithography.

Extracellular matrix mimetic hydrogels which hybridize synthetic and natural polymers offer molecularly-tailored, bioactive properties and tunable mechanical strength. In addition, 3D bioprinting by stereolithography allows fabrication of internal pores and defined macroscopic shapes. In this study, we formulated a hybrid biocompatible resin using natural and synthetic polymers (chitosan and polyethylene glycol diacrylate (PEGDA), respectively) by controlling molecular weight of chitosan, feed-ratios, and photo-initiator concentration. Ear-shaped, hybrid scaffolds were fabricated by a stereolithographic method using a 405nm laser. Hybrid hydrogel scaffolds of chitosan (50-190kDa) and PEGDA (575Da) were mixed at varying feed-ratios. Some of the cationic, amino groups of chitosan were neutralized by dialysis in acidic solution containing chitosan in excess of sodium acetate solution to inhibit quenching of newly formed photoradicals. A feed-ratio of 1:7.5 was found to be the most appropriate of the formulations considered in this study in terms of mechanical properties, cell adhesion, and printability. The biofabricated hybrid scaffold showed interconnected, homogeneous pores with a nominal pore size of 50µm and an elastic modulus of~400kPa. Moreover, long-term cell viability and cell spreading was observed via actin filament staining. Printability of the biocompatible resin was confirmed by printing thresholded MR images of an ear and the feed ratio of 1:7.5 provided the most faithful reproduction of the shape. To the best of our knowledge, this is the first report of stereolithographic printing hybridizing cell-adhesive properties of chitosan with mechanical robustness of PEG in scaffolds suitable for repair of complex tissue geometries, such as those of the human ear.

Simulation study on the effect of tissue geometries to fluence composition for non-invasive fetal pulse oximetry.

Transabdominal fetal pulse oximetry is a method to estimate the state of oxygenation of a fetus in-utero, utilizing the principle of reflection pulse oximetry. The extraction of fetal related information from a mixed fetal-maternal signal is elementary. Minimizing the ratio of purely maternal components of the signal at the detector side obviously facilitates signal separation. In this paper we analyze the influence of tissue geometries to the fluence composition at the surface of the abdomen. Monte-Carlo method is used to compute photon propagation in spherical layered tissue models. Spatial fluence distributions at the surface of the models are visualized and discussed. Our results show the characteristic effects of the distance between the fetus and the surface and the radius of the abdomen to the fluence composition at the detector. Further, the simulations indicate suitable source-detector configurations considering various anatomical conditions. We conclude that an adoption of the source-detector configuration to the individual tissue geometry at hand is necessary to achieve a proper signal composition and quality. Utilizing simulations for sensor design enhances the understanding of photon distributions in complex tissue geometries and supports a successful implementation of transabdominal fetal pulse oximetry.

Strain-induced Collagen Organization at the Micro-level in Fibrin-based Engineered Tissue Constructs

Full understanding of strain-induced collagen organization in complex tissue geometries to create tissues with predefined collagen architecture has not been achieved. This is mainly due to our limited knowledge of collagen remodeling in developing tissues. Here we investigate strain-induced collagen (re)organization in fibrin based engineered tissues using nondestructive time-lapse imaging. The tissues start from a biaxially constrained myofibroblast-populated fibrin gel and are used to study: (A) remodeling from a static equi-biaxial loading condition to static uniaxial loading; and (B) remodeling of a biaxially constrained tissue under uniaxial cyclic straining before and after a change in strain direction. Under static conditions, collagen oriented parallel to the direction of strain, whereas under cyclic conditions the orientation in the constrained tissue was perpendicular to the direction of strain. It is concluded that due to the biaxial constraints the uniaxially, cyclically strained cells can exert forces in two directions and strain shield themselves. A subsequent change in the direction of cyclic straining resulted in a rapid reorientation of collagen at the tissue surface. Reorientation was significantly slower in deeper tissue layers, where tissue remodeling was dominated by contact guidance provided by the endogenous matrix. These findings emphasize the relevance of achieving a functional collagen organization right from the start of tissue culture.

A Macro Finite-Element Formulation for Cardiac Electrophysiology Simulations Using Hybrid Unstructured Grids

Electrical activity in cardiac tissue can be described by the bidomain equations whose solution for large-scale simulations still remains a computational challenge. Therefore, improvements in the discrete formulation of the problem, which decrease computational and/or memory demands are highly desirable. In this study, we propose a novel technique for computing shape functions of finite elements (FEs). The technique generates macro FEs (MFEs) based on the local decomposition of elements into tetrahedral subelements with linear shape functions. Such an approach necessitates the direct use of hybrid meshes (HMs) composed of different types of elements. MFEs are compared to classic standard FEs with respect to accuracy and RAM memory usage under different scenarios of cardiac modeling, including bidomain and monodomain simulations in 2-D and 3-D for simple and complex tissue geometries. In problems with analytical solutions, MFEs displayed the same numerical accuracy of standard linear triangular and tetrahedral elements. In propagation simulations, conduction velocity and activation times agreed very well with those computed with standard FEs. However, MFEs offer a significant decrease in memory requirements. We conclude that HMs composed of MFEs are well suited for solving problems in cardiac computational electrophysiology.

Novel Macromolecular Crosslinking Hydrogel to Reduce Intra-Abdominal Adhesions

The aim of this study was to compare the anti-adhesion efficacy of a biodegradable, in situ, macromolecular cross-linking hydrogel made from oxidized dextran/N-carboxyethyl chitosan (Odex/CEC) with a commercially available carboxymethylcellulose/modified hyaluronan barrier film (Seprafilm; Genzyme Corporation, Cambridge, MA) in a rat cecum abrasion model. The rat model utilized a cecal abrasion and abdominal wall insult surgical protocol. The 2% Odex/CEC hydrogel treatment was applied by syringe to coat both the cecal and the abdominal wall insults, while other animals were treated with Seprafilm applied to the cecal injury only. Control animals did not receive any treatment. Animals were sacrificed after post operative day 21 and adhesion severity was quantitatively graded using a whole number scale from 0 - 3. Histological analysis was also performed for animals receiving Odex/CEC hydrogel treatment and no treatment (control). Mean adhesion score was 2.09+/-1.22 for control animals, 1.00+/-1.00 for 2% Odex/CEC hydrogel animals, and 1.25+/-1.22 for Seprafilm animals. Hydrogel treated animals showed significantly lower adhesion scores than control animals (P<0.05), while Seprafilm demonstrated a marginally lower adhesion score (P<0.1) compared with the controls. Histological analysis of an Odex/CEC treated rat showed tissue repair and small fragments of hydrogel inside both healed abdominal and cecal surfaces. Both Seprafilm and the 2% Odex/CEC hydrogel showed a significantly decreased adhesion score compared with the control. However, the hydrogel, compared with Seprafilm, offers ease of application and ability to conform to complex tissue geometries that could provide surgeons with another prophylactic treatment to prevent abdominal adhesions.

Light Dosimetry for Intraperitoneal Photodynamic Therapy in a Murine Xenograft Model of Human Epithelial Ovarian Carcinoma

Few studies have been published to date measuring spatially resolved fluence rates in complex tissue geometries. Here the light distributions of three different intraperitoneal light delivery geometries in a murine ovarian cancer model were investigated to assess their influence on the tumorcidal efficacy of photodynamic therapy (PDT). In vivo fluence rate measurements in the peritoneal cavities of mice, with the light intensity being mapped in three transverse planes, were performed using fiber-optic detectors. Three different source fiber designs and placements were tested for their ability to provide uniform irradiation of the peritoneal cavity. The biological response to a PDT protocol comprising three separate treatments administered at 72 h intervals, each consisting of a 0.25 mg kg-1 intraperitoneal injection of benzoporphyrin derivative-mono acid ring A followed 90 min later by delivery of 15 J of 690 nm light, was measured. The tissue response was evaluated by measuring the number of remaining visible lesions and the total residual tumor mass. Fluence rate measurements showed large variations in the fluence rate distribution for similar intended treatments. The most uniform and reproducible illumination was achieved using two 18 mm long cylindrical emitting optical fibers. The biological response was comparable to that produced when a flat-cleaved end optical fiber is used to illuminate the four quadrants of the abdomen sequentially. While a good reproducibility in tumor induction in this animal model exists, no correlation was found between the fluence rate distribution measured in one group of animals and the biological response in a separate group of similarly treated animals. Due to the large intra-animal variability in fluence rate distribution, representative fluence rate mapping in complex tissue geometries is of limited value when applied to an individual PDT treatment. Thus, surveillance of the fluence rate during individual treatments will be required for acceptable PDT dosimetry. To improve the versatility of this particular animal model for PDT research, a large number of extended sources are required to increase uniformity of the illumination in order to reduce unwanted cytotoxic side effects resulting from foci of high fluence rates. In this way, subsequent increase of the total energy delivered to the tumor may be possible.