bims-orenst 2022-07-17 papers

Issue of 2022–07–17
six papers selected by

Biochip J. 2022 Jul 08. 1-12

Biological Influence of Pulmonary Disease Conditions Induced by Particulate Matter on Microfluidic Lung Chips.

Faiza Jabbar, Young-Su Kim, Sang Ho Lee.

Particulate matter (PM10)-induced respiratory illnesses are difficult to investigate in trans-well culture systems. Microphysiological systems offer the capacity to mimic these phenomena to analyze any possible hazards that PM10 exposure poses to respiratory system of Humans. This study proposes an on-chip healthy human lung distal airway model that efficiently reconstitutes in vivo-like environmental conditions in a microfluidic device. The lung-on-chip model comprises a TEER sensor chip and portable microscope for continuous monitoring. To determine the efficacy of our model, we assessed the response to exposure to three PM environmental conditions (mild, average, and severe) and analyzed the relevant in vivo physiological and toxicological data using the airway model. Our results revealed significant increases in the levels of the IL-13, IL-6, and MUC5AC pathological biomarkers, which indicate increased incidences of on-chip asthma and chronic obstructive pulmonary disease conditions. Overall, we deduced that this model will facilitate the identification of potential therapeutics and the prevention of chronic life-threatening toxicities and pandemics such as COVID-19. The proposed system provides basic data for producing an improved in organ-on-chip technology.
Supplementary Information: The online version contains supplementary material available at 10.1007/s13206-022-00068-x.

Keywords: Distal airways; IL-13; IL-6; Microfluidic; Particulate matter; Real-time monitoring; TEER

DOI: https://doi.org/10.1007/s13206-022-00068-x

Lab Chip. 2022 Jul 14.

Application of a gut-liver-on-a-chip device and mechanistic modelling to the quantitative in vitro pharmacokinetic study of mycophenolate mofetil.

Nicoló Milani, Neil Parrott, Daniela Ortiz Franyuti, Patricio Godoy, Aleksandra Galetin, Michael Gertz, Stephen Fowler.

Microphysiological systems (MPS) consisting of multiple linked organ-on-a-chip (OoC) components are highly promising tools with potential to provide more relevant in vitro to in vivo translation of drug disposition, efficacy and toxicity. A gut-liver OoC system was employed with Caco2 cells in co-culture with HT29 cells in the intestinal compartment and single donor primary hepatocytes in the hepatic compartment for the investigation of intestinal permeability, metabolism (intestinal and hepatic) and potential interplay of those processes. The prodrug mycophenolate mofetil was tested for quantitative evaluation of the gut-liver OoC due to the contribution of both gut and liver in its metabolism. Conversion of mycophenolate mofetil to active drug mycophenolic acid and further metabolism to a glucuronide metabolite was assessed over time in the gut apical, gut basolateral and liver compartments. Mechanistic modelling of experimental data was performed to estimate clearance and permeability parameters for the prodrug, active drug and glucuronide metabolite. Integration of gut-liver OoC data with in silico modelling allowed investigation of the complex combination of intestinal and hepatic processes, which is not possible with standard single tissue in vitro systems. A comprehensive evaluation of the mechanistic model, including structural model and parameter identifiability and global sensitivity analysis, enabled a robust experimental design and estimation of in vitro pharmacokinetic parameters. We propose that similar methodologies may be applied to other multi-organ microphysiological systems used for drug metabolism studies or wherever quantitative knowledge of changing drug concentration with time enables better understanding of biological effect.

DOI: https://doi.org/10.1039/d2lc00276k

Front Toxicol. 2022 ;4 840606

A New Immortalized Human Alveolar Epithelial Cell Model to Study Lung Injury and Toxicity on a Breathing Lung-On-Chip System.

Arunima Sengupta, Nuria Roldan, Mirjam Kiener, Laurène Froment, Giulia Raggi, Theo Imler, Lea de Maddalena, Aude Rapet, Tobias May, Patrick Carius, Nicole Schneider-Daum, Claus-Michael Lehr, Marianna Kruithof-de Julio, Thomas Geiser, Thomas Michael Marti, Janick D Stucki, Nina Hobi, Olivier T Guenat.

The evaluation of inhalation toxicity, drug safety and efficacy assessment, as well as the investigation of complex disease pathomechanisms, are increasingly relying on in vitro lung models. This is due to the progressive shift towards human-based systems for more predictive and translational research. While several cellular models are currently available for the upper airways, modelling the distal alveolar region poses several constraints that make the standardization of reliable alveolar in vitro models relatively difficult. In this work, we present a new and reproducible alveolar in vitro model, that combines a human derived immortalized alveolar epithelial cell line (AXiAEC) and organ-on-chip technology mimicking the lung alveolar biophysical environment (AXlung-on-chip). The latter mimics key features of the in vivo alveolar milieu: breathing-like 3D cyclic stretch (10% linear strain, 0.2 Hz frequency) and an ultrathin, porous and elastic membrane. AXiAECs cultured on-chip were characterized for their alveolar epithelial cell markers by gene and protein expression. Cell barrier properties were examined by TER (Transbarrier Electrical Resistance) measurement and tight junction formation. To establish a physiological model for the distal lung, AXiAECs were cultured for long-term at air-liquid interface (ALI) on-chip. To this end, different stages of alveolar damage including inflammation (via exposure to bacterial lipopolysaccharide) and the response to a profibrotic mediator (via exposure to Transforming growth factor β1) were analyzed. In addition, the expression of relevant host cell factors involved in SARS-CoV-2 infection was investigated to evaluate its potential application for COVID-19 studies. This study shows that AXiAECs cultured on the AXlung-on-chip exhibit an enhanced in vivo-like alveolar character which is reflected into: 1) Alveolar type 1 (AT1) and 2 (AT2) cell specific phenotypes, 2) tight barrier formation (with TER above 1,000 Ω cm2) and 3) reproducible long-term preservation of alveolar characteristics in nearly physiological conditions (co-culture, breathing, ALI). To the best of our knowledge, this is the first time that a primary derived alveolar epithelial cell line on-chip representing both AT1 and AT2 characteristics is reported. This distal lung model thereby represents a valuable in vitro tool to study inhalation toxicity, test safety and efficacy of drug compounds and characterization of xenobiotics.

Keywords: AT1 and AT2; SARS-CoV-2; alveolar epithelial cells; cyclic stretch; distal lung; lung inflammation; lung toxicity; lung-on-a-chip

DOI: https://doi.org/10.3389/ftox.2022.840606

Front Bioeng Biotechnol. 2022 ;10 868857

Human Cholangiocytes Form a Polarized and Functional Bile Duct on Hollow Fiber Membranes.

Zhenguo Wang, João Faria, Luc J W van der Laan, Louis C Penning, Rosalinde Masereeuw, Bart Spee.

Liver diseases affect hundreds of millions of people worldwide; most often the hepatocytes or cholangiocytes are damaged. Diseases of the biliary tract cause severe patient burden, and cholangiocytes, the cells lining the biliary tract, are sensitive to numerous drugs. Therefore, investigations into proper cholangiocyte functions are of utmost importance, which is restricted, in vitro, by the lack of primary human cholangiocytes allowing such screening. To investigate biliary function, including transepithelial transport, cholangiocytes must be cultured as three-dimensional (3D) ductular structures. We previously established murine intrahepatic cholangiocyte organoid-derived cholangiocyte-like cells (CLCs) and cultured them onto polyethersulfone hollow fiber membranes (HFMs) to generate 3D duct structures that resemble native bile ducts at the structural and functional level. Here, we established an efficient, stepwise method for directed differentiation of human intrahepatic cholangiocyte organoids (ICOs) into CLCs. Human ICO-derived CLCs showed key characteristics of cholangiocytes, such as the expression of structural and functional markers, formation of primary cilia, and P-glycoprotein-mediated transport in a polarized fashion. The organoid cultures exhibit farnesoid X receptor (FXR)-dependent functions that are vital to liver bile acid homeostasis in vivo. Furthermore, human ICO-derived CLCs cultured on HFMs in a differentiation medium form tubular architecture with some tight, confluent, and polarized monolayers that better mimic native bile duct characteristics than differentiated cultures in standard 2D or Matrigel-based 3D culture plates. Together, our optimized differentiation protocol to obtain CLC organoids, when applied on HFMs to form bioengineered bile ducts, will facilitate studying cholangiopathies and allow developing therapeutic strategies.

Keywords: bioengineered bile duct; cholangiocytes; hollow fiber membrane; intrahepatic cholangiocyte organoids; monolayer; perfusable; polarity

DOI: https://doi.org/10.3389/fbioe.2022.868857

IEEE Open J Eng Med Biol. 2022 ;3 86-95

Assessment of CAR-T Cell-Mediated Cytotoxicity in 3D Microfluidic Cancer Co-Culture Models for Combination Therapy.

Karla Paterson, Sarah Paterson, Theresa Mulholland, Seth B Coffelt, Michele Zagnoni.

Chimeric antigen receptor (CAR)-T cell therapy is efficacious against many haematological malignancies, but challenges remain when using this cellular immunotherapy for treating solid tumours. Classical 2D in vitro models fail to recapitulate the complexity of the tumour microenvironment, whilst in vivo models, such as patient-derived xenografts, are costly and labour intensive. Microfluidic technologies can provide miniaturized solutions to assess CAR-T therapies in 3D complex preclinical models of solid tumours. Here, we present a novel microfluidic immunoassay for the evaluation of CAR-T cell cytotoxicity and targeting specificity on 3D spheroids containing cancer cells and stromal cells. Monitoring the interaction between CAR-T cells and spheroid co-cultures, we show that CAR-T cells home towards target-expressing cancer cells and elicit a cytotoxic effect. Testing CAR-T cells in combination therapies, we show that CAR-T cell cytotoxicity is enhanced with anti-PD-L1 therapy and carboplatin chemotherapy. We propose this proof-of-concept microfluidic immunoassay as a material-saving, pre-clinical screening tool for quantification of cell therapy efficacy.

Keywords: Immunotherapy; lab-on-a-chip; solid tumour microenvironment; three-dimensional in vitro complex model

DOI: https://doi.org/10.1109/OJEMB.2022.3178302

Sci Rep. 2022 Jul 12. 12(1): 11838

Utilizing novel TBI-on-a-chip device to link physical impacts to neurodegeneration and decipher primary and secondary injury mechanisms.

Edmond A Rogers, Timothy Beauclair, Andrew Thyen, Riyi Shi.

While clinical observations have confirmed a link between the development of neurodegenerative diseases and traumatic brain injuries (TBI), there are currently no treatments available and the underlying mechanisms remain elusive. In response, we have developed an in vitro pendulum trauma model capable of imparting rapid acceleration injuries to neuronal networks grown on microelectrode arrays within a clinically relevant range of g forces, with real-time electrophysiological and morphological monitoring. By coupling a primary physical insult with the quantification of post-impact levels of known biochemical pathological markers, we demonstrate the capability of our system to delineate and investigate the primary and secondary injury mechanisms leading to post-impact neurodegeneration. Specifically, impact experiments reveal significant, force-dependent increases in the pro-inflammatory, oxidative stress marker acrolein at 24 h post-impact. The elevation of acrolein was augmented by escalating g force exposures (30-200 g), increasing the number of rapidly repeated impacts (4-6 s interval, 3, 5 and 10×), and by exposing impacted cells to 40 mM ethanol, a known comorbidity of TBI. The elevated levels of acrolein following multiple impacts could be reduced by increasing time-intervals between repeated hits. In addition, we show that conditioned media from maximally-impacted cultures can cause cellular acrolein elevation when introduced to non-impact, control networks, further solidifying acrolein's role as a diffusive-factor in post-TBI secondary injuries. Finally, morphological data reveals post-impact acrolein generation to be primarily confined to soma, with some emergence in cellular processes. In conclusion, this novel technology provides accurate, physical insults with a unique level of structural and temporal resolution, facilitating the investigation of post-TBI neurodegeneration.

DOI: https://doi.org/10.1038/s41598-022-14937-w