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Cells
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In Vitro Model for Micro and Nano Technologies
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In Vitro Model for Micro and Nano Technologies

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Cellular Biophysics".

Deadline for manuscript submissions: closed (30 June 2023) | Viewed by 20381

Special Issue Editor

Dr. Tania Limongi

E-Mail Website
Guest Editor
Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy
Interests: nanoparticles; extracellular vesicles; drug delivery; nanomedicine; in vitro study; nanotoxicology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

By now, most of mankind accepts living in the era of technology, which boasts benefits in terms of goods, spaces, freedom, and certainly an improved quality of life.

Among the technologies available, micro- and nano-technology (MNT) includes solutions as devices, materials, and methods ranging from nanometers to microns. Innovations in MNT have allowed for surprising outcomes in the resolution of serious and long-standing unresolved issues for tissue engineering, theranostics, and drug delivery applications. Micro- and nano-structured lab-on-chips, smart hybrid devices, and a wide collection of drug delivery solutions, including extracellular vesicles, liposomes, microemulsions, nanoparticles, and quantum dots, undoubtedly need precise, reliable, and reproducible biomedical validation certifying their effectiveness and healthiness before their application in diagnostic and/or clinical contests. In vitro and in vivo studies are used the most for MNT validation. Although in vivo experiments are widely used for radiology, nuclear medicine, medical imaging, acute toxicity, and reproductive and developmental toxicity assays, every scientist should keep in mind the fundamental principles of the 3Rs (replacement, reduction, and refinement) proposed by Russel and Burch in 1959. In this perspective, substantial reductions in animal use in research can be achieved by improving in vitro study designs and combining new in silico solutions with high throughput “omics” approaches. The newest in vitro testing solutions include models for several human anatomical districts, such as the blood–brain barrier, placenta, kidneys, lungs, and skin, as well the organ-on-a-chip microfluidic systems, and can be considered ethical, quick, and in many cases cheaper alternatives to in vivo testing. Although they are not alone, they are certainly the first to represent fast and versatile systems to verify the cytotoxicity, hemocompatibility, and efficacy of a whole series of micro- and nano-devices born from the collaboration between medical physicists, chemical engineers, biologists, and clinicians.

In light of the above, this Special Issue of Cells will consist of a selection of high-level research articles, reviews, and communications focusing on the various in vitro systems used for the biomedical validation of the latest innovative micro- and nano-technology solutions.

Dr. Tania Limongi
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cells is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • in vitro cells culture
  • micro- and nano-technology
  • extracellular vesicles
  • nanoparticles
  • quantum dots
  • biocompatibility
  • hemocompatibility
  • diagnosis
  • drug delivery
  • tissue engineering

Published Papers (7 papers)

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Research

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22 pages, 8464 KiB  
Article
Towards the Standardization of Intestinal In Vitro Advanced Barrier Model for Nanoparticles Uptake and Crossing: The SiO2 Case Study
by Olimpia Vincentini, Valentina Prota, Serena Cecchetti, Lucia Bertuccini, Antonella Tinari, Francesca Iosi and Isabella De Angelis
Cells 2022, 11(21), 3357; https://doi.org/10.3390/cells11213357 - 25 Oct 2022
Cited by 2 | Viewed by 2402
Abstract
Increasing interest is being addressed to the development of a reliable, reproducible and relevant in vitro model of intestinal barrier, mainly for engineered nanomaterials hazard and risk assessment, in order to meet regulatory and scientific demands. Starting from the consolidated Caco-2 cell model, [...] Read more.
Increasing interest is being addressed to the development of a reliable, reproducible and relevant in vitro model of intestinal barrier, mainly for engineered nanomaterials hazard and risk assessment, in order to meet regulatory and scientific demands. Starting from the consolidated Caco-2 cell model, widely used for determining translocation of drugs and chemicals, the establishment of an advanced intestinal barrier model with different level of complexity is important for overcoming Caco-2 monoculture limitations. For this purpose, a tri-culture model, consisting of two human intestinal epithelial cells (Caco-2 and HT29-MTX) and a human lymphocyte B cell (Raji B), was developed by several research groups to mimic the in vivo intestinal epithelium, furnishing appropriate tools for nanotoxicological studies. However, tri-culture model shows high levels of variability in ENM uptake/translocation studies. With the aim of implementing the standardization and optimization of this tri-culture for ENM translocation studies, the present paper intends to identify and discuss such relevant parameters involved in model establishment as: tri-culture condition set-up, barrier integrity evaluation, mucus characterization, M-cell induction. SiO2 fluorescent nanoparticles were used to compare the different models. Although a low level of SiO2 translocation is reported for all the different culture conditions. a relevant role of mucus and M-cells in NPs uptake/translocation has been highlighted. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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14 pages, 1649 KiB  
Article
Formation of Lymphoma Hybrid Spheroids and Drug Testing in Real Time with the Use of Fluorescence Optical Tweezers
by Kamila Duś-Szachniewicz, Katarzyna Gdesz-Birula, Emilia Nowosielska, Piotr Ziółkowski and Sławomir Drobczyński
Cells 2022, 11(13), 2113; https://doi.org/10.3390/cells11132113 - 05 Jul 2022
Viewed by 2317
Abstract
Interactions between stromal and lymphoma cells in the bone marrow are closely related to drug resistance and therapy failure. Physiologically relevant pre-clinical three-dimensional (3D) models recapitulating lymphoma microenvironmental complexity do not currently exist. In this study, we proposed a scheme for optically controlled [...] Read more.
Interactions between stromal and lymphoma cells in the bone marrow are closely related to drug resistance and therapy failure. Physiologically relevant pre-clinical three-dimensional (3D) models recapitulating lymphoma microenvironmental complexity do not currently exist. In this study, we proposed a scheme for optically controlled hybrid lymphoma spheroid formation with the use of optical tweezers (OT). Following the preparation of stromal spheroids using agarose hydrogel, two aggressive non-Hodgkin lymphoma B-cell lines, Ri-1 (DLBCL) and Raji (Burkitt lymphoma), were used to conduct multi-cellular spheroid formation driven by in-house-developed fluorescence optical tweezers. Importantly, the newly formed hybrid spheroid preserved the 3D architecture for the next 24 h. Our model was successfully used for the evaluation of the influence of the anticancer agents doxorubicin (DOX), ibrutinib (IBR), and AMD3100 (plerixafor) on the adhesive properties of lymphoma cells. Importantly, our study revealed that a co-treatment of DOX and IBR with AMD3100 affects the adhesion of B-NHL lymphoma cells. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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11 pages, 2871 KiB  
Article
Genotoxicity Response of Fibroblast Cells and Human Epithelial Adenocarcinoma In Vitro Model Exposed to Bare and Ozone-Treated Silica Microparticles
by Sabrina Colafarina, Piero Di Carlo, Osvaldo Zarivi, Massimo Aloisi, Alessandra Di Serafino, Eleonora Aruffo, Lorenzo Arrizza, Tania Limongi and Anna Poma
Cells 2022, 11(2), 226; https://doi.org/10.3390/cells11020226 - 11 Jan 2022
Cited by 1 | Viewed by 2202
Abstract
Indoor air pollutants (IAP), which can pose a serious risk to human health, include biological pollutants, nitric oxide (NO), nitrogen dioxide (NO2), volatile organic compounds (VOC), sulfur dioxide (SO2), carbon monoxide (CO), carbon dioxide (CO2), silica, metals, [...] Read more.
Indoor air pollutants (IAP), which can pose a serious risk to human health, include biological pollutants, nitric oxide (NO), nitrogen dioxide (NO2), volatile organic compounds (VOC), sulfur dioxide (SO2), carbon monoxide (CO), carbon dioxide (CO2), silica, metals, radon, and particulate matter (PM). The aim of our work is to conduct a multidisciplinary study of fine silica particles (<2.5 μm) in the presence or absence of ozone (O3), and evaluate their potential cytotoxicity using MTS, micronucleus, and the comet test in two cell lines. We analyzed A549 (human basal alveolar epithelial cell adenocarcinoma) and Hs27 (human normal fibroblasts) exposed to dynamic conditions by an IRC simulator under ozone flow (120 ppb) and in the presence of silica particles (40 μg/h). The viability of A549 and Hs27 cells at 48 and 72 h of exposure to silica or silica/ozone decreases, except at 72 h in Hs27 treated with silica/ozone. The micronucleus and comet tests showed a significant increase in the number of micronuclei and the % of DNA in the queue, compared to the control, in both lines in all treatments, even if in different cell times/types. We found that silica alone or with more O3 causes more pronounced genotoxic effects in A549 tumor cells than in normal Hs27 fibroblasts. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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18 pages, 7513 KiB  
Article
Assessment of Collagen-Based Nanostructured Biomimetic Systems with a Co-Culture of Human Bone-Derived Cells
by Giorgia Borciani, Giorgia Montalbano, Priscila Melo, Nicola Baldini, Gabriela Ciapetti and Chiara Vitale-Brovarone
Cells 2022, 11(1), 26; https://doi.org/10.3390/cells11010026 - 23 Dec 2021
Cited by 3 | Viewed by 3374
Abstract
Osteoporosis is a worldwide disease resulting in the increase of bone fragility and enhanced fracture risk in adults. In the context of osteoporotic fractures, bone tissue engineering (BTE), i.e., the use of bone substitutes combining biomaterials, cells, and other factors, is considered a [...] Read more.
Osteoporosis is a worldwide disease resulting in the increase of bone fragility and enhanced fracture risk in adults. In the context of osteoporotic fractures, bone tissue engineering (BTE), i.e., the use of bone substitutes combining biomaterials, cells, and other factors, is considered a potential alternative to conventional treatments. Innovative scaffolds need to be tested in in vitro systems where the simultaneous presence of osteoblasts (OBs) and osteoclasts (OCs), the two main players of bone remodeling, is required to mimic their crosstalk and molecular cooperation. To this aim, two composite materials were developed, based on type I collagen, and containing either strontium-enriched mesoporous bioactive glasses or rod-like hydroxyapatite nanoparticles. The developed nanostructured systems underwent genipin chemical crosslinking and were then tested with an indirect co-culture of human trabecular bone-derived OBs and buffy coat-derived OC precursors, for 2–3 weeks. The favorable structural and biological properties of the materials proved to successfully support the viability, adhesion, and differentiation of cells, encouraging a further investigation of the developed bioactive systems as biomaterial inks for the 3D printing of more complex scaffolds for BTE. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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Review

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56 pages, 10701 KiB  
Review
Advances in Concentration Gradient Generation Approaches in a Microfluidic Device for Toxicity Analysis
by Nicole M. E. Valle, Mariana P. Nucci, Arielly H. Alves, Luiz D. Rodrigues, Javier B. Mamani, Fernando A. Oliveira, Caique S. Lopes, Alexandre T. Lopes, Marcelo N. P. Carreño and Lionel F. Gamarra
Cells 2022, 11(19), 3101; https://doi.org/10.3390/cells11193101 - 01 Oct 2022
Cited by 5 | Viewed by 2586
Abstract
This systematic review aimed to analyze the development and functionality of microfluidic concentration gradient generators (CGGs) for toxicological evaluation of different biological organisms. We searched articles using the keywords: concentration gradient generator, toxicity, and microfluidic device. Only 33 of the 352 articles found [...] Read more.
This systematic review aimed to analyze the development and functionality of microfluidic concentration gradient generators (CGGs) for toxicological evaluation of different biological organisms. We searched articles using the keywords: concentration gradient generator, toxicity, and microfluidic device. Only 33 of the 352 articles found were included and examined regarding the fabrication of the microdevices, the characteristics of the CGG, the biological model, and the desired results. The main fabrication method was soft lithography, using polydimethylsiloxane (PDMS) material (91%) and SU-8 as the mold (58.3%). New technologies were applied to minimize shear and bubble problems, reduce costs, and accelerate prototyping. The Christmas tree CGG design and its variations were the most reported in the studies, as well as the convective method of generation (61%). Biological models included bacteria and nematodes for antibiotic screening, microalgae for pollutant toxicity, tumor and normal cells for, primarily, chemotherapy screening, and Zebrafish embryos for drug and metal developmental toxicity. The toxic effects of each concentration generated were evaluated mostly with imaging and microscopy techniques. This study showed an advantage of CGGs over other techniques and their applicability for several biological models. Even with soft lithography, PDMS, and Christmas tree being more popular in their respective categories, current studies aim to apply new technologies and intricate architectures to improve testing effectiveness and reduce common microfluidics problems, allowing for high applicability of toxicity tests in different medical and environmental models. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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16 pages, 1608 KiB  
Review
Microfluidics for 3D Cell and Tissue Cultures: Microfabricative and Ethical Aspects Updates
by Tania Limongi, Francesco Guzzi, Elvira Parrotta, Patrizio Candeloro, Stefania Scalise, Valeria Lucchino, Francesco Gentile, Luca Tirinato, Maria Laura Coluccio, Bruno Torre, Marco Allione, Monica Marini, Francesca Susa, Enzo Di Fabrizio, Giovanni Cuda and Gerardo Perozziello
Cells 2022, 11(10), 1699; https://doi.org/10.3390/cells11101699 - 20 May 2022
Cited by 6 | Viewed by 3675
Abstract
The necessity to improve in vitro cell screening assays is becoming ever more important. Pharmaceutical companies, research laboratories and hospitals require technologies that help to speed up conventional screening and therapeutic procedures to produce more data in a short time in a realistic [...] Read more.
The necessity to improve in vitro cell screening assays is becoming ever more important. Pharmaceutical companies, research laboratories and hospitals require technologies that help to speed up conventional screening and therapeutic procedures to produce more data in a short time in a realistic and reliable manner. The design of new solutions for test biomaterials and active molecules is one of the urgent problems of preclinical screening and the limited correlation between in vitro and in vivo data remains one of the major issues. The establishment of the most suitable in vitro model provides reduction in times, costs and, last but not least, in the number of animal experiments as recommended by the 3Rs (replace, reduce, refine) ethical guiding principles for testing involving animals. Although two-dimensional (2D) traditional cell screening assays are generally cheap and practical to manage, they have strong limitations, as cells, within the transition from the three-dimensional (3D) in vivo to the 2D in vitro growth conditions, do not properly mimic the real morphologies and physiology of their native tissues. In the study of human pathologies, especially, animal experiments provide data closer to what happens in the target organ or apparatus, but they imply slow and costly procedures and they generally do not fully accomplish the 3Rs recommendations, i.e., the amount of laboratory animals and the stress that they undergo must be minimized. Microfluidic devices seem to offer different advantages in relation to the mentioned issues. This review aims to describe the critical issues connected with the conventional cells culture and screening procedures, showing what happens in the in vivo physiological micro and nano environment also from a physical point of view. During the discussion, some microfluidic tools and their components are described to explain how these devices can circumvent the actual limitations described in the introduction. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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17 pages, 740 KiB  
Review
The Effective Combination between 3D Cancer Models and Stimuli-Responsive Nanoscale Drug Delivery Systems
by Federica Foglietta, Loredana Serpe and Roberto Canaparo
Cells 2021, 10(12), 3295; https://doi.org/10.3390/cells10123295 - 25 Nov 2021
Cited by 11 | Viewed by 2317
Abstract
Stimuli-responsive drug-delivery systems (DDSs) have emerged as a potential tool for applications in healthcare, mainly in the treatment of cancer where versatile nanocarriers are co-triggered by endogenous and exogenous stimuli. Two-dimensional (2D) cell cultures are the most important in vitro model used to [...] Read more.
Stimuli-responsive drug-delivery systems (DDSs) have emerged as a potential tool for applications in healthcare, mainly in the treatment of cancer where versatile nanocarriers are co-triggered by endogenous and exogenous stimuli. Two-dimensional (2D) cell cultures are the most important in vitro model used to evaluate the anticancer activity of these stimuli-responsive DDSs due to their easy manipulation and versatility. However, some limitations suggest that these in vitro models poorly predict the outcome of in vivo studies. One of the main drawbacks of 2D cell cultures is their inadequate representation of the 3D environment’s physiological complexity, which sees cells interact with each other and the extracellular matrix (ECM) according to their specific cellular organization. In this regard, 3D cancer models are a promising approach that can overcome the main shortcomings of 2D cancer cell cultures, as these in vitro models possess many peculiarities by which they mimic in vivo tumors, including physiologically relevant cell–cell and cell–ECM interactions. This is, in our opinion, even more relevant when a stimuli-responsive DDS is being investigated. In this review, we therefore report and discuss endogenous and exogenous stimuli-responsive DDSs whose effectiveness has been tested using 3D cancer cell cultures. Full article
(This article belongs to the Special Issue In Vitro Model for Micro and Nano Technologies)
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