Laminin and hyaluronan supplementation of collagen hydrogels enhances endothelial function and tight junction expression on three-dimensional cylindrical microvessel-on-a-chip.

Vasculature-on-chip (VoC) models have become a prominent tool in the study of microvasculature functions because of their cost-effective and ethical production process. These models typically use a hydrogel in which the three-dimensional (3D) microvascular structure is embedded. Thus, VoCs are directly impacted by the physical and chemical cues of the supporting hydrogel. Endothelial cell (EC) response in VoCs is critical, especially in organ-specific vasculature models, in which ECs exhibit specific traits and behaviors that vary between organs. Many studies customize the stimuli ECs perceive in different ways; however, customizing the hydrogel composition accordingly to the target organ's extracellular matrix (ECM), which we believe has great potential, has been rarely investigated. We explored this approach to organ-specific VoCs by fabricating microvessels (MVs) with either human umbilical vein ECs or human brain microvascular ECs in a 3D cylindrical VoC using a collagen hydrogel alone or one supplemented with laminin and hyaluronan, components found in the brain ECM. We characterized the physical properties of these hydrogels and analyzed the barrier properties of the MVs. Barrier function and tight junction (ZO-1) expression improved with the addition of laminin and hyaluronan in the composite hydrogel.

Protocol for fabricating and characterizing microvessel-on-a-chip for human umbilical vein endothelial cells.

Organ-on-a-chip technologies enable the fabrication of endothelial tissues, so-called microvessels (MVs), which emulate the endothelial barrier function in healthy or disease conditions. In this protocol, we describe the fabrication of perfusable open-chamber style MVs embedded in collagen gels. We then report a simple technology to characterize the MV barrier properties in static or under pressure based on fluorescence confocal imaging. Finally, we provide quantification techniques that enable us to infer the structure of MV paracellular pores. For complete details on the use and execution of this protocol, please refer to Cacheux et al.1.

Standalone cell culture microfluidic device-based microphysiological system for automated cell observation and application in nephrotoxicity tests.

Microphysiological systems (MPS) offer an alternative method for culturing cells on microfluidic platforms to model organ functions in pharmaceutical and medical sciences. Although MPS hardware has been proposed to maintain physiological organ function through perfusion culture, no existing MPS can automatically assess cell morphology and conditions online to observe cellular dynamics in detail. Thus, with this study, we aimed to establish a practical strategy for automating cell observation and improving cell evaluation functions with low temporal resolution and throughput in MPS experiments. We developed a versatile standalone cell culture microfluidic device (SCCMD) that integrates microfluidic chips and their peripherals. This device is compliant with the ANSI/SLAS standards and has been seamlessly integrated into an existing automatic cell imaging system. This integration enables automatic cell observation with high temporal resolution in MPS experiments. Perfusion culture of human kidney proximal tubule epithelial cells using the SCCMD improves cell function. By combining the proximal tubule MPS with an existing cell imaging system, nephrotoxicity studies were successfully performed to automate morphological and material permeability evaluation. We believe that the concept of building the ANSI/SLAS-compliant-sized MPS device proposed herein and integrating it into an existing automatic cell imaging system for the online measurement of detailed cell dynamics information and improvement of throughput by automating observation operations is a novel potential research direction for MPS research.

The Constituent Phases and Micromechanical Properties of Steel Corrosion Layers Generated by Hyperbaric-Oxygen Accelerated Corrosion Test.

Hyperbaric oxygen-accelerated corrosion testing (HOACT) is a newly developed method to study in the labor the corrosion behavior of steel bars in concrete. This work aimed to intensively investigate the mechanical properties and microstructures of HOACT-generated corrosion products by means of nano-indentation tests, Raman micro-spectrometry, and scanning electron microscopy. The local elastic modulus and nanohardness varied over wide ranges of 6.8-75.2 GPa and 0.38-4.44 GPa, respectively. Goethite, lepidocrocite, maghemite, magnetite, and akageneite phases were identified in the corrosion products. Most regions of the rust layer were composed of a complex and heterogeneous mix of different phases, while some regions were composed of maghemite or akageneite only. The relationship between the micromechanical properties and typical microstructural features is finally discussed at the micro-scale level. It was found that the porosity of corrosion products can significantly influence their micromechanical properties.

Enhanced podocyte differentiation and changing drug toxicity sensitivity through pressure-controlled mechanical filtration stress on a glomerulus-on-a-chip.

Podocytes, localized in the glomerulus, are a prognostic factor of proteinuria in kidney disease and are exposed to distinct physiological stimuli from basal to apical filtration flow. Research studies on drug discovery and disease modeling for glomerulopathy have developed a glomerulus-on-a-chip and studied podocyte mechanobiology to realize alternative methods to animal experiments. However, the effect of filtration stimulus on podocytes has remained unclear. Herein, we report the successful development of a user-friendly filtration culture device and system that can precisely control the filtration flow using air pressure control by incorporating a commercially available culture insert. It allows mouse podocytes to be cultured under filtration conditions for three days with a guarantee of maintaining the integrity of the podocyte layer. Using our system, this study demonstrated that podocyte damage caused by hyperfiltration resulting from glomerular hypertension, a common pathophysiology of many glomerulopathies, was successfully recapitulated and that filtration stimulus promotes the maturation of podocytes in terms of their morphology and gene expression. Furthermore, we demonstrated that filtration stimulus induced different drug responsiveness in podocytes than those seen under static conditions, and that the difference in drug responsiveness was dependent on the pharmacological mechanism. Overall, this study has revealed differentiating and pharmacodynamic properties of filtration stimulus and brings new insights into the research field of podocyte mechanobiology towards the realization of glomerulus-on-a-chip.

Glomerulus-on-a-Chip: Current Insights and Future Potential Towards Recapitulating Selectively Permeable Filtration Systems.

Glomerulopathy, characterized by a dysfunctional glomerular capillary wall, results in proteinuria, leading to end-stage renal failure and poor clinical outcomes, including renal death and increased overall mortality. Conventional glomerulopathy research, including drug discovery, has mostly relied on animal experiments because in-vitro glomerulus models, capable of evaluating functional selective permeability, was unavailable in conventional in-vitro cell culture systems. However, animal experiments have limitations, including time- and cost-consuming, multi-organ effects, unstable reproducibility, inter-species reliability, and the social situation in the EU and US, where animal experiments have been discouraged. Glomerulus-on-a-chip, a new in-vitro organ model, has recently been developed in the field of organ-on-a-chip research based on microfluidic device technology. In the glomerulus-on-a-chip, the podocytes and endothelial cells are co-cultured in a microfluidic device with physical stimuli that mimic the physiological environment to enhance cell function to construct a functional filtration barrier, which can be assessed by permeability assays using fluorescently labeled molecules including inulin and albumin. A combination of this glomerulus-on-a chip technology with the culture technology to induce podocytes and endothelial cells from the human pluripotent stem cells could provide an alternative organ model and solve the issue of animal experiments. Additionally, previous experiments have verified the difference in the leakage of albumin using differentiated podocytes derived from patients with Alport syndrome, such that it could be applied to intractable hereditary glomerulopathy models. In this review, we provide an overview of the features of the existing glomerulus-on-a-chip systems, focusing on how they can address selective permeability verification tests, and the challenges they involved. We finally discuss the future approaches that should be developed for solving those challenges and allow further improvement of glomerulus-on-a-chip technologies.

[A Case of Resected Pancreatic Adenosquamous Carcinoma with Early Hepatic Metastatic Recurrence].

A 73-year-old man was presented with epigastric pain and indicated high CA19-9 levels, and computed tomography detected a tumor in the uncinate process of the pancreas infiltrated duodenum and superior mesenteric artery. The patient was diagnosed with borderline resectable pancreatic carcinoma and received neoadjuvant chemotherapy with gemcitabine and S-1. During neoadjuvant chemotherapy, the patient also received radiotherapy to control duodenal bleeding. After neoadjuvant chemotherapy, stable disease(SD)was proven on the Response Evaluation Criteria in Solid Tumors(RECIST), and subtotal stomach-preserving pancreaticoduodenectomy was performed. The pathological findings showed pancreatic adenosquamous carcinoma. After 7 days postoperatively, hepatic metastasis was detected, and after 78 days postoperatively, the patient died.

A novel method for successful induction of interdigitating process formation in conditionally immortalized podocytes from mice, rats, and humans.

Formation of processes in podocytes is regarded as the hallmark of maturity and normal physical condition for the cell. There are many accumulated findings about molecular mechanisms that cause retraction of podocyte processes; however, there is little knowledge of the positive mechanisms that promote process formation in vitro, and most previous reports about this topic have been limited to low-density cultures. Here, we found that process formation can be induced in 100% confluent cultures of conditionally immortalized podocytes in mouse, rat, and human species by combining serum depletion and Y-27632 ROCK inhibitor supplementation on the scaffold of laminin-521(L521). We noted the cytoskeletal reorganization of the radial extension pattern of vimentin filaments and downregulation of actin stress fiber formation under that condition. We also found that additional standard amount of serum, depletion of ROCK inhibitor, or slight mismatch of the scaffold as laminin-511(L511) hinder process formation. These findings suggest that the combination of reduced serum, podocyte-specific scaffold, and intracellular signaling to reduce the overexpression of ROCK are required factors for process formation.

In vivo degradation and bone formation behaviors of hydroxyapatite-coated Mg alloys in rat femur.

Various coatings have been developed for biodegradable Mg alloys to control the degradation speed and to improve the bone conductivity. In this study, hydroxyapatite (HAp) coatings were formed on pure Mg, Mg-0.8mass% Ca (MgCa), Mg-4mass% Y-3mass% rare earth (RE) (WE43), Mg-3mass% RE-1mass% Y (EW31) and Mg-4mass% RE (RE4) alloy rods with a chemical solution deposition method. The HAp-coated and uncoated Mg/Mg alloy rods were implanted in the femurs of rats for 3-6 months, and the corrosion suppression and bone formation abilities of the HAp coating were examined using a scanning electron microscope. The corrosion rate of WE43 was suppressed by 1/3 with the HAp coating for 6 months, and the corrosion product showed very slow dissolution. The effect of the HAp coating for pure Mg and MgCa disappeared in 1-2 months with the thinning of the rods accompanying with the obvious dissolution of the corrosion products. The effect of the HAp coating for EW31 and RE4 was not stable due to the expansion and collapse of the corrosion products. The bone formation was enhanced on the HAp layers. Eventually, the HAp coating basically suppressed the corrosion initiation and corrosion progress of Mg substrates. The magnitude of the suppression effect depended mainly on the chemical and physical stability of the corrosion products.

Pathologically diagnosed superficial form of placenta accreta: a comparative analysis with invasive form and asymptomatic muscular adhesion.

Pathologically diagnosed placenta accreta is defined as villi adjacent to the myometrium without decidua. It is classified into the superficial (placental accreta vera [PAV]) and deep invasive (placenta increta [PI] and placenta percreta [PP]) types. Data on the clinicopathological characteristics of PAV are limited. Basal plate myometrium (BPMYO) is found in PAV or placentas in asymptomatic women, but its significance is still controversial. This retrospective study aimed to determine the clinicopathological characteristics of pathologically diagnosed PAV and the significance of BPMYO. We reviewed 84 cases of pathologically diagnosed placenta accreta (PAV, 54; PI, 16; and PP, 14), and compared them with controls (i.e., not pathologically diagnosed of any type of placenta accreta, n = 51). Among the PAV cases, the incidence of in vitro fertilization was high, while that of previous cesarean section or placenta previa was low. The incidence of maternal complications was also high in pathologically diagnosed PAV cases, but some PAV were asymptomatic. The rate of prenatal diagnosis of PAV was low, and a high proportion of patients required emergency transportation to central hospitals. Histologically, BPMYO was found in 7 (14%) of controls and 54 (100%) of PAV cases. PAV cases had a higher rate of advanced stages of BPMYO, larger muscle tissue, and more foci than controls. In conclusion, almost PAV is a clinically symptomatic condition but has distinct risk factors and clinical findings from advanced type placenta accreta. Histological evaluation of BPMYO is useful for the diagnosis of PAV.

Microbubble flows in superwettable fluidic channels.

The control of bubble adhesion underwater is important for various applications, yet the dynamics under flow conditions are still to be unraveled. Herein, we observed the wetting dynamics of an underwater microbubble stream in superwettable channels. The flow of microbubbles was generated by integrating a microfluidic device with an electrochemical system. The microbubble motions were visualized via tracing the flow using a high-speed camera. We show that a vortex is generated in the air layer of the superaerophilic surface under laminar conditions and that the microbubbles are transported on the superaerophilic surface under turbulent conditions driven by the dynamic motion of the air film. Furthermore, microbubbles oscillated backward and forward on the superaerophobic surface under turbulent conditions. This investigation contributes to our understanding of the principles of drag reduction through wettability control and bubble flow.

Degradation of Ti-6Al-4V alloy under cyclic loading in a simulated body environment with cell culturing.

The present study reports the corrosion fatigue of the Ti-6Al-4V alloy using cyclic deformation test in a simulated body fluid under cell culturing for the first time. Cyclic deformation tests were carried out using three types of specimens to reveal the effects of proteins and cells on the corrosion fatigue of the alloy. For the 1-day-immersed and 1-week-immersed specimens, tensile specimens were soaked in a simulated body fluid for 1 day and 1 week, respectively, before cyclic deformation test, whereas for the cell-cultured specimen, MC3T3-E1 osteoblast-like cells were seeded and then cultured on tensile specimens for 1 week. The incubation period for crack initiation was longer for the cell-cultured and 1-week-immersed specimens compared to that for the 1-day-immersed specimen. On the other hand, crack propagation period for the cell-cultured and 1-week-immersed specimens was shorter than that for the 1-day-immersed specimen. These results indicate that proteins and cells adhered on the alloy surface inhibit metal dissolution at newly created surface emerged by cyclic deformation to suppress crack initiation, whereas they accelerate crack propagation because dissolution at crack tip is accelerated in the occluded space formed under proteins and cells.