{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T23:05:42Z","timestamp":1788303942563,"version":"build-2803163510"},"reference-count":41,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T00:00:00Z","timestamp":1742169600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Unione europea- Next Generation EU","award":["CUP D53D23010830006"],"award-info":[{"award-number":["CUP D53D23010830006"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Self-assembling amphiphilic polymers represent highly promising materials with emerging applications across various fields. In these polymers, the presence of hydrophilic and hydrophobic segments within their structure drives the self-assembly process in aqueous environments, leading to organized structures capable of incorporating lipophilic drugs. Their high chemical versatility enables the design of tailored structures to meet specific requirements, such as the active targeting ability, thereby broadening their potential applications. In this work, a polyethylene glycol-phospholipid conjugate was employed to form nanocarriers loaded with a lipophilic derivative of gemcitabine. To achieve nano-assemblies actively targeted towards cancer cells overexpressing the hyaluronic acid (HA) receptor CD44, a HA-phospholipid conjugate was co-formulated in various molar ratios (1%, 10%, and 20%). All formulations exhibited a mean diameter below 130 nm, a negative zeta potential (approximately \u221230 mV), and a high encapsulation efficiency (above 90%). These nano-assemblies demonstrated stability during storage and effectively released the encapsulated drug in a cell culture medium. Upon incubation with cancer cells, the nano-assemblies were internalized via a CD44 endocytosis-mediated mechanism, with the extent of internalization depending on the HA conjugate content. Consistently, cell viability studies revealed that the nanocarriers decorated with higher amounts of HA exerted a higher cytotoxicity, enabling a fine tuning of the nano-assembly properties.<\/jats:p>","DOI":"10.3390\/molecules30061349","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T15:24:56Z","timestamp":1742225096000},"page":"1349","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Tailoring the Composition of HA\/PEG Mixed Nano-Assemblies for Anticancer Drug Delivery"],"prefix":"10.3390","volume":"30","author":[{"given":"Beatrice","family":"Zurletti","sequence":"first","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9457-3667","authenticated-orcid":false,"given":"Ilaria","family":"Andreana","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6043-3794","authenticated-orcid":false,"given":"Iris Chiara","family":"Salaroglio","sequence":"additional","affiliation":[{"name":"Department of Oncology, Interdepartmental Center of Molecular Biotechnology \u201cGuido Tarone\u201d, University of Turin, Via Nizza 44, 10126 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3862-4698","authenticated-orcid":false,"given":"Valeria","family":"Bincoletto","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4427-7939","authenticated-orcid":false,"given":"Maela","family":"Manzoli","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6138-1503","authenticated-orcid":false,"given":"Barbara","family":"Rolando","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4245-4161","authenticated-orcid":false,"given":"Paola","family":"Milla","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9787-4836","authenticated-orcid":false,"given":"Chiara","family":"Riganti","sequence":"additional","affiliation":[{"name":"Department of Oncology, Interdepartmental Center of Molecular Biotechnology \u201cGuido Tarone\u201d, University of Turin, Via Nizza 44, 10126 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8266-6604","authenticated-orcid":false,"given":"Barbara","family":"Stella","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Arpicco","sequence":"additional","affiliation":[{"name":"Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.jconrel.2021.02.031","article-title":"Polymeric Micelles in Drug Delivery: An Insight of the Techniques for Their Characterization and Assessment in Biorelevant Conditions","volume":"332","author":"Ghezzi","year":"2021","journal-title":"J. Control Release"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.jconrel.2021.02.016","article-title":"Polymeric Micelles in Cancer Therapy: State of the Art","volume":"332","author":"Ghosh","year":"2021","journal-title":"J. Control Release"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1016\/j.ejpb.2006.11.009","article-title":"A Review of the Formation and Classification of Amphiphilic Block Copolymer Nanoparticulate Structures: Micelles, Nanospheres, Nanocapsules and Polymersomes","volume":"65","author":"Letchford","year":"2007","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_4","first-page":"4185","article-title":"Application of Poly(Ethylene Glycol)- Distearoylphosphatidylethanolamine (PEG-DSPE) Block Copolymers and Their Derivatives as Nanomaterials in Drug Delivery","volume":"7","author":"Wang","year":"2012","journal-title":"Int. J. Nanomed."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1598","DOI":"10.2174\/1381612821666150115144003","article-title":"DSPE-PEG: A Distinctive Component in Drug Delivery System","volume":"21","author":"Che","year":"2015","journal-title":"Curr. Pharm. Des."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2058","DOI":"10.1021\/acsami.3c13948","article-title":"Exploring the Impact of Nanoparticle Stealth Coatings in Cancer Models: From PEGylation to Cell Membrane-Coating Nanotechnology","volume":"16","author":"Pedrosa","year":"2024","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1021\/acs.molpharmaceut.3c00774","article-title":"Impact of Anti-PEG IgM Induced via the Topical Application of a Cosmetic Product Containing PEG Derivatives on the Antitumor Effects of PEGylated Liposomal Antitumor Drug Formulations in Mice","volume":"21","author":"Gaballa","year":"2024","journal-title":"Mol. Pharm."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"106518","DOI":"10.1016\/j.jddst.2024.106518","article-title":"Smart Hyaluronated Micelles to Enhance a Gemcitabine Prodrug Efficacy","volume":"104","author":"Andreana","year":"2025","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.msec.2014.01.052","article-title":"Chemical Functionalization of Hyaluronic Acid for Drug Delivery Applications","volume":"38","author":"Vasi","year":"2014","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Bayer, I.S. (2020). Hyaluronic Acid and Controlled Release: A Review. Molecules, 25.","DOI":"10.3390\/molecules25112649"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"118006","DOI":"10.1016\/j.carbpol.2021.118006","article-title":"Hyaluronic Acid in Ocular Drug Delivery","volume":"264","author":"Zhang","year":"2021","journal-title":"Carbohydr. Polym."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"108706","DOI":"10.1016\/j.carres.2022.108706","article-title":"Hyaluronic Acid and Cholecalciferol Conjugate Based Nanomicelles: Synthesis, Characterization, and Cytotoxicity against MCF-7 Breast Cancer Cells","volume":"522","author":"Gao","year":"2022","journal-title":"Carbohydr. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2400513","DOI":"10.1002\/smtd.202400513","article-title":"Hyaluronic Acid Receptor-Mediated Nanomedicines and Targeted Therapy","volume":"8","author":"Ouyang","year":"2024","journal-title":"Small Methods"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.jconrel.2004.09.001","article-title":"Preparation, Characterization, Cytotoxicity and Pharmacokinetics of Liposomes Containing Lipophilic Gemcitabine Prodrugs","volume":"100","author":"Immordino","year":"2004","journal-title":"J. Control Release"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.jconrel.2011.08.015","article-title":"EGFR-Targeted Stearoyl Gemcitabine Nanoparticles Show Enhanced Anti-Tumor Activity","volume":"157","author":"Sandoval","year":"2012","journal-title":"J. Control Release"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Chiesa, E., Greco, A., Riva, F., Dorati, R., Conti, B., Modena, T., and Genta, I. (2022). CD44-Targeted Carriers: The Role of Molecular Weight of Hyaluronic Acid in the Uptake of Hyaluronic Acid-Based Nanoparticles. Pharmaceuticals, 15.","DOI":"10.3390\/ph15010103"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.ejpb.2013.06.003","article-title":"Hyaluronic Acid-Coated Liposomes for Active Targeting of Gemcitabine","volume":"85","author":"Arpicco","year":"2013","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.jconrel.2023.01.021","article-title":"Accelerated Clearance by Antibodies against Methoxy PEG Depends on Pegylation Architecture","volume":"354","author":"Lin","year":"2023","journal-title":"J. Control Release"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2749","DOI":"10.1021\/acs.biomac.3c01397","article-title":"Investigation of Basolateral Targeting Micelles for Drug Delivery Applications in Polycystic Kidney Disease","volume":"25","author":"Huang","year":"2024","journal-title":"Biomacromolecules"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1007\/BF02531316","article-title":"Two Dimensional Thin Layer Chromatographic Separation of Polar Lipids and Determination of Phospholipids by Phosphorus Analysis of Spots","volume":"5","author":"Rouser","year":"1970","journal-title":"Lipids"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1016\/0003-2697(62)90095-7","article-title":"A Modified Uronic Acid Carbazole Reaction","volume":"4","author":"Bitter","year":"1962","journal-title":"Anal. Biochem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1039\/C3MH00106G","article-title":"Protein Corona Formation around Nanoparticles\u2014From the Past to the Future","volume":"1","author":"Pino","year":"2014","journal-title":"Mater. Horiz."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6287","DOI":"10.1039\/C4CS00487F","article-title":"Nanoparticle Colloidal Stability in Cell Culture Media and Impact on Cellular Interactions","volume":"44","author":"Moore","year":"2015","journal-title":"Chem. Soc. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1208\/s12249-018-1275-x","article-title":"The Impact of Serum Proteins and Surface Chemistry on Magnetic Nanoparticle Colloidal Stability and Cellular Uptake in Breast Cancer Cells","volume":"20","author":"Mekseriwattana","year":"2019","journal-title":"AAPS PharmSciTech"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4150","DOI":"10.1021\/bm401312j","article-title":"Achieving Micelle Control through Core Crystallinity","volume":"14","author":"Glavas","year":"2013","journal-title":"Biomacromolecules"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2920","DOI":"10.1021\/acs.biomac.5b00826","article-title":"Decreased Core Crystallinity Facilitated Drug Loading in Polymeric Micelles without Affecting Their Biological Performances","volume":"16","author":"Gou","year":"2015","journal-title":"Biomacromolecules"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Sathishbabu, P., Uthaiah, C.A., and Hani, U. (2025). Comprehensive Evaluation of EGFR and AKT Targeting Efficacy of Resveratrol Loaded PEGylated Liposomes for the Glioblastoma Management: In Silico, in Vitro BBB Permeation Studies. Bioorg Chem, 154.","DOI":"10.1016\/j.bioorg.2024.108077"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"102756","DOI":"10.1016\/j.jddst.2021.102756","article-title":"Preparation and Evaluation of Gemcitabin and Cisplatin-Entrapped Folate-PEGylated Liposomes as Targeting Co-Drug Delivery System in Cancer Therapy","volume":"65","author":"Parivar","year":"2021","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1002\/pat.1702","article-title":"Crystallization, Rheological, and Mechanical Properties of PLLA\/PEG Blend with Multiwalled Carbon Nanotubes","volume":"22","author":"Li","year":"2011","journal-title":"Polym. Adv. Technol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"42120","DOI":"10.1039\/D0RA08699A","article-title":"Effect of Molecular Weight of Polyethylene Glycol on Crystallization Behaviors, Thermal Properties and Tensile Performance of Polylactic Acid Stereocomplexes","volume":"10","author":"Li","year":"2020","journal-title":"RSC Adv."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1140","DOI":"10.1021\/mp4005904","article-title":"Enhanced Tumor Delivery of Gemcitabine via PEG-DSPE\/TPGS Mixed Micelles","volume":"11","author":"Wang","year":"2014","journal-title":"Mol. Pharm."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.ijpharm.2014.08.030","article-title":"Synthesis and Optimization of a Novel Polymeric Micelle Based on Hyaluronic Acid and Phospholipids for Delivery of Paclitaxel, in Vitro and in-Vivo Evaluation","volume":"475","author":"Saadat","year":"2014","journal-title":"Int. J. Pharm."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1203","DOI":"10.1021\/acs.molpharmaceut.7b01035","article-title":"Hyaluronic Acid-Modified Micelles Encapsulating Gem-C <inf>12<\/Inf> and HNK for Glioblastoma Multiforme Chemotherapy","volume":"15","author":"Liu","year":"2018","journal-title":"Mol. Pharm."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Saadat, E., Amini, M., Dinarvand, R., and Dorkoosh, F.A. (2014). Polymeric Micelles Based on Hyaluronic Acid and Phospholipids: Design, Characterization, and Cytotoxicity. J. Appl. Polym. Sci., 131.","DOI":"10.1002\/app.40944"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3185","DOI":"10.1200\/JCO.2020.38.4_suppl.638","article-title":"Randomized Phase III Trial of Pegvorhyaluronidase Alfa with Nab-Paclitaxel plus Gemcitabine for Patients with Hyaluronan-High Metastatic Pancreatic Adenocarcinoma","volume":"38","author":"Tempero","year":"2020","journal-title":"J. Clin. Oncol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.jconrel.2018.02.024","article-title":"Anti-Tumor Efficacy of Hyaluronan-Based Nanoparticles for the Co-Delivery of Drugs in Lung Cancer","volume":"275","author":"Jeannot","year":"2018","journal-title":"J. Control Release"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.jconrel.2017.08.016","article-title":"A Hyaluronic Acid Conjugate Engineered to Synergistically and Sequentially Deliver Gemcitabine and Doxorubicin to Treat Triple Negative Breast Cancer","volume":"267","author":"Vogus","year":"2017","journal-title":"J. Control Release"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1186\/s13046-024-03028-5","article-title":"A Paclitaxel-Hyaluronan Conjugate (ONCOFID-P-BTM) in Patients with BCG-Unresponsive Carcinoma in Situ of the Bladder: A Dynamic Assessment of the Tumor Microenvironment","volume":"43","author":"Tosi","year":"2024","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.jcis.2010.10.024","article-title":"Curcumin-Loaded PLGA-PEG-PLGA Triblock Copolymeric Micelles: Preparation, Pharmacokinetics and Distribution in Vivo","volume":"354","author":"Song","year":"2011","journal-title":"J. Colloid Interface Sci."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Karsten, E., Breen, E., McCracken, S.A., Clarke, S., and Herbert, B.R. (2020). Red Blood Cells Exposed to Cancer Cells in Culture Have Altered Cytokine Profiles and Immune Function. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-64319-3"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"116798","DOI":"10.1016\/j.carbpol.2020.116798","article-title":"Synthesis of Defined Oligohyaluronates-Decorated Liposomes and Interaction with Lung Cancer Cells","volume":"248","author":"Cano","year":"2020","journal-title":"Carbohydr. Polym."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/6\/1349\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:55:21Z","timestamp":1760028921000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/30\/6\/1349"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,17]]},"references-count":41,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["molecules30061349"],"URL":"https:\/\/doi.org\/10.3390\/molecules30061349","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,17]]}}}