title: Optimization, formulation, and characterization of multiflavonoids-loaded flavanosome by bulk or sequential technique.
abstract: This study involves adaptation of bulk or sequential technique to load multiple flavonoids in a single phytosome, which can be termed as "flavonosome". Three widely established and therapeutically valuable flavonoids, such as quercetin (Q), kaempferol (K), and apigenin (A), were quantified in the ethyl acetate fraction of Moringa oleifera leaves extract and were commercially obtained and incorporated in a single flavonosome (QKA-phosphatidylcholine) through four different methods of synthesis - bulk (M1) and serialized (M2) co-sonication and bulk (M3) and sequential (M4) co-loading. The study also established an optimal formulation method based on screening the synthesized flavonosomes with respect to their size, charge, polydispersity index, morphology, drug-carrier interaction, antioxidant potential through in vitro 1,1-diphenyl-2-picrylhydrazyl kinetics, and cytotoxicity evaluation against human hepatoma cell line (HepaRG). Furthermore, entrapment and loading efficiency of flavonoids in the optimal flavonosome have been identified. Among the four synthesis methods, sequential loading technique has been optimized as the best method for the synthesis of QKA-phosphatidylcholine flavonosome, which revealed an average diameter of 375.93±33.61 nm, with a zeta potential of -39.07±3.55 mV, and the entrapment efficiency was >98% for all the flavonoids, whereas the drug-loading capacity of Q, K, and A was 31.63%±0.17%, 34.51%±2.07%, and 31.79%±0.01%, respectively. The in vitro 1,1-diphenyl-2-picrylhydrazyl kinetics of the flavonoids indirectly depicts the release kinetic behavior of the flavonoids from the carrier. The QKA-loaded flavonosome had no indication of toxicity toward human hepatoma cell line as shown by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide result, wherein even at the higher concentration of 200 µg/mL, the flavonosomes exert >85% of cell viability. These results suggest that sequential loading technique may be a promising nanodrug delivery system for loading multiflavonoids in a single entity with sustained activity as an antioxidant, hepatoprotective, and hepatosupplement candidate.
authors.affiliation: Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience.
authors.forename: Govindarajan
authors.initials: G
authors.lastname: Karthivashan
authors.affiliation: Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences.
authors.forename: Mas Jaffri
authors.initials: MJ
authors.lastname: Masarudin
authors.affiliation: Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience.
authors.forename: Aminu Umar
authors.initials: AU
authors.lastname: Kura
authors.affiliation: Department of Food Science, Faculty of Food Science and Technology; Laboratory of Natural Products, Institute of Bioscience.
authors.forename: Faridah
authors.initials: F
authors.lastname: Abas
authors.affiliation: Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience; Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
authors.forename: Sharida
authors.initials: S
authors.lastname: Fakurazi
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000737:chemistry; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004337:Drug Carriers; D005419:Flavonoids / Q000494:pharmacology*; D016166:Free Radical Scavengers / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry*; D029587:Moringa / Q000737:chemistry; D010851:Picrates / Q000737:chemistry; D010936:Plant Extracts / Q000494:pharmacology; D018515:Plant Leaves / Q000737:chemistry
chemical_list: D000975:Antioxidants; D001713:Biphenyl Compounds; D004337:Drug Carriers; D005419:Flavonoids; D016166:Free Radical Scavengers; D008081:Liposomes; D010851:Picrates; D010936:Plant Extracts; C004931:1,1-diphenyl-2-picrylhydrazyl
keywords: HepaRG cell line; antioxidant; apigenin; kaempferol; phytosome; quercetin