{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T02:38:42Z","timestamp":1790908722140,"version":"4.1.0"},"reference-count":41,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,12,20]],"date-time":"2022-12-20T00:00:00Z","timestamp":1671494400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Foundation for Cystic fibrosis","award":["FFC#17\/2016"],"award-info":[{"award-number":["FFC#17\/2016"]}]},{"name":"Italian Foundation for Cystic fibrosis","award":["I69C20000320002"],"award-info":[{"award-number":["I69C20000320002"]}]},{"name":"Regione Toscana","award":["FFC#17\/2016"],"award-info":[{"award-number":["FFC#17\/2016"]}]},{"name":"Regione Toscana","award":["I69C20000320002"],"award-info":[{"award-number":["I69C20000320002"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Development of inhalable formulations for delivering peptides to the conductive airways and shielding their interactions with airway barriers, thus enhancing peptide\/bacteria interactions, is an important part of peptide-based drug development for lung applications. Here, we report the construction of a biocompatible nanosystem where the antimicrobial peptide SET-M33 is encapsulated within polymeric nanoparticles of poly(lactide-co-glycolide) (PLGA) conjugated with polyethylene glycol (PEG). This system was conceived for better delivery of the peptide to the lungs by aerosol. The encapsulated peptide showed prolonged antibacterial activity, due to its controlled release, and much lower toxicity than the free molecule. The peptide-based nanosystem killed Pseudomonas aeruginosa in planktonic and sessile forms in a dose-dependent manner, remaining active up to 72 h after application. The encapsulated peptide showed no cytotoxicity when incubated with human bronchial epithelial cells from healthy individuals and from cystic fibrosis patients, unlike the free peptide, which showed an EC50 of about 22 \u00b5M. In vivo acute toxicity studies in experimental animals showed that the peptide nanosystem did not cause any appreciable side effects, and confirmed its ability to mitigate the toxic and lethal effects of free SET-M33.<\/jats:p>","DOI":"10.3390\/pharmaceutics15010003","type":"journal-article","created":{"date-parts":[[2022,12,20]],"date-time":"2022-12-20T04:45:53Z","timestamp":1671511553000},"page":"3","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Inhalable Polymeric Nanoparticles for Pulmonary Delivery of Antimicrobial Peptide SET-M33: Antibacterial Activity and Toxicity In Vitro and In Vivo"],"prefix":"10.3390","volume":"15","author":[{"given":"Laura","family":"Cresti","sequence":"first","affiliation":[{"name":"Laboratory of Clinical Pathology, Santa Maria alle Scotte University Hospital, 53100 Siena, Italy"},{"name":"SetLance srl, 53100 Siena, Italy"},{"name":"Department of Medical Biotechnology, University of Siena, 53100 Siena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8432-1519","authenticated-orcid":false,"given":"Gemma","family":"Conte","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u201cLuigi Vanvitelli\u201d, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Cappello","sequence":"additional","affiliation":[{"name":"SetLance srl, 53100 Siena, Italy"},{"name":"Department of Medical Biotechnology, University of Siena, 53100 Siena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8144-7186","authenticated-orcid":false,"given":"Jlenia","family":"Brunetti","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, University of Siena, 53100 Siena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1027-8482","authenticated-orcid":false,"given":"Chiara","family":"Falciani","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, University of Siena, 53100 Siena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0738-5746","authenticated-orcid":false,"given":"Luisa","family":"Bracci","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Pathology, Santa Maria alle Scotte University Hospital, 53100 Siena, Italy"},{"name":"Department of Medical Biotechnology, University of Siena, 53100 Siena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6223-0782","authenticated-orcid":false,"given":"Fabiana","family":"Quaglia","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Naples Federico II, 80131 Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0850-9533","authenticated-orcid":false,"given":"Francesca","family":"Ungaro","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, University of Naples Federico II, 80131 Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0444-0444","authenticated-orcid":false,"given":"Ivana","family":"d\u2019Angelo","sequence":"additional","affiliation":[{"name":"Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania \u201cLuigi Vanvitelli\u201d, 81100 Caserta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro","family":"Pini","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Pathology, Santa Maria alle Scotte University Hospital, 53100 Siena, Italy"},{"name":"Department of Medical Biotechnology, University of Siena, 53100 Siena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Murugaiyan, J., Kumar, P.A., Rao, G.S., Iskandar, K., Hawser, S., Hays, J.P., Mohsen, Y., Adukkadukkam, S., Awuah, W.A., and Jose, R.A.M. (2022). Progress in Alternative Strategies to Combat Antimicrobial Resistance: Focus on Antibiotics. Antibiotics, 11.","DOI":"10.3390\/antibiotics11020200"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"641","DOI":"10.2174\/138920371408131227155308","article-title":"The development of antimicrobial peptides as new antibacterial drugs","volume":"14","author":"Roscia","year":"2013","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2665","DOI":"10.1128\/AAC.49.7.2665-2672.2005","article-title":"Antimicrobial activity of novel dendrimeric peptides obtained by phage display selection and rational modification","volume":"49","author":"Pini","year":"2005","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1002\/psc.858","article-title":"Characterization of the branched antimicrobial peptide M6 by analyzing its mechanism of action and in vivo toxicity","volume":"13","author":"Pini","year":"2007","journal-title":"J. Pept. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1096\/fj.09-145474","article-title":"A novel tetrabranched antimicrobial peptide that neutralizes bacterial lipopolysaccharide and prevents septic shock in vivo","volume":"24","author":"Pini","year":"2010","journal-title":"FASEB J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1007\/s00726-011-1103-z","article-title":"Efficacy and toxicity of the antimicrobial peptide M33 produced with different counter-ions","volume":"43","author":"Pini","year":"2012","journal-title":"Amino Acids"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1111\/j.1747-0285.2007.00487.x","article-title":"Molecular basis of branched peptides resistance to enzyme proteolysis","volume":"69","author":"Falciani","year":"2007","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Falciani, C., Lozzi, L., Pollini, S., Luca, V., Carnicelli, V., Brunetti, J., Lelli, B., Bindi, S., Scali, S., and Di Giulio, A. (2012). Isomerization of an antimicrobial peptide broadens antimicrobial spectrum to gram-positive bacterial pathogens. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0046259"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1403","DOI":"10.1007\/s00726-014-1686-2","article-title":"Site-specific pegylation of an antimicrobial peptide increases resistance to Pseudomonas aeruginosa elastase","volume":"46","author":"Falciani","year":"2014","journal-title":"Amino Acids"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Brunetti, J., Falciani, C., Roscia, G., Pollini, S., Bindi, S., Scali, S., Arrieta, U.C., G\u00f3mez-Vallejo, V., Quercini, L., and Ibba, E. (2016). In vitro and in vivo efficacy, toxicity, bio-distribution and resistance selection of a novel antibacterial drug candidate. Sci. Rep., 6.","DOI":"10.1038\/srep26077"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"25742","DOI":"10.1074\/jbc.M116.750257","article-title":"Immunomodulatory and anti-inflammatory activity in vitro and in vivo of a novel antimicrobial candidate","volume":"291","author":"Brunetti","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"613","DOI":"10.2174\/1568026616666160713143017","article-title":"Models of In-Vivo Bacterial Infections for the Development of Antimicrobial Peptide-based Drugs","volume":"17","author":"Brunetti","year":"2017","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1796","DOI":"10.1016\/j.bbamem.2017.06.001","article-title":"Investigations into the killing activity of an antimicrobial peptide active against extensively antibiotic-resistant K. pneumoniae and P. aeruginosa","volume":"1859","author":"Brunetti","year":"2017","journal-title":"Biochim. Biophys. Acta Biomembr."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Brunetti, J., Carnicelli, V., Ponzi, A., Di Giulio, A., Lizzi, A.R., Cristiano, L., Cresti, L., Cappello, G., Pollini, S., and Mosconi, L. (2020). Antibacterial and anti-inflammatory activity of an antimicrobial peptide aynthesized with D amino acids. Antibiotics, 9.","DOI":"10.3390\/antibiotics9120840"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1096\/fj.201901896RR","article-title":"An antimicrobial molecule mitigates signs of sepsis in vivo and eradicates infections from lung tissue","volume":"34","author":"Quercini","year":"2020","journal-title":"FASEB J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.2147\/IJN.S218966","article-title":"Antimicrobial peptide-loaded nanoparticles as inhalation therapy for Pseudomonas aeruginosa infections","volume":"15","author":"Falciani","year":"2020","journal-title":"Int. J. Nanomed."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"57","DOI":"10.3109\/02652040902954729","article-title":"Improved delivery of angiogenesis inhibitors from PLGA:poloxamer blend micro- and nanoparticles","volume":"27","author":"Alonso","year":"2010","journal-title":"J. Microencapsul."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.colsurfb.2015.08.027","article-title":"Overcoming barriers in Pseudomonas aeruginosa lung infections: Engineered nanoparticles for local delivery of a cationic antimicrobial peptide","volume":"135","author":"Casciaro","year":"2015","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1876","DOI":"10.1021\/acs.biomac.8b01829","article-title":"Poly(lactide- co-glycolide) Nanoparticles for Prolonged Therapeutic Efficacy of Esculentin-1a-Derived Antimicrobial Peptides against Pseudomonas aeruginosa Lung Infection: In Vitro and in Vivo Studies","volume":"20","author":"Casciaro","year":"2019","journal-title":"Biomacromolecules"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.addr.2017.08.010","article-title":"PEGylation for enhancing nanoparticle diffusion in mucus","volume":"124","author":"Huckaby","year":"2018","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9726","DOI":"10.1002\/anie.200803526","article-title":"Addressing the PEG mucoadhesivity paradox to engineer nanoparticles that \"slip\" through the human mucus barrier","volume":"47","author":"Wang","year":"2008","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"365","DOI":"10.2217\/nnm.10.123","article-title":"Rapid transport of muco-inert nanoparticles in cystic fibrosis sputum treated with N-acetyl cysteine","volume":"6","author":"Suk","year":"2011","journal-title":"Nanomedicine"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1021\/mp8001254","article-title":"Controlled surface modification with poly(ethylene)glycol enhances diffusion of PLGA nanoparticles in human cervical mucus","volume":"6","author":"Cu","year":"2009","journal-title":"Mol. Pharm."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1073\/pnas.0608611104","article-title":"Rapid transport of large polymeric nanoparticles in fresh undiluted human mucus","volume":"104","author":"Lai","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3439","DOI":"10.1016\/j.biomaterials.2013.01.064","article-title":"Nanoparticle diffusion in respiratory mucus from humans without lung disease","volume":"34","author":"Schuster","year":"2013","journal-title":"Biomaterials"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"935","DOI":"10.2217\/nnm.12.129","article-title":"Transport of nanoparticles in cystic fibrosis sputum and bacterial biofilms by single-particle tracking microscopy","volume":"8","author":"Forier","year":"2013","journal-title":"Nanomedicine"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.ejpb.2018.07.025","article-title":"Polyester-based particles to overcome the obstacles of mucus and biofilms in the lung for tobramycin application under static and dynamic fluidic conditions","volume":"131","author":"Ernst","year":"2018","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.jconrel.2011.08.010","article-title":"Dry powders based on PLGA nanoparticles for pulmonary delivery of antibiotics: Modulation of encapsulation efficiency, release rate and lung deposition pattern by hydrophilic polymers","volume":"157","author":"Ungaro","year":"2012","journal-title":"J. Control Release"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/S0378-5173(02)00075-3","article-title":"Effects of mixed polyethyleneglycol modification on fixed aqueous layer thickness and antitumor activity of doxorubicin containing liposome","volume":"238","author":"Sadzuka","year":"2002","journal-title":"Int. J. Pharm."},{"key":"ref_30","first-page":"447","article-title":"Simulants of lung interstitial fluid","volume":"36","author":"Moss","year":"1979","journal-title":"Health Phys."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0021-9258(18)65874-0","article-title":"Acetylornithinase of Escherichia coli: Partial purification and some properties","volume":"218","author":"Vogel","year":"1956","journal-title":"J. Biol. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2773","DOI":"10.1007\/s00018-013-1291-7","article-title":"Esculentin(1-21), an amphibian skin membrane-active peptide with potent activity on both planktonic and biofilm cells of the bacterial pathogen Pseudomonas aeruginosa","volume":"70","author":"Luca","year":"2013","journal-title":"Cell Mol. Life Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/j.ejpb.2012.04.004","article-title":"In vitro and in vivo evaluation of a dry powder endotracheal insufflator device for use in dose-dependent preclinical studies in mice","volume":"81","author":"Duret","year":"2012","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.ejpb.2014.06.014","article-title":"In vitro evaluation of the DP-4M PennCentury insufflator","volume":"88","author":"Hoppentocht","year":"2014","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1126\/scitranslmed.3003453","article-title":"Mucus-penetrating nanoparticles for vaginal drug delivery protect against herpes simplex virus","volume":"4","author":"Ensign","year":"2012","journal-title":"Sci. Transl. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/j.jconrel.2013.05.035","article-title":"Scalable method to produce biodegradable nanoparticles that rapidly penetrate human mucus","volume":"170","author":"Xu","year":"2013","journal-title":"J. Control. Release"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Cano, A., Ettcheto, M., Espina, M., L\u00f3pez-Machado, A., Cajal, Y., Rabanal, F., S\u00e1nchez-L\u00f3pez, E., Camins, A., Garc\u00eda, M.L., and Souto, E.B. (2020). State-of-the-art polymeric nanoparticles as promising therapeutic tools against human bacterial infections. J. Nanobiotechnol., 18.","DOI":"10.1186\/s12951-020-00714-2"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"429","DOI":"10.2174\/1389203720666191203101947","article-title":"Inorganic Gold and Polymeric Poly(Lactide-co-glycolide) Nanoparticles as Novel Strategies to Ameliorate the Biological Properties of Antimicrobial Peptides","volume":"21","author":"Casciaro","year":"2020","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.jcf.2020.10.008","article-title":"Factors influencing the acquisition and eradication of early Pseudomonas aeruginosa infection in cystic fibrosis","volume":"20","author":"Jackson","year":"2021","journal-title":"J. Cyst. Fibros."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Cresti, L., Falciani, C., Cappello, G., Brunetti, J., Vailati, S., Melloni, E., Bracci, L., and Pini, A. (2022). Safety evaluations of a synthetic antimicrobial peptide administered intravenously in rats and dogs. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-23841-2"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"386","DOI":"10.2174\/1568026615666150108123256","article-title":"Pulmonary drug delivery: A role for polymeric nanoparticles?","volume":"15","author":"Conte","year":"2015","journal-title":"Curr. Top. Med. Chem."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/1\/3\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:44:35Z","timestamp":1760147075000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/15\/1\/3"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,20]]},"references-count":41,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["pharmaceutics15010003"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics15010003","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,20]]}}}