3D Human Model Research Articles

Protein structure and interactions elucidated with in-cell NMR for different cell cycle phases and in 3D human tissue models

Most of our knowledge of protein structure and function originates from experiments performed with purified proteins resuspended in dilute, buffered solutions. However, most proteins function in crowded intracellular environments with complex compositions. Significant efforts have been made to develop tools to study proteins in their native cellular settings. Among these tools, in-cell NMR spectroscopy has been the sole technique for characterizing proteins in the intracellular space of living cells at atomic resolution and physiological temperature. Nevertheless, due to technological constraints, in-cell NMR studies have been limited to asynchronous single-cell suspensions, precluding obtaining information on protein behavior in different cellular states. In this study, we present a methodology that allows for obtaining an atomically resolved NMR readout of protein structure and interactions in living human cells synchronized in specific cell cycle phases and within 3D models of human tissue. The described approach opens avenues for investigating how protein structure or drug recognition responds to cell-cell communication or changes in intracellular space composition during transitions among cell cycle phases.

Batryticatus bombyx improved atopic dermatitis in NC/Nga mice and impact on inflammation and recovery of skin barrier via STAT1/P38/NF-κB downregulation and HO-1/Nrf2 activation in HaCaT keratinocytes.

Batryticatus bombyx (BB) (Beauveria bassiana Vuill.) is used as an herbal medicine and food additive. Although BB has neuroprotective and anti-apoptotic effects, its impacts on atopic dermatitis (AD) are unknown. We evaluated the effects of BB in an NC/Nga mouse model of house dust mite (HDM)-induced AD, in TNF-α and IFN-γ (TI)-stimulated HaCaT keratinocytes, and in a 3D human skin model. NC/Nga mice were induced with HDM and treated with BB for 6 weeks. We assessed skin symptoms, serum levels of IgE, histamine, and IL-6, immune cell counts, and performed histological analysis of dorsal skin tissue. Additionally, we induced inflammation in HaCaT cells and a 3D human skin model using TNF-α/IFN-γ. We measured inflammatory cytokine levels, skin hydration factors, and delved into underlying mechanisms via ELISA, real-time PCR, and Western blot. BB improved skin lesions, reduced thickness, and inflammatory cell infiltration in HDM-induced mice. It alleviated scratching, improved moisture retention, and reduced IgE, histamine, and IL-6 levels. In HaCaT cells, BB inhibited thymic stromal lymphopoietin and increased hyaluronan content. It also upregulated aquaporin-3, hyaluronan synthase-1/3, filaggrin, involucrin, and occludin, enhancing skin hydration and tight junction stability. BB decreased STAT1, p38 MAPK, and NF-κB p65 levels in HaCaT cells and exhibited antioxidant effects by increasing HO-1/Nrf2 expression. BB may serve as a therapeutic alternative for AD treatment by inhibiting skin inflammation.

GP-HSI: Human-Scene Interaction with Geometric and Physical Constraints

With the rapid development of AR/VR technologies, achieving natural and seamless human-scene interactions has emerged as a critical challenge in computer vision. Existing methods suffer from low model placement accuracy and unnatural scene interactions. Therefore, we propose a framework called human-scene interaction with geometric and physical constraints (GP-HSI), which places a given pose of a 3D human model in an appropriate position within a 3D scene by establishing geometric and physical constraints, while ensuring interactive fidelity between the human and the scene. Specifically, first, we propose a pose-guided human contact semantic generation method, which generates human semantic labels by classifying the given human poses. Second, we propose a geometrically and semantically constrainted human model placement method, which determines the optimal position of the human model in the scene by constraining the geometric proximity and semantic consistency between models. Third, we propose an inverse kinematics based pose adjustment method, which finds the target human-scene interaction points by constructing a heterogeneous kinematic tree and solves the rotation matrix of human joints to obtain a physically plausible optimal human pose. At last, we develop an interactive system to visualize the generated human-scene interaction. The results of qualitative and quantitative experiments show that our approach is able to place human models at appropriate locations in the scene and generate plausible interactions.

Tattooed human in vitro skin model for testing the biocompatibility of tattoo inks and healing progression after tattooing

Tattoos are widespread in the population. Tattoo inks, which contain a variety of ingredients among them hazardous compounds such as polyaromatic hydrocarbons, heavy metals and nanoparticles and that are made for injection into the skin, are not dermatologically tested. New testing systems for evaluation of biocompatibility of tattoo inks as composite products and the tattooing process itself are needed. This paper describes an in vitro 3D human skin model that was tattooed with black and red ink. Biocompatibility including analysis of cytotoxicity, cytokine release, and gene expression patterns of proinflammatory cytokines, proliferation markers, growth factors and structural components was investigated over a period of 7 days. Tattooing of the 3D skin model resulted in a strong inflammatory reaction comparable to in vivo observations that subsided 4 days after treatment. The subsequent healing phase was detectable in the gene expression patterns. Tattooing with two different tattoo inks resulted in distinguishable inflammatory reactions. The described 3D skin model is a useful tool for evaluation of the biocompatibility of tattoo inks and the tattooing process itself and for characterizing the healing process after tattooing.

Using the appropriate modulus of elasticity of periodontal ligament matters in stress analysis of human first premolar tooth and periodontium structures

Although the modulus of elasticity of the human periodontal ligament (EPDL) values used in dentistry widely ranged from 0.01 to 175 MPa, the exact EPDL value has not been determined. This study aimed to verify whether and how EPDL values affect the stress distribution over the tooth and periodontium structures, and to determine the appropriate EPDL range. A 3D multi-component human first premolar model was created based on a cone-beam computed tomography dataset. Finite element analysis was performed to analyze stress distribution and deformation of the structures under an average Asian occlusal force with different EPDL values (0.0689–68.9 MPa). The low EPDL caused excessive PDL deformation, contributing to a non-uniform stress distribution and localized stress concentration, especially in the cementum, enamel, and dentin. With the low EPDL value, the stress magnitude was overestimated by 1,195%, potentially leading to erroneous conclusions regarding material failure and tooth movement. The EPDL value significantly affects the stress magnitude and distribution over the tooth and periodontium. The appropriate EPDL range of 0.964 ± 0.276 MPa is suggested for human first premolars to ensure accurate and reliable FEA simulations and help avoid misinterpretation of the stress results, which could compromise orthodontic planning and restoration design.

Evaluation of the Cardiotoxicity of Echinochrome A using Human Induced Pluripotent Stem Cell-derived Cardiac Organoids

Echinochrome A (EchA), a marine-derived natural product, has shown promise in treating cardiovascular and inflammatory diseases due to its antioxidant and anti-inflammatory properties. However, its cardiac safety remains underexplored. In this study, we utilized human induced pluripotent stem cell-derived cardiac organoids (hCOs) to validate their ability to model the cardiac safety profile of EchA in a human-relevant system. While EchA’s therapeutic effects have been reported, prior studies have not evaluated its cardiotoxicity or arrhythmogenic potential in a high-fidelity 3D human cardiac model. The hCOs, characterized by expression of key cardiac markers (cTnT) and functional ion channels (Cav1.2, Nav1.5, hERG), exhibited structural and electrophysiological properties reflective of human cardiac physiology. Using multi-electrode array (MEA) analysis, we assessed the effects of EchA at concentrations ranging from 0.1 to 30µM on electrophysiological parameters, including beat period, field potential amplitude, field potential duration, and spike slope. EchA treatment induced no significant changes in these parameters, confirming its non-toxic electrophysiological profile. Cellular viability and lactate dehydrogenase (LDH) assays revealed no cytotoxic effects of EchA across tested concentrations. Contractility assays further demonstrated that EchA did not affect contraction velocity, relaxation velocity, or time to 50% maximal contraction and relaxation. This study fills a critical gap and highlights the translational relevance of hCOs for cardiotoxicity assessment, demonstrating EchA's cardiac safety and supporting its potential therapeutic and environmental applications.

GaitAGE: Gait Age and Gender Estimation Based on an Age- and Gender-Specific 3D Human Model

GaitAGE: Gait Age and Gender Estimation Based on an Age- and Gender-Specific 3D Human Model

Cooperation of Cysteine, Ascorbic Acid, Pyridoxine, and α-Lipoic Acid in Antioxidant Effects on Human Keratinocytes and 3D Human Skin Models

Combinations of cysteine, ascorbic acid, and pyridoxine are frequently used in oral formulations. Although there have been many reports on the efficacy of each of these ingredients, little information is known about their combined effects on skin cells. The purpose of this study was to evaluate the combined effects of cysteine, ascorbic acid, and pyridoxine, as well as the effect of adding α-lipoic acid, on skin cells. Human keratinocytes were treated with individual antioxidants or combinations of them, and glutathione levels and expression of glutathione synthesis-related genes were measured. The combination of cysteine, ascorbic acid, and pyridoxine increased the expression of γ-GCSc and tended to increase glutathione levels compared to the controls. Interestingly, the addition of α-lipoic acid further increased glutathione levels by increasing the expression of CD98. Similarly, in 3D-human skin models, the combination of cysteine, ascorbic acid, and pyridoxine promoted glutathione synthesis, and this effect was enhanced by the addition of α-lipoic acid. Furthermore, melanin synthesis inhibition was shown to be dependent on increased glutathione levels. These results suggest that the addition of α-lipoic acid to the combination of cysteine, ascorbic acid, and pyridoxine enhances glutathione production and may have anti-aging effects through efficient oxidative stress reduction.

A 3D Model of the Human Lung Airway for Evaluating Permeability of Inhaled Drugs.

Current in vitro cell-based methods, relying on single cell types, have structural and functional limitations in determining lung drug permeability, which is a contributing factor affecting both local and systemic drug levels. To address this issue, we investigated a 3D human lung airway model generated using a cell culture insert, wherein primary human lung epithelial and endothelial cells were cocultured at an air-liquid interface (ALI). To ensure that the cell culture mimics the physiological and functional characteristics of airway tissue, the model was characterized by evaluating several parameters such as cellular confluency, ciliation, tight junctions, mucus-layer formation, transepithelial electrical resistance, and barrier function through assaying fluorescein isothiocyanate-dextran permeability. To understand how the characterized ALI quality attributes influenced the absorption of inhaled drugs through the epithelial-endothelial barrier, we measured the permeability and epithelial intracellular concentrations of albuterol sulfate (AL), formoterol fumarate (FO), and fluticasone furoate (FL). The presented characterization results overall demonstrate that this culture platform mimicked the airway-specific structure and barrier function. An apparent permeability (P app) of 5.7 × 10-6 cm/s and an intracellular concentration below 1% were quantified for AL over 3 h. The P app of FO was 8.5 × 10-6 cm/s, with an intracellular concentration of 3.8%. Due to its high lipophilicity, FL showed a higher intracellular concentration (17.4%) compared to AL and FO, but also a 73.1% loss of the compound over 3 h due to nonspecific binding, with a P app as low as 1.3 × 10-7 cm/s. While the model exhibited physiologically relevant properties, its utility in estimating the permeability of inhaled drugs may be drug-specific, warranting further optimization and study.

Application of Computerized 3D Human Model Training Technology in Sports Movement Correction

Application of Computerized 3D Human Model Training Technology in Sports Movement Correction

Comparative evaluation of antitumor effects of methionine-γ-lyase in <i>in vitro</i> 2D and 3D human tumor models

BACKGROUND: Promising antitumor drug screening results obtained from monolayer cultures are often poorly reproduced in the in vivo models. Using clinically relevant 3D in vitro human tumor models, such as spheroids, provides a more reliable framework for evaluating antitumor activity. This is particularly important when the drug’s mechanism of action targets cellular metabolism. AIM: To conduct a comparative study of the antitumor activity of methionine-γ-lyase (MGL) in 2D and 3D human tumor models. MATERIALS AND METHODS: Human fibroblast cell culture and tumor cell lines, e.g. MCF7 human breast cancer, HCT-116 colorectal cancer, PANC-1 human pancreatic cancer, Huh7 liver cancer, and LNCaP human prostate cancer were used to evaluate MGL cytotoxicity. Cultural plates with low-adhesive coating were used to produce the spheroids. Cell survival was assessed using the resazurin test. RESULTS: The spheroid model showed higher cell survival after 72-hour MGL exposure compared with the monolayer culture model in all tested cultures. Fibroblasts demonstrated the lowest sensitivity to MGL exposure in both 2D and 3D culture models, with IC50=2.2 and 9.1 IU/mL, respectively. The rapidly proliferating PANC-1 and HCT-116 cells showed the highest sensitivity to MGL in 2D and 3D models: IC50=0.23 and 1.5 IU/mL; IC50=0.83 and 1.43 IU/mL, respectively. CONCLUSION: The effect of MGL on cell survival in the spheroid systems is less pronounced than in monolayers. The viability of cells exposed to survival MGL in the spheroid culture system is independent of spheroid size or growth rate. Despite the maximum cytotoxic effect being observed in the fast-growing spheroid model of colon cancer HCT-116, and the lowest in the model of slowly dividing fibroblasts, this dependence was not so obvious for other cell types. Overall, the spheroid model has proven useful for testing the specific activity of enzyme-based antitumor drugs, as far as it overcomes the excessive intrinsic sensitivity observed in monolayer cancer models.

Harnessing 2D and 3D human endometrial cell culture models to investigate SARS-CoV-2 infection in early pregnancy.

Vertical transmission of SARS-CoV-2 during human pregnancy remains highly controversial as most studies have focused on the third trimester or the peripartum period. Given the lack of early trimester data, determining the prevalence of vertical transmission during early pregnancy and assessing the potential risks for fetal morbidity and mortality poses a challenge. Therefore, we analyzed the impact of SARS-CoV-2 infection on an endometrial 3D spheroid model system. The 3D spheroids are capable of decidualization and express ACE2 as well as TMPRSS2, rendering them susceptible to SARS-CoV-2 infection. Employing this 3D cell model, we identified that SARS-CoV-2 can infect both non-decidualized and decidualized endometrial spheroids. Infection significantly increased the chemokine MCP-1 compared to non-infected spheroids. Decidualized spheroids exhibited upregulated chemokine IL-8 levels. Furthermore, RNA sequencing revealed dysregulation of several genes involved in tissue-specific immune response, Fc receptor signalling, angiotensin-activated signalling and actin function. Gene expression changes varied between SARS-CoV-2 infected non-decidualized and decidualized spheroids and genes associated with the innate immune system (CCL20, CD38, LCN2 and NR4A3) were dysregulated as a potential mechanism for immune evasion of SARS-CoV-2. Altogether, our study demonstrates that endometrial spheroids are a useful model to examine the clinical implications of SARS-CoV-2 vertical transmission, warranting further investigations.

3D human model guided pose transfer via progressive flow prediction network

Human pose transfer is to transfer a conditional person image to a new target pose. The difficulty lies in modeling the large-scale spatial deformation from the conditional pose to the target one. However, the commonly used 2D data representations and one-step flow prediction scheme lead to unreliable deformation prediction because of the lack of 3D information guidance and the great changes in the pose transfer. Therefore, to bring the original 3D motion information into human pose transfer, we propose to simulate the generation process of real person image. We drive the 3D human model reconstructed from the conditional person image with the target pose and project it to the 2D plane. The 2D projection thereby inherits the 3D information of the poses which can guide the flow prediction. Furthermore, we propose a progressive flow prediction network consisting of two streams. One stream is to predict the flow by decomposing the complex pose transformation into multiple sub-transformations. The other is to generate the features of the target image according to the predicted flow. Besides, to enhance the reliability of the generated invisible regions, we use the target pose information which contains structural information from the flow prediction stream as the supplementary information to the feature generation. The synthesized images with accurate depth information and sharp details demonstrate the effectiveness of the proposed method.

The Effects of Electronic Cigarettes on Oral Microbiome and Metabolome in 3D Tissue-Engineered Models.

Recent studies have shown that electronic cigarettes (ECs) use disrupts the oral microbiome composition and diversity, impairing the metabolic pathways of the mucosal cells. However, to date, no reports have evaluated the role of EC exposure in the context of oral metabolome. Hence, the aim of this study was to investigate the role of EC aerosol exposure in the dysregulation of the oral microbiome and metabolome profile using in vitro 3D organotypic models of human oral mucosa. 3D tissue-engineered human oral mucosa models were generated and infected with oral microbes obtained from saliva of a healthy donor. The epithelial surface of the oral mucosal models was exposed directly to the EC aerosol (flavoured; with and without nicotine) as it came out of a simulated activated device that mimicked the clinical situation. A comprehensive assessment of oral microbiome community composition by bacterial 16S rRNA gene sequencing was performed. A gas chromatography-based mass spectrometry analysis was also conducted to identify the effect of vaping on the oral metabolome profile. A higher alpha diversity in flavoured EC with nicotine groups was observed compared to controls, with notable differences in bacterial taxa abundance. Metabolomics analysis further demonstrated distinct clustering of control, EC with flavoured nicotine, and flavoured EC groups, confirming 13 metabolites that were statistically higher in levels in flavoured EC with nicotine group, indicating the adverse effects of nicotine on the oral mucosa model. Altered metabolites were mainly enriched in pathways associated with oral cancer progression. This study underscores the significant impact of EC use on oral health, highlighting alterations in the oral microbiome, bacterial composition, and metabolite profiles via a clinically relevant in vitro 3D organotypic model of human oral mucosa.

Implementing enclosed sterile integrated robotic platforms to improve cell-based screening for drug discovery.

At GSK, we have implemented custom integrated robotics platforms housed in bespoke biosafety enclosures to augment our capabilities in advanced cellular screening. Here we present and discuss the impact of one such system, the Cellular Automated Screening Platform (CASPer). We evaluate the benefits of implementing specific processes on CASPer that include increasing the throughput of safety screening assays and improving data integrity when testing complex in vitro 3D primary human hepatocyte models. This article provides an overview of the platforms installed and offers insight into their utilisation by presenting example workflows and quality control solutions which have been implemented. We offer perspective on the advantages of such custom integrated systems and their limitations in cellular screening for early drug discovery.

A Review of the State of the Art 3D Generative Models and Their Applications

Ever since 2022, there has been a large number of 3D generative models that have been devised and published, such as AvatarGen, CityDreamer, and HOLOFUSION. Generally speaking, these models can perform tasks such as generating a 3D human model, creating an unbounded city scene, and constructing a 3D object. And it is not a surprise that 3D generative models are very popular these years because there has been a witness of huge need for 3D models in the global market and the models themselves also serve as both convenient and productive tools for the relevant industries. For instance, 3D generative models can utilize a combination of Generative Adversarial Network (GAN) and Multi-Layer Perceptron (MLP) or Neural Radiance Field (NeRF) or Diffusion Model to produce 3D human model; Autoregressive Model or Feature Extraction + Volume Rendering to generate 3D scenes; Diffusion Model or GAN + MLP to produce 3D objects. This paper tries to present a taxonomy of the main 3D generative models from the angle of the kinds of outputs and strategies employed by different models.

Multicellular 3D models to study myocardial ischemia-reperfusion injury.

Coronary heart disease is a major global health threat, with acute myocardial ischemia-reperfusion injury (IRI) being a major contributor to myocardial damage following an ischemic event. IRI occurs when blood flow to ischemic tissues is restored and exacerbates the cellular damage caused by ischemia/hypoxia. Although animal studies investigating IRI have provided valuable insights, their translation into clinical outcomes has been limited, and translation into medical practice remains cumbersome. Recent advancements in engineered three-dimensional human in vitro models could offer a promising avenue to bridge the "therapeutic valley of death" from bench to bedside, enhancing the understanding of IRI pathology. This review summarizes the current state-of-the-art cardiovascular 3D models, including spheroids, organoids, engineered cardiac microtissues, and organ-on-a-chip systems. We provide an overview of their advantages and limitations in the context of IRI, with a particular emphasis on the crucial roles of cell-cell communication and the multi-omics approaches to enhance our understanding of the pathophysiological processes involved in IRI and its treatment. Finally, we discuss currently available multicellular human 3D models of IRI.

Lower hepatotoxicity risk in Xelaglifam, a novel GPR40 agonist, compared to Fasiglifam for type 2 diabetes therapy

Fasiglifam, a candidate targeting GPR40, showed efficacy in clinical trials for type 2 diabetes but exerted liver toxicity. This study investigated the drug-induced liver injury (DILI) risk of Xelaglifam, a new GPR40 agonist, based on the potential toxicity mechanism of Fasiglifam; transporter inhibition, mitochondrial dysfunction, reactive metabolite formation, and covalent binding to proteins. In the hepatobiliary transporter assay, Xelaglifam showed a broader safety margin (>10-fold) against bile acid transporters, suggesting its less likelihood to cause bile acids accumulation, unlike Fasiglifam (<10-fold safety margin). Moreover, Xelaglifam showed no effect on glycocholic acid accumulation at higher concentrations than the estimated Cmax in the 3D human liver model, whereas Fasiglifam affected the accumulation. In the HepaRG spheroids 3D model, the AC50 values of Xelaglifam for mitochondrial function-related parameters were higher than Fasiglifam. Unlike Fasiglifam, none of the cell parameters for Xelaglifam were below the estimated 5x Cmax. Additionally, the glucuronide metabolite of Xelaglifam was negligible (<1 % of the parent) in the Safety Testing, indicating a limited contribution to DILI. Fasiglifam activated genes related to liver disease, whereas Xelaglifam had no effect; instead, it increased FXR activity, a bile acid regulator. Notably, toxicity studies in rats and monkeys showed no adverse liver effects at higher exposure levels than the effective human blood concentration. Overall, these results support a low risk of DILI for Xelaglifam treatment and the justification for its long-term use for treating type 2 diabetes.

TMOD-20. RAPID PROCESSING AND BIOBANKING OF RESECTED BRAIN TUMOR TISSUE TO GENERATE PATIENT-DERIVED BRAIN TUMOR EXPLANT ORGANOIDS (GBOS) FOR PRECLINICAL STUDIES

Abstract Improved preclinical tools are urgently needed to translate new brain cancer treatments. Patient-derived brain tumor explant organoids (GBOs) offer promise for studying tumor cells in a relevant human tumor microenvironment and predicting patient responses to therapy. However, generating GBOs is time-consuming, costly, and technically challenging, hindering the creation of comprehensive biobanks covering the spectrum of brain tumor heterogeneity. This work presents a semi-automated method for producing GBOs, involving tumor tissue processing, size selection, and same-day cryopreservation, reducing generation time to less than 1 hour. We established a biobank of 24 GBOs from 33 samples (11 males and 13 females) with primary (n=13, GBO Yield (GY)&amp;gt;90%) and recurrent (n=5, GY&amp;gt;90%) glioblastomas, high-grade gliomas (HGG, n=2, GY≤50%), and low-grade gliomas (LGG, n=2 with GY&amp;gt;90%, n=2 with GY≤50%). Using GBO size and propidium iodide as readouts, we used this biobank to compare anticancer activities of novel idronoxil-conjugated benzopyran compounds (NX786, NX904/904E1) against Bortezomib (100% cell death reference). NX786 and NX904/904E1 reduced GBO growth (≥50%) in all primary glioblastoma and one (of two) recurrent glioblastoma GBOs with milder effects in LGG GBOs. Two (of four) primary glioblastoma, one (of two) recurrent glioblastoma, and one LGG GBOs exhibited significant cell death (&amp;gt;60%) in response to NX904/904E1. In contrast, NX786 induced cell death in one (of three) primary glioblastoma (34% cell death) and one (of two) recurrent glioblastoma (71% cell death) GBOs but not in LGG GBOs, together showcasing varied responses across different brain tumors. Highly-passaged GBOs lacking the non-malignant tumor microenvironment were more susceptible to these treatments, underlining the tumor microenvironment’s critical role in responses to anticancer agents. We provide a new method for efficient processing and cryopreserving GBOs, enabling the establishment of large biobanks for patient-specific preclinical drug testing across brain tumor subgroups in a clinically relevant human 3D model.

Standalone single- and bi-layered human skin 3D models supported by recombinant silk feature native spatial organization

Physiologically relevant human skin models that include key skin cell types can be used forin vitrodrug testing, skin pathology studies, or clinical applications such as skin grafts. However, there is still no golden standard for such a model. We investigated the potential of a recombinant functionalized spider silk protein, FN-silk, for the construction of a dermal, an epidermal, and a bilayered skin equivalent (BSE). Specifically, two formats of FN-silk (i.e. 3D network and nanomembrane) were evaluated. The 3D network was used as an elastic ECM-like support for the dermis, and the thin, permeable nanomembrane was used as a basement membrane to support the epidermal epithelium. Immunofluorescence microscopy and spatially resolved transcriptomics analysis demonstrated the secretion of key ECM components and the formation of microvascular-like structures. Furthermore, the epidermal layer exhibited clear stratification and the formation of a cornified layer, resulting in a tight physiologic epithelial barrier. Our findings indicate that the presented FN-silk-based skin models can be proposed as physiologically relevant standalone epidermal or dermal models, as well as a combined BSE.