title: Development of particulate drug formulation against C. parvum: Formulation, characterization and in vivo efficacy.
abstract: This research aims towards developing an alternative therapy against Cryptosporidium parvum using bioadhesive paromomycin and diloxanide furoate-loaded microspheres. Microspheres were prepared using chitosan and poly(vinyl alcohol) and two types of cyclodextrins (β-CD and DM-β-CD) for the potential use of treating cryptosporidiosis. This pathogen is associated with gastrointestinal illness in humans and animals. Microparticle formulations were characterized in terms of size, surface charge, drug release and morphology. In vivo bioadhesion properties of CHI/PVA microspheres were also evaluated in mice. Finally, the in vivo efficacy of CHI/PVA microspheres against C. parvum was tested in neonatal mouse model. In this work, microspheres prepared by spray-drying showed spherical shape, diameters between 6.67±0.11 and 18.78±0.07μm and positively surface charged. The bioadhesion studies demonstrated that MS remained attached at +16h (post-infection) to the intestinal cells as detected by fluorescence. This finding was crucial taking use of the fact that the parasite multiplication occurs between 16 and 20h post-infection. The efficacy of treatment was determined by calculating the number of oocysts recovered from the intestinal tract of mice after 7days of post-infection. Mice receiving orally administered microspheres with and without drug exhibited significantly lower parasite loads compared with the control mice. Ultrastructural observations by TEM bring to light the uptake of smallest particles by enterocytes associated with conspicuous changes in enterocytic cells. Completely recovery of cell morphology was detected after 24h of first inoculation with MS. CHI/PVA microspheres appear to be a safe and simple system to be used in an anticryptosporidial treatment. The distinctive features of neonatal mice requires further work to determine the suppressive effect of this particulate delivery system on C. parvum attachment in other animal models.
authors.affiliation: Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Ciencias, 27002, Universidad de Santiago de Compostela, Lugo, Spain.
authors.forename: Estefanía
authors.initials: E
authors.lastname: Blanco-García
authors.affiliation: Departamento de Anatomía y Producción Animal, Facultad de Veterinaria, 27002 Lugo, Universidad de Santiago de Compostela, Spain.
authors.forename: Florentina
authors.initials: F
authors.lastname: Guerrero-Callejas
authors.affiliation: Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Ciencias, 27002, Universidad de Santiago de Compostela, Lugo, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Blanco-Méndez
authors.affiliation: Departamento de Microbiología y Parasitología, Facultad de Farmacia, 15782, Universidad de Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Hipólito
authors.initials: H
authors.lastname: Gómez-Couso
authors.affiliation: Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Ciencias, 27002, Universidad de Santiago de Compostela, Lugo, Spain. Electronic address: asteriam.luzardo@usc.es.
authors.forename: Asteria
authors.initials: A
authors.lastname: Luzardo-Álvarez
mesh_terms: D000268:Adhesiveness; D000818:Animals; D000831:Animals, Newborn; D000981:Antiprotozoal Agents / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D048271:Chitosan / Q000737:chemistry; D003457:Cryptosporidiosis / Q000188:drug therapy* / Q000469:parasitology; D016785:Cryptosporidium parvum / Q000187:drug effects / Q000302:isolation & purification; D003505:Cyclodextrins / Q000737:chemistry; D004339:Drug Compounding; D016503:Drug Delivery Systems; D065546:Drug Liberation; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005663:Furans / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D007422:Intestines / Q000737:chemistry; D051379:Mice; D008863:Microspheres; D033661:Oocysts / Q000187:drug effects; D059208:Parasite Load; D010303:Paromomycin / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D011142:Polyvinyl Alcohol / Q000737:chemistry
chemical_list: D000981:Antiprotozoal Agents; D003505:Cyclodextrins; D005663:Furans; D010303:Paromomycin; D011142:Polyvinyl Alcohol; D048271:Chitosan; D016650:Fluorescein-5-isothiocyanate; C004932:diloxanide furoate
keywords: Bioadhesion; Cryptosporidium parvum; Cylodextrins; Diloxanide furoate; Microspheres; Paromomycin