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An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle

The development of novel nanoconstructs for biomedical applications requires the assessment of their biodistribution, metabolism and clearance in single cells, organs and entire organisms in a living environment. To reduce the number of in vivo experiments performed and to refine the methods used, i...

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Autores principales: Calderan, Laura, Carton, Flavia, Andreana, Ilaria, Bincoletto, Valeria, Arpicco, Silvia, Stella, Barbara, Malatesta, Manuela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PAGEPress Publications, Pavia, Italy 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827424/
https://www.ncbi.nlm.nih.gov/pubmed/36546417
http://dx.doi.org/10.4081/ejh.2023.3596
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author Calderan, Laura
Carton, Flavia
Andreana, Ilaria
Bincoletto, Valeria
Arpicco, Silvia
Stella, Barbara
Malatesta, Manuela
author_facet Calderan, Laura
Carton, Flavia
Andreana, Ilaria
Bincoletto, Valeria
Arpicco, Silvia
Stella, Barbara
Malatesta, Manuela
author_sort Calderan, Laura
collection PubMed
description The development of novel nanoconstructs for biomedical applications requires the assessment of their biodistribution, metabolism and clearance in single cells, organs and entire organisms in a living environment. To reduce the number of in vivo experiments performed and to refine the methods used, in accordance with the 3Rs principle, this work proposes an ex vivo experimental system to monitor, using fluorescence microscopy, the distribution of nanoparticles in explanted murine skeletal muscle maintained in a bioreactor that can preserve the structural and functional features of the organ for long periods of time. Fluorescently-labelled liposomes and poly(lactide-co-glycolide) (PLGA)-based nanoparticles were injected into the intact soleus muscle (in the distal region close to the tendon) immediately after explants, and their distribution was analysed at increasing incubation times in cross cryosections from the proximal region of the belly. Both nanocarriers were clearly recognized in the muscle and were found to enter and migrate inside the myofibres, whereas their migration in the connective tissue seemed to be limited. In addition, some fluorescence signals were observed inside the macrophages, demonstrating the physiological clearance of the nanocarriers that did not enter the myofibres. Our ex vivo system therefore provides more information than previous in vitro experiments on cultured muscle cells, highlighting the need for the appropriate functionalization of nanocarriers if myofibre targeting is to be improved.
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spelling pubmed-98274242023-01-10 An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle Calderan, Laura Carton, Flavia Andreana, Ilaria Bincoletto, Valeria Arpicco, Silvia Stella, Barbara Malatesta, Manuela Eur J Histochem Technical Note The development of novel nanoconstructs for biomedical applications requires the assessment of their biodistribution, metabolism and clearance in single cells, organs and entire organisms in a living environment. To reduce the number of in vivo experiments performed and to refine the methods used, in accordance with the 3Rs principle, this work proposes an ex vivo experimental system to monitor, using fluorescence microscopy, the distribution of nanoparticles in explanted murine skeletal muscle maintained in a bioreactor that can preserve the structural and functional features of the organ for long periods of time. Fluorescently-labelled liposomes and poly(lactide-co-glycolide) (PLGA)-based nanoparticles were injected into the intact soleus muscle (in the distal region close to the tendon) immediately after explants, and their distribution was analysed at increasing incubation times in cross cryosections from the proximal region of the belly. Both nanocarriers were clearly recognized in the muscle and were found to enter and migrate inside the myofibres, whereas their migration in the connective tissue seemed to be limited. In addition, some fluorescence signals were observed inside the macrophages, demonstrating the physiological clearance of the nanocarriers that did not enter the myofibres. Our ex vivo system therefore provides more information than previous in vitro experiments on cultured muscle cells, highlighting the need for the appropriate functionalization of nanocarriers if myofibre targeting is to be improved. PAGEPress Publications, Pavia, Italy 2022-12-22 /pmc/articles/PMC9827424/ /pubmed/36546417 http://dx.doi.org/10.4081/ejh.2023.3596 Text en ©Copyright: the Author(s) https://creativecommons.org/licenses/by-nc/4.0/This article is distributed under the terms of the Creative Commons Attribution Noncommercial License (by-nc 4.0) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Technical Note
Calderan, Laura
Carton, Flavia
Andreana, Ilaria
Bincoletto, Valeria
Arpicco, Silvia
Stella, Barbara
Malatesta, Manuela
An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
title An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
title_full An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
title_fullStr An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
title_full_unstemmed An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
title_short An ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
title_sort ex vivo experimental system to track fluorescent nanoparticles inside skeletal muscle
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9827424/
https://www.ncbi.nlm.nih.gov/pubmed/36546417
http://dx.doi.org/10.4081/ejh.2023.3596
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