title: The interaction of protamine nanocapsules with the intestinal epithelium: A mechanistic approach.
abstract: Single-layer protamine and double layer polysialic acid (PSA)/protamine nanocapsules (NCs) were designed in order to be used as carriers to facilitate the transport of macromolecules across the intestinal epithelium. The rational for the design of these NCs was based on that protamine is a non-toxic yet potent cell-penetrating peptide, capable of translocating protein cargos through cell membranes, while PSA is a low molecular weight polysaccharide used to enhance the stability of macromolecules and nanocarriers. The aim of this work was to study in vitro the mechanism of interaction of these NCs with different intestinal cell models (Caco-2, Caco-2/Raji mimicking follicle associated epithelium and Caco-2/HT29-MTX to study the effect of mucus). For this, a fluorescent marker, TAMRA was covalently linked to protamine. The interaction and transport of the NCs with the Caco-2 cells was found to be concentration, temperature and size dependent. In all cases, the double layer PSA-protamine NCs exhibited a significantly higher transport compared to protamine NCs. On the other hand, the transport of the NCs was significantly higher in the co-culture (Caco-2/Raji monolayer) compared to the monoculture model (Caco-2 monolayer), implying that M cells are involved in the transport of these nanosystems. The formulations, administered intra-jejunally to healthy rats (4h fasting) resulted in a moderate reduction of the glucose levels (20% reduction), which lasted for up to 4h. This work raises prospects that protamine-based nanocapsules may have the potential as oral peptide delivery nanocarriers.
authors.affiliation: Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Faculty of Pharmacy, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain; Université catholique de Louvain, Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, 1200 Brussels, Belgium.
authors.forename: Lungile Nomcebo
authors.initials: LN
authors.lastname: Thwala
authors.affiliation: Université catholique de Louvain, Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, 1200 Brussels, Belgium.
authors.forename: Ana
authors.initials: A
authors.lastname: Beloqui
authors.affiliation: Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Faculty of Pharmacy, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain.
authors.forename: Noemi Stefania
authors.initials: NS
authors.lastname: Csaba
authors.affiliation: Biomedical Research Group, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain.
authors.forename: David
authors.initials: D
authors.lastname: González-Touceda
authors.affiliation: Biomedical Research Group, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain.
authors.forename: Sulay
authors.initials: S
authors.lastname: Tovar
authors.affiliation: Biomedical Research Group, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Dieguez
authors.affiliation: Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Faculty of Pharmacy, Universidad de Santiago de Compostela, 15706 Santiago de Compostela, Spain. Electronic address: mariaj.alonso@usc.es.
authors.forename: Maria Jose
authors.initials: MJ
authors.lastname: Alonso
authors.affiliation: Université catholique de Louvain, Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, 1200 Brussels, Belgium. Electronic address: veronique.preat@uclouvain.be.
authors.forename: Véronique
authors.initials: V
authors.lastname: Préat
mesh_terms: D000818:Animals; D001692:Biological Transport; D018938:Caco-2 Cells; D002626:Chemistry, Pharmaceutical / Q000379:methods; D018920:Coculture Techniques; D004337:Drug Carriers / Q000737:chemistry*; D016503:Drug Delivery Systems; D006801:Humans; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000378:metabolism; D008297:Male; D053769:Nanocapsules; D010316:Particle Size; D010539:Permeability; D011479:Protamines / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012794:Sialic Acids / Q000737:chemistry*; D013696:Temperature
chemical_list: D004337:Drug Carriers; D053769:Nanocapsules; D011479:Protamines; D012794:Sialic Acids; C021319:polysialic acid
keywords: Caco-2 cells; Endocytosis; M cells; Mucus; Nanocapsules; Peptide delivery