Traditional 2D Model Research Articles

3D Models Currently Proposed to Investigate Human Skin Aging and Explore Preventive and Reparative Approaches: A Descriptive Review.

Skin aging is influenced by intrinsic and extrinsic factors that progressively impair skin functionality over time. Investigating the skin aging process requires thorough research using innovative technologies. This review explores the use of in vitro human 3D culture models, serving as valuable alternatives to animal ones, in skin aging research. The aim is to highlight the benefits and necessity of improving the methodology in analyzing the molecular mechanisms underlying human skin aging. Traditional 2D models, including monolayers of keratinocytes, fibroblasts, or melanocytes, even if providing cost-effective and straightforward methods to study critical processes such as extracellular matrix degradation, pigmentation, and the effects of secretome on skin cells, fail to replicate the complex tissue architecture with its intricated interactions. Advanced 3D models (organoid cultures, "skin-on-chip" technologies, reconstructed human skin, and 3D bioprinting) considerably enhance the physiological relevance, enabling a more accurate representation of skin aging and its peculiar features. By reporting the advantages and limitations of 3D models, this review highlights the importance of using advanced in vitro systems to develop practical anti-aging preventive and reparative approaches and improve human translational research in this field. Further exploration of these technologies will provide new opportunities for previously unexplored knowledge on skin aging.

MODL-01. IN VITRO MODELING OF AND AUTOMATED DRUG SCREENING IN EMBRYONAL BRAIN TUMORS USING TUMOR-BRAIN ORGANOIDS

Abstract BACKGROUND Embryonal brain tumors (EBTs) such as medulloblastoma, atypical teratoid rhabdoid tumor (AT/RT), and embryonal tumor with multilayered rosettes (ETMR) uniquely arise in the developing brain. Traditional 2D models often fall short in capturing the tumoral microenvironment (TME) and intratumoral heterogeneity, which may limit their translational potential. METHODS To address this, we co-aggregated EBT cells from AT/RT (Chla02 and SHH310) and ETMR (BT183 and murine GEMM cells) with forebrain organoids derived from human induced pluripotent stem cells and developed an automated, screening-ready in vitro model (EBT-FBO). We used immunohistochemistry, whole-mount immunostaining, and single-cell RNA sequencing to characterize our model and compared it with a developmental forebrain atlas. We further used our model for cell type-specific drug screening and validated the results in vivo. RESULTS Our EBT-FBO model mimicked the differentiation states of a mid-gestational fetal forebrain, providing a natural immature neuronal environment for supratentorial EBT cells. This environment allowed the EBT cells from both AT/RT and ETMR to mimic in vivo tumor cell states and avoided the cycling cell selection common in other models. We screened over 160 drugs in ETMR-FBO and identified antracyclines and Triptolide as having anti-tumor, yet non-neurotoxic effects. Our in-depth transcriptomic analysis identified new therapeutic targets, including PDGFR-signaling for ETMR. Building on this insight, we disrupted tumor-TME interactions and observed significant anti-tumor effects both in vitro and in vivo. CONCLUSIONS EBT-FBO provide embryonal brain tumors like ETMR or AT/RT with their intrinsic, immature neuronal microenvironment essential for recapitulating natural tumor heterogeneity. This marks a significant advancement towards targeted therapy and patient-specific models in precision medicine.

Construction of dentin-on-a-chip based on microfluidic technology and tissue engineering

AimThree-dimensional (3D) cell culture systems perform better in resembling tissue or organism structures compared with traditional 2D models. Organs-on-chips (OoCs) are becoming more efficient 3D models. This study aimed to create a novel simplified dentin-on-a-chip using microfluidic chip technology and tissue engineering for screening dental materials. MethodologyA microfluidic device with three channels was designed for creating 3D dental tissue constructs using stem cells from the apical papilla (SCAP) and gelatin methacrylate (GelMA). The study investigated the effect of varying cell densities and GelMA concentrations on the layer features formed within the microfluidic chip. Cell viability and distribution were evaluated through live/dead staining and nuclei/F-actin staining. The osteo/odontogenic potential was assessed through ALP staining and Alizarin red staining. The impact of GelMA concentrations (5 %, 10 %) on the osteo/odontogenic differentiation trajectory of SCAP was also studied. ResultsThe 3D tissue constructs maintained high viability and favorable spreading within the microfluidic chip for 3–7 days. A cell seeding density of 2 × 104 cells/μL was found to be the most optimal choice, ensuring favorable cell proliferation and even distribution. GelMA concentrations of 5 % and 10 % proved to be most effective for promoting cell growth and uniform distribution. Within the 5 % GelMA group, SCAP demonstrated higher osteo/odontogenic differentiation than that in the 10 % GelMA group. ConclusionIn 3D culture, GelMA concentration was found to regulate the osteo/odontogenic differentiation of SCAP. The study recommends a seeding density of 2 × 104 cells/μL of SCAP within 5 % GelMA for constructing simplified dentin-on-a-chip. Clinical SignificanceThis study built up the 3D culture protocol, and induced odontogenic differentiation of SCAP, thus forming the simplified dentin-on-a-chip and paving the way to be used as a well-defined biological model for regenerative endodontics. It may serve as a potential testing platform for cell differentiation.

Gas-Phase Fractionation Data-Independent Acquisition Analysis of 3D Cocultured Spheroid Tumor Model Reveals Altered Translational Processes and Signaling Using Proteomics.

Colorectal cancer (CRC) contains considerable heterogeneity; therefore, models of the disease must also reflect the multifarious components. Compared to traditional 2D models, 3D cellular models, such as tumor spheroids, have the utility to determine the drug efficacy of potential therapeutics. Monoculture spheroids are well-known to recapitulate gene expression, cell signaling, and pathophysiological gradients of avascularized tumors. However, they fail to mimic the stromal cell influence present in CRC, which is known to perturb drug efficacy and is associated with metastatic, late-stage colorectal cancer. This study seeks to develop a cocultured spheroid model using carcinoma and noncancerous fibroblast cells. We characterized the proteomic profile of cocultured spheroids in comparison to monocultured spheroids using data-independent acquisition with gas-phase fractionation. Specifically, we determined that proteomic differences related to translation and mTOR signaling are significantly increased in cocultured spheroids compared to monocultured spheroids. Proteins related to fibroblast function, such as exocytosis of coated vesicles and secretion of growth factors, were significantly differentially expressed in the cocultured spheroids. Finally, we compared the proteomic profiles of both the monocultured and cocultured spheroids against a publicly available data set derived from solid CRC tumors. We found that the proteome of the cocultured spheroids more closely resembles that of the patient samples, indicating their potential as tumor mimics.

Modeling extrusion process and layer deformation in 3D concrete printing via smoothed particle hydrodynamics

We developed a computational framework to simulate the intricate processes of extrusion and layer deformation in 3D concrete printing (3DCP). A novel two-dimensional (2D) virtual printing scheme is proposed that enables direct prediction of cross-sectional shape, releasing simulations from the constraint of rectangular nozzles inherent in traditional 2D models. The proposed scheme accurately captures significant lateral deformation, previously ignored by conventional 2D models. It is noteworthy that, for the first time, the effect of time-dependent yield stress on layer deformation is considered by introducing the structuration rate. Further, to leverage advantages in avoiding mesh generation and additional interface tracking, a weakly compressible smoothed particle hydrodynamics (SPH) method incorporating the regularized Bingham model is equipped in the proposed framework. The simulated cross-sectional shapes exhibit excellent consistency with experimental results and outperform existing numerical results across various nozzle heights, printing velocities, and extrusion velocities. Our exploration of rheological parameters reveals that the final layer deformation is influenced by the yield stress, while its deformation rate is affected by the plastic viscosity. The proposed virtual printing framework emerges as a promising tool for enhancing predictability and efficiency in the printing process.

Two Dimensional Hydrodynamics Model with Evaporation for Coastal Systems

Introduction. The use of two-dimensional (2D) hydrodynamic models is relevant, despite the development of numerical methods of marine hydrodynamics focused on the use of three-dimensional spatial models. This is due to the modelling of hydrodynamic processes in shallow and coastal systems in solving practically important problems of predicting the transport of pollutants in suspended and dissolved forms. Evaporation for the Southern of Russia marine coastal systems (the Azov Sea, the Northern Caspian, etc.), and even more so in the coastal areas of the Red Sea, is a significant factor that affects not only the balance of water masses, but also makes changes in the momentum of the system and the distribution of the velocity vector of the aquatic environment. This effect is significant for coastal currents and shallow-water systems.Materials and Methods. The traditional method of converting the terms of the Navier-Stokes equations containing differentiation by horizontal spatial variables was used, involving the rearrangement of differentiation operations by horizontal spatial coordinates and integration by vertical coordinate when constructing a spatially two-dimensional model of hydrodynamics of marine coastal systems when integrated by vertical coordinate. This made it possible to avoid the appearance of non-physical sources of energy and momentum in the spatially two-dimensional model, which can be essential in traditional 2D models with significant depth differences characteristic of coastal systems. The implementation of the analogue of the law of conservation of the total mechanical energy of the system for the constructed 2D model is investigated.Results. Using the correct transformation of the 3D model (integration of the Navier-Stokes equations and continuity along a vertical coordinate, taking into account evaporation from a free surface), spatially two-dimensional models of hydrodynamics are constructed, for which the basic conservation laws, including mass and total mechanical energy of the system, are fulfilled. The implementation of an analogue of the law of conservation of total mechanical energy for various types of boundary conditions, including at the bottom, is investigated. The evaporation from the free surface is correctly accounted for not only in the continuity equation, but also in the equations of motion taking into account wind and waves.Discussion and Conclusion. 2D model of hydrodynamics has been constructed and studied, taking into account evaporation not only in the mass balance equation (continuity), but also in the Navier-Stokes equations of motion. The proposed model can be used for predictive modelling of hydrophysical processes, including the spread of pollutants in the aquatic environment of coastal systems and shallow reservoirs in relation to marine systems such as the Sea of Azov, the Northern Caspian Sea, coastal areas of the Red Sea, etc. Spatially two-dimensional models of marine hydrodynamics, without replacing three-dimensional models, can serve as a model basis for operational forecasting of situations in coastal systems and shallow-water objects using computing systems with relatively low performance and a moderate amount of RAM (5–10 Tflops, 2–4 TB, respectively).

Cancer 3D Models for Metallodrug Preclinical Testing.

Despite being standard tools in research, the application of cellular and animal models in drug development is hindered by several limitations, such as limited translational significance, animal ethics, and inter-species physiological differences. In this regard, 3D cellular models can be presented as a step forward in biomedical research, allowing for mimicking tissue complexity more accurately than traditional 2D models, while also contributing to reducing the use of animal models. In cancer research, 3D models have the potential to replicate the tumor microenvironment, which is a key modulator of cancer cell behavior and drug response. These features make cancer 3D models prime tools for the preclinical study of anti-tumoral drugs, especially considering that there is still a need to develop effective anti-cancer drugs with high selectivity, minimal toxicity, and reduced side effects. Metallodrugs, especially transition-metal-based complexes, have been extensively studied for their therapeutic potential in cancer therapy due to their distinctive properties; however, despite the benefits of 3D models, their application in metallodrug testing is currently limited. Thus, this article reviews some of the most common types of 3D models in cancer research, as well as the application of 3D models in metallodrug preclinical studies.

The response and resistance to drugs in ovarian cancer cell lines in 2D monolayers and 3D spheroids

Ovarian cancer is the most common type of gynecologic cancer. One of the leading causes of high mortality is chemoresistance, developed primarily or during treatment. Different mechanisms of drug resistance appear at the cellular and cancer tissue organization levels. We examined the differences in response to the cytotoxic drugs CIS, MTX, DOX, VIN, PAC, and TOP using 2D (two-dimensional) and 3D (three-dimensional) culture methods. We tested the drug-sensitive ovarian cancer cell line W1 and established resistant cell lines to appropriate cytotoxic drugs. The following qualitative and quantitative methods were used to assess: 1) morphology - inverted microscope and hematoxylin & eosin staining; 2) viability - MTT assay; 3) gene expression – a quantitative polymerase chain reaction; 4) identification of proteins – immunohistochemistry, and immunofluorescence. Our results indicate that the drug-sensitive and drug-resistant cells cultured in 3D conditions exhibit stronger resistance than the cells cultured in 2D conditions. A traditional 2D model shows that drug resistance of cancer cells is caused mainly by changes in the expression of genes encoding ATP-binding cassette transporter proteins, components of the extracellular matrix, “new” established genes related to drug resistance in ovarian cancer cell lines, and universal marker of cancer stem cells. Whereas in a 3D model, the drug resistance in spheroids can be related to other mechanisms such as the structure of the spheroid (dense or loose), the cell type (necrotic, quiescent, proliferating cells), drug concentrations or drug diffusion into the dense cellular/ECM structure.

The Utilisation of Hydrogels for iPSC-Cardiomyocyte Research.

Cardiac fibroblasts' (FBs) and cardiomyocytes' (CMs) behaviour and morphology are influenced by their environment such as remodelling of the myocardium, thus highlighting the importance of biomaterial substrates in cell culture. Biomaterials have emerged as important tools for the development of physiological models, due to the range of adaptable properties of these materials, such as degradability and biocompatibility. Biomaterial hydrogels can act as alternative substrates for cellular studies, which have been particularly key to the progression of the cardiovascular field. This review will focus on the role of hydrogels in cardiac research, specifically the use of natural and synthetic biomaterials such as hyaluronic acid, polydimethylsiloxane and polyethylene glycol for culturing induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). The ability to fine-tune mechanical properties such as stiffness and the versatility of biomaterials is assessed, alongside applications of hydrogels with iPSC-CMs. Natural hydrogels often display higher biocompatibility with iPSC-CMs but often degrade quicker, whereas synthetic hydrogels can be modified to facilitate cell attachment and decrease degradation rates. iPSC-CM structure and electrophysiology can be assessed on natural and synthetic hydrogels, often resolving issues such as immaturity of iPSC-CMs. Biomaterial hydrogels can thus provide a more physiological model of the cardiac extracellular matrix compared to traditional 2D models, with the cardiac field expansively utilising hydrogels to recapitulate disease conditions such as stiffness, encourage alignment of iPSC-CMs and facilitate further model development such as engineered heart tissues (EHTs).

Viral and non-viral gene therapy using 3D (bio)printing.

The overall success in launching discovered drugs is tightly restricted to the high rate of late-stage failures, which ultimately inhibits the distribution of medicines in markets. As a result, it is imperative that methods reliably predict the effectiveness and, more critically, the toxicity of medicine early in the drug development process before clinical trials be continuously innovated. We must stay up to date with the fast appearance of new infections and diseases by rapidly developing the requisite vaccinations and medicines. Modern in vitro models of disease may be used as an alternative to traditional disease models, and advanced technology can be used for the creation of pharmaceuticals as well as cells, drugs, and gene delivery systems to expedite the drug discovery procedure. Furthermore, in vitro models that mimic the spatial and chemical characteristics of native tissues, such as a 3D bioprinting system or other technologies, have proven to be more effective for drug screening than traditional 2D models. Viral and non-viral gene delivery vectors are a hopeful tool for combinatorial gene therapy, suggesting a quick way of simultaneously deliver multiple genes. A 3D bioprinting system embraces an excellent potential for gene delivery into the different cells or tissues for different diseases, in tissue engineering and regeneration medicine, in which the precise nucleic acid is located in the 3D printed tissues and scaffolds. Non-viral nanocarriers, in combination with 3D printed scaffolds, are applied to their delivery of genes and controlled release properties. There remains, however, a big obstacle in reaching the full potential of 3D models because of a lack of in vitro manufacturing of live tissues. Bioprinting advancements have made it possible to create biomimetic constructions that may be used in various drug discovery research applications. 3D bioprinting also benefits vaccinations, medicines, and relevant delivery methods because of its flexibility and adaptability. This review discusses the potential of 3D bioprinting technologies for pharmaceutical studies.

SPATIAL ABILITY IN MEDICAL AND DENTAL EDUCATION: SCOPING REVIEW

Background: the ability to mentally manipulate 3D objects and understanding the relationships between structures is essential in many fields of medical and dental specialties. This scoping review of the literature on spatial ability in medical and dental education was conducted to provide a map of the literature and identify where gaps still exist for future research.Methods: the study was conducted using a scoping review method with guidelines from the Arksey and O'Malley framework to identify literatures related to the research topic. Searches was performed from February to May 2021 in PubMed, Cochrane, CINAHL and Google Scholar using keywords related to spatial ability, medical education and dental education. Literatures was also identified using snowballing technique. PRISMA-ScR analysis for study selection was performed.Results: fifteen articles were selected for review. Spatial ability correlates with performance in studying anatomical knowledge of medicine and dentistry, and surgery. 3D anatomical models are more effective when conveying complex spatial relationships than traditional 2D models. The Mental Rotation Test (MRT) is the most widely used test for measuring spatial abilities. The results of the measurement of spatial ability showed the superiority of males over females.Conclusion: spatial ability and learning in medical and dental education provide mutual benefits for both aspects. Additional spatial abilities module may be added to medical and dental education curricula to improve learning success

3D modeling in cancer studies.

The tumor microenvironment contributes significantly to tumor initiation, progression, and resistance to chemotherapy. Much of our understanding of the tumor and its microenvironment is developed using various methods of cell culture. Throughout the last two decades, research has increasingly shown that 3D cell culture systems can remarkably recapitulate the complexity of tumor architecture and physiology compared to traditional 2D models. Unlike the flat culture system, these novel models enabled more cell-cell and cell-extracellular matrix interactions. By mimicking in vivo microenvironment, 3D culture systems promise to become accurate tools ready to be used in diagnosis, drug screening, and personalized medicine. In this review, we discussed the importance of 3D culture in simulating the tumor microenvironment and focused on the effects of cancer cell-microenvironment interactions on cancer behavior, resistance, proliferation, and metastasis. Finally, we assessed the role of 3D cell culture systems in the contexts of drug screening. 2D culture system is used to study cancer cell growth, progression, behavior, and drug response. It provides contact between cells and supports paracrine crosstalk between host cells and cancer cells. However, this system fails to simulate the architecture and the physiological aspects of in vivo tumor microenvironment due to the absence of cell-cell/ cell-ECM interactions as well as unlimited access to O2 and nutrients, and the absence of tumor heterogeneity. Recently advanced research has led researchers to generate 3D culture system that can better recapitulate the in vivo environment by providing hypoxic medium, facilitating cell-cell and cell-ECM, interactions, and recapitulating heterogeneity of the tumor. Several approaches are used to maintain and expand cancer cells in 3D culture systems such as tumor spheroids (cell aggregate that mimics the in vivo growth of tumor cells), scaffold-based approaches, bioreactors, microfluidic derives, and organoids. 3D systems are currently used for disease modeling and pre-clinical drug testing.

Advances on colorectal cancer 3D models: The needed translational technology for nanomedicine screening.

Colorectal cancer (CRC) is a heterogeneous and molecularly complex disease, associated with high mortality worldwide, exposing the urgent need for novel therapeutic approaches. Their development and translation to the clinic have been hampered, partially due to the absence of reliable cellular models that resemble key features of the human disease. While traditional 2D models are not able to provide consistent and predictive responses about the in vivo efficiency of the formulation, animal models frequently fail to recapitulate cancer progression and to reproduce adverse effects. On its turn, multicellular 3D systems, by mimicking key genetic, physical and mechanical cues of the tumor microenvironment, constitute a promising tool in cancer research. In addition, they constitute more physiological and relevant environment for anticancer drugs screening and for predicting patient's response towards personalized approaches, bridging the gap between simplified 2D models and unrepresentative animal models. In this review, we provide an overview of CRC 3D models for translational research, with focus on their potential for nanomedicines screening.

Microfluidic systems to study tissue barriers to immunotherapy.

Immunotherapies have been heavily explored in the last decade, ranging from new treatments for cancer to allergic diseases. These therapies target the immune system, a complex organ system consisting of tissues with intricate structures and cells with a multitude of functions. To better understand immune functions and develop better therapeutics, many cellular and 2-dimensional (2D) tissue models have been developed. However, research has demonstrated that the 3-dimensional (3D) tissue structure can significantly affect cellular functions, and this is not recapitulated by more traditional 2D models. Microfluidics has been used to design 3D tissue models that allow for intricate arrangements of cells and extracellular spaces, thus allowing for more physiologically relevant in vitro model systems. Here, we summarize the multitude of microfluidic devices designed to study the immune system with the ultimate goal to improve existing and design new immunotherapies. We have included models of the different immune organs, including bone marrow and lymph node (LN), models of immunity in diseases such as cancer and inflammatory bowel disease, and therapeutic models to test or engineer new immune-modulatory treatments. We particularly emphasize research on how microfluidic devices are used to better understand different physiological states and how interactions within the immune microenvironment can influence the efficacy of immunotherapies.

3D Cocultures of Osteoblasts and Staphylococcus aureus on Biomimetic Bone Scaffolds as a Tool to Investigate the Host-Pathogen Interface in Osteomyelitis.

Osteomyelitis (OM) is an infectious disease of the bone primarily caused by the opportunistic pathogen Staphylococcus aureus (SA). This Gram-positive bacterium has evolved a number of strategies to evade the immune response and subvert bone homeostasis, yet the underlying mechanisms remain poorly understood. OM has been modeled in vitro to challenge pathogenetic hypotheses in controlled conditions, thus providing guidance and support to animal experimentation. In this regard, traditional 2D models of OM inherently lack the spatial complexity of bone architecture. Three-dimensional models of the disease overcome this limitation; however, they poorly reproduce composition and texture of the natural bone. Here, we developed a new 3D model of OM based on cocultures of SA and murine osteoblastic MC3T3-E1 cells on magnesium-doped hydroxyapatite/collagen I (MgHA/Col) scaffolds that closely recapitulate the bone extracellular matrix. In this model, matrix-dependent effects were observed in proliferation, gene transcription, protein expression, and cell–matrix interactions both of the osteoblastic cell line and of bacterium. Additionally, these had distinct metabolic and gene expression profiles, compared to conventional 2D settings, when grown on MgHA/Col scaffolds in separate monocultures. Our study points to MgHA/Col scaffolds as biocompatible and bioactive matrices and provides a novel and close-to-physiology tool to address the pathogenetic mechanisms of OM at the host–pathogen interface.

Fetal weight estimation by automated three-dimensional limb volume model in late third trimester compared to two-dimensional model: a cross-sectional prospective observational study

BackgroundAccurate estimation of fetal weight is important for prenatal care and for detection of fetal growth abnormalities. Prediction of fetal weight entails the indirect measurement of fetal biometry by ultrasound that is then introduced into formulae to calculate the estimated fetal weight. The aim of our study was to evaluate the accuracy of fetal weight estimation of Chinese fetuses in the third trimester using an automated three-dimensional (3D) fractional limb volume model, and to compare this model with the traditional two-dimensional (2D) model.MethodsProspective 2D and 3D ultrasonography were performed among women with singleton pregnancies 7 days before delivery to obtain 2D data, including fetal biparietal diameter, abdominal circumference and femur length, as well as 3D data, including the fractional arm volume (AVol) and fractional thigh volume (TVol). The fetal weight was estimated using the 2D model and the 3D fractional limb volume model respectively. Percentage error was defined as (estimated fetal weight - actual birth weight) divided by actual birth weight and multiplied by 100. Systematic errors (accuracy) were evaluated as the mean percentage error (MPE). Random errors (precision) were calculated as ±1 SD of percentage error. The intraclass correlation coefficient (ICC) was used to analyze the inter-observer reliability of the 3D ultrasound measurements of fractional limb volume.ResultsUltrasound examination was performed on 56 fetuses at 39.6 ± 1.4 weeks’ gestation. The average birth weight of the newborns was 3393 ± 530 g. The average fetal weight estimated by the 2D model was 3478 ± 467 g, and the MPE was 3.2 ± 8.9. The average fetal weights estimated by AVol and TVol of the 3D model were 3268 ± 467 g and 3250 ± 485 g, respectively, and the MPEs were − 3.3 ± 6.6 and − 3.9 ± 6.1, respectively. For the 3D TVol model, the proportion of fetuses with estimated error ≤ 5% was significantly higher than that of the 2D model (55.4% vs. 33.9%, p < 0.05). For fetuses with a birth weight < 3500 g, the accuracy of the AVol and TVol models were better than the 2D model (− 0.8 vs. 7.0 and − 2.8 vs. 7.0, both p < 0.05). Moreover, for these fetuses, the proportions of estimated error ≤ 5% of the AVol and TVol models were 58.1 and 64.5%, respectively, significantly higher than that of the 2D model (19.4%) (both p < 0.05). The inter-observer reliability of measuring fetal AVol and TVol were high, with the ICCs of 0.921 and 0.963, respectively.ConclusionIn this cohort, the automated 3D fractional limb volume model improves the accuracy of weight estimation in most third-trimester fetuses. Prediction accuracy of the 3D model for neonatal BW, particularly < 3500 g was higher than that of the traditional 2D model.

Simulations of residual offset of Five-Contact vertical Hall devices with slim waist

As an important part of three-dimensional (3D) magnetic sensors, the vertical Hall devices directly determine the overall performance of the sensors. However, the traditional vertical Hall devices (5CVHD) have large offset and low sensitivity due to an inherent electrical asymmetry. In order to solve this problem, the traditional five-contact vertical Hall devices are optimized to obtain the five-contact vertical Hall devices with slim waist (SW-5CVHD) in this paper, and an anti-parallel coupling method which is suitable for the devices is applied. In addition, based on TCAD software, this paper gives a new 3D process simulation model that is more accurate than the traditional 2D model. The 3D simulation results show that the SW-5CVHD have higher sensitivity and lower initial offset compared to the traditional 5CVHD. After applying the anti-parallel coupling method, the novel devices achieve a higher symmetry by double identical SW-5CVHD interconnected into what we call the anti-parallel coupling double SW-5CVHD (APSW-5CVHD). As the structure of APSW-5CVHD is fully symmetrical, which can be better combined with the spinning current technology for offset cancellation. As a simulation result, with a bias voltage of 1 V and after spinning current a mean equivalent residual offset of 60.0µT with a standard deviation of 74.4µT is achieved among 15 samples. Moreover, the Hall voltage sensitivity is approximately 21.7 mV/VT at 1 V bias voltage.

Consistent and reproducible cultures of large-scale 3D mammary epithelial structures using an accessible bioprinting platform

BackgroundStandard three-dimensional (3D) in vitro culture techniques, such as those used for mammary epithelial cells, rely on random distribution of cells within hydrogels. Although these systems offer advantages over traditional 2D models, limitations persist owing to the lack of control over cellular placement within the hydrogel. This results in experimental inconsistencies and random organoid morphology. Robust, high-throughput experimentation requires greater standardization of 3D epithelial culture techniques.MethodsHere, we detail the use of a 3D bioprinting platform as an investigative tool to control the 3D formation of organoids through the “self-assembly” of human mammary epithelial cells. Experimental bioprinting procedures were optimized to enable the formation of controlled arrays of individual mammary organoids. We define the distance and cell number parameters necessary to print individual organoids that do not interact between print locations as well as those required to generate large contiguous organoids connected through multiple print locations.ResultsWe demonstrate that as few as 10 cells can be used to form 3D mammary structures in a single print and that prints up to 500 μm apart can fuse to form single large structures. Using these fusion parameters, we demonstrate that both linear and non-linear (contiguous circles) can be generated with sizes of 3 mm in length/diameter. We confirm that cells from individual prints interact to form structures with a contiguous lumen. Finally, we demonstrate that organoids can be printed into human collagen hydrogels, allowing for all-human 3D culture systems.ConclusionsOur platform is adaptable to different culturing protocols and is superior to traditional random 3D culture techniques in efficiency, reproducibility, and scalability. Importantly, owing to the low-cost accessibility and computer numerical control–driven platform of our 3D bioprinter, we have the ability to disseminate our experiments with absolute precision to interested laboratories.

Development of a Novel 3D Tumor-tissue Invasion Model for High-throughput, High-content Phenotypic Drug Screening

While much progress has been made in the war on cancer, highly invasive cancers such as pancreatic cancer remain difficult to treat and anti-cancer clinical trial success rates remain low. One shortcoming of the drug development process that underlies these problems is the lack of predictive, pathophysiologically relevant preclinical models of invasive tumor phenotypes. While present-day 3D spheroid invasion models more accurately recreate tumor invasion than traditional 2D models, their shortcomings include poor reproducibility and inability to interface with automated, high-throughput systems. To address this gap, a novel 3D tumor-tissue invasion model which supports rapid, reproducible setup and user-definition of tumor and surrounding tissue compartments was developed. High-cell density tumor compartments were created using a custom-designed fabrication system and standardized oligomeric type I collagen to define and modulate ECM physical properties. Pancreatic cancer cell lines used within this model showed expected differential invasive phenotypes. Low-passage, patient-derived pancreatic cancer cells and cancer-associated fibroblasts were used to increase model pathophysiologic relevance, yielding fibroblast-mediated tumor invasion and matrix alignment. Additionally, a proof-of-concept multiplex drug screening assay was applied to highlight this model’s ability to interface with automated imaging systems and showcase its potential as a predictive tool for high-throughput, high-content drug screening.

In Vitro 3‐Dimensional Modeling of Tumor Microenvironment Reveals Host/Tumor Influence on Progression in Non‐Small Cell Lung Carcinoma

BackgroundLung cancer is the leading cause of cancer related death in the United States and worldwide. Five‐year survival rates for non‐small cell lung carcinoma patients have not improved in decades, and a vast majority of patients succumb to metastasis. Identification of therapeutic targets for advanced stage disease is essential for the development of novel therapies to halt the progression of this devastating disease. However, current 2‐dimensional (2D) in vitro models are not reflective of the tumor microenvironment and are inadequate in identifying these targets. We previously reported that lung carcinoma cells grown on a human lung fibroblast derived 3‐dimensional (3D) extracellular matrix (ECM) display differences in cell morphology, proliferation, migration, and survival as compared to when they are grown in 2D. Our 3D in vitro model provides the capability to study the interaction of lung carcinoma cells with the tumor microenvironment and characterize host microenvironment effects on cancer progression.ObjectiveDetermine how ECM interactions with lung adenocarcinoma cells can promote hallmarks of cancer metastasis in an in vitro model.MethodsHuman WI‐38 fibroblasts and mouse primary fibroblasts were maintained for 8 days and then decellularized to produce ECM. Human lung adenocarcinoma cell lines A549 and mouse KrasLSL‐G12D/+;p53flox/flox lung adenocarcinoma cell lines were then grown on these matrices (3D) or plastic (2D). ECM to cell interactions were disrupted via inhibitors of ‘outside‐in’ signaling including integrins, focal adhesion kinase, src family kinases, and cytoskeletal remodeling. Cancer cell growth was assessed by AlamarBlue and markers of cancer progression were assessed by western blot analysis. Migration and invasion were assessed by matrigel scratch wound assay.ResultsCarcinoma cells grown on 3D matrices are significantly more invasive as compared to 2D. The increased invasion of A549 cells grown in 3D ECM was attenuated with inhibitors of Src family kinases and focal adhesion kinases. The altered phenotype observed when A549 cells are grown on 3D ECM was not specific to the source of the matrix as it was also observed when ECM was generated by primary fibroblasts derived from mice. Growth of cancer cells on mouse derived ECM with cancer conditioned media altered expression of known markers of EMT.ConclusionsUsing our 3D ECM model, we demonstrate that we are able to study ECM‐to‐cell interactions in vitro in a way that is not possible for traditional 2D models. These interactions play a pivotal role in determining the fate of cancer cells. Additionally, our model is expandable to primary fibroblasts derived from mice, which allows manipulation of the host microenvironment to study mechanisms of cancer progression in a physiologically relevant model. An understanding of how remodeling of the tumor microenvironment is involved in late stage disease will allow development of preventative and curative therapeutic strategies to halt tumor progression and metastasis.Support or Funding InformationThis research was supported by funds from National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [Grant R01‐DK093462, to L.J.S], National Cancer Institute [Grant R01‐CA193220, to L.J.B], and startup funds from James Graham Brown Cancer Center, University of Louisville School of Medicine (to L.J.B. and L.J.S.).