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Res."],"published-print":{"date-parts":[[2025,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Gene therapy holds significant promise as a therapeutic approach for addressing a diverse range of diseases through the suppression of overexpressed proteins and the restoration of impaired cell functions. Developing a nanocarrier that can efficiently load and release genetic material into cells remains a challenge. The primary goal of this study is to develop formulations aimed to enhance the therapeutic potential of GapmeRs through technological approaches. To this end, lipid-polymeric hybrid nanoparticles (LPHNPs) with PLGA, DC-cholesterol, and DOPE-mPEG\n                    <jats:sub>2000<\/jats:sub>\n                    were produced by conventional single-step nanoprecipitation (SSN) and microfluidic (MF) methods. The optimized nanoparticles by SSN have a size of 149.9\u2009\u00b1\u200918.07\u00a0nm, a polydispersity index (PdI) of 0.23\u2009\u00b1\u20090.02, and a zeta potential of (ZP) of 29.34\u2009\u00b1\u20092.44 mV, while by MF the size was 179.8\u2009\u00b1\u20096.3, a PdI of 0.24\u2009\u00b1\u20090.01, and a ZP of 32.25\u2009\u00b1\u20091.36 mV. Furthermore, LPHNPs prepared with GapmeR-protamine by both methods exhibit a high encapsulation efficiency of approximately 90%. The encapsulated GapmeR is completely released in 24\u00a0h. The LPHNP suspensions are stable for up to 6\u00a0h in 10% FBS at pH 5.4 and 7.4. By contrast, LPHNPs remain stable in suspension in 4.5% albumin at pH 7.4 for 24\u00a0h. Additionally, LPHNPs were successfully freeze-dried using trehalose in the range of 2.5-5% as cryoprotectant The LPHNPs produced by MF and SSN increase, 6 and 12 fold respectively, GapmeR cell uptake, and both of them reduce by 60\u201370% expression of Tob1 in 48\u00a0h.Our study demonstrates the efficacy of the developed LPHNPs as carriers for oligonucleotide delivery, offering valuable insights for their scale up production from a conventional bulk methodology to a high-throughput microfluidic technology.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical Abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1007\/s13346-024-01644-4","type":"journal-article","created":{"date-parts":[[2024,6,13]],"date-time":"2024-06-13T14:05:22Z","timestamp":1718287522000},"page":"908-924","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Conventional and microfluidic methods: Design and optimization of lipid-polymeric hybrid nanoparticles for gene therapy"],"prefix":"10.1007","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-1083-2251","authenticated-orcid":false,"given":"Daniel","family":"Gonz\u00e1lez-Garc\u00eda","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0802-7975","authenticated-orcid":false,"given":"Olga","family":"Tapia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9347-4672","authenticated-orcid":false,"given":"Carmen","family":"\u00c9vora","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4801-4404","authenticated-orcid":false,"given":"Patricia","family":"Garc\u00eda-Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7045-6933","authenticated-orcid":false,"given":"Araceli","family":"Delgado","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,6,13]]},"reference":[{"key":"1644_CR1","doi-asserted-by":"crossref","unstructured":"Hosseinkhani H, Domb AJ, Sharifzadeh G, Nahum V. 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