scholarly journals Implementation of a Dynamic Co-Culture Model Abated Silver Nanoparticle Interactions and Nanotoxicological Outcomes In Vitro

Nanomaterials ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1807
Author(s):  
Nicholas J. Braun ◽  
Rachel M. Galaska ◽  
Maggie E. Jewett ◽  
Kristen A. Krupa
Keyword(s):  
In Vitro Models ◽  
Engineered Nanoparticles ◽  
Cellular Interactions ◽  
Dynamic Flow ◽  
In Vivo Models ◽  
Alveolar Epithelial ◽  
Biological Responses ◽  
Silver Nps

The incorporation of engineered nanoparticles (NPs) into everyday consumer goods, products, and applications has given rise to the field of nanotoxicology, which evaluates the safety of NPs within biological environments. The unique physicochemical properties of NPs have made this an insurmountable challenge, as their reactivity and variable behavior have given rise to discrepancies between standard cell-based in vitro and animal in vivo models. In this study, enhanced in vitro models were generated that retained the advantages of traditional cell cultures, but incorporated the modifications of (1) inclusion of an activated immune element and (2) the presence of physiologically-relevant dynamic flow. Following verification that the human alveolar epithelial and macrophage (A549/U937) co-culture could be successfully sustained under both static and dynamic conditions, these cultures, in addition to a standard A549 static model, were challenged with 10 nm citrate coated silver NPs (AgNPs). This work identified a reshaping of the AgNP-cellular interface and differential biological responses following exposure. The presence of dynamic flow modified cellular morphology and reduced AgNP deposition by approximately 20% over the static exposure environments. Cellular toxicity and stress endpoints, including reactive oxygen species, heat shock protein 70, and secretion of pro-inflammatory cytokines, were found to vary as a function of both cellular composition and flow conditions; with activated macrophages and fluid flow both mitigating the severity of AgNP-dependent bioeffects. This work highlights the possibility of enhanced in vitro systems to assess the safety of engineered NPs and demonstrates their effectiveness in elucidating novel NP-cellular interactions and toxicological profiles.

scholarly journals Characterizing the Role of Biologically Relevant Fluid Dynamics on Silver Nanoparticle Dependent Oxidative Stress in Adherent and Suspension In Vitro Models

Antioxidants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 832
Author(s):  
Katherine E. Burns ◽  
Robert F. Uhrig ◽  
Maggie E. Jewett ◽  
Madison F. Bourbon ◽  
Kristen A. Krupa
Keyword(s):  
Oxidative Stress ◽  
Fluid Dynamics ◽  
In Vitro Models ◽  
Poor Correlation ◽  
Cellular Interactions ◽  
Dynamic Flow ◽  
In Vivo Models ◽  
And Oxidative Stress

Silver nanoparticles (AgNPs) are being employed in numerous consumer goods and applications; however, they are renowned for inducing negative cellular consequences including toxicity, oxidative stress, and an inflammatory response. Nanotoxicological outcomes are dependent on numerous factors, including physicochemical, biological, and environmental influences. Currently, NP safety evaluations are carried out in both cell-based in vitro and animal in vivo models, with poor correlation between these mechanisms. These discrepancies highlight the need for enhanced exposure environments, which retain the advantages of in vitro models but incorporate critical in vivo influences, such as fluid dynamics. This study characterized the effects of dynamic flow on AgNP behavior, cellular interactions, and oxidative stress within both adherent alveolar (A549) and suspension monocyte (U937) models. This study determined that the presence of physiologically relevant flow resulted in substantial modifications to AgNP cellular interactions and subsequent oxidative stress, as assessed via reactive oxygen species (ROS), glutathione levels, p53, NFκB, and secretion of pro-inflammatory cytokines. Within the adherent model, dynamic flow reduced AgNP deposition and oxidative stress markers by roughly 20%. However, due to increased frequency of contact, the suspension U937 cells were associated with higher NP interactions and intracellular stress under fluid flow exposure conditions. For example, the increased AgNP association resulted in a 50% increase in intracellular ROS and p53 levels. This work highlights the potential of modified in vitro systems to improve analysis of AgNP dosimetry and safety evaluations, including oxidative stress assessments.

scholarly journals Bioreactors as physiologicallike in vitro models

2020 ◽  
Author(s):  
Sara Mantero ◽  
Federica Boschetti
Keyword(s):  
Culture Conditions ◽  
In Vitro Models ◽  
In Vivo Models ◽  
In Vitro Development ◽  
Culture Cells ◽  
Vitro Development ◽  
Tissue Models ◽  
Mechanical Stretching

Bioreactors are powerful tools for in vitro development of engineered substitutes through controlled biological, physical, and mechanical culture conditions: bioreactor technology allows a closer in vitro replication of native tissues. One of bioreactors applications is the design of in vitro 3D tissue models as a bridge between 2D and in vivo models, allowing the application of 3R (replacement, reduction, refinement) principle. To this aim, bioreactors can be used to culture cells seeded on engineered scaffolds under in vivo-like conditions. Another key use of bioreactors is for perfusion decellularization of tissues and organs to be used as scaffolds. This contribution describes a dynamic stretching. bioreactor, imposing a mechanical stretching to the cultured constructs, allowing the development of skeletal muscle engineered constructs, and a decellularization bioreactor, designed for decellularization of blood vessels.

scholarly journals Experimental Models in Rheumatoid Arthritis:

2020 ◽  
Vol 8 (1) ◽  
pp. 119-134
Author(s):  
Verônica Assalin Zorgetto-Pinheiro ◽  
Alexandre Meira de Vasconcelos ◽  
Rafael Sanaiotte Pinheiro ◽  
Danielle Bogo ◽  
Iandara Schettert Silva
Keyword(s):  
Rheumatoid Arthritis ◽  
Drug Testing ◽  
Pathological Condition ◽  
In Vitro Models ◽  
Experimental Models ◽  
In Vivo Models ◽  
Methodological Tool ◽  
Class 1

Rheumatoid arthritis is an autoimmune and chronic pathological condition characterized by an inflammatory process of the joints It is a complex and multifactorial, involving genetic, epigenetic and environmental factors and the use of experimental models is required to better understand its pathology and for drug testing. The aim of this study was to perform a systematic literature review on experimental models in rheumatoid arthritis using IRAMUTEQ, a software that analysis, qualitatively and quantitatively, text fragments, as a methodological tool. After searching for articles published in the last five years on Scopus database and applying the exclusion criteria, we ended with 84 articles. The most commonly employed experimental models was the arthritis induction by inoculation of the Complete Freund's Adjuvant (CFA), followed by the use of combined methodologies and the collagen-induced arthritis (CIA). The analyses of abstracts by the IRAMUTEQ software provided a classification according to their textual elements in four classes, which were grouped into three main themes: in vivo models (class 1), clinical practice and traditional medicine (classes 2 and 3) and in vitro models (class 4) and it was also possible to build a similarity tree of the terms present in the abstracts and a word cloud with the most cited terms. Thus, the use of the IRAMUTEQ software as a methodological tool has been satisfactory, since it was possible to identify the main experimental models used, keywords, pathological processes and molecules involved in the pathogenesis of rheumatoid arthritis free of the researchers’ bias, in addition to being a tool for visual and intuitive results.

scholarly journals CADASIL from Bench to Bedside: Disease Models and Novel Therapeutic Approaches

2021 ◽  
Author(s):  
Arianna Manini ◽  
Leonardo Pantoni
Keyword(s):  
Therapeutic Option ◽  
In Vitro Models ◽  
Monogenic Disease ◽  
Therapeutic Approaches ◽  
In Vivo Models ◽  
Human Phenotype ◽  
Knock Out ◽  
Novel Therapeutic

AbstractCerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a monogenic disease caused by NOTCH3 mutations and characterized by typical clinical, neuroradiological, and pathological features. NOTCH3 belongs to a family of highly conserved transmembrane receptors rich of epidermal growth factor repeats, mostly expressed in vascular smooth muscle cells and pericytes, which perform essential developmental functions and are involved in tissues maintenance and renewal. To date, no therapeutic option for CADASIL is available except for few symptomatic treatments. Novel in vitro and in vivo models are continuously explored with the aim to investigate underlying pathogenic mechanisms and to test novel therapeutic approaches. In this scenario, knock-out, knock-in, and transgenic mice studies have generated a large amount of information on molecular and biological aspects of CADASIL, despite that they incompletely reproduce the human phenotype. Moreover, the field of in vitro models has been revolutionized in the last two decades by the introduction of induced pluripotent stem cells (iPSCs) technology. As a consequence, novel therapeutic approaches, including immunotherapy, growth factors administration, and antisense oligonucleotides, are currently under investigation. While waiting that further studies confirm the promising results obtained, the data reviewed suggest that our therapeutic approach to the disease could be transformed, generating new hope for the future.

IN VITRO MODELS OF MYCOBACTERIUM TUBERCULOSIS DORMANCY, AND IN VIVO MODELS OF LATENT TUBERCULOSIS INFECTION

2019 ◽  
Vol 64 (5) ◽  
pp. 299-307
Author(s):  
M. V. Fursov ◽  
I. A. Dyatlov ◽  
V. D. Potapov
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Latent Tuberculosis Infection ◽  
Latent Tuberculosis ◽  
In Vitro Models ◽  
Tuberculosis Infection ◽  
Published Data ◽  
In Vivo Models ◽  
Dormant State

Modeling of tuberculosis infection is carried out in order to clarify various aspects of the tuberculosis pathogenesis, as well as the testing of new anti-tuberculosis drugs. The characteristic of in vitro models (n = 16) for Mycobacterium tuberculosis dormant state and in vivo models (n = 14) for the latent tuberculosis infection involving several animal species published to date are presented in this review. A brief description of the models and the results obtained by the authors are presented. The analysis of the published data reflects the list of methodological procedures that allow researchers to study the mechanism of the transition of M. tuberculosis cells to a dormant state and reverse to metabolically active state, as well as the process of conversion of active tuberculosis infection to a latent tuberculosis and reactivation.

Imaging Microphysiological Systems: A Review

2020 ◽  
Author(s):  
Samantha Peel ◽  
Mark Jackman
Keyword(s):  
Drug Discovery ◽  
Animal Studies ◽  
Imaging Techniques ◽  
Quantitative Information ◽  
In Vitro Models ◽  
Microscopy Imaging ◽  
In Vivo Models ◽  
Microphysiological Systems

Microphysiological Systems (MPS), often referred to as 'organ-on-chips' are microfluidic-based in vitro models that aim to recapitulate the dynamic chemical and mechanical microenvironment of living organs. MPS promise to bridge the gap between in vitro and in vivo models, and ultimately improve the translation from pre-clinical animal studies to clinical trials. However, despite the explosion of interest in recent years, and the obvious rewards for such models which could improve R&D efficiency and reduce drug attrition in the clinic, the pharmaceutical industry has been slow to fully adopt this technology. The ability to extract robust, quantitative information from MPS at scale is a key requirement if these models are to impact drug discovery and the subsequent drug development process. Microscopy imaging remains a core technology that enables the capture of information at the single cell level and with subcellular resolution. Furthermore, such imaging techniques can be automated, increasing throughput, enabling compound screening. In this review we discuss a range of imaging techniques that have been applied to MPS of varying focus, such as organoids and organ-chip-type models. We outline the opportunities these technologies can bring in terms of understanding mechanistic biology, but also how they could be used in higher-throughput screens, widening the scope of their impact in drug discovery. We discuss the associated challenges of imaging these complex models and the steps required to enable full exploitation. Finally, we discuss the requirements for MPS, if they are to be applied at a scale necessary to support drug discovery projects.

scholarly journals Assessment of In Vitro Bioaccessibility and In Vivo Oral Bioavailability as Complementary Tools to Better Understand the Effect of Cooking on Methylmercury, Arsenic, and Selenium in Tuna

Toxics ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 27
Author(s):  
Tania Charette ◽  
Danyel Bueno Dalto ◽  
Maikel Rosabal ◽  
J. Jacques Matte ◽  
Marc Amyot
Keyword(s):  
Oral Bioavailability ◽  
Fish Muscle ◽  
In Vitro Models ◽  
In Vivo Studies ◽  
In Vivo Models ◽  
Exposure Pathway ◽  
In Vitro Bioaccessibility ◽  
Kinetics Of

Fish consumption is the main exposure pathway of the neurotoxicant methylmercury (MeHg) in humans. The risk associated with exposure to MeHg may be modified by its interactions with selenium (Se) and arsenic (As). In vitro bioaccessibility studies have demonstrated that cooking the fish muscle decreases MeHg solubility markedly and, as a consequence, its potential absorption by the consumer. However, this phenomenon has yet to be validated by in vivo models. Our study aimed to test whether MeHg bioaccessibility can be used as a surrogate to assess the effect of cooking on MeHg in vivo availability. We fed pigs raw and cooked tuna meals and collected blood samples from catheters in the portal vein and carotid artery at: 0, 30, 60, 90, 120, 180, 240, 300, 360, 420, 480 and 540 min post-meal. In contrast to in vitro models, pig oral bioavailability of MeHg was not affected by cooking, although the MeHg kinetics of absorption was faster for the cooked meal than for the raw meal. We conclude that bioaccessibility should not be readily used as a direct surrogate for in vivo studies and that, in contrast with the in vitro results, the cooking of fish muscle did not decrease the exposure of the consumer to MeHg.

scholarly journals Melatonin as a Reducer of Neuro- and Vasculotoxic Oxidative Stress Induced by Homocysteine

Antioxidants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1178
Author(s):  
Kamil Karolczak ◽  
Cezary Watala
Keyword(s):  
Oxidative Stress ◽  
Structural Changes ◽  
Clinical Evidence ◽  
Neuronal Degeneration ◽  
Antioxidant Properties ◽  
In Vitro Models ◽  
In Vivo Models ◽  
Initial Research

The antioxidant properties of melatonin can be successfully used to reduce the effects of oxidative stress caused by homocysteine. The beneficial actions of melatonin are mainly due to its ability to inhibit the generation of the hydroxyl radical during the oxidation of homocysteine. Melatonin protects endothelial cells, neurons, and glia against the action of oxygen radicals generated by homocysteine and prevents the structural changes in cells that lead to impaired contractility of blood vessels and neuronal degeneration. It can be, therefore, assumed that the results obtained in experiments performed mainly in the in vitro models and occasionally in animal models may clear the way to clinical applications of melatonin in patients with hyperhomocysteinemia, who exhibit a higher risk of developing neurodegenerative diseases (e.g., Parkinson’s disease or Alzheimer’s disease) and cardiovascular diseases of atherothrombotic etiology. However, the results that have been obtained so far are scarce and have seldom been performed on advanced in vivo models. All findings predominately originate from the use of in vitro models and the scarcity of clinical evidence is huge. Thus, this mini-review should be considered as a summary of the outcomes of the initial research in the field concerning the use of melatonin as a possibly efficient attenuator of oxidative stress induced by homocysteine.

scholarly journals Assessment of CAR-T mediated and targeted cytotoxicity in 3D microfluidic TBNC co-culture models for combination therapy

2021 ◽  
Author(s):  
Karla Anna Findlay Paterson ◽  
Sarah Paterson ◽  
Theresa Mulholland ◽  
Seth B Coffelt ◽  
Michele Zagnoni
Keyword(s):  
T Cell ◽  
In Vitro Models ◽  
T Cell Therapy ◽  
In Vivo Models ◽  
Car T Cell ◽  
Effective Combination ◽  
Car T ◽  
Culture Models

Chimeric antigen receptor (CAR)-T cell therapy is efficacious against many haematological malignancies; however, their therapeutic application to treat solid tumours presents further challenges. A better understanding of how the solid TME impacts CAR-T anti-tumour effects would enable the selection of effective combination therapies to decipher the optimal course of treatment for patients and to better engineer CAR-Ts. Classical 2D in vitro models do not provide sufficient recapitulation of the native human TME, and in vivo models, such as patient-derived xenografts, are costly, complex and labour intensive. Here, we present a novel 3D, miniaturised assay for the evaluation of EGFR-targeted CAR-T cell cytotoxicity and specificity on tumour-stroma triple-negative breast cancer models in microfluidic devices. CAR-T cells were shown to home towards EGFR-expressing cancer cells to elicit a cytotoxic effect, whilst leaving low EGFR-expressing fibroblasts viable, an effect which was enhanced through combination anti-PD-L1 therapy and carboplatin chemotherapy. Hence, we propose this proof-of-concept immunoassay as a future preclinical screening tool for the development of novel immunotherapeutics and for use in personalized medicine.

scholarly journals ModelingIn VitroCellular Responses to Silver Nanoparticles

2014 ◽  
Vol 2014 ◽  
pp. 1-13 ◽  
Author(s):  
Dwaipayan Mukherjee ◽  
Steven G. Royce ◽  
Srijata Sarkar ◽  
Andrew Thorley ◽  
Stephan Schwander ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Cell Types ◽  
Engineered Nanoparticles ◽  
Mrna Levels ◽  
Cellular Interactions ◽  
Cellular Mechanisms ◽  
Nanoparticle Transport ◽  
Cell Culture Systems

Engineered nanoparticles (NPs) have been widely demonstrated to induce toxic effects to various cell types.In vitrocell exposure systems have high potential for reliable, high throughput screening of nanoparticle toxicity, allowing focusing on particular pathways while excluding unwanted effects due to other cells or tissue dosimetry. The work presented here involves a detailed biologically based computational model of cellular interactions with NPs; it utilizes measurements performed in human cell culture systemsin vitro, to develop a mechanistic mathematical model that can support analysis and prediction ofin vivoeffects of NPs. The model considers basic cellular mechanisms including proliferation, apoptosis, and production of cytokines in response to NPs. This new model is implemented for macrophages and parameterized usingin vitromeasurements of changes in cellular viability and mRNA levels of cytokines: TNF, IL-1b, IL-6, IL-8, and IL-10. The model includesin vitrocellular dosimetry due to nanoparticle transport and transformation. Furthermore, the model developed here optimizes the essential cellular parameters based onin vitromeasurements, and provides a “stepping stone” for the development of more advancedin vivomodels that will incorporate additional cellular and NP interactions.

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