Case report: Diagnostic challenges in an adolescent case of autistic catatonia.

Catatonia is a complex neuropsychiatric syndrome involving a constellation of psychomotor disturbances including catalepsy, waxy flexibility, stupor, mutism, negativism, agitation, posturing, stereotypes, mannerisms, grimacing, echolalia, and echopraxia. Catatonia occurs in several conditions including psychotic, affective and neurodevelopmental disorders such as autism spectrum disorder (ASD). ASD is a neurodevelopmental disorder characterized by persistent deficits in communication, social interaction, restricted interests, repetitive behaviours and sensory sensitivities. Catatonia can occur in response to life stressors such as extreme fear or threat, interpersonal conflict, tragic events or following significant loss. Those with ASD may be particularly vulnerable to the negative impact of stressors and the link between catatonia and ASD is being increasingly recognized. The overlapping features of catatonia and ASD make it difficult to differentiate often resulting in delayed or missed diagnosis. Catatonia in ASD remains a significant clinical challenge; it is difficult to diagnose and can pose debilitating difficulties for those affected. Catatonia is a treatable condition and prompt recognition is vital in securing the best possible outcome. We report a complex and unique case of a 15-year-old boy who presented with severe cognitive and functional decline with a background history of significant bullying and deterioration in his mental state. This case posed a diagnostic conundrum leading to a diagnosis of underlying ASD, anxiety and trauma.

Microengineering 3D Collagen Matrices with Tumor-Mimetic Gradients in Fiber Alignment.

Collagen fibers in the 3D tumor microenvironment (TME) exhibit complex alignment landscapes that are critical in directing cell migration through a process called contact guidance. Previous in vitro work studying this phenomenon has focused on quantifying cell responses in uniformly aligned environments. However, the TME also features short-range gradients in fiber alignment that result from cell-induced traction forces. Although the influence of graded biophysical taxis cues is well established, cell responses to physiological alignment gradients remain largely unexplored. In this work, fiber alignment gradients in biopsy samples are characterized and recreated using a new microfluidic biofabrication technique to achieve tunable sub-millimeter to millimeter scale gradients. This study represents the first successful engineering of continuous alignment gradients in soft, natural biomaterials. Migration experiments on graded alignment show that HUVECs exhibit increased directionality, persistence, and speed compared to uniform and unaligned fiber architectures. Similarly, patterned MDA-MB-231 aggregates exhibit biased migration toward increasing fiber alignment, suggesting a role for alignment gradients as a taxis cue. This user-friendly approach, requiring no specialized equipment, is anticipated to offer new insights into the biophysical cues that cells interpret as they traverse the extracellular matrix, with broad applicability in healthy and diseased tissue environments.

Prolonged HPA axis dysregulation in postpartum depression associated with adverse early life experiences: A cross-species translational study.

Childhood and adolescent stress increase the risk of postpartum depression (PPD), often providing an increased probability of treatment refractoriness. Nevertheless, the mechanisms linking childhood/adolescent stress to PPD remain unclear. Our study investigated the longitudinal effects of adolescent stress on the hypothalamic-pituitary-adrenal (HPA) axis and postpartum behaviors in mice and humans. Adolescent social isolation prolonged glucocorticoid elevation, leading to long-lasting postpartum behavioral changes in female mice. These changes were unresponsive to current PPD treatments but improved with post-delivery glucocorticoid receptor antagonist treatment. Childhood/adolescent stress significantly impacted HPA axis dysregulation and PPD in human females. Repurposing glucocorticoid receptor antagonists for some cases of treatment-resistant PPD may be considered.

Chemistry Aspects and Designing Strategies of Flexible Materials for High-Performance Flexible Lithium-Ion Batteries.

In recent years, flexible and wearable electronics such as smart cards, smart fabrics, bio-sensors, soft robotics, and internet-linked electronics have impacted our lives. In order to meet the requirements of more flexible and adaptable paradigm shifts, wearable products may need to be seamlessly integrated. A great deal of effort has been made in the last two decades to develop flexible lithium-ion batteries (FLIBs). The selection of suitable flexible materials is important for the development of flexible electrolytes self-supported and supported electrodes. This review is focused on the critical discussion of the factors that evaluate the flexibility of the materials and their potential path toward achieving the FLIBs. Following this analysis, we present how to evaluate the flexibility of the battery materials and FLIBs. We describe the chemistry of carbon-based materials, covalent-organic frameworks (COFs), metal-organic frameworks (MOFs), and MXene-based materials and their flexible cell design that represented excellent electrochemical performances during bending. Furthermore, the application of state-of-the-art solid polymer and solid electrolytes to accelerate the development of FLIBs is introduced. Analyzing the contributions and developments of different countries has also been highlighted in the past decade. In addition, the prospects and potential of flexible materials and their engineering are also discussed, providing the roadmap for further developments in this fast-evolving field of FLIB research.

Immunotherapy: cancer immunotherapy and its combination with nanomaterials and other therapies.

Immunotherapy is a new type of tumor treatment after surgery, radiotherapy and chemotherapy, and can be used to manage and destroy tumor cells through activating or strengthening the immune response. Immunotherapy has the benefits of a low recurrence rate and high specificity compared to traditional treatment methods. Immunotherapy has developed rapidly in recent years and has become a research hotspot. Currently, chimeric antigen receptor T-cell immunotherapy and immune checkpoint inhibitors are the most effective tumor immunotherapies in clinical practice. While tumor immunotherapy brings hope to patients, it also faces some challenges and still requires continuous research and progress. Combination therapy is the future direction of anti-tumor treatment. In this review, the main focus is on an overview of the research progress of immune checkpoint inhibitors, cellular therapies, tumor vaccines, small molecule inhibitors and oncolytic virotherapy in tumor treatment, as well as the combination of immunotherapy with other treatments.

Implementation of World Health Organization behaviorally anchored rating scale and checklist utilization: promising results for LMICs.

Background: Operating teams can decrease the likelihood of patient risk by using the WHO Surgical Safety Checklist. To ascertain the impact of demographic factors on behaviorally anchored ratings and investigate operating room (OR) staff attitudes toward checklist administration, we set out to better understand how OR personnel use the checklist in a tertiary care hospital in Pakistan. Materials and methods: A monocentric sequential mixed-methods study employing a quantitative approach of using World Health Organization Behaviorally Anchored Rating Scale (WHOBARS) assessments of surgical cases by OR personnel and two independent observers, who were certified surgeons having extensive experience in the rating of the WHOBARS scale for more than 1 year, followed by a qualitative approach of staff interviews were carried out in a tertiary care setting. In June and July 2022, over the period of 8 weeks, an intervention (training delivery) was implemented and evaluated. The information, skills, and behavior adjustments required to apply the checklist were taught in the course using lectures, videos, small group breakouts, participant feedback, and simulations. Results: After the introduction of WHOBARS, 50.81% of respondents reported always using the checklist, with another 30.81% using it in part. Participants' years in practice, hospital size, or surgical volume did not predict checklist use. Checklist use was associated with always counting instruments (51.08%), patient identity (67.83%), difficult intubation risk (39.72%), the risk of blood loss (51.08%), prophylactic administration of an antibiotic (52.43%), and the use of pulse oximeter (46.75%). Interviewees felt that the checklist could promote teamwork and a safe culture, particularly enabling speaking up. Senior staff were of key importance in setting the appropriate tone. Conclusion: The use of a multi-disciplinary course for checklist implementation resulted in 50.81% of participants always using the checklist and an increase in counting surgical instruments. Successful checklist implementation was not predicted by the participant's length of medical service, hospital size, or surgical volume. If reproducible in other countries, widespread implementation in LMICs becomes a realistic possibility.

Contemporaneous and upcoming trends in immunotherapy for prostate cancer: review.

Prostate cancer (PCa) is the most common cancer in men worldwide. It affects more than 1.4 million men worldwide and kills up to 37 5000 people. PCa is routinely managed with chemotherapy and androgen deprivation therapy, but the success rate of these treatments is unsatisfactory. Immunotherapy is a novel method of treating different types of cancers, and it utilizes the body's own immune system to fight cancer. Different types of cancer respond differently to immunotherapy, with some showing excellent responses, while others do not show very satisfactory responses. PCa is known to be an immunologically cold tumor, such that conventional immunotherapy does not work as effectively as it works in other cancers. In the past decade, multiple studies and trials have been conducted to test different types of therapies, ranging from immune checkpoint inhibitors to anticancer vaccines to anticancer cytokines. Even after many studies, there is still a drug to be discovered that can completely cure any stage of PCa. Recent immunotherapeutic drug trials have started using immunotherapy in conjunction with chemotherapy and radiotherapy and have shown promising results. In this paper, the authors present a comprehensive overview of the currently used immunotherapeutic drugs as well as emerging immunotherapies, including modalities of combination immunotherapy with radiotherapy and chemotherapy. This review can help readers gain the latest knowledge about emerging trends in the current immunotherapy landscape for the treatment of PCa, as well as a general overview of the already used immunotherapy drugs for PCa.

Microengineering 3D Collagen Matrices with Tumor-Mimetic Gradients in Fiber Alignment.

In the tumor microenvironment (TME), collagen fibers facilitate tumor cell migration through the extracellular matrix. Previous studies have focused on studying the responses of cells on uniformly aligned or randomly aligned collagen fibers. However, the in vivo environment also features spatial gradients in alignment, which arise from the local reorganization of the matrix architecture due to cell-induced traction forces. Although there has been extensive research on how cells respond to graded biophysical cues, such as stiffness, porosity, and ligand density, the cellular responses to physiological fiber alignment gradients have been largely unexplored. This is due, in part, to a lack of robust experimental techniques to create controlled alignment gradients in natural materials. In this study, we image tumor biopsy samples and characterize the alignment gradients present in the TME. To replicate physiological gradients, we introduce a first-of-its-kind biofabrication technique that utilizes a microfluidic channel with constricting and expanding geometry to engineer 3D collagen hydrogels with tunable fiber alignment gradients that range from sub-millimeter to millimeter length scales. Our modular approach allows easy access to the microengineered gradient gels, and we demonstrate that HUVECs migrate in response to the fiber architecture. We provide preliminary evidence suggesting that MDA-MB-231 cell aggregates, patterned onto a specific location on the alignment gradient, exhibit preferential migration towards increasing alignment. This finding suggests that alignment gradients could serve as an additional taxis cue in the ECM. Importantly, our study represents the first successful engineering of continuous gradients of fiber alignment in soft, natural materials. We anticipate that our user-friendly platform, which needs no specialized equipment, will offer new experimental capabilities to study the impact of fiber-based contact guidance on directed cell migration.

BBSF: Blockchain-Based Secure Weather Forecasting Information through Routing Protocol in Vanet.

A vehicular ad hoc network (VANET) is a technique that uses vehicles with the ability to sense data from the environment and use it for their safety measures. Flooding is a commonly used term used for sending network packets. VANET may cause redundancy, delay, collision, and the incorrect receipt of the messages to their destination. Weather information is one of the most important types of information used for network control and provides an enhanced version of the network simulation environments. The network traffic delay and packet losses are the main problems identified inside the network. In this research, we propose a routing protocol which can transmit the weather forecasting information on demand based on source vehicle to destination vehicles, with the minimum number of hop counts, and provide significant control over network performance parameters. We propose a BBSF-based routing approach. The proposed technique effectively enhances the routing information and provides the secure and reliable service delivery of the network performance. The results taken from the network are based on hop count, network latency, network overhead, and packet delivery ratio. The results effectively show that the proposed technique is reliable in reducing the network latency, and that the hop count is minimized when transferring the weather information.

Effective Strategies for Developing Potent, Broad-Spectrum Antibacterial and Wound Healing Promotion from Short-Chain Antimicrobial Peptides.

Traumatic multidrug resistant bacterial infections are the most lethal threat to wound healing. Antimicrobial peptides have been widely used in the antimicrobial field for their good biocompatibility and resistance to multidrug-resistant bacteria. In this work, bacterial membranes of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were extracted and immobilized on homemade silica microspheres to make a bacterial membrane chromatography stationary phase in order to quickly screen for peptides with antibacterial effects. The antimicrobial peptide was then successfully screened using bacterial membrane chromatography from a library of peptides synthesized by the one-bead-one-compound method. The antimicrobial peptide was effective in better shielding both Gram-positive and Gram-negative bacteria. Based on this antimicrobial peptide (RWPIL), we have developed an antimicrobial hydrogel with a backbone of this antimicrobial peptide and oxidized dextran (ODEX). Owing to the interlinkage between the aldehyde group in oxidized dextran and the amine group from the trauma tissue, the hydrogel extends over the irregular obverse of the skin defect and promotes epithelial cell adhesion. Based on the histomorphological analysis, we confirmed that the RWPIL-ODEX hydrogel exerts a powerful therapeutic effect in a wound infection model. In conclusion, we have developed a new antimicrobial peptide, RWPIL, and a hydrogel based on the peptide that kills multidrug-resistant bacteria parasitic on wounds and promotes wound healing.

The lymphatic endothelium-derived follistatin: activin A axis regulates neutrophil motility in response to Pseudomonas aeruginosa.

The lymphatic system plays an active role during infection, however the role of lymphatic-neutrophil interactions in host-defense responses is not well understood. During infection with pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus and Yersinia pestis, neutrophils traffic from sites of infection through the lymphatic vasculature, to draining lymph nodes to interact with resident lymphocytes. This process is poorly understood, in part, due to the lack of in vitro models of the lymphatic system. Here we use a 3D microscale lymphatic vessel model to examine neutrophil-lymphatic cell interactions during host defense responses to pathogens. In previous work, we have shown that follistatin is secreted at high concentrations by lymphatic endothelial cells during inflammation. Follistatin inhibits activin A, a member of the TGF-ß superfamily, and, together, these molecules form a signaling pathway that plays a role in regulating both innate and adaptive immune responses. Although follistatin and activin A are constitutively produced in the pituitary, gonads and skin, their major source in the serum and their effects on neutrophils are poorly understood. Here we report a microfluidic model that includes both blood and lymphatic endothelial vessels, and neutrophils to investigate neutrophil-lymphatic trafficking during infection with P. aeruginosa. We found that lymphatic endothelial cells produce secreted factors that increase neutrophil migration toward P. aeruginosa, and are a significant source of both follistatin and activin A during Pseudomonas infection. We determined that follistatin produced by lymphatic endothelial cells inhibits activin A, resulting in increased neutrophil migration. These data suggest that the follistatin:activin A ratio influences neutrophil trafficking during infection with higher ratios increasing neutrophil migration.

Mortality Rates and autopsy findings in fat embolism syndrome complicating sickle cell disease.

Fat embolism syndrome is a rare but underdiagnosed complication of sickle cell disease associated with high morbidity and mortality. It affects predominantly patients with a previously mild course of their illness and those of non-SS genotypes while there is possibly an association with infection with human parvovirus B19 (HPV B19). Here, we present the mortality rates and autopsy findings of all reported cases to date. A systematic review has revealed 99 published cases in the world literature with a mortality rate of 46%. Mortality varied greatly according to the time of reported cases with no survivors in the 1940s, 1950s or 1960s and no deaths since 2020. 35% of cases had previously undiagnosed sickle cell disease and the latter was only identified at autopsy after developing fat embolism with a fatal outcome. 20% of cases reported after 1986 tested positive for HPV B19 with an associated mortality of 63% whereas in cases that have not documented HPV B19 infection the mortality was 32%. The organs most often staining positive for fat were the kidneys, lungs, brain and heart whereas ectopic haematopoietic tissue was found in 45% of the examined lung specimens.

IoT-fog-based healthcare 4.0 system using blockchain technology.

Real-time tracking and surveillance of patients' health has become ubiquitous in the healthcare sector as a result of the development of fog, cloud computing, and Internet of Things (IoT) technologies. Medical IoT (MIoT) equipment often transfers health data to a pharmaceutical data center, where it is saved, evaluated, and made available to relevant stakeholders or users. Fog layers have been utilized to increase the scalability and flexibility of IoT-based healthcare services, by providing quick response times and low latency. Our proposed solution focuses on an electronic healthcare system that manages both critical and non-critical patients simultaneously. Fog layer is distributed into two halves: critical fog cluster and non-critical fog cluster. Critical patients are handled at critical fog clusters for quick response, while non-critical patients are handled using blockchain technology at non-critical fog cluster, which protects the privacy of patient health records. The suggested solution requires little modification to the current IoT ecosystem while decrease the response time for critical messages and offloading the cloud infrastructure. Reduced storage requirements for cloud data centers benefit users in addition to saving money on construction and operating expenses. In addition, we examined the proposed work for recall, accuracy, precision, and F-score. The results show that the suggested approach is successful in protecting privacy while retaining standard network settings. Moreover, suggested system and benchmark are evaluated in terms of system response time, drop rate, throughput, fog, and cloud utilization. Evaluated results clearly indicate the performance of proposed system is better than benchmark.

Efficient photocatalytic degradation of Rhodamine B dye using solar light-driven La-Mn co-doped Fe2O3 nanoparticles.

This work aims to develop a highly efficient solar light-induced photocatalyst based on La-Mn co-doped Fe2O3 nanoparticles. Pure Fe2O3 and La-Mn co-doped Fe2O3 nanoparticles were fabricated by a simple co-precipitation method. The photocatalysts were analyzed for their morphological, structural, and magnetic characteristics. Scanning electron microscopy analysis demonstrated the formation of semi-spherical nanoparticles along with small aggregations. The size of nanoparticles was measured using a transmission electron microscope and found in the range of 42-49 nm. The crystalline nature and geometry of synthesized nanoparticles were investigated using X-ray diffraction analysis. Due to the incorporation of La-Mn, the saturation magnetization and remanent magnetization of the nanoparticles decreased from 6.17 to 2.89 emu/g and 1.15 to 0.52 emu/g, respectively, while the coercivity was reduced from 756.72 to 756.67 Oe. The surface area of nanoparticles was increased from 77.93 to 87.45 m2/g as a result of La-Mn co-doping. The photocatalytic performance of the Fe2O3, La0.1Mn0.3Fe1.6O3, and La0.2Mn0.2Fe1.6O3 catalysts was assessed by their capability to degrade Rhodamine B (RhB) under solar light illumination. La0.2Mn0.2Fe1.6O3 displayed exceptional degradation performance, degrading RhB to 91.78% in 240 min, in comparison to La0.1Mn0.3Fe1.6O3 (71.09%) and pristine Fe2O3 (58.21%) under specified reaction conditions ((RhB) = 50 ppm; (catalyst) = 40 mg/L; pH = 7; T = 25 °C)). RhB degradation was affected by changing pH, catalytic dosage, dye concentration, and temperature. The degradation of RhB was found to be pseudo-1st order kinetics. The exceptional photocatalytic performance of La0.2Mn0.2Fe1.6O3 catalysts showed that the synthesized nanoparticles could be effectively utilized to remove organic pollutants from industrial wastewater.

Yolk-shell Co3 O4 @Fe3 O4 /C Nanocomposites as a Heterogeneous Fenton-like Catalyst for Organic Dye Removal.

The yolk-shell Co3 O4 @Fe3 O4 /C nanocomposites with Co3 O4 as the core, Fe3 O4 /C as the shell, and a cavity structure were synthesized by the hard template method. The physical and chemical properties of the composites were characterized by SEM, TEM, XRD, TGA, XPS, BET, and VSM. The specific surface area of yolk-shell Co3 O4 @Fe3 O4 /C nanocomposites is 175.9 m2  g-1 , showing superparamagnetic properties. The yolk-shell Co3 O4 @Fe3 O4 /C nanocomposites were used as heterogeneous Fenton catalysts to activate peroxymonosulfate (PMS) to degrade MB, which showed high catalytic degradation performance. The degradation rate of MB reached 100 % within 30 min under the circumstances of the yolk-shell Co3 O4 @Fe3 O4 /C nanocomposites dosage of 0.1 g L-1 , the PMS dosage of 1.0 g L-1 , the initial MB concentration of 100 mg L-1 , an initial pH of 5.5, and a temperature of 30±2 °C. The enhanced catalytic performance of the yolk-shell Co3 O4 @Fe3 O4 /C nanocomposites can be attributed to the synergistic effect of the two catalytically active materials and the middle cavity. The effects of different operating parameters and co-existing anion species on MB degradation were also investigated. Electron paramagnetic resonance (EPR) analysis and quenching experiments confirmed that the formation of SO4 ⋅- in the yolk-shell Co3 O4 @Fe3 O4 /C/PMS system contributes to MB degradation. In addition, yolk-shell Co3 O4 @Fe3 O4 /C nanocomposites can be easily separated from the pollutant solution under the action of an external magnetic field, and the degradation rate of MB can still reach 98 % after five cycles, indicating that it has good stability and reusability and has broad application prospects in the field of water purification.

Minocycline Ameliorates Chronic Unpredictable Mild Stress-Induced Neuroinflammation and Abnormal mPFC-HIPP Oscillations in Mice.

Stress-induced neuroinflammation is a hallmark of modern society and has been linked to various emotional disorders, including anxiety. However, how microglia-associated neuroinflammation under chronic unpredictable mild stress (CUMS) alters mitochondrial function and subsequent medial prefrontal cortex-hippocampus (mPFC-HIPP) connectivity remains obscure. We speculated that CUMS might induce neuroinflammation, which involves altered mitochondrial protein levels, blockade of neuroinflammation by a microglial modulator, minocycline, protects against CUMS-induced alterations. Mice were exposed to CUMS for 3 weeks and received minocycline (50 mg/kg) intraperitoneally for 7 consecutive days during the 3rd week of CUMS. Novelty-suppressed feeding test and contextual anxiety test assessed anxiety-like behavior. Western blotting and immunofluorescent staining were employed to evaluate levels of proteins involved in neuroinflammation and mitochondrial function. In vivo dual-site extracellular recordings of local field potential (LFP) were conducted to evaluate the oscillatory activity and brain connectivity in mPFC-HIPP circuitry. We show that CUMS results in excessive microglial activation accompanied by aberrant levels of mitochondrial proteins, such as ATP-5A and the fission protein, Drp-1, increased oxidative stress indicated by elevated levels of nitrotyrosine, and decreased Nrf-2 levels. Furthermore, CUMS causes downregulation of α1 subunit of GABAAR, vesicular GABA transporter (Vgat), and glutamine synthetase (GS), leading to impaired LFP and connectivity of the mPFC-HIPP circuitry. Strikingly, blockage of microglial activation by minocycline ameliorates CUMS-induced aberrant levels of mitochondrial and GABAergic signaling proteins and prevents CUMS-induced anxiety-like behavior in mice. To the end, the study revealed that microglia is critically involved in stress-induced neuroinflammation, which may underlie the molecular mechanism of CUMS-induced anxiety behavior.

Microengineering 3D Collagen Hydrogels with Long-Range Fiber Alignment.

Aligned collagen I (COL1) fibers guide tumor cell motility, influence endothelial cell morphology, control stem cell differentiation, and are a hallmark of cardiac and musculoskeletal tissues. To study cell response to aligned microenvironments in vitro, several protocols have been developed to generate COL1 matrices with defined fiber alignment, including magnetic, mechanical, cell-based, and microfluidic methods. Of these, microfluidic approaches offer advanced capabilities such as accurate control over fluid flows and the cellular microenvironment. However, the microfluidic approaches to generate aligned COL1 matrices for advanced in vitro culture platforms have been limited to thin "mats" (<40 µm in thickness) of COL1 fibers that extend over distances less than 500 µm and are not conducive to 3D cell culture applications. Here, we present a protocol to fabricate 3D COL1 matrices (130-250 µm in thickness) with millimeter-scale regions of defined fiber alignment in a microfluidic device. This platform provides advanced cell culture capabilities to model structured tissue microenvironments by providing direct access to the micro-engineered matrix for cell culture.

The Modular µSiM Reconfigured: Integration of Microfluidic Capabilities to Study In Vitro Barrier Tissue Models under Flow.

Microfluidic tissue barrier models have emerged to address the lack of physiological fluid flow in conventional "open-well" Transwell-like devices. However, microfluidic techniques have not achieved widespread usage in bioscience laboratories because they are not fully compatible with traditional experimental protocols. To advance barrier tissue research, there is a need for a platform that combines the key advantages of both conventional open-well and microfluidic systems. Here, a plug-and-play flow module is developed to introduce on-demand microfluidic flow capabilities to an open-well device that features a nanoporous membrane and live-cell imaging capabilities. The magnetic latching assembly of this design enables bi-directional reconfiguration and allows users to conduct an experiment in an open-well format with established protocols and then add or remove microfluidic capabilities as desired. This work also provides an experimentally-validated flow model to select flow conditions based on the experimental needs. As a proof-of-concept, flow-induced alignment of endothelial cells and the expression of shear-sensitive gene targets are demonstrated, and the different phases of neutrophil transmigration across a chemically stimulated endothelial monolayer under flow conditions are visualized. With these experimental capabilities, it is anticipated that both engineering and bioscience laboratories will adopt this reconfigurable design due to the compatibility with standard open-well protocols.

A miniaturized 3D printed pressure regulator (µPR) for microfluidic cell culture applications.

Well-defined fluid flows are the hallmark feature of microfluidic culture systems and enable precise control over biophysical and biochemical cues at the cellular scale. Microfluidic flow control is generally achieved using displacement-based (e.g., syringe or peristaltic pumps) or pressure-controlled techniques that provide numerous perfusion options, including constant, ramped, and pulsed flows. However, it can be challenging to integrate these large form-factor devices and accompanying peripherals into incubators or other confined environments. In addition, microfluidic culture studies are primarily carried out under constant perfusion conditions and more complex flow capabilities are often unused. Thus, there is a need for a simplified flow control platform that provides standard perfusion capabilities and can be easily integrated into incubated environments. To this end, we introduce a tunable, 3D printed micro pressure regulator (µPR) and show that it can provide robust flow control capabilities when combined with a battery-powered miniature air pump to support microfluidic applications. We detail the design and fabrication of the µPR and: (i) demonstrate a tunable outlet pressure range relevant for microfluidic applications (1-10 kPa), (ii) highlight dynamic control capabilities in a microfluidic network, (iii) and maintain human umbilical vein endothelial cells (HUVECs) in a multi-compartment culture device under continuous perfusion conditions. We anticipate that our 3D printed fabrication approach and open-access designs will enable customized µPRs that can support a broad range of microfluidic applications.

Local extensional flows promote long-range fiber alignment in 3D collagen hydrogels.

Randomly oriented type I collagen (COL1) fibers in the extracellular matrix are reorganized by biophysical forces into aligned domains extending several millimeters and with varying degrees of fiber alignment. These aligned fibers can transmit traction forces, guide tumor cell migration, facilitate angiogenesis, and influence tissue morphogenesis. To create aligned COL1 domains in microfluidic cell culture models, shear flows have been used to align thin COL1 matrices (<50µm in height) in a microchannel. However, there has been limited investigation into the role of shear flows in aligning 3D hydrogels (>130µm). Here, we show that pure shear flows do not induce fiber alignment in 3D atelo COL1 hydrogels, but the simple addition of local extensional flow promotes alignment that is maintained across several millimeters, with a degree of alignment directly related to the extensional strain rate. We further advance experimental capabilities by addressing the practical challenge of accessing a 3D hydrogel formed within a microchannel by introducing a magnetically coupled modular platform that can be released to expose the microengineered hydrogel. We demonstrate the platform's capability to pattern cells and fabricate multi-layered COL1 matrices using layer-by-layer fabrication and specialized modules. Our approach provides an easy-to-use fabrication method to achieve advanced hydrogel microengineering capabilities that combine fiber alignment with biofabrication capabilities.