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Research Article

LIPOSOME-INCORPORATED SANTOLINA INSULARIS ESSENTIAL OIL: PREPARATION, CHARACTERIZATION AND IN VITRO ANTIVIRAL ACTIVITY

, , , , , , & show all
Pages 73-90 | Published online: 30 Apr 2001
 

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

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