{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:55:01Z","timestamp":1787028901946,"version":"build-2736575974"},"reference-count":57,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,7,25]],"date-time":"2022-07-25T00:00:00Z","timestamp":1658707200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Council for the Improvement of Higher Education"},{"name":"S\u00e3o Paulo Research Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Nowadays, the therapeutic efficiency of small interfering RNAs (siRNA) is still limited by the efficiency of gene therapy vectors capable of carrying them inside the target cells. In this study, siRNA nanocarriers based on low molecular weight chitosan grafted with increasing proportions (5 to 55%) of diisopropylethylamine (DIPEA) groups were developed, which allowed precise control of the degree of ionization of the polycations at pH 7.4. This approach made obtaining siRNA nanocarriers with small sizes (100\u2013200 nm), positive surface charge and enhanced colloidal stability (up to 24 h) at physiological conditions of pH (7.4) and ionic strength (150 mmol L\u22121) possible. Moreover, the PEGylation improved the stability of the nanoparticles, which maintained their colloidal stability and nanometric sizes even in an albumin-containing medium. The chitosan-derivatives displayed non-cytotoxic effects in both fibroblasts (NIH\/3T3) and macrophages (RAW 264.7) at high N\/P ratios and polymer concentrations (up to 0.5 g L\u22121). Confocal microscopy showed a successful uptake of nanocarriers by RAW 264.7 macrophages and a promising ability to silence green fluorescent protein (GFP) in HeLa cells. These results were confirmed by a high level of tumor necrosis factor-\u03b1 (TNF\u03b1) knockdown (higher than 60%) in LPS-stimulated macrophages treated with the siRNA-loaded nanoparticles even in the FBS-containing medium, findings that reveal a good correlation between the degree of ionization of the polycations and the physicochemical properties of nanocarriers. Overall, this study provides an approach to enhance siRNA condensation by chitosan-based carriers and highlights the potential of these nanocarriers for in vivo studies.<\/jats:p>","DOI":"10.3390\/md20080476","type":"journal-article","created":{"date-parts":[[2022,7,25]],"date-time":"2022-07-25T21:20:59Z","timestamp":1658784059000},"page":"476","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions"],"prefix":"10.3390","volume":"20","author":[{"given":"Andr\u00e9 Miguel","family":"Martinez Junior","sequence":"first","affiliation":[{"name":"Department of Chemistry and Environmental Sciences, IBILCE, S\u00e3o Paulo State University\u2014UNESP, S\u00e3o Jos\u00e9 do Rio Preto 15054-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aline Margarete Furuyama","family":"Lima","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Environmental Sciences, IBILCE, S\u00e3o Paulo State University\u2014UNESP, S\u00e3o Jos\u00e9 do Rio Preto 15054-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grazieli Olinda","family":"Martins","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Environmental Sciences, IBILCE, S\u00e3o Paulo State University\u2014UNESP, S\u00e3o Jos\u00e9 do Rio Preto 15054-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vera Aparecida de Oliveira","family":"Tiera","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Environmental Sciences, IBILCE, S\u00e3o Paulo State University\u2014UNESP, S\u00e3o Jos\u00e9 do Rio Preto 15054-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed","family":"Benderdour","sequence":"additional","affiliation":[{"name":"Orthopedic Research Laboratory, H\u00f4pital du Sacr\u00e9-Coeur de Montr\u00e9al, Universit\u00e9 de Montr\u00e9al-Canada, Montreal, QC H3C3J7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julio Cesar","family":"Fernandes","sequence":"additional","affiliation":[{"name":"Orthopedic Research Laboratory, H\u00f4pital du Sacr\u00e9-Coeur de Montr\u00e9al, Universit\u00e9 de Montr\u00e9al-Canada, Montreal, QC H3C3J7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcio Jos\u00e9","family":"Tiera","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Environmental Sciences, IBILCE, S\u00e3o Paulo State University\u2014UNESP, S\u00e3o Jos\u00e9 do Rio Preto 15054-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1038\/s41392-020-0207-x","article-title":"Therapeutic SiRNA: State of the Art","volume":"5","author":"Hu","year":"2020","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"de Brito e Cunha, D., Frederico, A., Azamor, T., Melga\u00e7o, J., da Costa Neves, P., Bom, A., Tilli, T., and Missailidis, S. (2022). Biotechnological Evolution of SiRNA Molecules: From Bench Tool to the Refined Drug. Pharmaceuticals, 15.","DOI":"10.3390\/ph15050575"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1038\/s41435-021-00136-6","article-title":"Gene Therapy Avenues and COVID-19 Vaccines","volume":"22","year":"2021","journal-title":"Genes Immun."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Zamboulis, A., Nanaki, S., Michailidou, G., Koumentakou, I., Lazaridou, M., Ainali, N.M., Xanthopoulou, E., and Bikiaris, D.N. (2020). Chitosan and Its Derivatives for Ocular Delivery Formulations: Recent Advances and Developments. Polymers, 12.","DOI":"10.3390\/polym12071519"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Cao, Y., Tan, Y.F., Wong, Y.S., Liew, M.W.J., and Venkatraman, S. (2019). Recent Advances in Chitosan-Based Carriers for Gene Delivery. Mar. Drugs, 17.","DOI":"10.3390\/md17060381"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/j.ijbiomac.2019.02.036","article-title":"Optimization of Chitosan Nanoparticles as an Anti-HIV SiRNA Delivery Vehicle","volume":"129","author":"Modarressi","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_7","first-page":"433","article-title":"Self-Crosslinkable Chitosan-Hyaluronic Acid Dialdehyde Nanoparticles for CD44-Targeted SiRNA Delivery to Treat Bladder Cancer","volume":"6","author":"Liang","year":"2021","journal-title":"Bioact. Mater."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"102119","DOI":"10.1016\/j.nano.2019.102119","article-title":"Enriched Chitosan Nanoparticles Loaded with SiRNA are Effective in Lowering Huntington\u2019s Disease Gene Expression Following Intranasal Administration","volume":"24","author":"Sava","year":"2020","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1007\/s11051-019-4623-1","article-title":"Targeting Tuberculosis Infection in Macrophages Using Chitosan Oligosaccharide Nanoplexes","volume":"21","author":"Koli","year":"2019","journal-title":"J. Nanoparticle Res."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Moraru, C., Mincea, M., Menghiu, G., and Ostafe, V. (2020). Understanding the Factors Influencing Chitosan-Based Nanoparticles-Protein Corona Interaction and Drug Delivery Applications. Molecules, 25.","DOI":"10.3390\/molecules25204758"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Pathomthongtaweechai, N., and Muanprasat, C. (2021). Potential Applications of Chitosan-Based Nanomaterials to Surpass the Gastrointestinal Physiological Obstacles and Enhance the Intestinal Drug Absorption. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13060887"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"23859","DOI":"10.1038\/srep23859","article-title":"Novel Polymer Micelle Mediated Co-Delivery of Doxorubicin and P-Glycoprotein SiRNA for Reversal of Multidrug Resistance and Synergistic Tumor Therapy","volume":"6","author":"Zhang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"21422","DOI":"10.1038\/srep21422","article-title":"Transdermal Delivery of SiRNA through Microneedle Array","volume":"6","author":"Deng","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.carbpol.2015.08.053","article-title":"Target Gene Delivery from Targeting Ligand Conjugated Chitosan-PEI Copolymer for Cancer Therapy","volume":"135","author":"Nam","year":"2016","journal-title":"Carbohydr. Polym."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1016\/j.carbpol.2019.03.098","article-title":"Amphipathic Chitosans Improve the Physicochemical Properties of SiRNA-Chitosan Nanoparticles at Physiological Conditions","volume":"216","author":"Martins","year":"2019","journal-title":"Carbohydr. Polym."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Martinez Junior, A.M., de Souza, R.H.F.V., Petr\u00f4nio, M.S., Martins, G.O., Fernandes, J.C., Benderdour, M., Tiera, V.A.d.O., and Tiera, M.J. (2022). Double-Grafted Chitosans as SiRNA Nanocarriers: Effects of Diisopropylethylamine Substitution and Labile-PEG Coating. J. Nanostructure Chem., 1\u201320.","DOI":"10.1007\/s40097-022-00487-0"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.ijbiomac.2018.07.072","article-title":"Diethylaminoethyl-Chitosan as an Efficient Carrier for SiRNA Delivery: Improving the Condensation Process and the Nanoparticles Properties","volume":"119","author":"Picola","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1021\/acs.biomac.7b01297","article-title":"SiRNA Delivery with Chitosan: Influence of Chitosan Molecular Weight, Degree of Deacetylation, and Amine to Phosphate Ratio on In Vitro Silencing Efficiency, Hemocompatibility, Biodistribution, and In Vivo Efficacy","volume":"19","author":"Alameh","year":"2018","journal-title":"Biomacromolecules"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"12203","DOI":"10.1021\/acsami.6b16556","article-title":"Impact of PEG Chain Length on the Physical Properties and Bioactivity of PEGylated Chitosan\/SiRNA Nanoparticles In Vitro and In Vivo","volume":"9","author":"Yang","year":"2017","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"454","DOI":"10.3934\/bioeng.2016.4.454","article-title":"Increased Poly(Ethylene Glycol) Density Decreases Transfection Efficacy of SiRNA\/Poly(Ethylene Imine) Complexes","volume":"3","author":"Rheiner","year":"2016","journal-title":"AIMS Bioeng."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7803","DOI":"10.1021\/ja300176w","article-title":"Multicolored pH-Tunable and Activatable Fluorescence Nanoplatform Responsive to Physiologic PH Stimuli","volume":"134","author":"Zhou","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.ijbiomac.2021.01.010","article-title":"Characterization of Chitosan with Different Degree of Deacetylation and Equal Viscosity in Dissolved and Solid State\u2013Insights by Various Complimentary Methods","volume":"171","author":"Vehlow","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4698","DOI":"10.1016\/j.polymer.2010.08.034","article-title":"NMR and FT Raman Characterisation of Regioselectively Sulfated Chitosan Regarding the Distribution of Sulfate Groups and the Degree of Substitution","volume":"51","author":"Zhang","year":"2010","journal-title":"Polymer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1696","DOI":"10.1039\/c3tb00498h","article-title":"Functionalized-Chitosan\/Quantum Dot Nano-Hybrids for Nanomedicine Applications: Towards Biolabeling and Biosorbing Phosphate Metabolites","volume":"1","author":"Mansur","year":"2013","journal-title":"J. Mater. Chem. B"},{"key":"ref_25","first-page":"12","article-title":"Structural Characteristics of Chitin and Chitosan Isolated from the Biomass of Cultivated Rotifer, Brachionus Rotundiformis","volume":"3","author":"Rumengan","year":"2014","journal-title":"Int. J. Fish. Aquat. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1399","DOI":"10.3390\/ma4081399","article-title":"Deacetylation of Chitosan: Material Characterization and In Vitro Evaluation via Albumin Adsorption and Pre-Osteoblastic Cell Cultures","volume":"4","author":"Yuan","year":"2011","journal-title":"Materials"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"055101","DOI":"10.1088\/0957-4484\/24\/5\/055101","article-title":"Synthesis and Evaluation of Diethylethylamine-Chitosan for Gene Delivery: Composition Effects on the In Vitro Transfection Efficiency","volume":"24","author":"Oliveira","year":"2013","journal-title":"Nanotechnology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1186\/s11671-018-2728-6","article-title":"Insight into Cellular Uptake and Intracellular Trafficking of Nanoparticles","volume":"13","author":"Foroozandeh","year":"2018","journal-title":"Nanoscale Res. Lett."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3067","DOI":"10.1039\/C6PY00221H","article-title":"Synthesis of Polysarcosine from Air and Moisture Stable N-Phenoxycarbonyl-N-Methylglycine Assisted by Tertiary Amine Base","volume":"7","author":"Doriti","year":"2016","journal-title":"Polym. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4082","DOI":"10.1021\/mp400467x","article-title":"Overcoming Nonviral Gene Delivery Barriers: Perspective and Future","volume":"10","author":"Jones","year":"2013","journal-title":"Mol. Pharm."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2627","DOI":"10.1039\/c2sm06624f","article-title":"pH-Responsive Dendrimers Interacting with Lipid Membranes","volume":"8","author":"Ma","year":"2012","journal-title":"Soft Matter"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.addr.2016.08.014","article-title":"Investigation of Endosome and Lysosome Biology by Ultra pH-Sensitive Nanoprobes","volume":"113","author":"Wang","year":"2017","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6383","DOI":"10.2147\/IJN.S142060","article-title":"Intracellular Disposition of Chitosan Nanoparticles in Macrophages: Intracellular Uptake, Exocytosis, and Intercellular Transport","volume":"12","author":"Jiang","year":"2017","journal-title":"Int. J. Nanomed."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1021\/acsnano.8b05151","article-title":"Super-Resolution Imaging of Proton Sponge-Triggered Rupture of Endosomes and Cytosolic Release of Small Interfering RNA","volume":"13","author":"Wojnilowicz","year":"2019","journal-title":"ACS Nano"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1038\/nrm2820","article-title":"Sensors and Regulators of Intracellular pH","volume":"11","author":"Casey","year":"2010","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1615","DOI":"10.3109\/21691401.2015.1129624","article-title":"Polyelectrolyte Complexes: Mechanisms, Critical Experimental Aspects, and Applications","volume":"44","author":"Kulkarni","year":"2016","journal-title":"Artif. Cells Nanomed. Biotechnol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7923","DOI":"10.1021\/acs.chemmater.1c01832","article-title":"Nucleic Acids Based Polyelectrolyte Complexes: Their Complexation Mechanism, Morphology, and Stability","volume":"33","author":"Kim","year":"2021","journal-title":"Chem. Mater."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1727","DOI":"10.3390\/polym6061727","article-title":"Low Molecular Weight Chitosan (LMWC)-Based Polyplexes for pDNA Delivery: From Bench to Bedside","volume":"6","author":"Agirre","year":"2014","journal-title":"Polymers"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1038\/nmat2442","article-title":"Understanding Biophysicochemical Interactions at the Nano-Bio Interface","volume":"8","author":"Nel","year":"2009","journal-title":"Nat. Mater."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"288","DOI":"10.2174\/156652311796150408","article-title":"Polycation-Based Gene Therapy: Current Knowledge and New Perspectives","volume":"11","author":"Tiera","year":"2011","journal-title":"Curr. Gene Ther."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1080\/17458080.2011.602120","article-title":"Effect of Ionic Strength Solution on the Stability of Chitosan-DNA Nanoparticles","volume":"8","author":"Picola","year":"2013","journal-title":"J. Exp. Nanosci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1016\/j.apsb.2021.02.019","article-title":"Exploration and Insights into the Cellular Internalization and Intracellular Fate of Amphiphilic Polymeric Nanocarriers","volume":"11","author":"Mazumdar","year":"2021","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"6844","DOI":"10.1021\/acs.chemrev.8b00199","article-title":"Block Copolymer Micelles in Nanomedicine Applications","volume":"118","author":"Cabral","year":"2018","journal-title":"Chem. Rev."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3253","DOI":"10.1021\/nn3059295","article-title":"Polymer-Coated Nanoparticles Interacting with Proteins and Cells: Focusing on the Sign of the Net Charge","volume":"7","author":"Kantner","year":"2013","journal-title":"ACS Nano"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"9316","DOI":"10.1039\/c1sm05992k","article-title":"pH-Triggered Block Copolymer Micelles Based on a PH-Responsive PDPA (Poly [2-(Diisopropylamino)Ethyl Methacrylate]) Inner Core and a PEO (Poly(Ethylene Oxide)) Outer Shell as a Potential Tool for the Cancer Therapy","volume":"7","author":"Giacomelli","year":"2011","journal-title":"Soft Matter"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1007\/s10973-009-0040-5","article-title":"Isothermal Titration Calorimetric Studies of the pH Induced Conformational Changes of Bovine Serum Albumin","volume":"96","author":"Kun","year":"2009","journal-title":"J. Therm. Anal. Calorim."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"344","DOI":"10.3109\/17435390.2014.930533","article-title":"Inhibition of Autophagy Protects against PAMAM Dendrimers-Induced Hepatotoxicity","volume":"9","author":"Li","year":"2015","journal-title":"Nanotoxicology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1186\/1479-5876-9-46","article-title":"Comparisons of Three Polyethyleneimine-Derived Nanoparticles as a Gene Therapy Delivery System for Renal Cell Carcinoma","volume":"9","author":"Xu","year":"2011","journal-title":"J. Transl. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3307","DOI":"10.1038\/s41467-019-11190-0","article-title":"Highly Branched Poly(\u03b2-Amino Ester) Delivery of Minicircle DNA for Transfection of Neurodegenerative Disease Related Cells","volume":"10","author":"Liu","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Raik, S.V., Andranovit\u0161, S., Petrova, V.A., Xu, Y., Lam, J.K.W., Morris, G.A., Brodskaia, A.V., Casettari, L., Kritchenkov, A.S., and Skorik, Y.A. (2018). Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy. Polymers, 10.","DOI":"10.3390\/polym10040442"},{"key":"ref_51","unstructured":"(2022, July 11). BCRJ: Banco de C\u00e9lulas do Rio de Janeiro (RAW 264.7|APABCAM). Available online: https:\/\/bcrj.org.br\/celula\/0212."},{"key":"ref_52","unstructured":"(2022, July 11). BCRJ: Banco de C\u00e9lulas do Rio de Janeiro (HeLa\/GFP|APABCAM). Available online: https:\/\/bcrj.org.br\/celula\/HELA-GFP-CERVIX-ADENOCARCINOMA."},{"key":"ref_53","unstructured":"(2022, July 11). ATCC\u00ae (NIH\/3T3). Available online: https:\/\/www.atcc.org\/products\/crl-1658."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"3151","DOI":"10.1021\/bm060381u","article-title":"Synthesis and Characterization of Phosphorylcholine-Substituted Chitosans Soluble in Physiological pH Conditions","volume":"7","author":"Tiera","year":"2006","journal-title":"Biomacromolecules"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1016\/j.carbpol.2007.10.004","article-title":"Precise Derivatization of Structurally Distinct Chitosans with Rhodamine B Isothiocyanate","volume":"72","author":"Ma","year":"2008","journal-title":"Carbohydr. Polym."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.jcis.2009.02.069","article-title":"Physicochemical Characterization of Nanoparticles Formed between DNA and Phosphorylcholine Substituted Chitosans","volume":"336","author":"Picola","year":"2009","journal-title":"J. Colloid Interface Sci."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Dragicevic, N., and Maibach, H.I. (2016). Percutaneous Penetration Enhancers Chemical Methods in Penetration Enhancement: Nanocarriers, Springer.","DOI":"10.1007\/978-3-662-47862-2"}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/20\/8\/476\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:56:14Z","timestamp":1760140574000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/20\/8\/476"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,25]]},"references-count":57,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["md20080476"],"URL":"https:\/\/doi.org\/10.3390\/md20080476","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,25]]}}}