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Article ; Online: High Concentrations of Cannabidiol Induce Neurotoxicity in Neurosphere Culture System.

Romariz, Simone A A / Sanabria, Viviam / da Silva, Karina Ribeiro / Quintella, Miguel L / de Melo, Bruna A G / Porcionatto, Marimélia / de Almeida, Danilo Candido / Longo, Beatriz M

Neurotoxicity research

2024 Volume 42, Issue 1, Page(s) 14

Abstract: Recent studies have demonstrated that cannabinoids are potentially effective in the treatment of various neurological conditions, and cannabidiol (CBD), one of the most studied compounds, has been proposed as a non-toxic option. However, the adverse ... ...

Abstract	Recent studies have demonstrated that cannabinoids are potentially effective in the treatment of various neurological conditions, and cannabidiol (CBD), one of the most studied compounds, has been proposed as a non-toxic option. However, the adverse effects of CBD on neurodevelopmental processes have rarely been studied in cell culture systems. To better understand CBD's influence on neurodevelopment, we exposed neural progenitor cells (NPCs) to different concentrations of CBD (1 µM, 5 µM, and 10 µM). We assessed the morphology, migration, differentiation, cell death, and gene expression in 2D and 3D bioprinted models to stimulate physiological conditions more effectively. Our results showed that CBD was more toxic at higher concentrations (5 µM and 10 µM) and affected the viability of NPCs than at lower concentrations (1 µM), in both 2D and 3D models. Moreover, our study revealed that higher concentrations of CBD drastically reduced the size of neurospheres and the number of NPCs within neurospheres, impaired the morphology and mobility of neurons and astrocytes after differentiation, and reduced neurite sprouting. Interestingly, we also found that CBD alters cellular metabolism by influencing the expression of glycolytic and β-oxidative enzymes in the early and late stages of metabolic pathways. Therefore, our study demonstrated that higher concentrations of CBD promote important changes in cellular functions that are crucial during CNS development.
MeSH term(s)	Humans ; Cannabidiol/toxicity ; Neurotoxicity Syndromes ; Neurons ; Astrocytes ; Carbidopa
Chemical Substances	Cannabidiol (19GBJ60SN5) ; Carbidopa (MNX7R8C5VO)
Language	English
Publishing date	2024-02-13
Publishing country	United States
Document type	Journal Article
ZDB-ID	2036826-4
ISSN	1476-3524 ; 1029-8428
ISSN (online)	1476-3524
ISSN	1029-8428
DOI	10.1007/s12640-024-00692-5
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Article ; Online: 3D Bioprinting of Murine Cortical Astrocytes for Engineering Neural-Like Tissue.

de Melo, Bruna A G / Cruz, Elisa M / Ribeiro, Taís N / Mundim, Mayara V / Porcionatto, Marimelia A

Journal of visualized experiments : JoVE

2021 , Issue 173

Abstract: Astrocytes are glial cells with an essential role in the central nervous system (CNS), including neuronal support and functionality. These cells also respond to neural injuries and act to protect the tissue from degenerative events. In vitro studies of ... ...

Abstract	Astrocytes are glial cells with an essential role in the central nervous system (CNS), including neuronal support and functionality. These cells also respond to neural injuries and act to protect the tissue from degenerative events. In vitro studies of astrocytes' functionality are important to elucidate the mechanisms involved in such events and contribute to developing therapies to treat neurological disorders. This protocol describes a method to biofabricate a neural-like tissue structure rich in astrocytes by 3D bioprinting astrocytes-laden bioink. An extrusion-based 3D bioprinter was used in this work, and astrocytes were extracted from C57Bl/6 mice pups' brain cortices. The bioink was prepared by mixing cortical astrocytes from up to passage 3 to a biomaterial solution composed of gelatin, gelatin-methacryloyl (GelMA), and fibrinogen, supplemented with laminin, which presented optimal bioprinting conditions. The 3D bioprinting conditions minimized cell stress, contributing to the high viability of the astrocytes during the process, in which 74.08% ± 1.33% of cells were viable right after bioprinting. After 1 week of incubation, the viability of astrocytes significantly increased to 83.54% ± 3.00%, indicating that the 3D construct represents a suitable microenvironment for cell growth. The biomaterial composition allowed cell attachment and stimulated astrocytic behavior, with cells expressing the specific astrocytes marker glial fibrillary acidic protein (GFAP) and possessing typical astrocytic morphology. This reproducible protocol provides a valuable method to biofabricate 3D neural-like tissue rich in astrocytes that resembles cells' native microenvironment, useful to researchers that aim to understand astrocytes' functionality and their relation to the mechanisms involved in neurological diseases.
MeSH term(s)	Animals ; Astrocytes ; Bioprinting ; Gelatin ; Mice ; Printing, Three-Dimensional ; Tissue Engineering ; Tissue Scaffolds
Chemical Substances	Gelatin (9000-70-8)
Language	English
Publishing date	2021-07-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/62691
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Article: Simple and efficient protocol to isolate and culture brain microvascular endothelial cells from newborn mice.

Nicolicht-Amorim, Priscila / Delgado-Garcia, Lina M / Nakamura, Thabatta Karollynne Estevam / Courbassier, Natália Rodrigues / Mosini, Amanda Cristina / Porcionatto, Marimelia A

Frontiers in cellular neuroscience

2022 Volume 16, Page(s) 949412

Abstract: The neurovascular unit (NVU) is a multicellular structure comprising of neurons, glial cells, and non-neural cells, and it is supported by a specialized extracellular matrix, the basal lamina. Astrocytes, brain microvascular endothelial cells (BMECs), ... ...

Abstract	The neurovascular unit (NVU) is a multicellular structure comprising of neurons, glial cells, and non-neural cells, and it is supported by a specialized extracellular matrix, the basal lamina. Astrocytes, brain microvascular endothelial cells (BMECs), pericytes, and smooth muscle cells constitute the blood-brain barrier (BBB). BMECs have a mesodermal origin and invade the nervous system early in neural tube development, forming the BBB anatomical core. BMECs are connected by adherent junction complexes composed of integral membrane and cytoplasmic proteins.
Language	English
Publishing date	2022-10-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2452963-1
ISSN	1662-5102
ISSN	1662-5102
DOI	10.3389/fncel.2022.949412
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Article ; Online: Temporal pattern of Fos and Jun families expression after mitogenic stimulation with FGF-2 in rat neural stem cells and fibroblasts.

Mosini, A C / Mazzonetto, P C / Calió, M L / Pompeu, C / Massinhani, F H / Nakamura, T K E / Pires, J M / Silva, C S / Porcionatto, M A / Mello, L E

Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas

2023 Volume 56, Page(s) e12546

Abstract: Intense stimulation of most living cells triggers the activation of immediate early genes, such as Fos and Jun families. These genes are important in cellular and biochemical processes, such as mitosis and cell death. The present study focused on ... ...

Abstract	Intense stimulation of most living cells triggers the activation of immediate early genes, such as Fos and Jun families. These genes are important in cellular and biochemical processes, such as mitosis and cell death. The present study focused on determining the temporal expression pattern of Fos and Jun families in fibroblasts and neural stem cells of cerebellum, hippocampus, and subventricular zone (SVZ) of rats of different ages at 0, 0.5, 1, 3, and 6 h after stimulation with fibroblast growth factor (FGF)-2. In neonates, a similar expression pattern was observed in all cells analyzed, with lower expression in basal condition, peak expression at 0.5 h after stimulation, returning to baseline values between 1 and 3 h after stimulation. On the other hand, cells from adult animals only showed Fra1 and JunD expression after stimulation. In fibroblasts and hippocampus, Fra1 reached peak expression at 0.5 h after stimulation, while in the SVZ, peak level was observed at 6 h after stimulation. JunD in fibroblasts presented two peak expressions, at 0.5 and 6 h after stimulation. Between these periods, the expression observed was at a basal level. Nevertheless, JunD expression in SVZ and hippocampus was low and without significant changes after stimulation. Differences in mRNA expression in neonate and adult animals characterize the significant differences in neurogenesis and cell response to stimulation at different stages of development. Characterizing these differences might be important for the development of cell cultures, replacement therapy, and the understanding of the physiological response profile of different cell types.
MeSH term(s)	Animals ; Rats ; Fibroblast Growth Factor 2 ; Mitogens ; Cell Proliferation ; Fibroblasts ; Neural Stem Cells
Chemical Substances	Fibroblast Growth Factor 2 (103107-01-3) ; Mitogens
Language	English
Publishing date	2023-09-08
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	786234-9
ISSN	1414-431X ; 0100-879X
ISSN (online)	1414-431X
ISSN	0100-879X
DOI	10.1590/1414-431X2023e12546
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Article ; Online: 3D culture models to study SARS-CoV-2 infectivity and antiviral candidates: From spheroids to bioprinting.

de Melo, Bruna A G / Benincasa, Julia C / Cruz, Elisa M / Maricato, Juliana Terzi / Porcionatto, Marimelia A

Biomedical journal

2020 Volume 44, Issue 1, Page(s) 31–42

Abstract: The pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is receiving worldwide attention, due to the severity of the disease (COVID-19) that resulted in more than a million global deaths so far. The urgent need for vaccines ... ...

Abstract	The pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is receiving worldwide attention, due to the severity of the disease (COVID-19) that resulted in more than a million global deaths so far. The urgent need for vaccines and antiviral drugs is mobilizing the scientific community to develop strategies for studying the mechanisms of SARS-CoV-2 infection, replication kinetics, pathogenesis, host-virus interaction, and infection inhibition. In this work, we review the strategies of tissue engineering in the fabrication of three-dimensional (3D) models used in virology studies, which presented many advantages over conventional cell cultures, such as complex cytoarchitecture and a more physiological microenvironment. Scaffold-free (spheroids and organoids) and scaffold-based (3D scaffolding and 3D bioprinting) approach allow the biofabrication of more realistic models relevant to the pandemic, to be used as in vitro platforms for the development of new vaccines and therapies against COVID-19.
MeSH term(s)	Angiotensin-Converting Enzyme 2/physiology ; Animals ; Antiviral Agents/pharmacology ; Bioprinting ; Humans ; Organoids ; SARS-CoV-2/drug effects ; SARS-CoV-2/pathogenicity ; Spheroids, Cellular ; Tissue Engineering/methods ; Tissue Scaffolds
Chemical Substances	Antiviral Agents ; ACE2 protein, human (EC 3.4.17.23) ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
Language	English
Publishing date	2020-11-21
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2698541-X
ISSN	2320-2890 ; 2320-2890
ISSN (online)	2320-2890
ISSN	2320-2890
DOI	10.1016/j.bj.2020.11.009
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Article ; Online: Therapeutic Potential of Targeting Prokineticin Receptors in Diseases.

Vincenzi, Martina / Kremić, Amin / Jouve, Appoline / Lattanzi, Roberta / Miele, Rossella / Benharouga, Mohamed / Alfaidy, Nadia / Migrenne-Li, Stephanie / Kanthasamy, Anumantha G / Porcionatto, Marimelia / Ferrara, Napoleone / Tetko, Igor V / Désaubry, Laurent / Nebigil, Canan G

Pharmacological reviews

2023 Volume 75, Issue 6, Page(s) 1167–1199

Abstract: The prokineticins (PKs) were discovered approximately 20 years ago as small peptides inducing gut contractility. Today, they are established as angiogenic, anorectic, and proinflammatory cytokines, chemokines, hormones, and neuropeptides involved in ... ...

Abstract	The prokineticins (PKs) were discovered approximately 20 years ago as small peptides inducing gut contractility. Today, they are established as angiogenic, anorectic, and proinflammatory cytokines, chemokines, hormones, and neuropeptides involved in variety of physiologic and pathophysiological pathways. Their altered expression or mutations implicated in several diseases make them a potential biomarker. Their G-protein coupled receptors, PKR1 and PKR2, have divergent roles that can be therapeutic target for treatment of cardiovascular, metabolic, and neural diseases as well as pain and cancer. This article reviews and summarizes our current knowledge of PK family functions from development of heart and brain to regulation of homeostasis in health and diseases. Finally, the review summarizes the established roles of the endogenous peptides, synthetic peptides and the selective ligands of PKR1 and PKR2, and nonpeptide orthostatic and allosteric modulator of the receptors in preclinical disease models. The present review emphasizes the ambiguous aspects and gaps in our knowledge of functions of PKR ligands and elucidates future perspectives for PK research. SIGNIFICANCE STATEMENT: This review provides an in-depth view of the prokineticin family and PK receptors that can be active without their endogenous ligand and exhibits "constitutive" activity in diseases. Their non- peptide ligands display promising effects in several preclinical disease models. PKs can be the diagnostic biomarker of several diseases. A thorough understanding of the role of prokineticin family and their receptor types in health and diseases is critical to develop novel therapeutic strategies with safety concerns.
MeSH term(s)	Humans ; Receptors, G-Protein-Coupled/metabolism ; Neuropeptides/metabolism ; Peptides ; Neoplasms/drug therapy ; Biomarkers
Chemical Substances	Receptors, G-Protein-Coupled ; Neuropeptides ; Peptides ; Biomarkers
Language	English
Publishing date	2023-09-08
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	209898-2
ISSN	1521-0081 ; 0031-6997
ISSN (online)	1521-0081
ISSN	0031-6997
DOI	10.1124/pharmrev.122.000801
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Article ; Online: Neuroprotective effect of heparin Trisulfated disaccharide on ischemic stroke.

Chiarantin, Gabrielly M D / Delgado-Garcia, Lina M / Zamproni, Laura N / Lima, Marcelo A / Nader, Helena B / Tersariol, Ivarne L S / Porcionatto, Marimélia

Glycoconjugate journal

2021 Volume 38, Issue 1, Page(s) 35–43

Abstract: Cells undergoing hypoxia experience intense cytoplasmic calcium ( ... ...

Abstract	Cells undergoing hypoxia experience intense cytoplasmic calcium (Ca
MeSH term(s)	Animals ; Calcium/metabolism ; Cell Death/drug effects ; Cell Hypoxia/drug effects ; Cell Survival/drug effects ; Disaccharides/chemistry ; Disaccharides/pharmacology ; Heparin/analogs & derivatives ; Heparin/chemistry ; Heparin/pharmacology ; Ischemic Stroke/metabolism ; Ischemic Stroke/pathology ; Ischemic Stroke/prevention & control ; Male ; Mice, Inbred C57BL ; Neurons/drug effects ; Neurons/pathology ; Neuroprotective Agents/chemistry ; Neuroprotective Agents/pharmacology ; Thapsigargin/pharmacology ; Thiourea/analogs & derivatives ; Thiourea/pharmacology ; Mice
Chemical Substances	2-(2-(4-(4-nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate ; Disaccharides ; Neuroprotective Agents ; heparin disaccharide ; Thapsigargin (67526-95-8) ; Heparin (9005-49-6) ; Thiourea (GYV9AM2QAG) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2021-01-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	283770-5
ISSN	1573-4986 ; 0282-0080
ISSN (online)	1573-4986
ISSN	0282-0080
DOI	10.1007/s10719-020-09966-4
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Zs.A 2171: Show issues

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Article: The extracellular matrix provides directional cues for neuronal migration during cerebellar development.

Porcionatto, M A

Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas

2006 Volume 39, Issue 3, Page(s) 313–320

Abstract: Normal central nervous system development relies on accurate intrinsic cellular programs as well as on extrinsic informative cues provided by extracellular molecules. Migration of neuronal progenitors from defined proliferative zones to their final ... ...

Abstract	Normal central nervous system development relies on accurate intrinsic cellular programs as well as on extrinsic informative cues provided by extracellular molecules. Migration of neuronal progenitors from defined proliferative zones to their final location is a key event during embryonic and postnatal development. Extracellular matrix components play important roles in these processes, and interactions between neurons and extracellular matrix are fundamental for the normal development of the central nervous system. Guidance cues are provided by extracellular factors that orient neuronal migration. During cerebellar development, the extracellular matrix molecules laminin and fibronectin give support to neuronal precursor migration, while other molecules such as reelin, tenascin, and netrin orient their migration. Reelin and tenascin are extracellular matrix components that attract or repel neuronal precursors and axons during development through interaction with membrane receptors, and netrin associates with laminin and heparan sulfate proteoglycans, and binds to the extracellular matrix receptor integrins present on the neuronal surface. Altogether, the dynamic changes in the composition and distribution of extracellular matrix components provide external cues that direct neurons leaving their birthplaces to reach their correct final location. Understanding the molecular mechanisms that orient neurons to reach precisely their final location during development is fundamental to understand how neuronal misplacement leads to neurological diseases and eventually to find ways to treat them.
MeSH term(s)	Cell Adhesion Molecules, Neuronal/physiology ; Cell Movement/physiology ; Cerebellum/embryology ; Extracellular Matrix/physiology ; Extracellular Matrix Proteins/physiology ; Humans ; Nerve Tissue Proteins/physiology ; Neurons/physiology ; Signal Transduction/physiology
Chemical Substances	Cell Adhesion Molecules, Neuronal ; Extracellular Matrix Proteins ; Nerve Tissue Proteins
Language	English
Publishing date	2006-02-22
Publishing country	Brazil
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	786234-9
ISSN	0100-879X
ISSN	0100-879X
DOI	10.1590/s0100-879x2006000300001
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Article ; Online: Strategies to use fibrinogen as bioink for 3D bioprinting fibrin-based soft and hard tissues.

de Melo, Bruna A G / Jodat, Yasamin A / Cruz, Elisa M / Benincasa, Julia C / Shin, Su Ryon / Porcionatto, Marimelia A

Acta biomaterialia

2020 Volume 117, Page(s) 60–76

Abstract: Fibrin gel has been widely used for engineering various types of tissues due to its biocompatible nature, biodegradability, and tunable mechanical and nanofibrous structural properties. Despite their promising regenerative capacity and extensive ... ...

Abstract	Fibrin gel has been widely used for engineering various types of tissues due to its biocompatible nature, biodegradability, and tunable mechanical and nanofibrous structural properties. Despite their promising regenerative capacity and extensive biocompatibility with various tissue types, fibrin-based biomaterials are often notoriously known as burdensome candidates for 3D biofabrication and bioprinting. The high viscosity of fibrin (crosslinked form) hinders proper ink extrusion, and its pre-polymer form, fibrinogen, is not capable of maintaining shape fidelity. To overcome these limitations and empower fibrinogen-based bioinks for fibrin biomimetics and regenerative applications, different strategies can be practiced. The aim of this review is to report the strategies that bring fabrication compatibility to these bioinks through mixing fibrinogen with printable biomaterials, using supporting bath supplemented with crosslinking agents, and crosslinking fibrin in situ. Moreover, the review discusses some of the recent advances in 3D bioprinting of biomimetic soft and hard tissues using fibrinogen-based bioinks, and highlights the impacts of these strategies on fibrin properties, its bioactivity, and the functionality of the consequent biomimetic tissue. Statement of Significance Due to its biocompatible nature, biodegradability, and tunable mechanical and nanofibrous structural properties, fibrin gel has been widely employed in tissue engineering and more recently, used as in 3D bioprinting. The fibrinogen's poor printable properties make it difficult to maintain the 3D shape of bioprinted constructs. Our work describes the strategies employed in tissue engineering to allow the 3D bioprinting of fibrinogen-based bioinks, such as the combination of fibrinogen with printable biomaterials, the in situ fibrin crosslinking, and the use of supporting bath supplemented with crosslinking agents. Further, this review discuss the application of 3D bioprinting technology to biofabricate fibrin-based soft and hard tissues for biomedical applications, and discuss current limitations and future of such in vitro models.
MeSH term(s)	Bioprinting ; Fibrin ; Fibrinogen ; Printing, Three-Dimensional ; Tissue Engineering ; Tissue Scaffolds
Chemical Substances	Fibrin (9001-31-4) ; Fibrinogen (9001-32-5)
Language	English
Publishing date	2020-09-16
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2173841-5
ISSN	1878-7568 ; 1742-7061
ISSN (online)	1878-7568
ISSN	1742-7061
DOI	10.1016/j.actbio.2020.09.024
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Article ; Online: 3D culture models to study SARS-CoV-2 infectivity and antiviral candidates

Bruna A.G. de Melo / Julia C. Benincasa / Elisa M. Cruz / Juliana Terzi Maricato / Marimelia A. Porcionatto

Biomedical Journal, Vol 44, Iss 1, Pp 31-

From spheroids to bioprinting

2021 Volume 42

Abstract	The pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is receiving worldwide attention, due to the severity of the disease (COVID-19) that resulted in more than a million global deaths so far. The urgent need for vaccines and antiviral drugs is mobilizing the scientific community to develop strategies for studying the mechanisms of SARS-CoV-2 infection, replication kinetics, pathogenesis, host–virus interaction, and infection inhibition. In this work, we review the strategies of tissue engineering in the fabrication of three-dimensional (3D) models used in virology studies, which presented many advantages over conventional cell cultures, such as complex cytoarchitecture and a more physiological microenvironment. Scaffold-free (spheroids and organoids) and scaffold-based (3D scaffolding and 3D bioprinting) approach allow the biofabrication of more realistic models relevant to the pandemic, to be used as in vitro platforms for the development of new vaccines and therapies against COVID-19.
Keywords	SARS-CoV-2 ; COVID-19 ; Tissue engineering ; Organoids ; 3D bioprinting ; Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
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