title: Formulation and cytotoxicity evaluation of new self-emulsifying multiple W/O/W nanoemulsions.
abstract: Three multiple water-in-oil-in-water (W/O/W) nanoemulsions have been designed for potential inclusion of either lipophilic or hydrophilic drugs using a two-step emulsification process exclusively based on low-energy self-emulsification. The W/O primary emulsion was constituted by a blend of oil (medium chain triglyceride), a mixture (7:3) of two surfactants, and a 10% water phase. The surfactants were a mixture of Polysorbate-85/Labrasol(®), Polysorbate-85/Cremophor(®) EL or glycerol/Polysorbate-85. The final W/O/W nanoemulsions were obtained by the addition of water, with a weight ratio nanoemulsion/water of 1:2. The multiple emulsion stability was found to increase from 24 hours to 2 and 6 months with Labrasol, glycerol, and Cremophor, respectively. Cytotoxicity was found for formulations including Labrasol and Cremophor EL. The concentration of emulsion inhibiting 50% cell viability (IC(50)) was determined using the alamarBlue(®) test, giving after 24 hours of incubation, IC(50) = 10.2 mg/mL for the Labrasol formulation and IC(50) = 11.8 mg/mL for the Cremophor EL formulation. Corresponding calculated IC(50) values for surfactants were 0.51 mg/mL for Labrasol and 0.59 mg/mL for Cremophor EL. In both cases, cytotoxicity was due to an apoptotic mechanism, evidenced by chromatin condensation and P2X7 cell death receptor activation. The formulation including glycerol, investigated between 1 and 100 mg/mL concentration of nanoemulsion, did not affect cell viability. Moreover, neither chromatin condensation nor P2X7 activation was found between the 10 and 30 mg/mL final concentration of the emulsion. This last formulation would therefore be of major interest for further developments.
authors: Chemical, Genetic and Imaging Pharmacology Laboratory; INSERM U1022, CNRS UMR8151, Chimie ParisTech, Faculty of Pharmacy, Paris Descartes University, Sorbone Paris Cité, Paris, France.
authors: Estelle
authors: E
authors: Sigward
authors: Nathalie
authors: N
authors: Mignet
authors: Patrice
authors: P
authors: Rat
authors: Mélody
authors: M
authors: Dutot
authors: Saleh
authors: S
authors: Muhamed
authors: Jean-Michel
authors: JM
authors: Guigner
authors: Daniel
authors: D
authors: Scherman
authors: Denis
authors: D
authors: Brossard
authors: Sylvie
authors: S
authors: Crauste-Manciet
mesh_terms: D000704:Analysis of Variance; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004355:Drug Stability; D004655:Emulsions / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects; D019573:Epithelium, Corneal / Q000166:cytology / Q000187:drug effects; D005989:Glycerides; D005990:Glycerol / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry* / Q000633:toxicity; D009930:Organic Chemicals / Q000008:administration & dosage / Q000737:chemistry; D014677:Pharmaceutical Vehicles / Q000008:administration & dosage / Q000737:chemistry* / Q000633:toxicity; D011136:Polysorbates / Q000008:administration & dosage / Q000737:chemistry; D013501:Surface-Active Agents / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity; D014867:Water / Q000737:chemistry
chemical_list: D004655:Emulsions; D005989:Glycerides; D009930:Organic Chemicals; D014677:Pharmaceutical Vehicles; D011136:Polysorbates; D013501:Surface-Active Agents; C440220:Labrasol; D014867:Water; C000515:cremophor EL; D005990:Glycerol
keywords: Cremophor EL; Labrasol; P2X7 receptor; apoptosis; glycerol; polysorbate
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