Patient-derived xenografts (PDXs) provide biologically relevant models and potential platforms for the development of treatment strategies for precision medicine in pancreatic cancer. Furthermore, circulating epithelial tumor cells (CETCs/CTCs) are released into the bloodstream by solid tumors and a rare subpopulation—circulating cancer stem cells (cCSCs) – is considered to be responsible for recurrence and plays a key role in metastasis. For the identification of cCSCs, an innovative in vitro assay to generate tumorspheres was established in this study. The number of tumorspheres and CETCs/CTCs was analyzed perioperatively in 25 pancreatic cancer patients. Additionally, an individual in vivo chorioallantoic membrane (CAM) culture system was used to generate PDXs from these tumorspheres. While overall correlations of CETCs/CTCs with clinicopathological parameters did not reach statistical significance, a significant difference in the number of tumorspheres was observed between patient subgroups with lower and higher UICC stages. This finding underscores their potential as biomarkers, providing valuable insights into clinical decision-making and tumor progression. The application of tumorspheres on the CAM successfully established PDXs within 7 days. These xenografts closely resembled the histological features of the primary tumor. Hence, this model represents a novel and fast option for individualized testing of new therapies for PDAC.

Cerebral cavernous malformations (CCMs) are clusters of thin-walled enlarged blood vessels in the central nervous system that are prone to recurrent hemorrhage and can occur in both sporadic and familial forms. The familial form results from loss-of-function variants in the CCM1, CCM2, or CCM3 gene. Despite a better understanding of CCM pathogenesis in recent years, it is still unclear why CCM3 mutations often lead to a more aggressive phenotype than CCM1 or CCM2 variants. By combining high-throughput differentiation of blood vessel organoids from human induced pluripotent stem cells (hiPSCs) with a CCM1, CCM2, or CCM3 knockout, single-cell RNA sequencing, and high-content imaging, we uncovered both shared and distinct functions of the CCM proteins. While there was a significant overlap of differentially expressed genes in fibroblasts across all three knockout conditions, inactivation of CCM1, CCM2, or CCM3 also led to specific gene expression patterns in neuronal, mesenchymal, and endothelial cell populations, respectively. Taking advantage of the different fluorescent labels of the hiPSCs, we could also visualize the abnormal expansion of CCM1 and CCM3 knockout cells when differentiated together with wild-type cells into mosaic blood vessel organoids. In contrast, CCM2 knockout cells showed even reduced proliferation. These observations may help to explain the less severe clinical course in individuals with a pathogenic variant in CCM2 and to decode the molecular and cellular heterogeneity in CCM disease. Finally, the excellent scalability of blood vessel organoid differentiation in a 96-well format further supports their use in high-throughput drug discovery and other biomedical research studies.

Bei Lymphozelen handelt es sich um eine Ansammlung von klarer Lymphflüssigkeit, in einem anatomisch dafür nicht vorgesehenen Raum. Zur Bildung von Lymphozelen kommt es beispielsweise nach Lymphknotendissektionen oder gefäßchirurgischen Eingriffen. Zur Behandlung von Lymphozelen wurden unterschiedliche Therapien beschrieben, wozu konservative Therapie, niedrig-dosierte Radiotherapie, sklerosierend wirkende Substanzen und die sehr erfolgversprechende Therapieoption der (supra-)mikrochirurgischen Resektion der Lymphozelen mit Einsatz von Indocyanin-Grün-Lymphographie zählen.

Abstract Background: Pancreatic cancer carries a poor prognosis in the majority of patients, with an overall 5-year survival rate of less than 10%. Therefore, the development of new therapies and preclinical models is crucial and urgently needed. Circulating cancer stem cells (cCSCs) are a subset of circulating cancer cells that have stem cell-like properties and play a significant role in tumor progression, metastasis, and treatment resistance. Patient-derived xenografts (PDX) reflect the genotypic and phenotypic landscape of patients' cancers and are useful for drug screening, biomarker development, and preclinical evaluation of personalized medicine strategies. In this study, we describe a fast, simple, and cost effective method to generate PDX on Chorioallantoic Membrane (CAM) from circulating cancer stem cells. Methods: In this study, primary cultures from circulating cancer stem cells were established using sphere-forming assays from 30 patients with newly diagnosed pancreatic cancer. Subsequently, tumorspheres were transplanted onto the CAM membrane of fertilized chicken eggs to form patient- derived xenografts. Results: For the identification of cCSCs from peripheral blood of pancreatic cancer patients, an innovative in vitro sphere-forming assay was established. The number of tumorspheres was correlated with clinicopathological parameters like tumor size, lymph node involvement and grading status of the primary tumor. Implantation of tumorspheres onto the CAM was successful in ~90 % of the applied samples. The histological analysis suggests that the PDX tumors were morphologically and pathologically identical to primary tumor tissue. Conclusions: The number of tumorspheres was associated with clinicopathological parameters. Further follow-up is needed to evaluate the prognostic significance of tumorsphere detection in pancreatic cancer. Generation of PDX from cCSCs on CAM has a high success rate and may become an integral part of the drug development arena, including drug screening and biomarker development. Citation Format: Monika Pizon, Dorothea Schott, Andreas Ettner-Sittner, Benedikt J. Wagner, Katharina Pachmann, Thiha Aung, Christina Hackl, Silke Haerteis. Patient-derived xenografts from circulating cancer stem cells and their potential utility in personalized medicine of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6900.

Textile-based microfluidic biosensors represent an innovative fusion of various multidisciplinary fields, including bioelectronics, material sciences, and microfluidics. Their potential in biomedicine is significant as they leverage textiles to achieve high demands of biocompatibility with the human body and conform to the irregular surfaces of the body. In the field of microfluidics, fabric coated with hydrophobic materials serves as channels through which liquids are transferred in precise amounts to the sensing element, which in this case is a biosensor. This paper presents a condensed overview of the current developments in textile-based microfluidics and biosensors in biomedical applications over the past 20 years (2005–2024). A literature search was performed using the Scopus database. The fabrication techniques and materials used are discussed in this paper, as these will be key in various modifications and advancements in textile-based microfluidics. Furthermore, we also address the gaps in the application of textile-based microfluidic analytical devices in biomedicine and discuss the potential solutions. Advances in textile-based microfluidics are enabled by various printing and fabric manufacturing techniques, such as screen printing, embroidery, and weaving. Integration of these devices into everyday clothing holds promise for future vital sign monitoring, such as glucose, albumin, lactate, and ion levels, as well as early detection of hereditary diseases through gene detection. Although most testing currently takes place in a laboratory or controlled environment, this field is rapidly evolving and pushing the boundaries of biomedicine, improving the quality of human life.

Ultra high frequency (UHF) ultrasound enables the visualization of very small structures that cannot be detected by conventional ultrasound. The utilization of UHF imaging as a new imaging technique for the 3D-in-vivo chorioallantoic membrane (CAM) model can facilitate new insights into tissue perfusion and survival. Therefore, human renal cystic tissue was grafted onto the CAM and examined using UHF ultrasound imaging. Due to the unprecedented resolution of UHF ultrasound, it was possible to visualize microvessels, their development, and the formation of anastomoses. This enabled the observation of anastomoses between human and chicken vessels only 12 h after transplantation. These observations were validated by 3D reconstructions from a light sheet microscopy image stack, indocyanine green angiography, and histological analysis. Contrary to the assumption that the nutrient supply of the human cystic tissue and the gas exchange happens through diffusion from CAM vessels, this study shows that the vasculature of the human cystic tissue is directly connected to the blood vessels of the CAM and perfusion is established within a short period. Therefore, this in-vivo model combined with UHF imaging appears to be the ideal platform for studying the effects of intravenously applied therapeutics to inhibit renal cyst growth.

Dopamine (DA) neurotransmitter is a prominent component of the central nervous system and its deficiency contributes to several diseases, such as Parkinson's disease and schizophrenia, which significantly influence the quality of life of affected individuals. This paper introduces a circular interdigitated ring electrode, the three-dimensional carbon interdigitated ring array nanofibers (3D C-IDRA NFs), for the detection of DA using amperometric detection. The study utilized cyclic voltammetry and chronoamperometry to investigate DA. The integration of carbon nanofibers (CNFs) with 3D C-IDRA demonstrated enhanced surface area, increased current peak, and superior performance compared to the standalone 3D C-IDRA electrode, as evidenced by a redox amplification factor of 2.94 and a collection efficiency of 81.1%. The results highlight that integrating CNFs with 3D C-IDRA improves the electrochemical performance in detecting DA neurotransmitter. Thus, this discovery elucidates the advantageous potential of the 3D C-IDRA NF electrode in a wide range of applications.

Covarine, copper phthalocyanine, a novel tooth whitening ingredient, has been incorporated into various toothpaste formulations using diverse technologies such as larger flakes, two-phase pastes, and microbeads. In this study, we investigated the behavior of covarine microbeads (200 μm) in Colgate advanced white toothpaste when mixed with artificial and real saliva. Our analysis utilized a custom-designed microfluidic mixer with 400 μm wide channels arranged in serpentine patterns, featuring a Y-shaped design for saliva and toothpaste flow. The mixer, fabricated using stereolithography 3D printing technology, incorporated a flexible transparent resin (Formlabs' Flexible 80A resin) and PMMA layers. COMSOL simulations were performed by utilizing parameters extracted from toothpaste and saliva datasheets, supplemented by laboratory measurements, to enhance simulation accuracy. Experimental assessments encompassing the behavior of covarine particles were conducted using an optical profilometer. Viscosity tests and electrical impedance spectroscopy employing recently developed all-carbon electrodes were employed to analyze different toothpaste dilutions. The integration of experimental data from microfluidic chips with computational simulations offers thorough insights into the interactions of covarine particles with saliva and the formation of microfilms on enamel surfaces.

Molecular Communications (MC) research is increasingly focused on applications within the human body, such as health monitoring and drug delivery. These applications require testing in realistic and living environments. Thus, advancing experimental MC research to the next level requires the development of in vivo experimental testbeds. In this paper, we introduce the Chorioallantoic Membrane ( CAM ) model as a versatile 3D in vivo MC testbed. The CAM is a highly vascularized membrane formed in fertilized chicken eggs and has gained significance in various research fields, including bioengineering, cancer research, and drug development. Its versatility, reproducibility, and realistic biological properties make the CAM model perfectly suited for next-generation MC testbeds, facilitating the transition from proof-of-concept systems to practical applications. We provide a comprehensive introduction to the CAM model, its properties, and its applications in practical research. Additionally, we present a characterization of the CAM model as an MC system. As a preliminary experimental study, we investigate the distribution of fluorescent molecules in the closed-loop vascular system of the CAM model. We also derive an approximate analytical model for the propagation of molecules in closed-loop systems, and show that the proposed model is able to approximate molecule propagation in the CAM model.

This article presents a low-cost 2x2 metasurface-based rectenna array operating at the 2.45 GHz industrial, scientific, and medical (ISM) band, designed to power low-power wearable medical sensors (WMS). The proposed antenna is a novel textile-based metasurface interlayer patch (meta-patch) antenna fabricated on a felt fabric substrate, which replaces the conventional ground plane with a grounded metasurface consisting of a 3x3 unit cell array. A single excitation port is used for both the patch antenna and the metasurface to improve gain, efficiency, and bandwidth. The meta-patch antenna achieved a high gain of 7 dB, an efficiency of 77%, and an enhanced bandwidth of 120 MHz. The rectifier employs a seven-stage Cockcroft-Walton Voltage Multiplier (CWVM) topology, fabricated on a rigid substrate. To evaluate the performance of the meta-patch rectenna array and the effect of adding more elements, the proposed meta-patch rectenna was tested in single-element, 2x1 array, and 2x2 array configurations under continuous RF power. The single-element, 2x1 rectenna array, and 2x2 rectenna array achieved power conversion efficiencies (PCE) of 52%, 53%, and 56%, with DC output powers of 414 mu W, 429 mu W, and 450 mu W, respectively, at an input power of -1 dBm. The results demonstrate the potential of the proposed rectenna arrays for efficiently energizing low-power WMS.

PurposeThis study aims to use an additive process for the first time to develop a microfluidic device that uses centrifugal technique for precise and repeatable generation of microdroplets. Droplets have versatile applications in life sciences, but so far centrifugal devices for their production have been made mainly using standard subtractive techniques. This study focused on evaluating the applicability of 3D printing technology in the development of centrifugal microfluidic devices and investigating their properties and future applications.Design/methodology/approachFirst, the background of this interdisciplinary research, including the principle of droplet microfluidics and the centrifugal technique, is explained. The developed device has the form of a disc (similar to an audio CD), containing an integrated microfluidic system for droplet generation. The disc is rotated at a specific spin profile to induce controlled liquid flow and accurate production of oil-in-water microdroplets. The device was fabricated using material jetting technology. The design, operation principles, printing process parameters and post-processing steps are explained in detail.FindingsThe device was thoroughly characterised, including its mechanical properties, the impact of chemical treatment and the flow measurement of the liquids. The study confirms that the disc can be applied to produce various emulsions using centrifugal force alone. 3D printing technology enables potential mass production and other applications of the device.Originality/valueThe 3D printing process allowed for easy design, fabrication and duplication of the device. Compared to standard PMMA discs, a simpler fabrication protocol and a more flexible and monolithic structure were obtained. The device can be adapted to other microfluidic processes in a lab with high potential for point-of-care applications.

This study presents a systematic redesign of a microfluidic compact disc (CD) that showcases six distinct chamber subsystems. This type of a subsystem is specifically designed to facilitate concurrent sample loading, mixing, and analysis. Departing from the original focus on saliva as a diagnostic medium, this study shifts towards the utilization of sweat. Modifications are made to the electrode designs to ensure accurate comparison and electrochemical analysis. The primary aim of this investigation is to explore the variations in electrochemical properties that occur within dilution of sweat samples as well as varying pH levels. This is important as specimens can vary greatly in their composition and complexity. By contrasting the electrochemical impedance characteristics of sweat samples with different pH levels, this study aims to shed light on potential foundations for the early diagnosis and monitoring of skin diseases. Preliminary findings from this extension of the study reveal distinct conductance patterns. These findings provide valuable insights and further support the notion that the redesigned microfluidic CD platform possesses the capability for multiplex processes. This, in turn, paves the way for future research in the field of theranostics and the electrochemical analysis of diverse sweat compositions at the point of care.

Although significant improvements have been made in the treatment of pancreatic cancer, its prognosis remains poor with an overall 5-year survival rate of less than 10%. New experimental approaches are necessary to develop novel therapeutics. In this study, the investigation of pancreatic cancer tissue growth in the chorioallantoic membrane (CAM) model and the subsequent use of indocyanine green (ICG) injections for the verification of intratumoral perfusion was conducted. ICG was injected into the CAM vasculature to visualize the perfusion of the tumor tissue. The presence of metastasis was investigated through PCR for the human-specific ALU element in the liver of the chicken embryo. Additionally, the usage of cryopreserved pancreatic tumors was established. Intratumoral perfusion of tumor tissue on the CAM was observed in recently obtained and cryopreserved tumors. ALU-PCR detected metastasis in the chick embryos’ livers. After cryopreservation, the tissue was still vital, and the xenografts generated from these tumors resembled the histological features of the primary tumor. This methodology represents the proof of principle for intravenous drug testing of pancreatic cancer in the CAM model. The cryopreserved tumors can be used for testing novel therapeutics and can be integrated into the molecular tumor board, facilitating personalized tumor treatment.

BackgroundOral health is closely linked to systemic conditions, particularly non-communicable diseases (NCDs), which can exacerbate oral issues. Essential oils (EOs) have emerged as potential alternatives for oral health due to their antibacterial, anti-inflammatory, and antioxidant properties. Among these, rosemary essential oil (REO) shows promise due to its various biological activities. This study investigates the potential of REO in dental applications using microfluidic devices and electrochemical impedance spectroscopy (EIS) to analyze the electrical properties of REO in artificial saliva (AS) mixtures.ResultsThe study demonstrated significant variations in impedance across different REO concentrations and their mixtures with AS. Higher impedance was observed in REO mixtures, particularly at lower frequencies, indicating distinct electrical properties compared to pure AS. The impedance of REO was influenced by its concentration, with a 1% REO solution showing higher impedance than a 4% solution, possibly due to micelle formation and changes in dielectric properties. Additionally, microfluidic devices enabled precise control over fluid interactions and real-time monitoring, offering valuable insights into REO's behavior in a simulated oral environment. The impedance data demonstrated significant differences in REO-AS mixtures, highlighting potential interactions critical for oral care applications.ConclusionsRosemary essential oil exhibits unique electrical properties, making it a promising candidate for dental applications, particularly in preventing and treating oral diseases. Microfluidic devices enhance the accuracy and reliability of studying REO's interactions with AS, providing a robust platform for future dental research. The findings suggest that REO could be effectively incorporated into oral care products, offering a natural alternative for combating oral pathogens, reducing inflammation, and protecting against oxidative stress. Future research should focus on clinical trials to validate these findings and explore the synergistic effects of REO with other essential oils.

Osteosarcomas are the most common primary malignant bone tumors and mostly affect children, adolescents, and young adults. Despite current treatment options such as surgery and polychemotherapy, the survival of patients with metastatic disease remains poor. In recent studies, punicalagin has reduced the cell viability, angiogenesis, and invasion in cell culture trials. The aim of this study was to examine the effects of punicalagin on osteosarcomas in a 3D in vivo tumor model. Human osteosarcoma biopsies and SaOs-2 and MG-63 cells, were grown in a 3D in vivo chorioallantoic membrane (CAM) model. After a cultivation period of up to 72 h, the tumors received daily treatment with punicalagin for 4 days. Weight measurements of the CAM tumors were performed, and laser speckle contrast imaging (LSCI) and a deep learning-based image analysis software (CAM Assay Application v.3.1.0) were used to measure angiogenesis. HE, Ki-67, and Caspase-3 staining was performed after explantation. The osteosarcoma cell lines SaOs-2 and MG-63 and osteosarcoma patient tissue displayed satisfactory growth patterns on the CAM. Treatment with punicalagin decreased tumor weight, proliferation, and tumor-induced angiogenesis, and the tumor tissue showed pro-apoptotic characteristics. These results provide a robust foundation for the implementation of further studies and show that punicalagin offers a promising supplementary treatment option for osteosarcoma patients. The 3D in vivo tumor model represents a beneficial model for the testing of anti-cancer therapies.

Background: This study explored the integration of conductive threads into a microfluidic compact disc (CD), developed using the xurographic method, for a potential sweat biosensing platform. Material/Methods: The microfluidic CD platform, fabricated using the xurographic method with PVC films, included venting channels and conductive threads linked to copper electrodes. With distinct microfluidic sets for load and metering, flow control, and measurement, the CD's operation involved spinning for sequential liquid movement. Impedance analysis using HIOKI IM3590 was conducted for saline and artificial sweat solutions on 4 identical CDs, ensuring reliable conductivity and measurements over a 1 kHz to 200 kHz frequency range. Results: Significant differences in |Z| values were observed between saline and artificial sweat treatments. 27.5 ilL of saline differed significantly from 27.5 ilL of artificial sweat, 72.5 ilL of saline from 72.5 ilL of artificial sweat, and 192.5 ilL of saline from 192.5 ilL of sweat. Significant disparities in |Z| values were observed between dry fibers and Groups 2, 3, and 4 (varying saline amounts). No significant differences emerged between dry fibers and Groups 6, 7, and 8 (distinct artificial sweat amounts). These findings underscore variations in fiber characteristics between equivalent exposures, emphasizing the nuanced response of the microfluidic CD platform to different liquid compositions. Conclusions: This study shows the potential of integrating conductive threads in a microfluidic CD platform for sweat sensing. Challenges in volume control and thread coating degradation must be addressed for transformative biosensing devices in personalized healthcare.

Diagnosing oral diseases at an early stage may lead to better preventive treatments, thus reducing treatment burden and costs. This paper introduces a systematic design of a microfluidic compact disc (CD) consisting of six unique chambers that run simultaneously from sample loading, holding, mixing and analysis. In this study, the electrochemical property changes between real saliva and artificial saliva mixed with three different types of mouthwashes (i.e. chlorhexidine-, fluoride- and essential oil (Listerine)-based mouthwashes) were investigated using electrical impedance analysis. Given the diversity and complexity of patient’s salivary samples, we investigated the electrochemical impedance property of healthy real saliva mixed with different types of mouthwashes to understand the different electrochemical property which could be a foundation for diagnosis and monitoring of oral diseases. On the other hand, electrochemical impedance property of artificial saliva, a commonly used moisturizing agent and lubricant for the treatment of xerostomia or dry mouth syndrome was also studied. The findings indicate that artificial saliva and fluoride-based mouthwash showed higher conductance values compared to real saliva and two other different types of mouthwashes. The ability of our new microfluidic CD platform to perform multiplex processes and detection of electrochemical property of different types of saliva and mouthwashes is a fundamental concept for future research on salivary theranostics using point-of-care microfluidic CD platform.

Intraoral theranostics, the integration of diagnostics and therapeutics within the oral cavity, is gaining significant traction. This pioneering approach primarily addresses issues like xerostomia (dry mouth), commonly resulting from cancer treatment, with a specific focus on monitoring temperature and humidity. This paper introduces the innovative Intra-Oral Portable Micro-Electronic (IOPM) fluidic theranostic device platform. It leverages conventional dental spoons by incorporating advanced sensors for precise measurements of oral temperature and humidity. Personalization options include a microfluidic chip and a tooth model, enabling targeted delivery of therapeutic agents to optimize treatment outcomes. The electronic control system simplifies the administration of fluid dosages, intelligently adjusted based on real-time oral cavity temperature and humidity readings. Rigorous experimental evaluations validate the platform's precision in delivering fluid volumes at predefined intervals. This platform represents a transformative advancement for individuals contending with oral health challenges such as xerostomia (dry mouth). Furthermore, it has the potential to elevate oral healthcare standards by providing advanced diagnostics and tailored therapeutic solutions, benefiting both patients and dental professionals alike.

Supp. Figure 1 (Detection of doxorubicin in the exosomes by HPLC and visualization of vesicular Transport); Supp. Figure 2 (DLBCL in ovo xenotransplantation model); Supp. Figure 3 (Increased cytotoxicity pixantrone and doxorubicin against DLBCL-ABC cell line OCI-Ly3 by pretreatment with indomethacin); Supp. Figure 4 (Celecoxib and omeprazole do not alter doxorubicin-susceptibility in SU-DHL-4 and OCI-Ly1 cells).

Tumor angiogenesis is one essential aspect for the growth and metastasis of cancer cells, which means that adequate in vivo angiogenesis models are of utmost importance for the investigation of such diseases. The chick chorioallantoic membrane (CAM) model is one established method for this purpose and has already been used for research on multiple cancer types. One important part of the evaluation of tumors grafted onto the CAM is the measurement of tumor-induced angiogenesis. In order to address this central aspect, we utilized the novel PeriCam perfusion speckle imager (PSI) system high resolution (HR) model (Perimed AB, Järfälla, Sweden), which is based on laser speckle contrast imaging (LSCI) for the semiquantitative measurement of blood flow in the CAM model. This method enables a fast and accurate analysis of the angiogenesis of cell line tumors and primary tumors that are grafted onto the CAM. The proposed model can be regarded as a precursor model for personalized cancer therapy.