title: Nanostructured lipid carriers loaded with resveratrol modulate human dendritic cells.
abstract: Dendritic cells (DCs) are promising targets for drug delivery, as they can induce immunity or tolerance. The current study aims to examine the potential of using nanostructured lipid carriers (NLC) as delivery systems for human DC by evaluating nanoparticle internalization, cell labeling, and drug activity. NLC were formulated incorporating the fluorochrome fluorescein isothiocyanate (FITC-NLC) or the natural anti-inflammatory molecule resveratrol (rsv-NLC). Primary human DCs were differentiated from peripheral blood monocytes, and the innovative imaging flow cytometry technique was used to examine FITC-NLC internalization. The capacity of rsv-NLC to inhibit DC activation in response to proinflammatory cytokine tumor necrosis factor-α (TNF- α) was investigated by conventional flow cytometry. A combination of imaging and conventional flow cytometry was used to assess NLC cytotoxicity. The results obtained indicate that both NLC formulations were stable over time, with mean diameter <200 nm and highly negative zeta potential (about -30 mV). When DCs were placed in contact with NLC, imaging flow cytometry clearly showed that DCs efficiently internalized FITC-NLC, with nearly 100% of cells internalizing nanoparticles upon 1 hour of incubation. Both immature and mature DCs internalized NLC to high and comparable levels, and without cytotoxicity. Stimulating DC with TNF-α in the presence of rsv-NLC revealed that, using these nanoparticles, very small concentrations of rsv were sufficient to significantly decrease surface expression of activation marker CD83 (5 µM) and major histocompatibility complex-class II molecule human leukocyte antigen - antigen D related (10 µM), both upregulated in response to TNF-α stimulation. Rsv-NLC were compared with free rsv; at 5 µM, rsv-NLC were able to inhibit nuclear factor κ beta phosphorylation and significantly decrease the level of interleukin-12/23, both upregulated in response to TNF-α, while 10 µM free rsv were needed to promote a similar effect. Taken together, the results presented show that NLC are suitable carriers of fluorescent labels or bioactive molecules for human DCs, leading to inflammation modulation.
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Portugal; UCIBIO, REQUIMTE, Chemical Sciences Department, Faculty of Pharmacy, University of Porto, Portugal.
authors.forename: João P
authors.initials: JP
authors.lastname: Barbosa
authors.affiliation: UCIBIO, REQUIMTE, Chemical Sciences Department, Faculty of Pharmacy, University of Porto, Portugal.
authors.forename: Ana R
authors.initials: AR
authors.lastname: Neves
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal.
authors.forename: Andreia M
authors.initials: AM
authors.lastname: Silva
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal.
authors.forename: Mário A
authors.initials: MA
authors.lastname: Barbosa
authors.affiliation: UCIBIO, REQUIMTE, Chemical Sciences Department, Faculty of Pharmacy, University of Porto, Portugal.
authors.forename: M Salette
authors.initials: MS
authors.lastname: Reis
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Portugal.
authors.forename: Susana G
authors.initials: SG
authors.lastname: Santos
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism; D004337:Drug Carriers / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D007108:Immune Tolerance / Q000187:drug effects; D007136:Immunoglobulins / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D008055:Lipids / Q000737:chemistry / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D049329:Nanostructures / Q000008:administration & dosage / Q000737:chemistry*; D000077185:Resveratrol; D013267:Stilbenes / Q000008:administration & dosage / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D004337:Drug Carriers; D016650:Fluorescein-5-isothiocyanate; D007136:Immunoglobulins; D018664:Interleukin-12; D008055:Lipids; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D000077185:Resveratrol; D013267:Stilbenes; D014409:Tumor Necrosis Factor-alpha; D000097802:CD83 Antigen
keywords: TNF-α; dendritic cell; imaging flow cytometry; immunomodulation; resveratrol; solid lipid nanoparticle