{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T06:34:57Z","timestamp":1789626897437,"version":"build-2803163510"},"reference-count":38,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,8,11]],"date-time":"2021-08-11T00:00:00Z","timestamp":1628640000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Melittin is a promising antitumor substance; however, it is a nonspecific cytolytic peptide, which limits its clinical application. In this study, melittin liposomes (Mel-Lip) and hyaluronic acid (HA)-modified Mel-Lip (Mel-HA-Lip) were designed to reduce the toxicity and increase the anti-tumor effects of melittin. The optimal preparation procedure was evaluated using a uniform design based on the single factor method, and the concentration of HA was determined based on the cellular uptake of coumarin 6 labeled HA-Lip. Liposomes and HA-modified liposomes were evaluated in vitro by assessing cytotoxicity, cellular uptake, and release behavior. Liposomes prepared in the optimum formulation improved stability, with a particle size of 132.7 \u00b1 1.55 nm, zeta potential of \u221211.5 \u00b1 1.51 mV, entrapment efficiency of 86.25 \u00b1 1.28%, and drug-loading efficiency of 3.91 \u00b1 0.49%. Cellular uptake tests revealed that the uptake of nanoparticles significantly increased with HA modification, suggesting that HA modification enhanced the internalization of liposomes within cells, which was consistent with the results of the cytotoxicity analysis. Furthermore, in vitro release experiments showed that Mel-HA-Lip possessed a stronger sustained-release effect compared with Mel-Lip. The results of this experiment provide insight into the potential tumor-targeting effects of melittin.<\/jats:p>","DOI":"10.3390\/pharmaceutics13081235","type":"journal-article","created":{"date-parts":[[2021,8,11]],"date-time":"2021-08-11T04:56:22Z","timestamp":1628657782000},"page":"1235","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":44,"title":["Hyaluronic Acid Coating Reduces the Leakage of Melittin Encapsulated in Liposomes and Increases Targeted Delivery to Melanoma Cells"],"prefix":"10.3390","volume":"13","author":[{"given":"Yanyan","family":"Li","sequence":"first","affiliation":[{"name":"Department of Pharmaceutics, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuyao","family":"Ruan","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5303-2941","authenticated-orcid":false,"given":"Nianping","family":"Feng","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1427-4868","authenticated-orcid":false,"given":"Yongtai","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"115","DOI":"10.3322\/caac.21338","article-title":"Cancer statistics in China, 2015","volume":"66","author":"Chen","year":"2016","journal-title":"CA A Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA A Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1602567","DOI":"10.1002\/smll.201602567","article-title":"Safe and Effective Reversal of Cancer Multidrug Resistance Using Sericin-Coated Mesoporous Silica Nanoparticles for Lysosome-Targeting Delivery in Mice","volume":"13","author":"Liu","year":"2017","journal-title":"Small"},{"key":"ref_4","first-page":"747","article-title":"Research progress on drugs for treatment of nasopharyngeal carcinoma","volume":"32","author":"Chen","year":"2017","journal-title":"Drugs Clinic."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.canlet.2018.02.004","article-title":"Osimertinib resistance in non-small cell lung cancer: Mechanisms and therapeutic strategies","volume":"420","author":"Tang","year":"2018","journal-title":"Cancer Lett."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.canlet.2017.05.010","article-title":"Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy","volume":"402","author":"Rady","year":"2017","journal-title":"Cancer Lett."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1016\/j.pharmthera.2007.04.004","article-title":"Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds","volume":"115","author":"Son","year":"2007","journal-title":"Pharmacol. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1007\/s00280-016-3160-1","article-title":"Application of bee venom and its main constituent melittin for cancer treatment","volume":"78","author":"Liu","year":"2016","journal-title":"Cancer Chemother. Pharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1126\/science.177.4046.314","article-title":"Bee and wasp venoms","volume":"177","author":"Habermann","year":"1972","journal-title":"Science"},{"key":"ref_10","first-page":"931","article-title":"The anti-tumour effect of Mel and its role in autophagy in human hepatocellular carcinoma cells","volume":"11","author":"Lv","year":"2019","journal-title":"Am. J. Transl. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"240","DOI":"10.2174\/1389203719666180612084615","article-title":"Anti-Tumor Effects of Melittin and Its Potential Applications in Clinic","volume":"20","author":"Lyu","year":"2019","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1615\/ForumImmunDisTher.2014008125","article-title":"Possible Molecular Targets of Bee Venom in the Treatment of Cancer: Application and Perspectives","volume":"4","year":"2013","journal-title":"Forum Immunopathol. Dis. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1007\/s00249-005-0033-7","article-title":"A molecular dynamics study of the bee venom melittin in aqueous solution, in methanol, and inserted in a phospholipid bilayer","volume":"35","author":"Glattli","year":"2006","journal-title":"Eur. Biophys. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1522","DOI":"10.1016\/j.chembiol.2014.09.008","article-title":"The membrane-lytic peptides K8L9 and melittin enter cancer cells via receptor endocytosis following subcytotoxic exposure","volume":"21","author":"Kohno","year":"2014","journal-title":"Chem. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3186","DOI":"10.3748\/wjg.v22.i11.3186","article-title":"Melittin induces human gastric cancer cell apoptosis via activation of mitochondrial pathway","volume":"22","author":"Kong","year":"2016","journal-title":"World J. Gastroenterol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2759","DOI":"10.1007\/s00726-014-1833-9","article-title":"Melittin peptides exhibit different activity on different cells and model membranes","volume":"46","author":"Jamasbi","year":"2014","journal-title":"Amino Acids."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s13045-017-0442-y","article-title":"A novel melittin nano-liposome exerted excellent anti-hepatocellular carcinoma efficacy with better biological safety","volume":"10","author":"Mao","year":"2017","journal-title":"J. Hematol. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.bbrc.2021.03.073","article-title":"Enhancement of target toxin neutralization effect in vivo by PEGylation of multifunctionalized lipid nanoparticles","volume":"555","author":"Koide","year":"2021","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Gu, Z., Da Silva, C.G., Van der Maaden, K., Ossendorp, F., and Cruz, L.J. (2020). Liposome-Based Drug Delivery Systems in Cancer Immunotherapy. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12111054"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2419","DOI":"10.1529\/biophysj.104.043596","article-title":"Interaction of melittin with membrane cholesterol: A fluorescence approach","volume":"87","author":"Raghuraman","year":"2004","journal-title":"Biophys. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.addr.2015.11.011","article-title":"Hyaluronic acid for anticancer drug and nucleic acid delivery","volume":"97","author":"Dosio","year":"2016","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Bellei, B., Caputo, S., Migliano, E., Lopez, G., Marcaccini, V., Cota, C., and Picardo, M. (2021). Simultaneous Targeting Tumor Cells and Cancer-Associated Fibroblasts with a Paclitaxel-Hyaluronan Bioconjugate: In Vitro Evaluation in Non-Melanoma Skin Cancer. Biomedicines, 9.","DOI":"10.3390\/biomedicines9060597"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.jcis.2021.04.056","article-title":"Redox-responsive hyaluronic acid-based nanoparticles for targeted photodynamic therapy\/chemotherapy against breast cancer","volume":"598","author":"Wang","year":"2021","journal-title":"J. Colloid Interface Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5258","DOI":"10.1016\/j.biomaterials.2010.03.018","article-title":"Target specific tumor treatment by VEGF siRNA complexed with reducible polyethyleneimine-hyaluronic acid conjugate","volume":"31","author":"Park","year":"2010","journal-title":"Biomaterials"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.semcancer.2008.03.015","article-title":"Involvement of CD44, a molecule with a thousand faces, in cancer dissemination","volume":"18","author":"Naor","year":"2008","journal-title":"Semin. Cancer Biol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Koltai, T., Reshkin, S.J., Carvalho, T.M.A., and Cardone, R.A. (2021). Targeting the Stromal Pro-Tumoral Hyaluronan-CD44 Pathway in Pancreatic Cancer. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22083953"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1062","DOI":"10.1021\/mp800215d","article-title":"Lipoplexes Targeting the CD44 Hyaluronic Acid Receptor for Efficient Transfection of Breast Cancer Cells","volume":"6","author":"Surace","year":"2009","journal-title":"Mol. Pharm."},{"key":"ref_28","first-page":"2592","article-title":"Liposome-encapsulated doxorubicin targeted to CD44: A strategy to kill CD44-overexpressing tumor cells","volume":"61","author":"Eliaz","year":"2001","journal-title":"Cancer Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1016\/j.ijbiomac.2020.11.039","article-title":"Study on the anti-infection ability of vancomycin cationic liposome combined with polylactide fracture internal fixator","volume":"167","author":"Cai","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_30","first-page":"117","article-title":"Preparation and characterization of hyaluronic acid-modified dexamethasone core-shell liponanoparticles","volume":"44","author":"Jing","year":"2013","journal-title":"J. China Pharm. Univ."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"48","DOI":"10.7150\/thno.29715","article-title":"CD44 Assists the Topical Anti-Psoriatic Efficacy of Curcumin-Loaded Hyaluronan-Modified Ethosomes: A New Strategy for Clustering Drug in Inflammatory Skin","volume":"9","author":"Zhang","year":"2019","journal-title":"Theranostics"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5977","DOI":"10.2147\/IJN.S259938","article-title":"Preparation, Characterization and in vivo Study of Borneol-Baicalin-Liposomes for Treatment of Cerebral Ischemia-Reperfusion Injury","volume":"15","author":"Zhang","year":"2020","journal-title":"Int. J. Nanomed."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"110337","DOI":"10.1016\/j.colsurfb.2019.06.066","article-title":"Improving systemic circulation of paclitaxel nanocrystals by surface hybridization of DSPE-PEG2000","volume":"182","author":"Wang","year":"2019","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.jconrel.2016.03.012","article-title":"Hyaluronic acid-coated liposomes for targeted delivery of paclitaxel, in-vitro characterization and in-vivo evaluation","volume":"229","author":"Ravar","year":"2016","journal-title":"J Control. Release"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.jconrel.2013.11.016","article-title":"Hyaluronic acid derivative-coated nanohybrid liposomes for cancer imaging and drug delivery","volume":"174","author":"Park","year":"2014","journal-title":"J. Control. Release"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/j.jconrel.2016.01.049","article-title":"Hyaluronic acid ion-pairing nanoparticles for targeted tumor therapy","volume":"225","author":"Li","year":"2016","journal-title":"J. Control. Release"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"15287","DOI":"10.3390\/ijms150915287","article-title":"Dual Targeting Biomimetic Liposomes for Paclitaxel\/DNA Combination Cancer Treatment","volume":"15","author":"Liu","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Sangboonruang, S., Kitidee, K., Chantawannakul, P., Tragoolpua, K., and Tragoolpua, Y. (2020). Melittin from Apis florea Venom as a Promising Therapeutic Agent for Skin Cancer Treatment. 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