eprintid: 10205433 rev_number: 7 eprint_status: archive userid: 699 dir: disk0/10/20/54/33 datestamp: 2025-02-28 11:27:01 lastmod: 2025-02-28 11:27:01 status_changed: 2025-02-28 11:27:01 type: article metadata_visibility: show sword_depositor: 699 creators_name: Ma, Cheng creators_name: Chow, Michael YT creators_name: Zhang, Chengyang creators_name: Goldbaum, Paulina creators_name: Hsieh, Jamie Chien-Ming creators_name: Lam, Jenny KW title: Robust peptide/RNA complexes prepared with microfluidic mixing for pulmonary delivery by nebulisation ispublished: inpress divisions: UCL divisions: B02 divisions: C08 divisions: D10 divisions: G08 keywords: Aerosolisation, Inhalation, mRNA, siRNA, Transfection, Vibrating mesh nebuliser note: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. abstract: Small interfering RNA (siRNA) and messenger RNA (mRNA) have drawn considerable attention in recent years due to their ability to modulate the expression of specific disease-related proteins. However, it is difficult to find safe, robust, and effective RNA delivery systems suitable for pulmonary delivery to treat lung diseases. In this study, two cationic peptides, namely LAH4-L1 and PEG12KL4, were employed as non-viral vectors for siRNA and mRNA delivery. Four formulations (i.e. LAH4-L1/siRNA; PEG12KL4/siRNA; LAH4-L1/mRNA and PEG12KL4/mRNA) were investigated. Microfluidic mixing method was utilised to fabricate RNA complexes in a controllable and reproducible manner. Upon optimisation of the microfluidic mixing protocol, a vibrating mesh nebuliser was employed to aerosolise the RNA complexes, and their transfection efficiency was evaluated on A549 and BEAS-2B cells. Following nebulisation, inhalable mist was generated for all RNA formulations with mass median aerodynamic diameter below 5 μm. Although the hydrodynamic particle sizes of the RNA complexes were significantly reduced to around 100 nm after nebulisation regardless of the original size of the complexes prior to nebulisation, the RNA binding efficiency and the in vitro RNA transfection ability of all the peptide formulations were successfully preserved with no significant differences compared to the same system before nebulisation. The current result indicates that both LAH4-L1 and PEG12KL4 hold significant potential for future clinical application for pulmonary siRNA and mRNA delivery through nebulisation. date: 2025-01-18 date_type: published publisher: SPRINGER HEIDELBERG official_url: https://doi.org/10.1007/s13346-024-01773-w oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2354404 doi: 10.1007/s13346-024-01773-w medium: Print-Electronic pii: 10.1007/s13346-024-01773-w lyricists_name: Lam, Ka Wing lyricists_id: KWLAM29 actors_name: Booker, Anthony actors_name: Harris, Jean actors_id: AJBOO99 actors_id: JAHAR68 actors_role: owner actors_role: impersonator funding_acknowledgements: [UCL Therapeutic Acceleration Support Scheme]; MR/X502984/1 [MRC IAA 2021 UCL]; [HKU Presidential PhD scholarship]; [DDL Career Development Grant] full_text_status: public publication: Drug Delivery and Translational Research pages: 14 event_location: United States issn: 2190-393X citation: Ma, Cheng; Chow, Michael YT; Zhang, Chengyang; Goldbaum, Paulina; Hsieh, Jamie Chien-Ming; Lam, Jenny KW; (2025) Robust peptide/RNA complexes prepared with microfluidic mixing for pulmonary delivery by nebulisation. Drug Delivery and Translational Research 10.1007/s13346-024-01773-w <https://doi.org/10.1007/s13346-024-01773-w>. (In press). Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10205433/1/s13346-024-01773-w.pdf