title: The effect of two drug delivery systems in ropivacaine cytotoxicity and cytokine release by human keratinocytes and fibroblasts.
abstract: OBJECTIVE
  Modified drug delivery systems have been developed to improve pharmacological properties of local anaesthetics. However, the inflammatory potential of these formulations was not investigated. This study compared the in-vitro effects of ropivacaine (ropi) in plain, liposomal (MLV) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) formulations on cell viability, apoptosis and cytokine (IL-1α, TNF-α, IL-6 and IL-10) release.
  
  
  METHODS
  Human immortalized keratinocytes (HaCaT) and human immortalized gingival fibroblasts (HGF) were exposed to 1-100 μm ropi concentrations. The cell viability was measured by XTT and LIVE/DEAD assay. Apoptosis was performed by flow cytometry, and cytokine release was measured by ELISA assay.
  
  
  RESULTS
  Human immortalized keratinocyte viability was reduced by ropi and both drug delivery systems. However, none of the formulations induced apoptosis. Results showed a differential regulation of IL-1α TNF-α, IL-6 and IL-10 by HaCaT and HGF. Ropi-HP-β-CD increased twofold the IL-6 release by HGF in comparison with the control, while 100 μm ropi-MLV led to an increased release of all pro-inflammatory cytokines by HGF.
  
  
  CONCLUSIONS
  The loss in cell viability was not related to cellular apoptosis. Ropi complexed with HP-β-CD showed a similar cytokine release pattern when compared to the plain formulation. Thus, the HP-β-CD form was a better drug carrier than the MLV form for ropivacaine drug delivery.
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas - UNICAMP - Piracicaba, São Paulo, Brazil.
authors.forename: Luiz Eduardo Nunes
authors.identifier: http://orcid.org/0000-0002-4882-5402
authors.initials: LE
authors.lastname: Ferreira
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas - UNICAMP - Piracicaba, São Paulo, Brazil.
authors.forename: Bruno Vilela
authors.initials: BV
authors.lastname: Muniz
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas - UNICAMP - Piracicaba, São Paulo, Brazil.
authors.forename: Jonny
authors.initials: J
authors.lastname: Burga-Sánchez
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas - UNICAMP - Piracicaba, São Paulo, Brazil.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Volpato
authors.affiliation: Department of Biochemistry, Biology Institute, University of Campinas - UNICAMP - Campinas, São Paulo, Brazil.
authors.forename: Eneida
authors.initials: E
authors.lastname: de Paula
authors.affiliation: Xenobiotics Research Unit, Laboratory of Stomatology, Biological and Health Sciences Center, The Pontifical Catholic University of Paraná - Curitiba, Paraná, Brazil.
authors.forename: Edvaldo Antonio Ribeiro
authors.initials: EA
authors.lastname: Rosa
authors.affiliation: Department of Physiological Sciences, Piracicaba Dental School, University of Campinas - UNICAMP - Piracicaba, São Paulo, Brazil.
authors.forename: Francisco Carlos
authors.initials: FC
authors.lastname: Groppo
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000577:Amides / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000009:adverse effects*; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D016753:Interleukin-10 / Q000378:metabolism; D053582:Interleukin-1alpha / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism*; D008081:Liposomes / Q000009:adverse effects; D000077212:Ropivacaine; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D047392:beta-Cyclodextrins / Q000009:adverse effects
chemical_list: D000577:Amides; D016207:Cytokines; C508609:IL10 protein, human; C508584:IL1A protein, human; D053582:Interleukin-1alpha; D015850:Interleukin-6; D008081:Liposomes; D014409:Tumor Necrosis Factor-alpha; D047392:beta-Cyclodextrins; D016753:Interleukin-10; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000077212:Ropivacaine
keywords: 2-hydroxypropyl-β-cyclodextrin; cell viability; cytokines; multilamellar liposome; ropivacaine