Tissue Mimic Research Articles

Meta-Learning Analysis of Ultrasonic Guided Waves for Coated Cortical Bone Characterization.

Due to its sensitivity to geometrical and mechanical properties of waveguides, ultrasonic guided waves (UGWs) propagating in cortical bones play an important role in the early diagnosis of osteoporosis. However, as impacts of overlaid soft tissues are complex, it remains challenging to retrieve bone properties accurately. Meta-learning, i.e., learning to learn, is capable of extracting transferable features from a few data and, thus, suitable to capture potential characteristics, leading to accurate bone assessment. In this study, we investigate the feasibility to apply the multichannel identification neural network (MCINN) to estimate the thickness and bulk velocities of coated cortical bone. It minimizes the effects of soft tissue by extracting specific features of UGW, which shares the same cortical properties, while the overlaid soft tissue varies. Distinguished from most reported methods, this work moves from the hand-design inversion scheme to data-driven assessment by automatically mapping features of UGW to the space of bone properties. The MCINN was trained and validated using simulated datasets produced by the finite-difference time-domain (FDTD) method and then applied to experimental data obtained from cortical bovine bone plates overlaid with soft tissue mimics. A good match was found between experimental trajectories and theoretical dispersion curves. The results demonstrated that the proposed method was feasible to assess the thickness of coated cortical bone plates.

Nonlinear Viscoelastic Properties of 3D-Printed Tissue Mimicking Materials and Metrics to Determine the Best Printed Material Match to Tissue Mechanical Behavior

3D-printed biomaterials have become ubiquitous for clinical applications including tissue-mimicking surgical/procedure planning models and implantable tissue engineering scaffolds. In each case, a fundamental hypothesis is that printed material mechanical properties should match those of the tissue being replaced or modeled as closely as possible. Evaluating these hypotheses requires 1) consistent nonlinear elastic/viscoelastic constitutive model fits of 3D-printed biomaterials and tissues and 2) metrics to determine how well 3D-printed biomaterial mechanical properties match a corresponding tissue. Here we utilize inverse finite element modeling to fit nonlinear viscoelastic models with Neo-Hookean kernels to 29 Polyjet 3D-printed tissue-mimicking materials. We demonstrate that the viscoelastic models fit well with R2 > 0.95. We also introduce three metrics ( least-squares difference, Kolmogorov–Smirnov statistics, and the area under stress/strain or load/displacement curve) to compare printed material properties to tissue properties. All metrics showed lower values for better matches between 3D-printed materials and tissues. These results provide a template for comparing 3D-printed material mechanical properties to tissue mechanical properties, and therefore, a basis for testing the fundamental hypotheses of 3D-printed tissue-mimicking materials.

Particulate ECM biomaterial ink is 3D printed and naturally crosslinked to form structurally-layered and lubricated cartilage tissue mimics

Articular cartilage is a layered tissue with a complex, heterogeneous structure and lubricated surface which is challenging to reproduce using traditional tissue engineering methods. Three-dimensional printing techniques have enabled engineering of complex scaffolds for cartilage regeneration, but constructs fail to replicate the unique zonal layers, and limited cytocompatible crosslinkers exist. To address the need for mechanically robust, layered scaffolds, we developed an extracellular matrix particle-based biomaterial ink (pECM biomaterial ink) which can be extruded, polymerizes via disulfide bonding, and restores layered tissue structure and surface lubrication. Our cartilage pECM biomaterial ink utilizes functionalized hyaluronan (HA), a naturally occurring glycosaminoglycan, crosslinked directly to decellularized tissue particles (ø40–100 µm). We experimentally determined that HA functionalized with thiol groups (t-HA) forms disulfide bonds with the ECM particles to form a 3D network. We show that two inks can be co-printed to create a layered cartilage scaffold with bulk compressive and surface (friction coefficient, adhesion, and roughness) mechanics approaching values measured on native cartilage. We demonstrate that our printing process enables the addition of macropores throughout the construct, increasing the viability of introduced cells by 10%. The delivery of these 3D printed scaffolds to a defect is straightforward, customizable to any shape, and adheres to surrounding tissue.

Method Matters: Exploring Alkoxysulfonate-Functionalized Poly(3,4-ethylenedioxythiophene) and Its Unintentional Self-Aggregating Copolymer toward Injectable Bioelectronics.

Injectable bioelectronics could become an alternative or a complement to traditional drug treatments. To this end, a new self-doped p-type conducting PEDOT-S copolymer (A5) was synthesized. This copolymer formed highly water-dispersed nanoparticles and aggregated into a mixed ion–electron conducting hydrogel when injected into a tissue model. First, we synthetically repeated most of the published methods for PEDOT-S at the lab scale. Surprisingly, analysis using high-resolution matrix-assisted laser desorption ionization-mass spectroscopy showed that almost all the methods generated PEDOT-S derivatives with the same polymer lengths (i.e., oligomers, seven to eight monomers in average); thus, the polymer length cannot account for the differences in the conductivities reported earlier. The main difference, however, was that some methods generated an unintentional copolymer P(EDOT-S/EDOT-OH) that is more prone to aggregate and display higher conductivities in general than the PEDOT-S homopolymer. Based on this, we synthesized the PEDOT-S derivative A5, that displayed the highest film conductivity (33 S cm–1) among all PEDOT-S derivatives synthesized. Injecting A5 nanoparticles into the agarose gel cast with a physiological buffer generated a stable and highly conductive hydrogel (1–5 S cm–1), where no conductive structures were seen in agarose with the other PEDOT-S derivatives. Furthermore, the ion-treated A5 hydrogel remained stable and maintained initial conductivities for 7 months (the longest period tested) in pure water, and A5 mixed with Fe3O4 nanoparticles generated a magnetoconductive relay device in water. Thus, we have successfully synthesized a water-processable, syringe-injectable, and self-doped PEDOT-S polymer capable of forming a conductive hydrogel in tissue mimics, thereby paving a way for future applications within in vivo electronics.

Hierarchically Structured Surfaces Prepared by Phase Separation: Tissue Mimicking Culture Substrate

The pseudo 3D hierarchical structure mimicking in vivo microenvironment was prepared by phase separation on tissue culture plastic. For surface treatment, time-sequenced dosing of the solvent mixture with various concentrations of polymer component was used. The experiments showed that hierarchically structured surfaces with macro, meso and micro pores can be prepared with multi-step phase separation processes. Changes in polystyrene surface topography were characterized by atomic force microscopy, scanning electron microscopy and contact profilometry. The cell proliferation and changes in cell morphology were tested on the prepared structured surfaces. Four types of cell lines were used for the determination of impact of the 3D architecture on the cell behavior, namely the mouse embryonic fibroblast, human lung carcinoma, primary human keratinocyte and mouse embryonic stem cells. The increase of proliferation of embryonic stem cells and mouse fibroblasts was the most remarkable. Moreover, the embryonic stem cells express different morphology when cultured on the structured surface. The acquired findings expand the current state of knowledge in the field of cell behavior on structured surfaces and bring new technological procedures leading to their preparation without the use of problematic temporary templates or additives.

Analysis of surgical needle insertion modeling and viscoelastic tissue material interaction for minimally invasive surgery (MIS)

Soft tissue modeling of surgical events is essential for both considering tissue reactions to surgical instrumentation, in addition, to conducting the design of the surgical instrument. Mainly with observes to surgical needle insertion for minimally invasive surgery (MIS) into soft biological tissue, one of the regularly performed surgical events, many works have been prepared in references to modeling how surgical needles insert the soft biological tissue material as they pass through in the direction of objects deep inside the living body. Deformation of biological soft tissue is one of the important issues, which impacts the surgical needle insert inside the living soft viscoelastic tissue. A needle insertion model is presented to calculate the insertion forces and tissue deformation phenomenon. The experimentations are done on the tissue mimic gel specimens, and the surgical needle insertion-based model for the deformation phenomenon is analyzed to investigate the force model, frictional stress and deflection phenomenon. This study can help to investigate the needle insertion process for analyzing the effect of the surgical needle insertion, which could advance the enactment of several surgical needle insertion-based procedures such as minimally invasive surgery (MIS) and brachytherapy.

An Investigation for Large Volume, Focal Blood-Brain Barrier Disruption with High-Frequency Pulsed Electric Fields.

The treatment of CNS disorders suffers from the inability to deliver large therapeutic agents to the brain parenchyma due to protection from the blood-brain barrier (BBB). Herein, we investigated high-frequency pulsed electric field (HF-PEF) therapy of various pulse widths and interphase delays for BBB disruption while selectively minimizing cell ablation. Eighteen male Fisher rats underwent craniectomy procedures and two blunt-tipped electrodes were advanced into the brain for pulsing. BBB disruption was verified with contrast T1W MRI and pathologically with Evans blue dye. High-frequency irreversible electroporation cell death of healthy rodent astrocytes was investigated in vitro using a collagen hydrogel tissue mimic. Numerical analysis was conducted to determine the electric fields in which BBB disruption and cell ablation occur. Differences between the BBB disruption and ablation thresholds for each waveform are as follows: 2-2-2 s (1028 V/cm), 5-2-5 s (721 V/cm), 10-1-10 s (547 V/cm), 2-5-2 s (1043 V/cm), and 5-5-5 s (751 V/cm). These data suggest that HF-PEFs can be fine-tuned to modulate the extent of cell death while maximizing peri-ablative BBB disruption. Furthermore, numerical modeling elucidated the diffuse field gradients of a single-needle grounding pad configuration to favor large-volume BBB disruption, while the monopolar probe configuration is more amenable to ablation and reversible electroporation effects.

Novel Chitosan-Silica Hybrid Hydrogels for Cell Encapsulation and Drug Delivery.

Hydrogels constructed from naturally derived polymers provide an aqueous environment that encourages cell growth, however, mechanical properties are poor and degradation can be difficult to predict. Whilst, synthetic hydrogels exhibit some improved mechanical properties, these materials lack biochemical cues for cells growing and have limited biodegradation. To produce hydrogels that support 3D cell cultures to form tissue mimics, materials must exhibit appropriate biological and mechanical properties. In this study, novel organic-inorganic hybrid hydrogels based on chitosan and silica were prepared using the sol-gel technique. The chemical, physical and biological properties of the hydrogels were assessed. Statistical analysis was performed using One-Way ANOVAs and independent-sample t-tests. Fourier transform infrared spectroscopy showed characteristic absorption bands including amide II, Si-O and Si-O-Si confirming formation of hybrid networks. Oscillatory rheometry was used to characterise the sol to gel transition and viscoelastic behaviour of hydrogels. Furthermore, in vitro degradation revealed both chitosan and silica were released over 21 days. The hydrogels exhibited high loading efficiency as total protein loading was released in a week. There were significant differences between TC2G and C2G at all-time points (p < 0.05). The viability of osteoblasts seeded on, and encapsulated within, the hydrogels was >70% over 168 h culture and antimicrobial activity was demonstrated against Pseudomonas aeruginosa and Enterococcus faecalis. The hydrogels developed here offer alternatives for biopolymer hydrogels for biomedical use, including for application in drug/cell delivery and for bone tissue engineering.

Applications of 3D Bioprinting in Tissue Engineering and Regenerative Medicine.

Regenerative medicine is an emerging field that centers on the restoration and regeneration of functional components of damaged tissue. Tissue engineering is an application of regenerative medicine and seeks to create functional tissue components and whole organs. Using 3D printing technologies, native tissue mimics can be created utilizing biomaterials and living cells. Recently, regenerative medicine has begun to employ 3D bioprinting methods to create highly specialized tissue models to improve upon conventional tissue engineering methods. Here, we review the use of 3D bioprinting in the advancement of tissue engineering by describing the process of 3D bioprinting and its advantages over other tissue engineering methods. Materials and techniques in bioprinting are also reviewed, in addition to future clinical applications, challenges, and future directions of the field.

Microscale characterisation of the time-dependent mechanical behaviour of brain white matter

Brain mechanics is a topic of deep interest because of the significant role of mechanical cues in both brain function and form. Specifically, capturing the heterogeneous and anisotropic behaviour of cerebral white matter (WM) is extremely challenging and yet the data on WM at a spatial resolution relevant to tissue components are sparse. To investigate the time-dependent mechanical behaviour of WM, and its dependence on local microstructural features when subjected to small deformations, we conducted atomic force microscopy (AFM) stress relaxation experiments on corpus callosum (CC), corona radiata (CR) and fornix (FO) of fresh ovine brain. Our experimental results show a dependency of the tissue mechanical response on axons orientation, with e.g. the stiffness of perpendicular and parallel samples is different in all three regions of WM whereas the relaxation behaviour is different for the CC and FO regions.An inverse modelling approach was adopted to extract Prony series parameters of the tissue components, i.e. axons and extra cellular matrix with its accessory cells, from experimental data. Using a bottom-up approach, we developed analytical and FEA estimates that are in good agreement with our experimental results.Our systematic characterisation of sheep brain WM using a combination of AFM experiments and micromechanical models provide a significant contribution for predicting localised time-dependent mechanics of brain tissue. This information can lead to more accurate computational simulations, therefore aiding the development of surgical robotic solutions for drug delivery and accurate tissue mimics, as well as the determination of criteria for tissue injury and predict brain development and disease progression.

4D Reconstruction and Identification of Carotid Artery Stenosis Utilizing a Novel Pulsatile Ultrasound Phantom.

As a major application focus of vascular ultrasonography, the carotid artery has long been the subject of phantom design and procedure focus. It is therefore important to devise procedures that are minimally invasive and informative, initially using a physiologically accurate anthropomorphic phantom to validate the methodology. In this article, a novel phantom design protocol is presented that enables the efficient production of a pulsatile ultrasound phantom consisting of soft and vascular tissue mimics, as well as a blood surrogate fluid. These components when combined give the phantom high acoustic compatibility and lifelike mechanical properties. The phantom was developed using "at-home" purchasable components and 3D printing technology. The phantom was subsequently used to develop a 4D reconstruction algorithm of the pulsing vessel in MATLAB. In pattern with recent developments in medical imaging, the 4D reconstruction enables clinicians to view vessel wall motion in a 3D space without the need for manual intervention. The reconstruction algorithm also produces measured inner luminal areas and vessel wall thickness, providing further information relating to structural properties and stenosis, as well as elastic properties such as arterial stiffness, which could provide helpful markers for disease diagnosis. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Constructing a pulsatile ultrasound phantom model Support Protocol: Creating a vascular mimic mold Basic Protocol 2: Creating a 4D reconstruction from ultrasound frames.

Lithographically patterned micro-nozzles for controlling fluid flow profiles for drug delivery and in vitro imaging applications.

Precisely controlling delivery of drugs and other reagents is important for intravital microscopy studies. In this work, photolithographic integration of micro-nozzles onto a microfluidic platform was performed to tune the fluid flow profile and depth of penetration into biological tissue mimics. Performance characteristics were measured by correlating the flow rate through the device to the applied pressure and/or delivery of dyes into solution and agarose gel-based phantom tissue. From these results, the implementation of micro-nozzles was demonstrated to significantly improve the lateral dispersion of delivered fluid and increase the depth of penetration into phantom tissue.

Biomimetic Strain‐Stiffening Hydrogel with Crimped Structure

AbstractSynthetic hydrogels are unique tissue mimics but rarely reproduce the strain‐stiffening properties of native tissues. This mechanical mismatch impairs the performance of hydrogels in practical applications. Inspired by the crimped structure of collagenous tissues, a series of strain‐stiffening hydrogels composed of curved parallel fibers are developed. These fibers are constructed from a bundle of intertwisted nanofibrils composed of short alkyl side chain‐modified polymer chains. This hierarchical organization enables exquisitely cascaded deformation that facilitates soft‐to‐firm and resilience‐to‐viscoelasticity transitions, thus synergically mimicking the strain‐adaptive stiffening and damping behaviors of natural tissues. Together with structural evolution and a constitutive model, rationally tuning the tortuosity and flexibility of the curved fibers produces a diverse combination of strain‐stiffening properties and unprecedented penetration into the regions of several tissues. The crimped structure and the resultant stiffening properties constitute major improvements to nanofiber‐based scaffolds for use in collagenous tissue repair.

Bacterial cellulose adhesive composites for oral cavity applications

Topical approaches to oral diseases require frequent dosing due to limited retention time. A mucoadhesive drug delivery platform with extended soft tissue adhesion capability of up to 7 days is proposed for on-site management of oral wound. Bacterial cellulose (BC) and photoactivated carbene-based bioadhesives (PDz) are combined to yield flexible film platform for interfacing soft tissues in dynamic, wet environments. Structure-activity relationships evaluate UV dose and hydration state with respect to adhesive strength on soft tissue mimics. The bioadhesive composite has an adhesion strength ranging from 7 to 17 kPa and duration exceeding 48 h in wet conditions under sustained shear forces, while other mucoadhesives based on hydrophilic macromolecules exhibit adhesion strength of 0.5–5 kPa and last only a few hours. The work highlights the first evaluation of BC composites for mucoadhesive treatments in the buccal cavity.

3D Bioprinting Human‐Induced Pluripotent Stem Cells and Drug‐Releasing Microspheres to Produce Responsive Neural Tissues

3D bioprinting can produce complex human tissue mimics using stem cells (SCs). Herein, cylindrical constructs containing human‐induced pluripotent stem cell (hiPSC)‐derived neural progenitor cells (NPCs) encapsulated in a fibrin‐based bioink containing polycaprolactone (PCL)–retinoic acid (RA) and purmorphamine (puro)‐releasing microspheres are bioprinted in a layer‐by‐layer fashion using the microfluidic‐based RX1 bioprinter to engineer responsive neural tissues. The differentiated constructs contain neurons expressing ChAT, GABA, and MAP2, astrocytes expressing GFAP, and oligodendrocytes expressing O4 as indicated by immunocytochemistry and flow cytometry analysis on days 30 and 45. The bioprinted tissues also respond to treatment with acetylcholine (Ach) and gamma‐aminobutyric acid (GABA) on days 30 and 45. The use of microsphere‐laden bioinks efficiently promotes neural tissue differentiation and maturation in situ using a lower amount of morphogens in comparison with using soluble drugs. This bioprinting strategy serves as a cost‐effective solution for engineering humanized neural tissues.

Microparticle-Based Biochemical Sensing Using Optical Coherence Tomography and Deep Learning.

Advancing continuous health monitoring beyond vital signs to biochemistry will revolutionize personalized medicine. Herein, we report a biosensing platform to achieve remote biochemical monitoring using microparticle-based biosensors and optical coherence tomography (OCT). Stimuli-responsive, polymeric microparticles were designed to serve as freely dispersible biorecognition units, wherein binding with a target biochemical induces volumetric changes of the microparticle. Analytical approaches to detect these submicron changes in 3D using OCT were devised by modeling the microparticle as an optical cavity, enabling estimations far below the resolution of the OCT system. As a proof of concept, we demonstrated the 3D spatiotemporal monitoring of glucose-responsive microparticles distributed throughout a tissue mimic in response to dynamically fluctuating levels of glucose. Deep learning was further implemented using 3D convolutional neural networks to automate the vast processing of the continuous stream of three-dimensional time series data, resulting in a robust end-to-end pipeline with immense potential for continuous in vivo biochemical monitoring.

A New Methodology for the Assessment of Very Low Concentrations of Cells in Serous Body Fluids Based on the Count of Ultrasound Echoes Backscattered From Cells.

A methodology for the assessment of cell concentration, in the range 5-100 cells/ [Formula: see text], suitable for in vivo analysis of serous body fluids is presented in this work. This methodology is based on the quantitative analysis of ultrasound images obtained from cell suspensions and considers applicability criteria, such as short analysis times, moderate frequency, and absolute concentration estimation, all necessary to deal with the variability of tissues among different patients. Numerical simulations provided the framework to analyze the impact of echo overlapping and the polydispersion of scatterer sizes on the cell concentration estimation. The cell concentration range that can be analyzed as a function of the transducer and emitted waveform used was also discussed. Experiments were conducted to evaluate the performance of the method using 7- [Formula: see text] and 12- [Formula: see text] polystyrene particles in water suspensions in the 5-100 particles/ [Formula: see text] range. A single scanning focused transducer working at a central frequency of 20 MHz was used to obtain ultrasound images. The method proposed to estimate the concentration proved to be robust for different particle sizes and variations of gain acquisition settings. The effect of tissues placed in the ultrasound path between the probe and the sample was also investigated using 3-mm-thick tissue mimics. Under this situation, the algorithm was robust for the concentration analysis of 12 [Formula: see text] particle suspensions, yet significant deviations were obtained for the smallest particles.

Helical toroid phantom for 3D flow imaging investigations

The medical physics community has hitherto lacked an effective calibration phantom to holistically evaluate the performance of three-dimensional (3D) flow imaging techniques. Here, we present the design of a new omnidirectional, three-component (3-C) flow phantom whose lumen is consisted of a helical toroid structure (4 mm lumen diameter; helically winded for 5 revolutions over a torus with 10 mm radius; 5 mm helix radius). This phantom’s intraluminal flow trajectory embraces all combinations of x, y, and z directional components, as confirmed using computational fluid dynamics (CFD) simulations. The phantom was physically fabricated via lost-core casting with polyvinyl alcohol cryogel (PVA) as the tissue mimic. 3D ultrasound confirmed that the phantom lumen expectedly resembled a helical toroid geometry. Pulsed Doppler measurements showed that the phantom, when operating under steady flow conditions (3 ml s−1 flow rate), yielded flow velocity magnitudes that agreed well with those derived from CFD at both the inner torus (−47.6 ± 5.7 versus −52.0 ± 2.2 cm s−1; mean ± 1 S.D.) and the outer torus (49.5 ± 4.2 versus 48.0 ± 1.7 cm s−1). Additionally, 3-C velocity vectors acquired from multi-angle pulsed Doppler showed good agreement with CFD-derived velocity vectors (<7% and 10° difference in magnitude and flow angle, respectively). Ultrasound color flow imaging further revealed that the phantom’s axial flow pattern was aligned with the CFD-derived flow profile. Overall, the helical toroid phantom has strong potential as an investigative tool in 3D flow imaging innovation endeavors, such as the development of flow vector estimators and visualization algorithms.

Bioprintability: Physiomechanical and Biological Requirements of Materials for 3D Bioprinting Processes.

Three-dimensional (3D) bioprinting is an additive manufacturing process that utilizes various biomaterials that either contain or interact with living cells and biological systems with the goal of fabricating functional tissue or organ mimics, which will be referred to as bioinks. These bioinks are typically hydrogel-based hybrid systems with many specific features and requirements. The characterizing and fine tuning of bioink properties before, during, and after printing are therefore essential in developing reproducible and stable bioprinted constructs. To date, myriad computational methods, mechanical testing, and rheological evaluations have been used to predict, measure, and optimize bioinks properties and their printability, but none are properly standardized. There is a lack of robust universal guidelines in the field for the evaluation and quantification of bioprintability. In this review, we introduced the concept of bioprintability and discussed the significant roles of various physiomechanical and biological processes in bioprinting fidelity. Furthermore, different quantitative and qualitative methodologies used to assess bioprintability will be reviewed, with a focus on the processes related to pre, during, and post printing. Establishing fully characterized, functional bioink solutions would be a big step towards the effective clinical applications of bioprinted products.

Characterization of 3-Dimensional Printing and Casting Materials for use in Computed Tomography and X-ray Imaging Phantoms.

Imaging phantoms are used to calibrate and validate the performance of medical computed tomography (CT) systems. Many new materials developed for three-dimensional (3D) printing processes may be useful in the direct printing or casting of biomimetic and geometrically accurate CT and X-ray phantoms. The X-ray linear attenuation coefficients of polymer samples were measured to discover materials for use as tissue mimics in phantoms. This study included a cohort of polymer compounds that were tested in cured form. The cohort consisted of 101 standardized polymer samples fabricated from: two-part silicones and polyurethanes used in commercial casting processes; one-part optically cured polyurethanes used in 3D printing; and fused deposition thermoplastics used in 3D printing. The testing was performed with a commercial micro-CT imaging system from 40 kVp to 140 kVp. The X-ray linear coefficients of the samples and human tissues were plotted with error bars to allow the reader to identify suitable mimics. The X-ray linear attenuation coefficients of the tested material samples spanned a wide range of values, with a small number of them overlapping established human tissue mimic values. Twenty 3D printer materials and one castable polyurethane tracked nylon and polymethyl methacrylate (PMMA) as established X-ray mimics for fat. Five 3D printer materials tracked water as an established X-ray mimic for muscle.