An investigation into the cytotoxic effects of microbubbles and their constituents on osteosarcoma and bone marrow stromal cells
An investigation into the cytotoxic effects of microbubbles and their constituents on osteosarcoma and bone marrow stromal cells
Background: ultrasound-responsive microbubbles offer a means of achieving minimally invasive, localised drug delivery in applications including regenerative medicine. To facilitate their use, however, it is important to determine any cytotoxic effects they or their constituents may have. The aim of this study was to test the hypothesis that phospholipid-shelled microbubbles are non-toxic to human bone-derived cells at biologically-relevant concentrations.
Methods: microbubbles were fabricated using combinations of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dibehenoyl-sn-glycero-3-phosphocholine (DBPC), polyoxyethylene(40) stearate (PEG40S) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene-glycol)-2000] (DSPE-PEG 2000). Microbubble size and concentration were measured as a function of time and temperature by optical microscopy. Effects on MG63 osteosarcoma and human bone marrow stromal cells (BMSCs) were measured for up to 72 h by assay for viability, metabolic activity and proliferation.
Results: DBPC:DSPE-PEG 2000 microbubbles were significantly more stable than DSPC:PEG40S microbubbles under all conditions tested. Serum-containing medium had no detrimental effect on microbubble stability, but storage at 37 °C compared to at 4 °C reduced stability for both preparations, with almost complete dissolution of microbubbles at times ≥24 h. DSPC:PEG40S microbubbles had greater inhibitory effects on cell metabolism and growth than DBPC:DSPE-PEG 2000 microbubbles, with PEG40S found to be the principle inhibitory component. These effects were only evident at high microbubble concentrations (≥20% (v/v)) or with prolonged culture (≥24 h). Increasing cell-microbubble contact by inversion culture in a custom-built device had no inhibitory effect on metabolism.
Conclusions: these data indicate that, over a broad range of concentrations and incubation times, DBPC:DSPE-PEG 2000 and DSPC:PEG40S microbubbles have little effect on osteoblastic cell viability and growth, and that PEG40S is the principle inhibitory component in the formulations investigated.
Bone, Microbubble, Phospholipids, Polyethylene glycol, Toxicity
Polydorou, A.E.
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May, J.P.
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Makris, K.
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Ferri, S.
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Wu, Q.
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Stride, E.
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Carugo, D.
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Evans, N.D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
13 October 2023
Polydorou, A.E.
0da7e4ca-bd4d-487f-8d9b-1dba058e3293
May, J.P.
b54a262b-9f32-4435-8866-3b9c495294f3
Makris, K.
1d7847d4-7ace-417c-8c5b-c0f892558d47
Ferri, S.
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Wu, Q.
a013aa15-2165-47d6-bbe2-f83ae858e275
Stride, E.
c0143e95-81fa-47c8-b9bc-5b4fc319bba6
Carugo, D.
4f0b78fb-683a-4191-8b0f-5d4e45a0c663
Evans, N.D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Polydorou, A.E., May, J.P., Makris, K., Ferri, S., Wu, Q., Stride, E., Carugo, D. and Evans, N.D.
(2023)
An investigation into the cytotoxic effects of microbubbles and their constituents on osteosarcoma and bone marrow stromal cells.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1867 (12), [130481].
(doi:10.1016/j.bbagen.2023.130481).
Abstract
Background: ultrasound-responsive microbubbles offer a means of achieving minimally invasive, localised drug delivery in applications including regenerative medicine. To facilitate their use, however, it is important to determine any cytotoxic effects they or their constituents may have. The aim of this study was to test the hypothesis that phospholipid-shelled microbubbles are non-toxic to human bone-derived cells at biologically-relevant concentrations.
Methods: microbubbles were fabricated using combinations of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dibehenoyl-sn-glycero-3-phosphocholine (DBPC), polyoxyethylene(40) stearate (PEG40S) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene-glycol)-2000] (DSPE-PEG 2000). Microbubble size and concentration were measured as a function of time and temperature by optical microscopy. Effects on MG63 osteosarcoma and human bone marrow stromal cells (BMSCs) were measured for up to 72 h by assay for viability, metabolic activity and proliferation.
Results: DBPC:DSPE-PEG 2000 microbubbles were significantly more stable than DSPC:PEG40S microbubbles under all conditions tested. Serum-containing medium had no detrimental effect on microbubble stability, but storage at 37 °C compared to at 4 °C reduced stability for both preparations, with almost complete dissolution of microbubbles at times ≥24 h. DSPC:PEG40S microbubbles had greater inhibitory effects on cell metabolism and growth than DBPC:DSPE-PEG 2000 microbubbles, with PEG40S found to be the principle inhibitory component. These effects were only evident at high microbubble concentrations (≥20% (v/v)) or with prolonged culture (≥24 h). Increasing cell-microbubble contact by inversion culture in a custom-built device had no inhibitory effect on metabolism.
Conclusions: these data indicate that, over a broad range of concentrations and incubation times, DBPC:DSPE-PEG 2000 and DSPC:PEG40S microbubbles have little effect on osteoblastic cell viability and growth, and that PEG40S is the principle inhibitory component in the formulations investigated.
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Accepted/In Press date: 2 October 2023
e-pub ahead of print date: 5 October 2023
Published date: 13 October 2023
Keywords:
Bone, Microbubble, Phospholipids, Polyethylene glycol, Toxicity
Identifiers
Local EPrints ID: 483432
URI: http://eprints.soton.ac.uk/id/eprint/483432
ISSN: 0304-4165
PURE UUID: de6cd5a3-3c13-4688-9b82-a7088244d4e4
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Date deposited: 30 Oct 2023 18:04
Last modified: 26 Mar 2024 02:55
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Contributors
Author:
K. Makris
Author:
Q. Wu
Author:
E. Stride
Author:
D. Carugo
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