Tracking cellular uptake, intracellular trafficking and fate of nanoclay particles in human bone marrow stromal cells
Tracking cellular uptake, intracellular trafficking and fate of nanoclay particles in human bone marrow stromal cells
Clay nanoparticles, in particular synthetic smectites, have generated interest in the field of tissue engineering and regenerative medicine due to their utility as cross-linkers for polymers in biomaterial design and as protein release modifiers for growth factor delivery. In addition, recent studies have suggested a direct influence on the osteogenic differentiation of responsive stem and progenitor cell populations. Relatively little is known however about the mechanisms underlying nanoclay bioactivity and in particular the cellular processes involved in nanoclay-stem cell interactions. In this study we employed confocal microscopy, inductively coupled plasma mass spectrometry and transmission electron microscopy to track the interactions between clay nanoparticles and human bone marrow stromal cells (hBMSCs). In particular we studied nanoparticle cellular uptake mechanisms and uptake kinetics, intracellular trafficking pathways and the fate of endocytosed nanoclay. We found that nanoclay particles present on the cell surface as μm-sized aggregates, enter hBMSCs through clathrin-mediated endocytosis, and their uptake kinetics follow a linear increase with time during the first week of nanoclay addition. The endocytosed particles were observed within the endosomal/lysosomal compartments and we found evidence for both intracellular degradation of nanoclay and exocytosis as well as an increase in autophagosomal activity. Inhibitor studies indicated that endocytosis was required for nanoclay upregulation of alkaline phosphatase activity but a similar dependency was not observed for autophagy. This study into the nature of nanoclay-stem cell interactions, in particular the intracellular processing of nanosilicate, may provide insights into the mechanisms underlying nanoclay bioactivity and inform the successful utilisation of clay nanoparticles in biomaterial design.
18457-18472
Mousa, Mohamed
f415be35-3a79-48c2-b6f5-e03fc564c5e2
Kim, Yang-Hee
de0d641b-c2cb-4e73-9ae2-e20d33689f5d
Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Oreffo, Richard O.C.
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Dawson, Jonathan I.
982bf202-af02-4a6c-9db0-2d4a72c1ea92
21 September 2023
Mousa, Mohamed
f415be35-3a79-48c2-b6f5-e03fc564c5e2
Kim, Yang-Hee
de0d641b-c2cb-4e73-9ae2-e20d33689f5d
Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Oreffo, Richard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Dawson, Jonathan I.
982bf202-af02-4a6c-9db0-2d4a72c1ea92
Mousa, Mohamed, Kim, Yang-Hee, Evans, Nicholas D., Oreffo, Richard O.C. and Dawson, Jonathan I.
(2023)
Tracking cellular uptake, intracellular trafficking and fate of nanoclay particles in human bone marrow stromal cells.
Nanoscale, 15 (45), .
(doi:10.1039/d3nr02447d).
Abstract
Clay nanoparticles, in particular synthetic smectites, have generated interest in the field of tissue engineering and regenerative medicine due to their utility as cross-linkers for polymers in biomaterial design and as protein release modifiers for growth factor delivery. In addition, recent studies have suggested a direct influence on the osteogenic differentiation of responsive stem and progenitor cell populations. Relatively little is known however about the mechanisms underlying nanoclay bioactivity and in particular the cellular processes involved in nanoclay-stem cell interactions. In this study we employed confocal microscopy, inductively coupled plasma mass spectrometry and transmission electron microscopy to track the interactions between clay nanoparticles and human bone marrow stromal cells (hBMSCs). In particular we studied nanoparticle cellular uptake mechanisms and uptake kinetics, intracellular trafficking pathways and the fate of endocytosed nanoclay. We found that nanoclay particles present on the cell surface as μm-sized aggregates, enter hBMSCs through clathrin-mediated endocytosis, and their uptake kinetics follow a linear increase with time during the first week of nanoclay addition. The endocytosed particles were observed within the endosomal/lysosomal compartments and we found evidence for both intracellular degradation of nanoclay and exocytosis as well as an increase in autophagosomal activity. Inhibitor studies indicated that endocytosis was required for nanoclay upregulation of alkaline phosphatase activity but a similar dependency was not observed for autophagy. This study into the nature of nanoclay-stem cell interactions, in particular the intracellular processing of nanosilicate, may provide insights into the mechanisms underlying nanoclay bioactivity and inform the successful utilisation of clay nanoparticles in biomaterial design.
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d3nr02447d
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Submitted date: 26 May 2023
Accepted/In Press date: 21 September 2023
e-pub ahead of print date: 21 September 2023
Published date: 21 September 2023
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Funding Information:
This study is supported by Dr Jonathan I. Dawson EPSRC fellowship (grant number EP/L010259/1). Dr Mohamed Mousa would like to acknowledge the Faculty of Engineering and the Environment, University of Southampton and BYK-ALTANA for funding his PhD. The results presented in this study are part of Dr Mohamed Mousa's PhD thesis which could be accessed through the University of Southampton Institutional Repository: https://eprints.soton.ac.uk/448875/1/PhD_Thesis_MM_28871499.pdf .
Funding Information:
This study is supported by Dr Jonathan I. Dawson EPSRC fellowship (grant number EP/L010259/1). Dr Mohamed Mousa would like to acknowledge the Faculty of Engineering and the Environment, University of Southampton and BYK-ALTANA for funding his PhD. The results presented in this study are part of Dr Mohamed Mousa's PhD thesis which could be accessed through the University of Southampton Institutional Repository: https://eprints.soton.ac.uk/448875/1/PhD_Thesis_MM_28871499.pdf.
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© 2023 The Royal Society of Chemistry.
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Local EPrints ID: 484962
URI: http://eprints.soton.ac.uk/id/eprint/484962
ISSN: 2040-3364
PURE UUID: 75971442-a6fd-40d8-90fb-b501a476dacd
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Date deposited: 27 Nov 2023 17:32
Last modified: 15 Aug 2024 01:47
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Author:
Mohamed Mousa
Author:
Jonathan I. Dawson
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