Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing
Impaired fracture healing impacts patients’ quality of life and imposes a financial burden on healthcare services. Up to 10% of bone fractures result in delayed/non-union fractures, for which new treatments are urgently required. However, systemic delivery of bone anabolic molecules is often sub-opt...
| Main Authors: | , , , , , , |
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| Format: | Conference item |
| Language: | English |
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MDPI
2020
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| _version_ | 1797110531220307968 |
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| author | May, JP Polydorou, A Ferri, S Wu, Q Stride, E Carugo, D Evans, ND |
| author_facet | May, JP Polydorou, A Ferri, S Wu, Q Stride, E Carugo, D Evans, ND |
| author_sort | May, JP |
| collection | OXFORD |
| description | Impaired fracture healing impacts patients’ quality of life and imposes a financial burden on healthcare services. Up to 10% of bone fractures result in delayed/non-union fractures, for which new treatments are urgently required. However, systemic delivery of bone anabolic molecules is often sub-optimal and can lead to significant side effects. In this study, we developed ultrasound (US) responsive nano-sized vehicles in the form of perfluorocarbon nanodroplets (NDs), as a means of targeting delivery of drugs to localised tissues. We tested the hypothesis that NDs could stably encapsulate BIO (GSK-3β inhibitor), which could then be released upon US stimulation to activate Wnt signalling and induce ossification. NDs (~280 nm) were prepared from phospholipids and liquid perfluorocarbon and their stability and drug loading was studied by NTA (Nano Tracking Analysis) and HPLC. ND cytotoxicity was assessed in patient-derived bone marrow stromal cells (BMSCs) with Alamar Blue (24 h), and in vitro bioactivity of BIO-NDs was evaluated in a 3T3 Wnt-pathway reporter cell line with luciferase readout. To investigate the acoustic behaviour of NDs, 2% agarose (LM) containing NDs was injected into a bespoke bone fracture model (Sawbones) of various geometries and stimulated by US (1 MHz, 5% duty cycle, 1 MPa, 30 s), allowing the simultaneous capture of optical images and acoustic emissions. Femoral bone hole defects (1–2 mm) were made in WT-MF1 mice (age: 8–12 wks) and DiR-labelled NDs (100 µL, 109 NDs/mL, i.v.) were injected post-fracture to determine biodistribution by IVIS imaging. NDs were stable (4 and 37 °C) and retained >90% BIO until US was applied, which caused ~100% release. ND exposure up to a concentration of 109 NDs/mL showed no cytotoxicity (24 h). BIO-loaded NDs induced Wnt pathway activation in a dose dependent manner. Biodistribution of DiR-NDs in a femoral bone hole defect model in mice demonstrated increased localisation at the fracture site (~2-fold relative to that found in healthy mice or contralateral femurs at 48 h). |
| first_indexed | 2024-03-07T07:56:10Z |
| format | Conference item |
| id | oxford-uuid:85c8cd53-894c-4bd9-aa83-dfe8cbcff681 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:56:10Z |
| publishDate | 2020 |
| publisher | MDPI |
| record_format | dspace |
| spelling | oxford-uuid:85c8cd53-894c-4bd9-aa83-dfe8cbcff6812023-08-22T12:37:30ZDevelopment of a nanodroplet formulation for triggered release of BIO for bone fracture healingConference itemhttp://purl.org/coar/resource_type/c_5794uuid:85c8cd53-894c-4bd9-aa83-dfe8cbcff681EnglishSymplectic ElementsMDPI2020May, JPPolydorou, AFerri, SWu, QStride, ECarugo, DEvans, NDImpaired fracture healing impacts patients’ quality of life and imposes a financial burden on healthcare services. Up to 10% of bone fractures result in delayed/non-union fractures, for which new treatments are urgently required. However, systemic delivery of bone anabolic molecules is often sub-optimal and can lead to significant side effects. In this study, we developed ultrasound (US) responsive nano-sized vehicles in the form of perfluorocarbon nanodroplets (NDs), as a means of targeting delivery of drugs to localised tissues. We tested the hypothesis that NDs could stably encapsulate BIO (GSK-3β inhibitor), which could then be released upon US stimulation to activate Wnt signalling and induce ossification. NDs (~280 nm) were prepared from phospholipids and liquid perfluorocarbon and their stability and drug loading was studied by NTA (Nano Tracking Analysis) and HPLC. ND cytotoxicity was assessed in patient-derived bone marrow stromal cells (BMSCs) with Alamar Blue (24 h), and in vitro bioactivity of BIO-NDs was evaluated in a 3T3 Wnt-pathway reporter cell line with luciferase readout. To investigate the acoustic behaviour of NDs, 2% agarose (LM) containing NDs was injected into a bespoke bone fracture model (Sawbones) of various geometries and stimulated by US (1 MHz, 5% duty cycle, 1 MPa, 30 s), allowing the simultaneous capture of optical images and acoustic emissions. Femoral bone hole defects (1–2 mm) were made in WT-MF1 mice (age: 8–12 wks) and DiR-labelled NDs (100 µL, 109 NDs/mL, i.v.) were injected post-fracture to determine biodistribution by IVIS imaging. NDs were stable (4 and 37 °C) and retained >90% BIO until US was applied, which caused ~100% release. ND exposure up to a concentration of 109 NDs/mL showed no cytotoxicity (24 h). BIO-loaded NDs induced Wnt pathway activation in a dose dependent manner. Biodistribution of DiR-NDs in a femoral bone hole defect model in mice demonstrated increased localisation at the fracture site (~2-fold relative to that found in healthy mice or contralateral femurs at 48 h). |
| spellingShingle | May, JP Polydorou, A Ferri, S Wu, Q Stride, E Carugo, D Evans, ND Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing |
| title | Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing |
| title_full | Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing |
| title_fullStr | Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing |
| title_full_unstemmed | Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing |
| title_short | Development of a nanodroplet formulation for triggered release of BIO for bone fracture healing |
| title_sort | development of a nanodroplet formulation for triggered release of bio for bone fracture healing |
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