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Abstract
The effect of liposomal inclusion on the stability and in vitro antiherpetic activity of Santolina insularis essential oil was investigated. In order to study the influence of vesicle structure on the liposome properties, multilamellar and unilamellar vesicles were prepared by the film method and sonication, respectively. Vesicles were obtained from hydrogenated soya phosphatydilcholine and cholesterol. Formulations were examined for their stability for over one year monitoring the drug leakage from vesicles and the average size distribution. The stability of the incorporated oil was verified by studying its quali-quantitative composition. The antiviral activity was studied against Herpes simplex virus type 1 (HSV-1) by plaque reduction and yield reduction assays. Results showed that Santolina insularis essential oil can be incorporated in high amounts in the prepared liposomes, which successfully prevented its degradation. Moreover, stability studies pointed out that vesicle dispersions were stable for at least one year and neither oil leakage nor vesicle size alteration occurred during this period. Antiviral activity assays demonstrated that Santolina insularis essential oil is effective in inactivating HSV-1 and that the activity is principally due to direct virucidal effects. Free essential oil proved to be more effective than liposomal oil and a different activity was discovered which related to the vesicular structure. The ED50 values, significantly lower when cells were pre-incubated with the essential oil before the virus adsorption, indicate an intracellular mechanism in the antiviral activity of Santolina insularis. Moreover, liposomal Santolina essential oil is non toxic in the range of the concentration tested.
| ABBREVIATIONS | ||
| HSV-1 | = | Herpes simplex virus type 1 |
| HSV-2 | = | Herpes simplex virus 2 |
| HIV-1 | = | Human Immunodeficiency Virus-type 1 |
| S. | = | Santolina |
| MPS | = | Mononuclear phagocityc system |
| HPC | = | Hydrogenated soya phosphatidylcholine |
| HPLC | = | High Performance Liquid Chromatography |
| CHOL | = | Cholesterol |
| FBS | = | Fetal bovine serum |
| MEM | = | Modified Eagle Medium |
| GC | = | Gas Chromatograghy |
| PFU | = | Plaque forming units |
| i.d. | = | internal diameter |
| MLV | = | Multilamellar Vesicles |
| SUV | = | Small Unilamellar Vesicles |
| TEM | = | Transmission electron microscopy |
| E% | = | Incorporation Efficiency |
| PBS | = | Phosphate buffered saline solution |
| IC50 | = | 50% inhibition concentration |
| PFU/cell | = | Plaque forming unit per cell |
| MOI 1 | = | Multiplicity of infection |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide |
| CMDA | = | 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene |
| Tm | = | Phase-transition temperature |
| EO | = | Essential oil |
| P.I. | = | Polidispersivity index |
| CG50 | = | 50% Cytotoxic Concentration |
| [n]20D | = | Refractive Index |
| [α]D | = | Specific Rotation |
| ABBREVIATIONS | ||
| HSV-1 | = | Herpes simplex virus type 1 |
| HSV-2 | = | Herpes simplex virus 2 |
| HIV-1 | = | Human Immunodeficiency Virus-type 1 |
| S. | = | Santolina |
| MPS | = | Mononuclear phagocityc system |
| HPC | = | Hydrogenated soya phosphatidylcholine |
| HPLC | = | High Performance Liquid Chromatography |
| CHOL | = | Cholesterol |
| FBS | = | Fetal bovine serum |
| MEM | = | Modified Eagle Medium |
| GC | = | Gas Chromatograghy |
| PFU | = | Plaque forming units |
| i.d. | = | internal diameter |
| MLV | = | Multilamellar Vesicles |
| SUV | = | Small Unilamellar Vesicles |
| TEM | = | Transmission electron microscopy |
| E% | = | Incorporation Efficiency |
| PBS | = | Phosphate buffered saline solution |
| IC50 | = | 50% inhibition concentration |
| PFU/cell | = | Plaque forming unit per cell |
| MOI 1 | = | Multiplicity of infection |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide |
| CMDA | = | 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene |
| Tm | = | Phase-transition temperature |
| EO | = | Essential oil |
| P.I. | = | Polidispersivity index |
| CG50 | = | 50% Cytotoxic Concentration |
| [n]20D | = | Refractive Index |
| [α]D | = | Specific Rotation |