Engineered Lego®-like microphysiological models of the human airway clearance phenomena

Mucociliary clearance is a crucial event that supports the elimination of inhaled particles, bacteria, pollution and hazardous agents from the human airways, and it also limits the diffusion of aerosolized drugs into the airway epithelium. In spite of its relevance, few in vitro models sufficiently address the cumulative effect of the steric and interactive barrier function of mucus on the one hand, and the dynamic mucus transport imposed by ciliary mucus propulsion on the other hand. Here, ad hoc mucus models of physiological and pathological mucus are combined with magnetic artificial cilia to model mucociliary transport in both physiological and pathological states. The Lego®-like concept adopted, in this study, enables the development of mucociliary clearance models with high versatility, since these can be easily modified to reproduce phenomena characteristic of healthy and diseased human airways, while allowing to determine the effect of each parameter and/or structure separately on the overall mucociliary transport. These Lego®-like airway models can be available off-the-shelf because they are exclusively made of readily available materials, thus ensuring reproducibility across different laboratories.

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State of the art on lung organoids in mammals

Veterinary Research ◽

10.1186/s13567-021-00946-6 ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Fabienne Archer ◽

Alexandra Bobet-Erny ◽

Maryline Gomes

Keyword(s):

Lung Development ◽

One Health ◽

State Of The Art ◽

Animal Health ◽

In Vitro Models ◽

Cancer Genetic ◽

3 Dimensional ◽

The One ◽

Species Specific

AbstractThe number and severity of diseases affecting lung development and adult respiratory function have stimulated great interest in developing new in vitro models to study lung in different species. Recent breakthroughs in 3-dimensional (3D) organoid cultures have led to new physiological in vitro models that better mimic the lung than conventional 2D cultures. Lung organoids simulate multiple aspects of the real organ, making them promising and useful models for studying organ development, function and disease (infection, cancer, genetic disease). Due to their dynamics in culture, they can serve as a sustainable source of functional cells (biobanking) and be manipulated genetically. Given the differences between species regarding developmental kinetics, the maturation of the lung at birth, the distribution of the different cell populations along the respiratory tract and species barriers for infectious diseases, there is a need for species-specific lung models capable of mimicking mammal lungs as they are of great interest for animal health and production, following the One Health approach. This paper reviews the latest developments in the growing field of lung organoids.

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Comparative physiology of rodent pulmonary macrophages: in vitro functional responses

Journal of Applied Physiology ◽

10.1152/jappl.1988.64.5.1953 ◽

1988 ◽

Vol 64 (5) ◽

pp. 1953-1959 ◽

Cited By ~ 19

Author(s):

D. B. Warheit ◽

M. A. Hartsky ◽

M. S. Stefaniak

Keyword(s):

Guinea Pig ◽

Phagocytic Activity ◽

Lung Cells ◽

Comparative Physiology ◽

Functional Responses ◽

Inhaled Particles ◽

Pulmonary Macrophages ◽

The One ◽

Formyl Peptides

Since toxicological testing of inhaled materials frequently requires utilization of several species, we have investigated pulmonary macrophage (PM) functional responses and compared the rat model with other rodents. Two strains of rats, three strains of mice, and one strain each of hamster and guinea pig were used in this study. The numbers of recovered cells by bronchoalveolar lavage generally correlated with animal body weight. The one exception was the Syrian Golden hamster from which increased numbers of macrophages were recovered. Cellular differential data obtained from lavaged cytocentrifuge preparations demonstrated that PM's account for greater than 97% of recoverable free lung cells for all species except the guinea pig, which contains a resident population of eosinophils. Cell morphology studies indicated that hamster PM exhibited the highest proportion of ruffled PM and demonstrated the highest phagocytic activity, whereas mouse PM phagocytic activity was significantly reduced compared with the other three species. In addition, chemotaxis studies showed that rat PM migrated best to zymosan-activated, complement-dependent chemoattractants, whereas hamster PM demonstrated an enhanced chemotactic response to N-formyl peptides. The results of these studies suggest that the rat may be the most efficient species for clearing inhaled particles, whereas hamsters and guinea pigs may best respond to bacteria.

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How to Achieve High-Quality Oocytes? The Key Role of Myo-Inositol and Melatonin

International Journal of Endocrinology ◽

10.1155/2016/4987436 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 36

Author(s):

Salvatore Giovanni Vitale ◽

Paola Rossetti ◽

Francesco Corrado ◽

Agnese Maria Chiara Rapisarda ◽

Sandro La Vignera ◽

...

Keyword(s):

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Oocyte Quality ◽

Fertilization Rate ◽

In Vitro Models ◽

The One ◽

High Level

Assisted reproductive technologies (ART) have experienced growing interest from infertile patients seeking to become pregnant. The quality of oocytes plays a pivotal role in determining ART outcomes. Although many authors have studied how supplementation therapy may affect this important parameter for both in vivo and in vitro models, data are not yet robust enough to support firm conclusions. Regarding this last point, in this review our objective has been to evaluate the state of the art regarding supplementation with melatonin and myo-inositol in order to improve oocyte quality during ART. On the one hand, the antioxidant effect of melatonin is well known as being useful during ovulation and oocyte incubation, two occasions with a high level of oxidative stress. On the other hand, myo-inositol is important in cellular structure and in cellular signaling pathways. Our analysis suggests that the use of these two molecules may significantly improve the quality of oocytes and the quality of embryos: melatonin seems to raise the fertilization rate, and myo-inositol improves the pregnancy rate, although all published studies do not fully agree with these conclusions. However, previous studies have demonstrated that cotreatment improves these results compared with melatonin alone or myo-inositol alone. We recommend that further studies be performed in order to confirm these positive outcomes in routine ART treatment.

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Adenosine A3 receptor-mediated potentiation of mucociliary transport and epithelial ciliary motility

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00360.2001 ◽

2002 ◽

Vol 282 (3) ◽

pp. L556-L562 ◽

Cited By ~ 7

Author(s):

Manako Taira ◽

Jun Tamaoki ◽

Kazuyuki Nishimura ◽

Junko Nakata ◽

Mitsuko Kondo ◽

...

Keyword(s):

Mucociliary Clearance ◽

Beat Frequency ◽

Blue Dye ◽

Dependent Manner ◽

Mucociliary Transport ◽

Concentration Dependent Manner ◽

Maximal Increase ◽

Ciliary Motility

To examine the effect of adenosine A3 receptor stimulation on airway mucociliary clearance, we measured transport of Evans blue dye in rabbit trachea in vivo and ciliary motility of epithelium by the photoelectric method in vitro. Mucociliary transport was enhanced dose dependently by the selective A3 agonist N 6-(3-iodobenzyl)-5′- N-methylcarbamoyladenosine (IB-MECA) and to a lesser extent by the less-selective N 6-2-(4-amino-3-iodophenyl)ethyladenosine, whereas the A1 agonist N-cyclopentyladenosine (CPA) and the A2 agonist CGS-21680 had no effect. The effect of IB-MECA was abolished by pretreatment with the selective A3 antagonist MRS-1220 but not by the A1 antagonist 1,3-dipropyly-8-cyclopentylxanthine or the A2 antagonist 3,7-dimethyl-l-propargylxanthine. Epithelial ciliary beat frequency was increased by IB-MECA in a concentration-dependent manner, the maximal increase being 33%, and this effect was inhibited by MRS-1220. The IB-MECA-induced ciliary stimulation was not altered by the Rp diastereomer of cAMP but was greatly inhibited by Ca2+-free medium containing BAPTA-AM. Incubation with IB-MECA increased intracellular Ca2+ contents. Therefore, A3 agonist enhances airway mucociliary clearance probably through Ca2+-mediated stimulation of ciliary motility of airway epithelium.

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In Vitro Ciliotoxicity and Cytotoxicity Testing of Repeated Chronic Exposure to Topical Nasal Formulations for Safety Studies

Pharmaceutics ◽

10.3390/pharmaceutics13111750 ◽

2021 ◽

Vol 13 (11) ◽

pp. 1750

Author(s):

Larisa Tratnjek ◽

Nadica Sibinovska ◽

Katja Kristan ◽

Mateja Erdani Kreft

Keyword(s):

Nasal Spray ◽

Mucociliary Clearance ◽

Beat Frequency ◽

Cumulative Effect ◽

In Vitro Assays ◽

Ciliary Activity ◽

Nasal Sprays ◽

Repeated Doses

Certain active drugs and excipients of nasal formulations may impair ciliary function and mucociliary clearance. The ciliary beat frequency (CBF) is a key parameter for determining mucociliary clearance rate, and in vitro assessments of CBF have proven to be accurate and reproducible. Since topical nasal formulations are applied with repeated doses, it is essential to elucidate their chronic, as opposed to acute, effect on mucociliary clearance and nasal mucosa. The aim of this study was to assess for the first time the ciliotoxicity and cytotoxicity of nasal sprays intended for chronic treatment (with repeated doses) using a previously designed set-up for CBF measurements. For 2 weeks, the 3D nasal MucilAir™ in vitro models were treated daily with undiluted or clinically relevant doses of mometasone nasal spray, placebo nasal spray, culture medium, or they were untreated. We demonstrated a dose-dependent and time-dependent (cumulative) effect of the nasal sprays on ciliary activity and cytotoxicity using CBF measurements and ultrastructural analysis, respectively. Our results indicate that repeated administration of clinically relevant doses of mometasone nasal spray is safe for in vivo use, which is in good agreement with a previous clinical study. Overall, our study suggests that such in vitro assays have great potential for topical nasal drug screening.

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Animal models of chronic pain. In vivo experiments

Regional Anesthesia and Acute Pain Management ◽

10.17816/1993-6508-2020-14-3-121-129 ◽

2021 ◽

Vol 14 (3) ◽

pp. 121-129

Author(s):

V. V. Khinovker ◽

E. V. Khinovker

Keyword(s):

Chronic Pain ◽

Population Aging ◽

In Vitro Models ◽

Medical Problems ◽

Rat Models ◽

In Vivo Experiments ◽

The One ◽

Different Levels

As a result of population aging and increasing of comorbide patients procent, chronic pain nowadays is the one of the most important medical problems and its treatment becoming one of progressive and popular researching topics. First part of acute or chronic pain research is a model that meets the specified criteria. First model of cronic pain sciatic nerve damaging was founded and proven more than 30 years ago. Nowadays there is a lot of in vivo and in vitro models that matched with different pathologies and also mechanisms of chronic nociceptive and neuropathic pain on different levels have been learned. In this article we review the most effective and often used rat models of chronic pain its mechanisms and assessment methods. The information based on the most citated articles for 10 years.

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An integrated screening method for evaluating the pulmonary toxicity of inhaled particulates: Role of microscopic techniques in assessing pulmonary macrophage clearance functions

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100161692 ◽

1990 ◽

Vol 48 (3) ◽

pp. 826-827

Author(s):

David B. Warheit ◽

Lena Achinko ◽

Mark A. Hartsky

Keyword(s):

Bronchoalveolar Lavage ◽

Pulmonary Toxicity ◽

Screening Method ◽

Pulmonary Response ◽

Inhaled Particles ◽

Cytochemical Analysis ◽

Pulmonary Macrophages ◽

Integrated Screening

There is a great need for the development of a rapid and reliable bioassay to evaluate the pulmonary toxicity of inhaled particles. A number of methods have been proposed, including lung clearance studies, bronchoalveolar lavage analysis, and in vitro cytotoxicity tests. These methods are often limited in scope inasmuch as they measure only one dimension of the pulmonary response to inhaled, instilled or incubated dusts. Accordingly, a comprehensive approach to lung toxicity studies has been developed.To validate the method, rats were exposed for 6 hours or 3 days to various concentrations of either aerosolized alpha quartz silica (Si) or carbonyl iron (CI) particles. Cells and fluids from groups of sham and dust-exposed animals were recovered by bronchoalveolar lavage (BAL). Alkaline phosphatase, LDH and protein values were measured in BAL fluids at several time points postexposure. Cells were counted and evaluated for viability, as well as differential and cytochemical analysis. In addition, pulmonary macrophages (PM) were cultured and studied for morphology, chemotaxis, and phagocytosis by scanning electron microscopy.

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