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

Submitted: 7 November 2022
Accepted: 14 December 2022
Published: 22 December 2022
Abstract Views: 679
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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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Citations

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Ethics Approval

This study was approved by the Italian Ministry of Health

Supporting Agencies

This work did not receive specific funding and was performed thanks to intramural funds to M.M.
Flavia Carton, Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of Verona

Present address: Department of Health Sciences, University of Piemonte Orientale “A. Avogadro”, Novara, Italy

How to Cite

Calderan, L. ., Carton, F. ., Andreana, I. ., Bincoletto, V. ., Arpicco, S. ., Stella, B. ., & Malatesta, M. (2022). An <em>ex vivo</em> experimental system to track fluorescent nanoparticles inside skeletal muscle. European Journal of Histochemistry, 67(1). https://doi.org/10.4081/ejh.2023.3596