{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,25]],"date-time":"2026-03-25T19:39:33Z","timestamp":1774467573289,"version":"3.50.1"},"reference-count":56,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,6,6]],"date-time":"2023-06-06T00:00:00Z","timestamp":1686009600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"JSPS KAKENHI","doi-asserted-by":"publisher","award":["23300193"],"award-info":[{"award-number":["23300193"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Strategies for gene and nucleic acid delivery to skeletal muscles have been extensively explored to treat Duchenne muscular dystrophy (DMD) and other neuromuscular diseases. Of these, effective intravascular delivery of naked plasmid DNA (pDNA) and nucleic acids into muscles is an attractive approach, given the high capillary density in close contact with myofibers. We developed lipid-based nanobubbles (NBs) using polyethylene-glycol-modified liposomes and an echo-contrast gas and found that these NBs could improve tissue permeability by ultrasound (US)-induced cavitation. Herein, we delivered naked pDNA or antisense phosphorodiamidate morpholino oligomers (PMOs) into the regional hindlimb muscle via limb perfusion using NBs and US exposure. pDNA encoding the luciferase gene was injected with NBs via limb perfusion into normal mice with application of US. High luciferase activity was achieved in a wide area of the limb muscle. DMD model mice were administered PMOs, designed to skip the mutated exon 23 of the dystrophin gene, with NBs via intravenous limb perfusion, followed by US exposure. The number of dystrophin-positive fibers increased in the muscles of mdx mice. Combining NBs and US exposure, which can be widely delivered to the hind limb muscles via the limb vein, could be an effective therapeutic approach for DMD and other neuromuscular disorders.<\/jats:p>","DOI":"10.3390\/pharmaceutics15061665","type":"journal-article","created":{"date-parts":[[2023,6,6]],"date-time":"2023-06-06T02:57:30Z","timestamp":1686020250000},"page":"1665","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Development of a Gene and Nucleic Acid Delivery System for Skeletal Muscle Administration via Limb Perfusion Using Nanobubbles and Ultrasound"],"prefix":"10.3390","volume":"15","author":[{"given":"Shohko","family":"Sekine","sequence":"first","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sayaka","family":"Mayama","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nobuaki","family":"Nishijima","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takuo","family":"Kojima","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6881-6044","authenticated-orcid":false,"given":"Yoko","family":"Endo-Takahashi","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuko","family":"Ishii","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hitomi","family":"Shiono","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saki","family":"Akiyama","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akane","family":"Sakurai","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sanae","family":"Sashida","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4731-0236","authenticated-orcid":false,"given":"Nobuhito","family":"Hamano","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0098-9587","authenticated-orcid":false,"given":"Rui","family":"Tada","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ryo","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Laboratory of Drug and Gene Delivery Research, Faculty of Pharma-Sciences, Teikyo University, 2-11-1 Kaga, Itabashi-ku, Tokyo 173-8605, Japan"},{"name":"Advanced Comprehensive Research Organization (ACRO), Teikyo University, Tokyo 173-8605, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuo","family":"Maruyama","sequence":"additional","affiliation":[{"name":"Advanced Comprehensive Research Organization (ACRO), Teikyo University, Tokyo 173-8605, Japan"},{"name":"Laboratory of Ultrasound Theranostics, Faculty of Pharma-Sciences, Teikyo University, Tokyo 173-8605, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoichi","family":"Negishi","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1016\/0092-8674(87)90579-4","article-title":"Dystrophin: The protein product of the Duchenne muscular dystrophy locus","volume":"51","author":"Hoffman","year":"1987","journal-title":"Cell"},{"key":"ref_2","first-page":"498","article-title":"The molecular basis for Duchenne versus Becker muscular dystrophy: Correlation of severity with type of deletion","volume":"45","author":"Koenig","year":"1989","journal-title":"Am. J. Hum. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/0888-7543(88)90113-9","article-title":"An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus","volume":"2","author":"Monaco","year":"1988","journal-title":"Genomics"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.1056\/NEJM198805263182104","article-title":"Characterization of dystrophin in muscle-biopsy specimens from patients with Duchenne\u2019s or Becker\u2019s muscular dystrophy","volume":"318","author":"Hoffman","year":"1988","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1006\/bbrc.1996.1375","article-title":"Induction of exon skipping of the dystrophin transcript in lymphoblastoid cells by transfecting an antisense oligodeoxynucleotide complementary to an exon recognition sequence","volume":"226","author":"Pramono","year":"1996","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1073\/pnas.98.1.42","article-title":"Antisense induced exon skipping and synthesis of dystrophin in the mdx mouse","volume":"98","author":"Mann","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1038\/nm897","article-title":"Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse","volume":"9","author":"Lu","year":"2003","journal-title":"Nat. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/nm1345","article-title":"Systemic delivery of morpholino oligonucleotide restores dystrophin expression body wide and improves dystrophic pathology","volume":"12","author":"Alter","year":"2006","journal-title":"Nat. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1801","DOI":"10.1093\/hmg\/ddg196","article-title":"Morpholino antisense oligonucleotide induced dystrophin exon 23 skipping in mdx mouse muscle","volume":"12","author":"Gebski","year":"2006","journal-title":"Hum. Mol. Genet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1038\/s41573-020-0075-7","article-title":"Advances in oligonucleotide drug delivery","volume":"19","author":"Roberts","year":"2020","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e2146","DOI":"10.1212\/WNL.0000000000005680","article-title":"Eteplirsen treatment for Duchenne muscular dystrophy: Exon skipping and dystrophin production","volume":"90","author":"Charleston","year":"2018","journal-title":"Neurology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"eaan0713","DOI":"10.1126\/scitranslmed.aan0713","article-title":"Systemic administration of the antisense oligonucleotide NS-065\/NCNP-01 for skipping of exon 53 in patients with Duchenne muscular dystrophy","volume":"10","author":"Komaki","year":"2018","journal-title":"Sci. Transl. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1089\/oli.1.1997.7.187","article-title":"Morphoolino antisense oligomers design, preparation, and properties","volume":"7","author":"Summerton","year":"1997","journal-title":"Antisense Nucleic Acid Drug Dev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/S0301-5629(98)00003-9","article-title":"Artificial cavitation nuclei significantly enhance acoustically induced cell transfection","volume":"24","author":"Greenleaf","year":"1998","journal-title":"Ultrasound Med. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"8463","DOI":"10.1073\/pnas.84.23.8463","article-title":"Transfection of mammalian cells with plasmid DNA by scrape loading and sonication loading","volume":"84","author":"Fechheimer","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.1038\/sj.gt.3300947","article-title":"Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA","volume":"6","author":"Liu","year":"1999","journal-title":"Gene Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/j.jconrel.2005.08.025","article-title":"Therapeutic ultrasound optimization for gene delivery: A key factor achieving nuclear DNA localization","volume":"108","author":"Machluf","year":"2005","journal-title":"J. Control. Release"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/S1525-0016(02)90715-X","article-title":"Transcutaneous ultrasound augments naked DNA transfection of skeletal muscle","volume":"6","author":"Schratzberger","year":"2002","journal-title":"Mol. Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1016\/S0022-5347(17)37368-8","article-title":"Altered neutrophil permeability following shock wave exposure in vitro","volume":"147","author":"Holmes","year":"1992","journal-title":"J. Urol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1161\/01.CIR.98.4.290","article-title":"Direct in vivo visualization of intravascular destruction of microbubbles by ultrasound and its local effects on tissue","volume":"98","author":"Skyba","year":"1998","journal-title":"Circulation"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1148\/radiol.2292020500","article-title":"Gene transfer with echo-enhanced contrast agents: Comparison between Albunex, Optison, and Levovist in mice\u2014Initial results","volume":"229","author":"Li","year":"2003","journal-title":"Radiology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1161\/hc1002.105228","article-title":"Local delivery of plasmid DNA into rat carotid artery using ultrasound","volume":"105","author":"Taniyama","year":"2002","journal-title":"Circulation"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1016\/j.addr.2003.12.006","article-title":"Therapeutic applications of lipid-coated microbubbles","volume":"56","author":"Unger","year":"2004","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.jconrel.2006.09.008","article-title":"Gene delivery by combination of novel liposomal bubbles with perfluoropropane and ultrasound","volume":"117","author":"Suzuki","year":"2007","journal-title":"J. Control. Release"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/j.jconrel.2008.08.019","article-title":"Delivery of siRNA into the cytoplasm by liposomal bubbles and ultrasound","volume":"132","author":"Negishi","year":"2008","journal-title":"J. Control. Release"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1007\/s11095-010-0286-4","article-title":"Delivery of an angiogenic gene into ischemic muscle by novel Bubble liposomes followed by ultrasound exposure","volume":"28","author":"Negishi","year":"2011","journal-title":"Pharm. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1834","DOI":"10.1021\/mp200554c","article-title":"Systemic delivery systems of angiogenic gene by novel Bubble liposomes containing cationic lipid and ultrasound exposure","volume":"9","author":"Negishi","year":"2012","journal-title":"Mol. Pharm."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1021\/mp4004755","article-title":"Bubble liposomes and ultrasound exposure improve localized morpholino oligomer delivery into the skeletal muscles of dystrophic mdx mice","volume":"11","author":"Negishi","year":"2014","journal-title":"Mol. Pharm."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"344","DOI":"10.3390\/pharmaceutics7030344","article-title":"Enhancement of blood\u2013brain barrier permeability and delivery of antisense oligonucleotides or plasmid DNA to the brain by the combination of Bubble Liposomes and high-intensity focused ultrasound","volume":"7","author":"Negishi","year":"2015","journal-title":"Pharmaceutics"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Endo-Takahashi, Y., Kurokawa, R., Sato, K., Takizawa, N., Katagiri, F., Hamano, N., Suzuki, R., Maruyama, K., Nomizu, M., and Takagi, N. (2021). Ternary complexes of pDNA, neuron-binding peptide, and PEGylated polyethyleneimine for brain delivery with nano-bubbles and ultrasound. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13071003"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1038\/mt.2011.79","article-title":"Pip5 transduction peptides direct high efficiency oligonucleotide-mediated dystrophin exon skipping in heart and phenotypic correction in mdx mice","volume":"19","author":"Yin","year":"2011","journal-title":"Mol. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1002\/mus.21527","article-title":"Use of Evans blue dye to compare limb muscles in exercised young and old mdx mice","volume":"41","author":"Wooddell","year":"2010","journal-title":"Muscle Nerve"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1365","DOI":"10.1096\/fj.05-3720fje","article-title":"Stabilized HIF-1alpha is superior to VEGF for angiogenesis in skeletal muscle via adeno-associated virus gene transfer","volume":"19","author":"Pajusola","year":"2005","journal-title":"FASEB J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1038\/nrd1662","article-title":"Healing sound: The use of ultrasound in drug delivery and other therapeutic applications","volume":"4","author":"Mitragotri","year":"2005","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1038\/mt.2008.225","article-title":"Transduction efficiency and immune response associated with the administration of AAV8 vector into dog skeletal muscle","volume":"17","author":"Ohshima","year":"2009","journal-title":"Mol. Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1465","DOI":"10.1126\/science.1690918","article-title":"Direct gene transfer into mouse muscle in vivo","volume":"247","author":"Wolff","year":"1990","journal-title":"Science"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1435","DOI":"10.1093\/hmg\/6.9.1435","article-title":"High expression of naked plasmid DNA in muscles of young rodents","volume":"6","author":"Danko","year":"1997","journal-title":"Hum. Mol. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"959","DOI":"10.1093\/jb\/118.5.959","article-title":"Visualization of dystrophic muscle fibers in mdx mouse by vital staining with Evans blue: Evidence of apoptosis in dystrophin-deficient muscle","volume":"118","author":"Matsuda","year":"1995","journal-title":"J. Biochem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1083\/jcb.139.2.375","article-title":"Animal models for muscular dystrophy show different patterns of sarcolemmal disruption","volume":"139","author":"Straub","year":"1997","journal-title":"J. Cell Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1038\/nbt.3659","article-title":"Applications of CRISPR technologies in research and beyond","volume":"34","author":"Barrangou","year":"2016","journal-title":"Nat. Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.cell.2015.12.035","article-title":"Biology and applications of CRISPR systems: Harnessing nature\u2019s toolbox for genome engineering","volume":"164","author":"Wright","year":"2016","journal-title":"Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1126\/science.aau1549","article-title":"Gene editing restores dystrophin expression in a canine model of Duchenne muscular dystrophy","volume":"362","author":"Amoasii","year":"2018","journal-title":"Science"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"eaay6812","DOI":"10.1126\/sciadv.aay6812","article-title":"Enhanced CRISPR-Cas9 correction of Duchenne muscular dystrophy in mice by a self-complementary AAV delivery system","volume":"19","author":"Zhang","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"eaav4324","DOI":"10.1126\/sciadv.aav4324","article-title":"CRISPRCas9 corrects Duchenne muscular dystrophy exon 44 deletion mutations in mice and human cells","volume":"6","author":"Min","year":"2019","journal-title":"Sci. Adv."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1093\/toxsci\/kfw220","article-title":"Viral vectors: The road to reducing genotoxicity","volume":"155","author":"David","year":"2017","journal-title":"Toxicol. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1007\/s13311-022-01197-9","article-title":"CRISPR-Cas9 Gene Therapy for Duchenne Muscular Dystrophy","volume":"19","author":"Lamothe","year":"2022","journal-title":"Neurotherapeutics"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Spunde, K., Korotkaja, K., and Zajakina, A. (2022). Recombinant Viral Vectors for Therapeutic Programming of Tumour Microenvironment: Advantages and Limitations. Biomedicines, 10.","DOI":"10.3390\/biomedicines10092142"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1016\/j.gendis.2022.05.009","article-title":"Prolonged Control of Insulin-Dependent Diabetes via Intramuscular Expression of Plasmid-Encoded Single-Strand Insulin Analogue","volume":"10","author":"Deng","year":"2023","journal-title":"Genes Dis."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2807","DOI":"10.1016\/j.biomaterials.2012.12.018","article-title":"PDNA-Loaded Bubble Liposomes as Potential Ultrasound Imaging and Gene Delivery Agents","volume":"34","author":"Negishi","year":"2013","journal-title":"Biomaterials"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3883","DOI":"10.1038\/srep03883","article-title":"Systemic Delivery of MiR-126 by MiRNA-Loaded Bubble Liposomes for the Treatment of Hindlimb Ischemia","volume":"4","author":"Negishi","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Endo-Takahashi, Y., and Negishi, Y. (2020). Microbubbles and Nanobubbles with Ultrasound for Systemic Gene Delivery. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12100964"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1038\/sj.gt.3300572","article-title":"The Efficient Expression of Intravascularly Delivered DNA in Rat Muscle","volume":"5","author":"Budker","year":"1998","journal-title":"Gene Ther."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1089\/hum.2009.133","article-title":"Functional Efficacy of Dystrophin Expression from Plasmids Delivered to Mdx Mice by Hydrodynamic Limb Vein Injection","volume":"21","author":"Zhang","year":"2010","journal-title":"Hum. Gene Ther."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e3150","DOI":"10.1002\/jgm.3150","article-title":"Gene Transfer to Skeletal Muscle Using Hydrodynamic Limb Vein Injection: Current Applications, Hurdles and Possible Optimizations","volume":"22","author":"Midoux","year":"2020","journal-title":"J. Gene Med."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"113998","DOI":"10.1016\/j.addr.2021.113998","article-title":"Ultrasonic Particles: An Approach for Targeted Gene Delivery","volume":"179","author":"Walsh","year":"2021","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"10179","DOI":"10.1073\/pnas.180294397","article-title":"Diagnostic Ultrasound Activation of Contrast Agent Gas Bodies Induces Capillary Rupture in Mice","volume":"97","author":"Miller","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/6\/1665\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:49:00Z","timestamp":1760125740000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/6\/1665"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,6]]},"references-count":56,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["pharmaceutics15061665"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics15061665","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,6]]}}}