An <em>ex vivo</em> 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,...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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PAGEPress Publications
2022-12-01
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| Series: | European Journal of Histochemistry |
| Subjects: | |
| Online Access: | https://ejh.it/index.php/ejh/article/view/3596 |
| _version_ | 1797979125048147968 |
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| author | Laura Calderan Flavia Carton Ilaria Andreana Valeria Bincoletto Silvia Arpicco Barbara Stella Manuela Malatesta |
| author_facet | Laura Calderan Flavia Carton Ilaria Andreana Valeria Bincoletto Silvia Arpicco Barbara Stella Manuela Malatesta |
| author_sort | Laura Calderan |
| collection | DOAJ |
| 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 fluorescent 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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| first_indexed | 2024-04-11T05:35:03Z |
| format | Article |
| id | doaj.art-b9908706fec14194891a35f355798bab |
| institution | Directory Open Access Journal |
| issn | 1121-760X 2038-8306 |
| language | English |
| last_indexed | 2024-04-11T05:35:03Z |
| publishDate | 2022-12-01 |
| publisher | PAGEPress Publications |
| record_format | Article |
| series | European Journal of Histochemistry |
| spelling | doaj.art-b9908706fec14194891a35f355798bab2022-12-22T14:52:44ZengPAGEPress PublicationsEuropean Journal of Histochemistry1121-760X2038-83062022-12-0167110.4081/ejh.2023.3596An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscleLaura Calderan0Flavia Carton1Ilaria Andreana2Valeria BincolettoSilvia Arpicco3Barbara Stella4Manuela Malatesta5Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of VeronaDepartment of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of VeronaDepartment of Drug Science and Technology, University of TurinDepartment of Drug Science and Technology, University of TurinDepartment of Drug Science and Technology, University of TurinDepartment of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of Verona 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 fluorescent 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. https://ejh.it/index.php/ejh/article/view/3596explanted muscleintramuscular injectionliposomesPLGA nanoparticlesbioreactorfluorescence microscopy |
| spellingShingle | Laura Calderan Flavia Carton Ilaria Andreana Valeria Bincoletto Silvia Arpicco Barbara Stella Manuela Malatesta An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle European Journal of Histochemistry explanted muscle intramuscular injection liposomes PLGA nanoparticles bioreactor fluorescence microscopy |
| title | An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle |
| title_full | An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle |
| title_fullStr | An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle |
| title_full_unstemmed | An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle |
| title_short | An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle |
| title_sort | em ex vivo em experimental system to track fluorescent nanoparticles inside skeletal muscle |
| topic | explanted muscle intramuscular injection liposomes PLGA nanoparticles bioreactor fluorescence microscopy |
| url | https://ejh.it/index.php/ejh/article/view/3596 |
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