title: Safety profile of solid lipid nanoparticles loaded with rosmarinic acid for oral use: in vitro and animal approaches.
abstract: Rosmarinic acid (RA) possesses several protective bioactivities that have attracted increasing interest by nutraceutical/pharmaceutical industries. Considering the reduced bioavailability after oral use, effective (and safe) delivery systems are crucial to protect RA from gastrointestinal degradation. This study aims to characterize the safety profile of solid lipid nanoparticles produced with Witepsol and Carnauba waxes and loaded with RA, using in vitro and in vivo approaches, focused on genotoxicity and cytotoxicity assays, redox status markers, hematological and biochemical profile, liver and kidney function, gut bacterial microbiota, and fecal fatty acids composition. Free RA and sage extract, empty nanoparticles, or nanoparticles loaded with RA or sage extract (0.15 and 1.5 mg/mL) were evaluated for cell (lymphocytes) viability, necrosis and apoptosis, and antioxidant/prooxidant effects upon DNA. Wistar rats were orally treated for 14 days with vehicle (control) and with Witepsol or Carnauba nanoparticles loaded with RA at 1 and 10 mg/kg body weight/d. Blood, urine, feces, and several tissues were collected for analysis. Free and loaded RA, at 0.15 mg/mL, presented a safe profile, while genotoxic potential was found for the higher dose (1.5 mg/mL), mainly by necrosis. Our data suggest that both types of nanoparticles are safe when loaded with moderate concentrations of RA, without in vitro genotoxicity and cytotoxicity and with an in vivo safety profile in rats orally treated, thus opening new avenues for use in nutraceutical applications.
authors.affiliation: Catholic University of Portugal, CBQF - Center for Biotechnology and Fine Chemistry - Associate Laboratory, Faculty of Biotechnology, Porto, Portugal.
authors.forename: Ana Raquel
authors.initials: AR
authors.lastname: Madureira
authors.affiliation: Laboratory of Pharmacology and Experimental Therapeutics, Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, and CNC.IBILI Consortium, University of Coimbra, Coimbra, Portugal.
authors.forename: Sara
authors.initials: S
authors.lastname: Nunes
authors.affiliation: Catholic University of Portugal, CBQF - Center for Biotechnology and Fine Chemistry - Associate Laboratory, Faculty of Biotechnology, Porto, Portugal.
authors.forename: Débora A
authors.initials: DA
authors.lastname: Campos
authors.affiliation: Laboratory of Pharmacology and Experimental Therapeutics, Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, and CNC.IBILI Consortium, University of Coimbra, Coimbra, Portugal.
authors.forename: João C
authors.initials: JC
authors.lastname: Fernandes
authors.affiliation: Department of Biochemistry, Faculty of Medicine, University of Porto, Porto, Portugal.
authors.forename: Cláudia
authors.initials: C
authors.lastname: Marques
authors.affiliation: Laboratory of Pharmacology and Experimental Therapeutics, Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, and CNC.IBILI Consortium, University of Coimbra, Coimbra, Portugal.
authors.forename: Monica
authors.initials: M
authors.lastname: Zuzarte
authors.affiliation: Catholic University of Portugal, CBQF - Center for Biotechnology and Fine Chemistry - Associate Laboratory, Faculty of Biotechnology, Porto, Portugal.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Gullón
authors.affiliation: Catholic University of Portugal, CBQF - Center for Biotechnology and Fine Chemistry - Associate Laboratory, Faculty of Biotechnology, Porto, Portugal.
authors.forename: Luís M
authors.initials: LM
authors.lastname: Rodríguez-Alcalá
authors.affiliation: Department of Biochemistry, Faculty of Medicine, University of Porto, Porto, Portugal; Center for Health Technology and Services Research (CINTESIS), Porto, Portugal.
authors.forename: Conceição
authors.initials: C
authors.lastname: Calhau
authors.affiliation: Department of Pharmaceutical Sciences, Institute of Health Sciences-North, CESPU, Gandra, Portugal; "I3S" Instituto de Investigação e Inovação em Saúde, University of Porto, Porto, Portugal; INEB, Institute of Biomedical Engineering, University of Porto, Porto, Portugal.
authors.forename: Bruno
authors.initials: B
authors.lastname: Sarmento
authors.affiliation: Catholic University of Portugal, CBQF - Center for Biotechnology and Fine Chemistry - Associate Laboratory, Faculty of Biotechnology, Porto, Portugal.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Gomes
authors.affiliation: Catholic University of Portugal, CBQF - Center for Biotechnology and Fine Chemistry - Associate Laboratory, Faculty of Biotechnology, Porto, Portugal.
authors.forename: Maria Manuela
authors.initials: MM
authors.lastname: Pintado
authors.affiliation: Laboratory of Pharmacology and Experimental Therapeutics, Institute for Biomedical Imaging and Life Sciences (IBILI), Faculty of Medicine, and CNC.IBILI Consortium, University of Coimbra, Coimbra, Portugal.
authors.forename: Flávio
authors.initials: F
authors.lastname: Reis
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002934:Cinnamates / Q000008:administration & dosage* / Q000009:adverse effects / Q000494:pharmacology*; D048749:Cytokinesis / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D053630:Depsides / Q000008:administration & dosage* / Q000009:adverse effects / Q000494:pharmacology*; D000069196:Gastrointestinal Microbiome / Q000187:drug effects; D008055:Lipids / Q000737:chemistry* / Q000633:toxicity; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D053758:Nanoparticles / Q000737:chemistry* / Q000633:toxicity / Q000648:ultrastructure; D010084:Oxidation-Reduction; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D014280:Triglycerides / Q000737:chemistry; D014885:Waxes / Q000737:chemistry; D000097123:Rosmarinic Acid
chemical_list: D002934:Cinnamates; D004247:DNA; D053630:Depsides; D008055:Lipids; D014280:Triglycerides; D014885:Waxes; D000097123:Rosmarinic Acid; C009683:witepsol; C026344:carnauba wax
keywords: Witepsol and Carnauba waxes; in vitro and animal toxicity; rosmarinic acid; safety profile; solid lipid nanoparticles