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Innovative In Vitro Model for Medical, Pharmaceutical and Diagnostic Applications

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: closed (20 April 2025) | Viewed by 8087

Special Issue Editors


E-Mail Website1 Website2
Guest Editor
1. Department of Drug and Health Sciences, Section of Biochemistry, University of Catania, 95125 Catania, Italy
2. Research Center on Nutraceuticals and Health Products (CERNUT), University of Catania, 95125 Catania, Italy
3. Molecular, Preclinical and Translational Imaging Research Centre (IMPRONTE), University of Catania, 95125 Catania, Italy
Interests: nutraceuticals; cancer biology; biomarkers; oxidative stress and natural compounds in cancer, aging, degenerative and metabolic disease; cancer epigenetics; DNA damage and comet assay; inflammation; nanosystem for drug delivery
Special Issues, Collections and Topics in MDPI journals

E-Mail Website
Guest Editor
INOVOTION, 38700 La Tronche, France
Interests: CAM assay; chicken embryo; immuno-oncology; in ovo; preclinical model; metastasis; in vivo efficacy and toxicity of drugs

Special Issue Information

Dear Colleagues,

New in vitro systems that mimic the physiopathological complexity and dynamics of native organ/tissue are useful tools to provide more accurate information about the cell-to-cell interaction, cell–microenvironment interplay, tumor characteristics, drug discovery, drug and nutraceutical testing, metabolic profiling, infection mechanism, and to study various types of diseases. To date, conventional 2D and specifically 3D models (e.g., spheroids and patient-derived organoids), scaffold-based or scaffold-free systems, microfluidic systems, and tissue engineering represent the models of choice to improve our knowledge about human organ homeostasis and its possible disfunctions being in balance between the extreme homogeneity of conventional cell cultures on one side and the extreme complexity of real organs on the other. Moreover, 3D technology methods have made it possible to obtain human in vitro models, which have proved to be a viable alternative to in vivo animal models in preclinical diagnostic imaging, thus overcoming the issue of strict regulation of animal studies.

This Special Issue of the International Journal of Molecular Sciences titled “Innovative Platforms for Medical, Pharmaceutical and Diagnostic Applications” will include a selection of scientific contributions aimed at presenting the development and the application of such innovative models or protocols in cell biology, toxicology, infection, diseases, and drug/nutraceuticals development and screening, as well as preclinical 3D imaging in order to study physiological or pathological conditions and to address personalized and regenerative medicine approaches.

Original research, review articles, preclinical studies, short communications, and letters on all topics in these research fields are invited.

This Special Issue is now open for submissions. If you are interested in contributing your work, please send a short abstract or tentative title to the Guest editor or Editorial Office.

Dr. Angelita Costantino () is serving as co-guest editor and will assist managing this Special Issue.

Prof. Dr. Barbara Tomasello
Dr. Rosaria Acquaviva
Dr. Jean Viallet
Guest Editors

Manuscript Submission Information

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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. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

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Keywords

  • in vitro 3D models
  • organoids
  • drug and nutraceutical screening
  • preclinical imaging
  • tissue engineering
  • precision and regenerative medicine

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Published Papers (4 papers)

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Research

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12 pages, 2418 KiB  
Article
A Dynamic Cellular Model as an Emerging Platform to Reproduce the Complexity of Human Vascular Calcification In Vitro
by Elisa Ceccherini, Elisa Persiani, Manuela Cabiati, Letizia Guiducci, Silvia Del Ry, Ilaria Gisone, Alessandra Falleni, Antonella Cecchettini and Federico Vozzi
Int. J. Mol. Sci. 2024, 25(13), 7427; https://doi.org/10.3390/ijms25137427 - 6 Jul 2024
Cited by 3 | Viewed by 1262
Abstract
Vascular calcification (VC) is a cardiovascular disease characterized by calcium salt deposition in vascular smooth muscle cells (VSMCs). Standard in vitro models used in VC investigations are based on VSMC monocultures under static conditions. Although these platforms are easy to use, the absence [...] Read more.
Vascular calcification (VC) is a cardiovascular disease characterized by calcium salt deposition in vascular smooth muscle cells (VSMCs). Standard in vitro models used in VC investigations are based on VSMC monocultures under static conditions. Although these platforms are easy to use, the absence of interactions between different cell types and dynamic conditions makes these models insufficient to study key aspects of vascular pathophysiology. The present study aimed to develop a dynamic endothelial cell–VSMC co-culture that better mimics the in vivo vascular microenvironment. A double-flow bioreactor supported cellular interactions and reproduced the blood flow dynamic. VSMC calcification was stimulated with a DMEM high glucose calcification medium supplemented with 1.9 mM NaH2PO4/Na2HPO4 (1:1) for 7 days. Calcification, cell viability, inflammatory mediators, and molecular markers (SIRT-1, TGFβ1) related to VSMC differentiation were evaluated. Our dynamic model was able to reproduce VSMC calcification and inflammation and evidenced differences in the modulation of effectors involved in the VSMC calcified phenotype compared with standard monocultures, highlighting the importance of the microenvironment in controlling cell behavior. Hence, our platform represents an advanced system to investigate the pathophysiologic mechanisms underlying VC, providing information not available with the standard cell monoculture. Full article
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12 pages, 5203 KiB  
Article
Microporous Polylactic Acid Scaffolds Enable Fluorescence-Based Perfusion Imaging of Intrinsic In Vivo Vascularization
by Christoph Koepple, Lukas Pollmann, Nicola Sariye Pollmann, Matthias Schulte, Ulrich Kneser, Norbert Gretz and Volker J. Schmidt
Int. J. Mol. Sci. 2023, 24(19), 14813; https://doi.org/10.3390/ijms241914813 - 1 Oct 2023
Cited by 4 | Viewed by 1637
Abstract
In vivo tissue engineering (TE) techniques like the AV loop model provide an isolated and well-defined microenvironment to study angiogenesis-related cell interactions. Functional visualization of the microvascular network within these artificial tissue constructs is crucial for the fundamental understanding of vessel network formation [...] Read more.
In vivo tissue engineering (TE) techniques like the AV loop model provide an isolated and well-defined microenvironment to study angiogenesis-related cell interactions. Functional visualization of the microvascular network within these artificial tissue constructs is crucial for the fundamental understanding of vessel network formation and to identify the underlying key regulatory mechanisms. To facilitate microvascular tracking advanced fluorescence imaging techniques are required. We studied the suitability of microporous polylactic acid (PLA) scaffolds with known low autofluorescence to form axial vascularized tissue constructs in the AV loop model and to validate these scaffolds for fluorescence-based perfusion imaging. Compared to commonly used collagen elastin (CE) scaffolds, the total number of vessels and cells in PLA scaffolds was lower. In detail, CE-based constructs exhibited significantly higher vessel numbers on day 14 and 28 (d14: 316 ± 53; d28: 610 ± 74) compared to the respective time points in PLA-based constructs (d14: 144 ± 18; d28: 327 ± 34; each p < 0.05). Analogously, cell counts in CE scaffolds were higher compared to corresponding PLA constructs (d14: 7661.25 ± 505.93 and 5804.04 ± 716.59; d28: 11211.75 + 1278.97 and 6045.71 ± 572.72, p < 0.05). CE scaffolds showed significantly higher vessel densities in proximity to the main vessel axis compared to PLA scaffolds (200–400 µm and 600–800 µm on day 14; 400–1000 µm and 1400–1600 µm on day 28). CE scaffolds had significantly higher cell counts on day 14 at distances from 800 to 2000 µm and at distances from 400 to 1600 µm on day 28. While the total number of vessels and cells in PLA scaffolds were lower, both scaffold types were ideally suited for axial vascularization techniques. The intravascular perfusion of PLA-based constructs with fluorescence dye MHI148-PEI demonstrated dye specificity against vascular walls of low- and high-order branches as well as capillaries and facilitated the fluorescence-based visualization of microcirculatory networks. Fluorophore tracking may contribute to the development of automated quantification methods after 3D reconstruction and image segmentation. These technologies may facilitate the characterization of key regulators within specific subdomains and add to the current understanding of vessel formation in axially vascularized tissue constructs. Full article
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17 pages, 2900 KiB  
Article
Monitoring Cultured Rat Hepatocytes Using RNA-Seq In Vitro
by Yung-Te Hou, Chia-Chun Wu, Wen-Ting Wang, Wen-Tse Yang, Ying-Hsiu Liao and Chien-Yu Chen
Int. J. Mol. Sci. 2023, 24(8), 7534; https://doi.org/10.3390/ijms24087534 - 19 Apr 2023
Cited by 3 | Viewed by 1843
Abstract
Compared to other techniques, RNA sequencing (RNA-Seq) has the advantage of having details of the expression abundance of all transcripts in a single run. In this study, we used RNA-Seq to monitor the maturity and dynamic characteristics of in vitro hepatocyte cultures. Hepatocytes, [...] Read more.
Compared to other techniques, RNA sequencing (RNA-Seq) has the advantage of having details of the expression abundance of all transcripts in a single run. In this study, we used RNA-Seq to monitor the maturity and dynamic characteristics of in vitro hepatocyte cultures. Hepatocytes, including mature hepatocytes and small hepatocytes, were analyzed in vitro using RNA-Seq and quantitative polymerase chain reaction (qPCR). The results demonstrated that the gene expression profiles measured by RNA-Seq showed a similar trend to the expression profiles measured by qPCR, and can be used to infer the success of in vitro hepatocyte cultures. The results of the differential analysis, which compared mature hepatocytes against small hepatocytes, revealed 836 downregulated and 137 upregulated genes. In addition, the success of the hepatocyte cultures could be explained by the gene list screened from the adopted gene enrichment test. In summary, we demonstrated that RNA-Seq could become an effective method for monitoring the whole transcriptome of hepatocyte cultures and provide a more comprehensive list of factors related to the differentiation of small hepatocytes into mature hepatocytes. This monitoring system not only shows high potential in medical applications but may also be a novel method for the clinical diagnosis of liver-related diseases. Full article
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Review

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13 pages, 1175 KiB  
Review
Three-Dimensional iPSC-Based In Vitro Cardiac Models for Biomedical and Pharmaceutical Research Applications
by Simona Bufi and Rosaria Santoro
Int. J. Mol. Sci. 2024, 25(19), 10690; https://doi.org/10.3390/ijms251910690 - 4 Oct 2024
Cited by 1 | Viewed by 1864
Abstract
Cardiovascular diseases are a major cause of death worldwide. Advanced in vitro models can be the key stone for a better understanding of the mechanisms at the basis of the different pathologies, supporting the development of novel therapeutic protocols. In particular, the implementation [...] Read more.
Cardiovascular diseases are a major cause of death worldwide. Advanced in vitro models can be the key stone for a better understanding of the mechanisms at the basis of the different pathologies, supporting the development of novel therapeutic protocols. In particular, the implementation of induced pluripotent stem cell (iPSC) technology allows for the generation of a patient-specific pluripotent cell line that is able to differentiate in several organ-specific cell subsets while retaining the patient genetic background, thus putting the basis for personalized in vitro modeling toward personalized medicine. The design of iPSC-based models able to recapitulate the complexity of the cardiac environment is a critical goal. Here, we review some of the published efforts to exploit three dimensional (3D) iPSC-based methods to recapitulate the relevant cardiomyopathies, including genetically and non-genetically determined cardiomyopathies and cardiotoxicity studies. Finally, we discuss the actual method limitations and the future perspectives in the field. Full article
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