Dynamic microphysiological system chip platform for high-throughput, customizable, and multi-dimensional drug screening

Drugs may undergo costly preclinical studies but still fail to demonstrate their efficacy in clinical trials, which makes it challenging to discover new drugs. Both in vitro and in vivo models are essential for disease research and therapeutic development. However, these models cannot simulate the physiological and pathological environment in the human body, resulting in limited drug detection and inaccurate disease modelling, failing to provide valid guidance for clinical application. Organs-on-chips (OCs) are devices that serve as a micro-physiological system or a tissue-on-a-chip; they provide accurate insights into certain functions and the pathophysiology of organs to precisely predict the safety and efficiency of drugs in the body. OCs are faster, more economical, and more precise. Thus, they are projected to become a crucial addition to, and a long-term replacement for, traditional preclinical cell cultures, animal studies, and even human clinical trials. This paper first outlines the nature of OCs and their significance, and then details their manufacturing-related materials and methodology. It also discusses applications of OCs in drug screening and disease modelling and treatment, and presents the future perspective of OCs.

A microphysiological system (MPS) holds great promise for drug screening and toxicological testing as an alternative to animal models. However, this platform faces several challenges in terms of the materials used (e.g. polydimethylsiloxane; PDMS). For instance, absorption of drug candidates and fluorescent dyes into PDMS, as well as the effect elicited by materials on cultured cells, can cause inaccurate or misleading results in cell assays. The use of PDMS also poses challenges for mass production and long-term storage of fabricated MPSs. Hence, to circumvent these issues, herein we describe the development of a cyclo olefin polymer (COP)-based MPS using photobonding processes and vacuum ultraviolet (VUV), designated as COP-VUV-MPS. COP is an amorphous polymer with chemical/physical stability, high purity and optical clarity. Due to the thermostability and high modulus of COP, the metal molding processes was applied for mass production of MPSs without deformation of microstructures and with quick fabrication cycle time (approx. 10 min/cycle). Moreover, VUV photobonding process with an excimer light at a 172nm wavelength allowed assembling COP materials without the use of additional solvents and tapes, which might cause cell damages. In comparison with the conventional MPS made of PDMS (PDMS-MPS), COP-VUV-MPS showed improved chemical resistance without causing molecule absorption. Moreover, COP-VUV-MPS maintained the stemness of environmentally sensitive human-induced pluripotent stem cells without causing undesired cellular phenotypes or gene expression. These results suggest that COP-VUV-MPS may be broadly applicable for the advancement of MPS and applications in drug development, as well as in vitro toxicological testing.

In recent years, many researchers have proposed biohybrid systems that integrate biological materials, especially cells or cultured tissues formed by in vitro cell culture, and microdevices to utilize the functions of cultured cellular tissues in an engineering way. In this paper, we focus on microphysiological systems composed of microfluidic channels or microdevices and cells or cultured tissues and introduce the functional characteristics of these microphysiological systems. In the microphysiological system, we can adjust cell culture conditions under mechanical stress control and bring out the dynamic performance of the cultured tissue. The systems can be applied not only to drug development to replace laboratory animals, but also to the development of advanced robots with cellular functions. Therefore, we believe that the biohybrid systems are a promising technology to apply cells and cultured tissues in various fields.

Using organoids and organ-on-a-chip (OoC) technologies as possible substitutes To understand the effects of fluidic shear stress on the primary inner medullary collecting duct (IMCD) cells from rats, Jang et al. developed a multi-layer microfluidic device consisting of two PDMS microfluidic channel layers and a polyester porous membrane with pore size of 400 nm. This allowed them to create a 3D in vivo cell culture model for the analysis of the renal tubule cells (Fig. 3(a)). 66 The extracellular matrix protein-coated porous membrane separated the device into two flow channels: an interstitial flow channel and a luminal flow channel. for animal testing in drug development, this study examined the changing field of Micro physiological Systems (MPS). Because of their special qualities, which include their high surface-to-volume ratio, nanoscale size, and potential for functionalization, nanoparticles (NPs) hold promise for a range of biological uses, such as targeted therapy, drug transport, and diagnostics. We go over the benefits and drawbacks of these platforms while highlighting their effective use in supporting drug development. We also go over how TCMs can be studied using these organotypic models. Lastly, we go at the prospects for integrated microphysiology. This paper looks at how AI is changing preclinical research, stressing its developments, difficulties, and crucial actions required to make AI a pillar of moral and effective medication developmentiscovery. Organs-onIn addition to many other organ systems in development, chips have been created for the liver, kidneys, gut, lungs, blood–brain barrier, blood vessels, skin, heart, muscles, lymphatics, eyes, and bone marrow.2. These models include prototypes, commercial products, and early proof-of-concepts in university labs. Nevertheless, additional development is required in order for these networks of interconnected organs-on-chips to replicate the various organ interactions and communication in order to simulate a "human-on-a-chip." International OoC programs and the availability of stem cells have led to the success of organ-on-chip (OoC) technology. To effectively advance OoC technology and realize its potential in drug research, disease modeling, and personalized medicine, a concerted effort to standardize, qualify, and independently test devices is required. Moving ahead can be guided by the OoC roadmap. With an emphasis on(1) their capacity to mimic the activities of human and animal organs, (2) applications in food safety and ecotoxicology, and (3) their usage in researching food components, this systematic review explores the promise of OoC technologies within the One Health paradigm and nutrition research. Organoids and organ-on-a-chip technologies are examples of Microphysiological Systems (MPS), which present viable substitutes for animal testing in drug development due to ethical, legal, and scientific reasons. Organoids and organ-on-a-chip technologies are examples of Microphysiological Systems (MPS), which present viable substitutes for animal testing in drug development due to ethical, legal, and scientific reasons.

Microphysiological systems (MPS) are drawing increasing interest from academia and from biomedical industry due to their improved capability to capture human physiology. MPS offer an advanced in vitro platform that can be used to study human organ and tissue level functions in health and in diseased states more accurately than traditional single cell cultures or even animal models. Key features in MPS include microenvironmental control and monitoring as well as high biological complexity of the target tissue. To reach these qualities, cross-disciplinary collaboration from multiple fields of science is required to build MPS. Here, we review different areas of expertise and describe essential building blocks of heart MPS including relevant cardiac cell types, supporting matrix, mechanical stimulation, functional measurements, and computational modelling. The review presents current methods in cardiac MPS and provides insights for future MPS development with improved recapitulation of human physiology.

To increase success rates of clinical studies, preclinical evaluation systems have been expected to improve human predictability. In addition, future preclinical studies need to become more sophisticated and efficient on the back ground of the adoption of FDA Modernization Act 2.0 and the 3R principle promotion of animal tests. In this review, we will discuss about the efficiency of in vivo imaging in preclinical studies taking 'an attempt to establishment of in vitro in vivo extraporation (IVIVE) model for seizure risk assessment using microphysiological system (MPS) and magnetic resonance imaging (MRI)', and 'an attempt to predict drug delivery to the alveoli' as examples. In the seizure risk assessment of new drugs so far, primary cultures of rodent neurons and in vivo behavioral observation have been mainly used, however, since the human induced pluripotent stem cell (iPSC) technology was reported, the need for IVIVE model is more and more increasing to improve human predictability. As an MPS, we here introduce microelectrode array (MEA) system recording of primary culture of rodent neurons, while as in vivo experiments, we here introduce the measurement of cerebrospinal fluid (CSF) concentrations and MRI imaging of forebrains of the rats i.p. injected with seizurogenic compounds. In case of inhalation drugs, it has been difficult to confirm whether or not the drugs surely reach alveoli. We visualized two-dimensional spatial localization of inhaled ciclesonide (CIC) in rat lungs after administration of a single dose of a CIC aerosol using by desorption electrospray ionization-time of flight mass spectrometry imaging (DESI-MSI).

In vitro 3D tissue models within microfluidic-based microphysiological systems (MPS) provide controlled and reproducible platforms for quantification of isolated cellular processes in response to biochemical or biophysical stimulus. This study demonstrates the development of a 3D MPS with a dual-chamber, closed-capillary circuit microfluidic culture platform to study chemotherapy drug efficacy in vitro for aggressive malignancies such as breast cancer and glioblastoma. This novel microfluidic system was used to model HER2 + breast cancer (BCTM-SKBR3) co-cultured with cardiac (CTM-AC16) tissue for proof-of-concept chemotherapy-induced cardiotoxicity studies. To further demonstrate the versatility of this system, a glioblastoma tissue model with chemotherapy efficacy studies was included. Additionally, implementation of a Python-based automated image analysis script (AIAPS) facilitated quantification of cell size within the tissue models from 3D fluorescence z-stack images. The results demonstrate maintenance of lineage-specific biomarker expression, physiologically relevant cell morphology and structural organization, and detectable changes in cell sizes with chemotherapy treatment within the 3D tissue models. These results demonstrated the system’s potential for use as a preclinical drug screening platform.

Innervated adrenomedullary microphysiological system to model nicotine and opioid exposure

Oral bioavailability is a crucial pharmacokinetic (PK) parameter optimized by drug developers working on new therapeutic compounds. The goal is to obtain adequate systemic exposure and to establish safe and effective therapeutic dosages. Critical to optimizing oral bioavailability is understanding a compound’s susceptibility to first-pass metabolism, determined by the intricate relationship between gut absorption, metabolism, and hepatic metabolism. Efforts to model this relationship in vitro have led to the emergence of microphysiological systems (MPSs) that consist of multiple, fluidically linked organs. Here, we describe an MPS that links the gut and liver, capable of simulating both oral and intravenous dosing routes and is made up of entirely primary human cells: a gut barrier tissue comprised of an intestinal epithelial monolayer derived from the human jejunum, and a liver microtissue, derived from primary human hepatocytes. Functionality of gut and liver tissues is maintained in coculture enabling the PK investigation of oral compounds. We combine the primary Gut/Liver MPS with a mechanistic mathematical model to generate organ-specific PK parameters and estimate human oral bioavailability and its components: the fraction absorbed (Fa), the fraction escaping gut wall elimination (Fg), and the fraction escaping hepatic elimination (Fh). We used the CYP3A-mediated compound midazolam, which is subjected to both intestinal and hepatic extraction, to demonstrate the transformative potential of the primary Gut/Liver MPS to mechanistically model the PKs of oral compounds in vitro.Significance StatementBioavailability underpins the success or when insufficient, can stall the development of oral therapeutics. The drug discovery process lacks in vitro assays that can profile the contribution to bioavailability by the gut and liver. The primary Gut/Liver MPS outlined in this study bridges this gap.

Microphysiological systems (MPSs) reconstitute tissue interfaces and organ functions, presenting a promising alternative to animal models in drug development. However, traditional materials like polydimethylsiloxane (PDMS) often interfere by absorbing hydrophobic molecules, affecting drug testing accuracy. Additive manufacturing, including 3D bioprinting, offers viable solutions. GlioFlow3D, a novel microfluidic platform combining extrusion bioprinting and stereolithography (SLA) is introduced. GlioFlow3D integrates primary human cells and glioblastoma (GBM) lines in hydrogel‐based microchannels mimicking vasculature, within an SLA resin framework using cost‐effective materials. The study introduces a robust protocol to mitigate SLA resin cytotoxicity. Compared to PDMS, GlioFlow3D demonstrated lower small molecule absorption, which is relevant for accurate testing of small molecules like Temozolomide (TMZ). Computational modeling is used to optimize a pumpless setup simulating interstitial fluid flow dynamics in tissues. Co‐culturing GBM with brain endothelial cells in GlioFlow3D showed enhanced CD133 expression and TMZ resistance near vascular interfaces, highlighting spatial drug resistance mechanisms. This PDMS‐free platform promises advanced drug testing, improving preclinical research and personalized therapy by elucidating complex GBM drug resistance mechanisms influenced by the tissue microenvironment.

Microphysiological systems (MPS), which include engineered organoids (EOs), single organ/tissue chips (TCs), and multiple organs interconnected to create miniature in vitro models of human physiological systems, are rapidly becoming effective tools for drug development and the mechanistic understanding of tissue physiology and pathophysiology. The second MPS thematic issue of Experimental Biology and Medicine comprises 15 articles by scientists and engineers from the National Institutes of Health, the IQ Consortium, the Food and Drug Administration, and Environmental Protection Agency, an MPS company, and academia. Topics include the progress, challenges, and future of organs-on-chips, dissemination of TCs into Pharma, children's health protection, liver zonation, liver chips and their coupling to interconnected systems, gastrointestinal MPS, maturation of immature cardiomyocytes in a heart-on-a-chip, coculture of multiple cell types in a human skin construct, use of synthetic hydrogels to create EOs that form neural tissue models, the blood-brain barrier-on-a-chip, MPS models of coupled female reproductive organs, coupling MPS devices to create a body-on-a-chip, and the use of a microformulator to recapitulate endocrine circadian rhythms. While MPS hardware has been relatively stable since the last MPS thematic issue, there have been significant advances in cell sourcing, with increased reliance on human-induced pluripotent stem cells, and in characterization of the genetic and functional cell state in MPS bioreactors. There is growing appreciation of the need to minimize perfusate-to-cell-volume ratios and respect physiological scaling of coupled TCs. Questions asked by drug developers are followed by an analysis of the potential value, costs, and needs of Pharma. Of highest value and lowest switching costs may be the development of MPS disease models to aid in the discovery of disease mechanisms; novel compounds including probes, leads, and clinical candidates; and mechanism of action of drug candidates. Impact statement Microphysiological systems (MPS), which include engineered organoids and both individual and coupled organs-on-chips and tissue chips, are a rapidly growing topic of research that addresses the known limitations of conventional cellular monoculture on flat plastic - a well-perfected set of techniques that produces reliable, statistically significant results that may not adequately represent human biology and disease. As reviewed in this article and the others in this thematic issue, MPS research has made notable progress in the past three years in both cell sourcing and characterization. As the field matures, currently identified challenges are being addressed, and new ones are being recognized. Building upon investments by the Defense Advanced Research Projects Agency, National Institutes of Health, Food and Drug Administration, Defense Threat Reduction Agency, and Environmental Protection Agency of more than $200 million since 2012 and sizable corporate spending, academic and commercial players in the MPS community are demonstrating their ability to meet the translational challenges required to apply MPS technologies to accelerate drug development and advance toxicology.

The lymphatic system—integral to fluid balance, immune surveillance, and lipid absorption—is frequently overlooked despite its vital roles. Traditional research modalities, including static two-dimensional cultures and animal models, have illuminated key molecular and cellular features but fall short in recapitulating human lymphatic function, due to limited physiological relevance, throughput, and mechanobiological complexity. Recent advances in microfluidic organ-on-a-chip systems offer biomimetic platforms that integrate three-dimensional architecture, fluid flow, and biomechanical stimuli alongside human lymphatic endothelial and supporting cells. These lymphatics-on-a-chip constructs faithfully reproduce dynamic behaviors such as fluid drainage, junction remodeling, and cell trafficking under physiological and pathological responses. This review highlights the foundational lymphatic biology and engineering principles behind these devices, their capacity for disease modeling and drug testing, and their potential to drive future innovation through induced pluripotent stem cell integration, organ-specific customization, and computational modeling. Merging bioengineering, cell biology, and machine learning, lymphatic microphysiological systems stand poised to significantly expand our understanding and treatment of lymphatic-related disorders.

Gastrointestinal diseases are a significant health care and economic burden. Prevention and treatment of these diseases have been limited by the available human biologic models. Microphysiological systems comprise organ-specific human cultures that recapitulate many structural, biological, and functional properties of the organ in smaller scale including aspects of flow, shear stress and chemical gradients. The development of intestinal microphysiological system platforms represents a critical component in improving our understanding, prevention, and treatment of gastrointestinal diseases. This minireview discusses: shortcomings of classical cell culture models of the gastrointestinal tract; human intestinal enteroids as a new model and their advantages compared to cell lines; why intestinal microphysiological systems are needed; potential functional uses of intestinal microphysiological systems in areas of drug development and modeling acute and chronic diseases; and current challenges in the development of intestinal microphysiological systems. Impact statement The development of a gastrointestinal MPS has the potential to facilitate the understanding of GI physiology. An ultimate goal is the integration of the intestinal MPS with other organ MPS. The development and characterization of nontransformed human intestinal cultures for use in MPS have progressed significantly since the inception of the MPS program in 2012, and these cultures are a key component of advancing MPS. Continued efforts are needed to optimize MPS to comprehensively and accurately recapitulate the complexity of the intestinal epithelium within intestinal tissue. These systems will need to include peristalsis, flow, and oxygen gradients, with incorporation of vascular, immune, and nerve cells. Regional cellular organization of crypt and villus areas will also be necessary to better model complete intestinal structure.

Liver microphysiological systems (MPSs) are promising models for predicting hepatic drug effects. Yet, after a decade since their introduction, MPSs are not routinely used in drug development due to lack of criteria for ensuring reproducibility of results. We characterized the feasibility of a liver MPS to yield reproducible outcomes of experiments assaying drug toxicity, metabolism, and intracellular accumulation. The ability of the liver MPS to reproduce hepatotoxic effects was assessed using trovafloxacin, which increased lactate dehydrogenase (LDH) release and reduced cytochrome P450 3A4 (CYP3A4) activity. These observations were made in two test sites and with different batches of Kupffer cells. Upon culturing equivalent hepatocytes in the MPS, spheroids, and sandwich cultures, differences between culture formats were detected in CYP3A4 activity and albumin production. Cells in all culture formats exhibited different sensitivities to hepatotoxicant exposure. Hepatocytes in the MPS were more functionally stable than those of other culture platforms, as CYP3A4 activity and albumin secretion remained prominent for greater than 18 days in culture, whereas functional decline occurred earlier in spheroids (12 days) and sandwich cultures (7 days). The MPS was also demonstrated to be suitable for metabolism studies, where CYP3A4 activity, troglitazone metabolites, diclofenac clearance, and intracellular accumulation of chloroquine were quantified. To ensure reproducibility between studies with the MPS, the combined use of LDH and CYP3A4 assays were implemented as quality control metrics. Overall results indicated that the liver MPS can be used reproducibly in general drug evaluation applications. Study outcomes led to general considerations and recommendations for using liver MPSs.Study HighlightsWHAT IS THE CURRENT KNOWLEDGE ON THE TOPIC?Microphysiological systems (MPSs) have been designed to recreate organ‐ or tissue‐specific characteristics of extracellular microenvironments that enhance the physiological relevance of cells in culture. Liver MPSs enable long‐lasting and stable culture of hepatic cells by culturing them in three‐dimensions and exposing them to fluid flow.WHAT QUESTION DID THIS STUDY ADDRESS?What is the functional performance relative to other cell culture platforms and the reproducibility of a liver MPS for assessing drug development and evaluation questions, such as toxicity, metabolism, and pharmacokinetics?WHAT DOES THIS STUDY ADD TO OUR KNOWLEDGE?The liver MPS systematically detected the toxicity of trovafloxacin. When compared with spheroids and sandwich cultures, this system had a more stable function and different sensitivity to troglitazone, tamoxifen, and digoxin. Quantifying phase II metabolism of troglitazone and intracellular accumulation of chloroquine demonstrated the potential use of the liver MPS for studying drug metabolism and pharmacokinetics. Quality control criteria for assessing chip function were key for reliably using the liver MPS.HOW MIGHT THIS CHANGE CLINICAL PHARMACOLOGY OR TRANSLATIONAL SCIENCE?Due to its functional robustness and physiological relevance (3D culture, cells expose to fluid flow and co‐culture of different cell types), the liver MPS can, in a reproducible manner: (i) detect inflammatory‐induced drug toxicity, as demonstrated with trovafloxacin, (ii) detect the toxicity of other drugs, such as troglitazone, tamoxifen, and digoxin, with different effects than those detected in spheroids and sandwich cultures, (iii) enable studies of hepatic function that rely on prolonged cellular activity, and (iv) detect phase II metabolites and drug accumulation to potentially support the interpretation of clinical data. The integration of MPSs in drug development will be facilitated by careful evaluation of performance and reproducibility as performed in this study.

Cancer microenvironment has a significant impact on the progression of the disease. In particular, hypoxia is the key driver of cancer survival, invasion, and chemoresistance. Although several in vitro models have been developed to study hypoxia-related cancer pathology, the complex interplay of the cancer microenvironment observedin vivo has not been reproduced yet owing to the lack of precise spatial control. Instead, 3D biofabrication approaches have been proposed to create microphysiological systems for better emulation of cancer ecology and accurate anticancer treatment evaluation. Herein, we propose a 3D cell-printing approach to fabricate a hypoxic cancer-on-a-chip. The hypoxia-inducing components in the chip were determined based on a computer simulation of the oxygen distribution. Cancer-stroma concentric rings were printed using bioinks containing glioblastoma cells and endothelial cells to recapitulate a type of solid cancer. The resulting chip realized central hypoxia and aggravated malignancy in cancer with the formation of representative pathophysiological markers. Overall, the proposed approach for creating a solid-cancer-mimetic microphysiological system is expected to bridge the gap between in vivo and in vitro models for cancer research.