Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles
Recently, bioinspired cell-derived nanovesicles (CDNs) have gained much interest in the field of nanomedicine due to the preservation of biomolecular structure characteristics derived from their parent cells, which impart CDNs with unique properties in terms of binding and uptake by target cells and...
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| Format: | Article |
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MDPI AG
2021-07-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/13/7/1052 |
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| author | Yub Raj Neupane Chenyuan Huang Xiaoyuan Wang Wei Heng Chng Gopalakrishnan Venkatesan Olga Zharkova Matthias Gerhard Wacker Bertrand Czarny Gerrit Storm Jiong-Wei Wang Giorgia Pastorin |
| author_facet | Yub Raj Neupane Chenyuan Huang Xiaoyuan Wang Wei Heng Chng Gopalakrishnan Venkatesan Olga Zharkova Matthias Gerhard Wacker Bertrand Czarny Gerrit Storm Jiong-Wei Wang Giorgia Pastorin |
| author_sort | Yub Raj Neupane |
| collection | DOAJ |
| description | Recently, bioinspired cell-derived nanovesicles (CDNs) have gained much interest in the field of nanomedicine due to the preservation of biomolecular structure characteristics derived from their parent cells, which impart CDNs with unique properties in terms of binding and uptake by target cells and intrinsic biological activities. Although the production of CDNs can be easily and reproducibly achieved with any kind of cell culture, application of CDNs for therapeutic purposes has been greatly hampered by their physical and chemical instability during long-term storage in aqueous dispersion. In the present study, we conceived a lyophilization approach that would preserve critical characteristics regarding stability (vesicles’ size and protein content), structural integrity, and biological activity of CDNs for enabling long-term storage in freeze-dried form. Compared to the lyoprotectant sucrose, trehalose-lyoprotected CDNs showed significantly higher glass transition temperature and lower residual moisture content. As assessed by ATR-FTIR and far-UV circular dichroism, lyophilization in the presence of the lyoprotectant effectively maintained the secondary structure of cellular proteins. After reconstitution, lyoprotected CDNs were efficiently associated with HeLa cells, CT26 cells, and bone marrow-derived macrophages at a rate comparable to the freshly prepared CDNs. In vivo, both lyoprotected and freshly prepared CDNs, for the first time ever reported, targeted the injured heart, and exerted intrinsic cardioprotective effects within 24 h, attributable to the antioxidant capacity of CDNs in a myocardial ischemia/reperfusion injury animal model. Taken together, these results pave the way for further development of CDNs as cell-based therapeutics stabilized by lyophilization that enabled long-term storage while preserving their activity. |
| first_indexed | 2024-03-10T09:28:19Z |
| format | Article |
| id | doaj.art-5e19a1463492410290d1b7727ef5226d |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T09:28:19Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-5e19a1463492410290d1b7727ef5226d2023-11-22T04:41:17ZengMDPI AGPharmaceutics1999-49232021-07-01137105210.3390/pharmaceutics13071052Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived NanovesiclesYub Raj Neupane0Chenyuan Huang1Xiaoyuan Wang2Wei Heng Chng3Gopalakrishnan Venkatesan4Olga Zharkova5Matthias Gerhard Wacker6Bertrand Czarny7Gerrit Storm8Jiong-Wei Wang9Giorgia Pastorin10Department of Pharmacy, National University of Singapore, Singapore 117559, SingaporeDepartment of Surgery, National University of Singapore, Singapore 119228, SingaporeDepartment of Surgery, National University of Singapore, Singapore 119228, SingaporeDepartment of Pharmacy, National University of Singapore, Singapore 117559, SingaporeDepartment of Pharmacy, National University of Singapore, Singapore 117559, SingaporeDepartment of Surgery, National University of Singapore, Singapore 119228, SingaporeDepartment of Pharmacy, National University of Singapore, Singapore 117559, SingaporeSchool of Materials, Science and Engineering & Lee Kong Chian School of Medicine (LKC Medicine), Nanyang Technological University, Singapore 308232, SingaporeDepartment of Surgery, National University of Singapore, Singapore 119228, SingaporeDepartment of Surgery, National University of Singapore, Singapore 119228, SingaporeDepartment of Pharmacy, National University of Singapore, Singapore 117559, SingaporeRecently, bioinspired cell-derived nanovesicles (CDNs) have gained much interest in the field of nanomedicine due to the preservation of biomolecular structure characteristics derived from their parent cells, which impart CDNs with unique properties in terms of binding and uptake by target cells and intrinsic biological activities. Although the production of CDNs can be easily and reproducibly achieved with any kind of cell culture, application of CDNs for therapeutic purposes has been greatly hampered by their physical and chemical instability during long-term storage in aqueous dispersion. In the present study, we conceived a lyophilization approach that would preserve critical characteristics regarding stability (vesicles’ size and protein content), structural integrity, and biological activity of CDNs for enabling long-term storage in freeze-dried form. Compared to the lyoprotectant sucrose, trehalose-lyoprotected CDNs showed significantly higher glass transition temperature and lower residual moisture content. As assessed by ATR-FTIR and far-UV circular dichroism, lyophilization in the presence of the lyoprotectant effectively maintained the secondary structure of cellular proteins. After reconstitution, lyoprotected CDNs were efficiently associated with HeLa cells, CT26 cells, and bone marrow-derived macrophages at a rate comparable to the freshly prepared CDNs. In vivo, both lyoprotected and freshly prepared CDNs, for the first time ever reported, targeted the injured heart, and exerted intrinsic cardioprotective effects within 24 h, attributable to the antioxidant capacity of CDNs in a myocardial ischemia/reperfusion injury animal model. Taken together, these results pave the way for further development of CDNs as cell-based therapeutics stabilized by lyophilization that enabled long-term storage while preserving their activity.https://www.mdpi.com/1999-4923/13/7/1052cell-derived nanovesiclesexosome mimeticsbionanotechnologylyophilizationtrehalosecardioprotection |
| spellingShingle | Yub Raj Neupane Chenyuan Huang Xiaoyuan Wang Wei Heng Chng Gopalakrishnan Venkatesan Olga Zharkova Matthias Gerhard Wacker Bertrand Czarny Gerrit Storm Jiong-Wei Wang Giorgia Pastorin Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles Pharmaceutics cell-derived nanovesicles exosome mimetics bionanotechnology lyophilization trehalose cardioprotection |
| title | Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles |
| title_full | Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles |
| title_fullStr | Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles |
| title_full_unstemmed | Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles |
| title_short | Lyophilization Preserves the Intrinsic Cardioprotective Activity of Bioinspired Cell-Derived Nanovesicles |
| title_sort | lyophilization preserves the intrinsic cardioprotective activity of bioinspired cell derived nanovesicles |
| topic | cell-derived nanovesicles exosome mimetics bionanotechnology lyophilization trehalose cardioprotection |
| url | https://www.mdpi.com/1999-4923/13/7/1052 |
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