title: A toxicological evaluation of inhaled solid lipid nanoparticles used as a potential drug delivery system for the lung.
abstract: Inhalation is a non-invasive approach for both local and systemic drug delivery. This study aimed to define the therapeutic window for solid lipid nanoparticles (SLNs) as a drug delivery system by inhalation from a toxicological point of view. To estimate the toxic dose of SLNs in vitro, A549 cells and murine precision-cut lung slices (PCLS) were exposed to increasing concentrations of SLNs. The cytotoxic effect of SLNs on A549 cells was evaluated by MTT and NRU assays. Viability of lung tissue was determined with WST assay and by life/dead staining using calcein AM/EthD-1 for confocal microscopy (CLSM) followed by quantitative analysis with IMARIS. Inflammation was assessed by measuring chemokine KC and TNF-alpha levels. The in vivo effects were determined in a 16-day repeated-dose inhalation toxicity study using female BALB/c mice, which were daily exposed to different concentrations of SLN30 aerosols (1-200 microg deposit dose). Local inflammatory effects in the respiratory tract were evaluated by determination of total protein content, LDH, chemokine KC, IL-6, and differential cell counts, performed on days 4, 8, 12, and 16 in bronchoalveolar lavage fluid. Additionally, a histopathological evaluation of toxicologically relevant organs was accomplished. The in vitro and ex vivo dose finding experiments showed toxic effects beginning at concentrations of about 500 microg/ml. Therefore, we used 1-200 microg deposit doses/animal for the in vivo experiments. Even after 16 days of challenge with a 200-microg deposit dose, SLNs induced no significant signs of inflammation. We observed no consistent increase in LDH release, protein levels, or other signs of inflammation such as chemokine KC, IL-6, or neutrophilia. In contrast, the particle control (carbon black) caused inflammatory and cytotoxic effects at corresponding concentrations. These results confirm that repeated inhalation exposure to SLN30 at concentrations lower than a 200-microg deposit dose is safe in a murine inhalation model.
authors.affiliation: Fraunhofer Institute for Toxicology and Experimental Medicine, Department of Immunology, Allergology and Immunotoxicology, Hannover, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Nassimi
authors.forename: C
authors.initials: C
authors.lastname: Schleh
authors.forename: H D
authors.initials: HD
authors.lastname: Lauenstein
authors.forename: R
authors.initials: R
authors.lastname: Hussein
authors.forename: H G
authors.initials: HG
authors.lastname: Hoymann
authors.forename: W
authors.initials: W
authors.lastname: Koch
authors.forename: G
authors.initials: G
authors.lastname: Pohlmann
authors.forename: N
authors.initials: N
authors.lastname: Krug
authors.forename: K
authors.initials: K
authors.lastname: Sewald
authors.forename: S
authors.initials: S
authors.lastname: Rittinghausen
authors.forename: A
authors.initials: A
authors.lastname: Braun
authors.forename: C
authors.initials: C
authors.lastname: Müller-Goymann
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems; D005260:Female; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000503:physiopathology; D008055:Lipids / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity*; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D053758:Nanoparticles / Q000633:toxicity*; D013997:Time Factors; D018675:Toxicity Tests
chemical_list: D008055:Lipids