Bracotti, Francesco

Butnarasu, Cosmin

FIORIO PLA, Alessandra

Visentin, Sonja

English

Institutional Research Information System University of Turin

MUCOSOMES a novel multi drug delivery platform bioinspired from mucin immunomodulatory and antimicrobial activityCosmin Butnarasu1, Paola Petrini2, Francesco Bracotti1, Livia Visai3,4, Giuseppe Guagliano2, Alessandra Fiorio Pla5, Elena Percivalle6, Alessandro Ferrari6, and Sonja Visentin11 University of Turin, Department of Molecular Biotechnology and Health Science, via Quarello 15, 10135 Torino, Italy2 Department of Chemistry, Materials and Chemical Engineering “Giulio Natta”, Politecnico di Milano, Italy3 Molecular Medicine Department (DMM), Centre for Health Technologies (CHT), UdR INSTM, University of Pavia, Pavia, Italy4 Medicina Clinica-Specialistica, UOR5 Laboratorio di Nanotecnologie, ICS Maugeri, IRCCS, 27100 Pavia, Italy5 Department of Life Sciences and Systems Biology, University of Torino, via Accademia Albertina 13, 10123 Torino, Italy6 Molecular Virology Unit, Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalySurface glycosylationBlank PEI primingMucin graftingMucosomes adsorptionWashing0500100015002000Δ M/A [ng/cm2 ]✱ns24 48 720.00.51.01.52.02.5Time (h)Cell viability (490 nm) a.u.✱ ✱✱✱✱✱✱✱✱✱✱24 48 72Time (h)✱✱✱✱✱✱✱✱✱✱✱✱✱✱✱✱CTRL0.250.500.75121020501000.25 0.5 1LPS051015log2 FCIL-1 *****0.25 0.5 1LPSTNF-a**0.25 0.5 1LPSIL-6*****Concentration (µg/mL)15 min 1 h 4 h 24 h 48 h(1) Lungs, (2) liver, (3) spleen, (4) kidney, (5) blood, (6) urine, (7) heart, (8) intestine84h1234567ControlTreated0 5 10 150.80.91.01.11.21.3Time (days)Normalized weightCTRLTreatedBloodHeartIntestineKidneyLiverLungsSpleenUrine024Average radiant efficiency(p/s/cm2 /sr)/(µW/cm2 ) 108 15 min1h4h24h48h4.80 mmMWDyesDrugOligonucleotideProtein0 20 40 60 80 100HSA-FITCPNA-FITCCy5.5CeftazidimeDexamethasoneFITCOseltamivirEncapsulation (%)10 100 10000.00.51.0Size  (nm)Concentration relative1 10 100 1000051015Size (nm)Intensity (a.u.)EXPERIMENTALSize and shapeBacteriumMicrobial lectinGalectinSelectinSiglecImmunecellGlycan GBPsMucin structureMucusMucin glycoproteins are the backbone of mucus which cover all the wet tissues of the human body. Carbohydrate groups on mucins such as N-acetylglucosamine, fucose, galactose and sialic acid, can be ligands for glycan-binding proteins (lectins) on both pathogens and immune cells (Fig. 1) [1]Mucosomes are nanoparticles of spherical shape and diameter of ≈200 nm. The size and shape were assessed by TEM, FESEM, DLS and NTA analysis (Fig. 3).Surface glycosylationMucoadhesive propertiesMucosomes are glycosylated nanoparticles. The presence of carbohydrates on the surface of mucosomes is demonstrated by a Periodic acid-Schiff (PAS) staining and by derivatization and fluorometric detection of sialic acid (Fig. 4).Mucosomes are mucoadhesive nanoparticles. Mucoadhesive properties were studied by QCM analysis. Nanoparticles at first adsorb over the BSM-PEI layer and remain adsorbed even after two washing cycles with PBS (Fig. 5). Mucoadhesivity was also investigated by a flow-through assay by measuring the retention time of FITC-loaded mucosomes on a cystic fibrosis mucus model (Fig. 6)In vitro & in vivo testingIn vitro tests showed cytocompatibility with HeLa cells (Fig. 7) and absence of immunogenic effects (Fig. 8).In vivo experiments showed that mucosomes mainly biodistributed in the lungs and liver without inducing localized or systemic toxicity (Fig. 9).Scan meInspired by the unique properties of mucins and mucus (our first line of defense), we developed acutting-edge nanoplatform that exploits mucus's natural mucoadhesive and binding capacity. We used mucin glycoproteins to synthesize a novel class of nanoparticles that have been named mucosomes [2, 3].Mucosomes production, functionalization with glycans, and drug loading occur via an easily scalable one-pot synthesis. The presence of surface glycans could mediate the engagement of lectins expressed by pathogens but also mammalian cells (Fig. 2).Immune cellImmunomodulatory activityM0 / M1BacteriumAntimicrobial activityGlycan-mediated interactionDrugMucinOne-potInflammationInfectionApplications & Take home messagesThe possibility to deliver active ingredients using mucosomes may offer several advantages over conventional systems in terms of mucoadhesive properties and targeted delivery, especially in pathological conditions where the mucus barrier represents an obstacle to effective treatment. Mucosomes can be loaded with small- and macromolecules (Fig. 10) which could be administered also by nasal administration (Fig. 11). Preliminary in vitro tests with antibacterial (Fig. 12) and antiviral (Fig. 13) drugs showed promising results in terms of activity.[1] Johansson, M. E. V. et al. Nat. Rev. Immunol. 16, 639–649 (2016)[2] EPO, WO2021260525A2 patent number[3] Butnarasu, C et al. Adv. Healthcare Mater. 2022, 2200340Figure 1Figure 2Figure 3Figure 4Figure 5Figure 6Figure 7Figure 8 Figure 9Figure 10 Figure 11Figure 12Figure 13-2 -1 0 1 2 3-10123log [API] (µg/mL)log survival rateS. aureus 24 h-2 -1 0 1 2 3-10123log [API] (µg/mL)P. aeruginosa 24 hAPI-mucosomesMucosomesAPI-8 -6 -4 -2 0050100log API (µg/mL)Neutralization (%)SARS-CoV-2 UK variant-8 -6 -4 -2 0050100log API (µg/mL)SARS-CoV-2 Brazilian variantMucosomes - PralatrexateMucosomesAPIcosminstefan.butnarasu@unito.it

Mucosomes: a novel multi drug delivery platform bioinspired from mucin immunomodulatory and antimicrobial activity

https://iris.unito.it/bitstream/2318/1879073/1/Butnarasu_IIDC_poster_compressed.pdf

Mucosomes: a novel multi drug delivery platform bioinspired from mucin immunomodulatory and antimicrobial activity

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