title: Intracellular Delivery of Poorly Soluble Polyphenols: Elucidating the Interplay of Self-Assembling Nanocarriers and Human Chondrocytes.
abstract: Increased molecular understanding of multifactorial diseases paves the way for novel therapeutic approaches requiring sophisticated carriers for intracellular delivery of actives. We designed and characterized self-assembling lipid-core nanocapsules for coencapsulation of two poorly soluble natural polyphenols curcumin and resveratrol. The polyphenols were identified as high-potential therapeutic candidates intervening in the intracellular inflammation cascade of chondrocytes during the progress of osteoarthritis. To elucidate the interplay between chondrocytes and nanocapsules and their therapeutic effect, we pursued a complementary analytical approach combining label-free visualization with biological assays. Primary human chondrocytes did not show any adverse effects upon nanocapsule application and coherent anti-Stokes Raman scattering images visualized their intracellular uptake. Further, by systematically blocking different uptake mechanisms, an energy independent uptake into the cells could be identified. Additionally, we tested the therapeutic effect of the polyphenol-loaded carriers on inflamed chondrocytes. Treatment with nanocapsules resulted in a major reduction of nitric oxide levels, a well-known apoptosis trigger during the course of osteoarthritis. For a more profound examination of this protective effect on joint cells, we pursued studies with atomic force microscopy investigations. Significant changes in the cell cytoskeleton as well as prominent dents in the cell membrane upon induced apoptosis were revealed. Interestingly, these effects could not be detected for chondrocytes which were pretreated with the nanocapsules. Overall, besides presenting a sophisticated carrier system for joint application, these results highlight the necessity of establishing combinatorial analytical approaches to elucidate cellular uptake, the interplay of codelivered drugs and their therapeutic effect on the subcellular level.
authors.affiliation: Saarland University , Department of Biopharmaceutics and Pharmaceutical Technology, Campus A4.1, 66123 Saarbruecken, Germany.|University of Twente , Optical Sciences Group, MESA+ Institute for Nanotechnology, P.O. Box 217, 7500 AE Enschede, The Netherlands.
authors.forename: Birthe
authors.initials: B
authors.lastname: Kann
authors.affiliation: Saarland University , Experimental Physics, Campus E2.9, 66123 Saarbruecken, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Spengler
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Federal University of Rio Grande do Sul (UFRGS) , 90610-000, Porto Alegre, Rio Grande do Sul, Brazil.
authors.forename: Karine
authors.initials: K
authors.lastname: Coradini
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Federal University of Rio Grande do Sul (UFRGS) , 90610-000, Porto Alegre, Rio Grande do Sul, Brazil.
authors.forename: Lucas A
authors.initials: LA
authors.lastname: Rigo
authors.affiliation: University of Twente , Nanobiophysics Group, MESA+ Institute for Nanotechnology, P.O. Box 217, 7500 AE Enschede, The Netherlands.
authors.forename: Martin L
authors.initials: ML
authors.lastname: Bennink
authors.affiliation: Saarland University , Experimental Physics, Campus E2.9, 66123 Saarbruecken, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Jacobs
authors.affiliation: University of Twente , Optical Sciences Group, MESA+ Institute for Nanotechnology, P.O. Box 217, 7500 AE Enschede, The Netherlands.
authors.forename: Herman L
authors.initials: HL
authors.lastname: Offerhaus
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Federal University of Rio Grande do Sul (UFRGS) , 90610-000, Porto Alegre, Rio Grande do Sul, Brazil.
authors.forename: Ruy C R
authors.initials: RC
authors.lastname: Beck
authors.affiliation: Saarland University , Department of Biopharmaceutics and Pharmaceutical Technology, Campus A4.1, 66123 Saarbruecken, Germany.|Helmholtz Centre for Infection Research and Helmholtz Institute for Pharmaceutical Research Saarland , Department of Drug Delivery, Campus E 8.1, 66123 Saarbruecken, Germany.|PharmBioTec GmbH , Department of Drug Delivery, Science Park 1, 66123 Saarbruecken, Germany.
authors.forename: Maike
authors.initials: M
authors.lastname: Windbergs
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000494:pharmacology; D019902:Chondrocytes / Q000378:metabolism*; D003474:Curcumin / Q000378:metabolism* / Q000494:pharmacology; D004337:Drug Carriers / Q000378:metabolism* / Q000494:pharmacology; D056604:Grape Seed Extract / Q000737:chemistry; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D018625:Microscopy, Atomic Force; D053769:Nanocapsules / Q000737:chemistry*; D000073758:Nonlinear Optical Microscopy; D010316:Particle Size; D059808:Polyphenols / Q000378:metabolism* / Q000494:pharmacology; D011136:Polysorbates / Q000737:chemistry; D000077185:Resveratrol; D013267:Stilbenes / Q000378:metabolism* / Q000494:pharmacology; D027843:Vitis
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D004337:Drug Carriers; D056604:Grape Seed Extract; D053769:Nanocapsules; D059808:Polyphenols; D011136:Polysorbates; D013267:Stilbenes; D003474:Curcumin; D000077185:Resveratrol