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Safety and anti-HIV assessments of natural vaginal cleansing products in an established topical microbicides in vitro testing algorithm.

Lackman-Smith CS, Snyder BA, Marotte KM, Osterling MC, Mankowski MK, Jones M, Nieves-Duran L, Richardson-Harman N, Cummins JE, Sanders-Beer BE - AIDS Res Ther (2010)

Bottom Line: These products were also tested for their effect on viability of cervico-vaginal cell lines, human cervical explant tissues, and beneficial Lactobacillus species.All three liquid products inhibited viability of beneficial Lactobacillus species associated with vaginal health.Lemon and lime juice and household vinegar do not fulfill the safety criteria mandated for a topical microbicide.

View Article: PubMed Central - HTML - PubMed

Affiliation: Southern Research Institute, Frederick, MD, USA. lackmansmith@southernresearch.org.

ABSTRACT

Background: At present, there is no effective vaccine or other approved product for the prevention of sexually transmitted human immunodeficiency virus type 1 (HIV-1) infection. It has been reported that women in resource-poor communities use vaginally applied citrus juices as topical microbicides. These easily accessible food products have historically been applied to prevent pregnancy and sexually transmitted diseases. The aim of this study was to evaluate the efficacy and cytotoxicity of these substances using an established topical microbicide testing algorithm. Freshly squeezed lemon and lime juice and household vinegar were tested in their original state or in pH neutralized form for efficacy and cytotoxicity in the CCR5-tropic cell-free entry and cell-associated transmission assays, CXCR4-tropic entry and fusion assays, and in a human PBMC-based anti-HIV-1 assay. These products were also tested for their effect on viability of cervico-vaginal cell lines, human cervical explant tissues, and beneficial Lactobacillus species.

Results: Natural lime and lemon juice and household vinegar demonstrated anti-HIV-1 activity and cytotoxicity in transformed cell lines. Neutralization of the products reduced both anti-HIV-1 activity and cytotoxicity, resulting in a low therapeutic window for both acidic and neutralized formulations. For the natural juices and vinegar, the IC50 was

Conclusions: Lemon and lime juice and household vinegar do not fulfill the safety criteria mandated for a topical microbicide. As a result of their unphysiological formulation for the vaginal tract, they exhibit cytotoxicity to human cell lines, human vaginal tissues, and beneficial vaginal Lactobacillus species.

No MeSH data available.


Related in: MedlinePlus

HIV-1 Replication Inhibition and Cytotoxicity with Increasing Concentrations of Lemon Juice, Lime Juice, and Vinegar in Cell-based Assays. CCR5-tropic cell-free HIV-1 entry assay (A), CCR5-tropic cell-associated HIV-1 transmission assay (B), CXCR4-tropic cell-free HIV-1 entry assay (C), CXCR4-tropic fusion assay (D), and PBMC antiviral assay (E). Virus growth (shown as % of virus control (VC), solid lines) and cell viability (shown as % viability of untreated cell control (CC), dashed lines) are presented for lemon juice, lime juice, and vinegar. Results are shown for both pH neutral (blue) and natural (red) formulations. Means ± standard deviations (SD) of replicate experiments are presented. The black, horizontal line indicates the level for 50% cell death (i.e., TC50) or 50% virus inhibition (i.e., IC50), respectively, for each assay. The number of experiments performed (n) is indicated within each figure for the pH neutralized (*) and the natural products (#). The concentration of juice or vinegar is expressed as percent (%) solution (v/v).
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Figure 1: HIV-1 Replication Inhibition and Cytotoxicity with Increasing Concentrations of Lemon Juice, Lime Juice, and Vinegar in Cell-based Assays. CCR5-tropic cell-free HIV-1 entry assay (A), CCR5-tropic cell-associated HIV-1 transmission assay (B), CXCR4-tropic cell-free HIV-1 entry assay (C), CXCR4-tropic fusion assay (D), and PBMC antiviral assay (E). Virus growth (shown as % of virus control (VC), solid lines) and cell viability (shown as % viability of untreated cell control (CC), dashed lines) are presented for lemon juice, lime juice, and vinegar. Results are shown for both pH neutral (blue) and natural (red) formulations. Means ± standard deviations (SD) of replicate experiments are presented. The black, horizontal line indicates the level for 50% cell death (i.e., TC50) or 50% virus inhibition (i.e., IC50), respectively, for each assay. The number of experiments performed (n) is indicated within each figure for the pH neutralized (*) and the natural products (#). The concentration of juice or vinegar is expressed as percent (%) solution (v/v).

Mentions: The antiviral and cytotoxic effects of natural and pH neutralized lemon and lime juices and house-hold vinegar in cell-free and cell-associated HIV-1 transmission inhibition assays are presented in Figure 1a-e. In the CCR5-tropic and CXCR4-tropic cell-free HIV-1 entry assays (Figure 1a and Table 1), the IC50s and TC50s of natural lemon and lime juices and vinegar ranged from 1.7 to 4.1% solution (v/v), resulting in a very low therapeutic index (0.9-2.3). Neutralization of the juices increased the therapeutic index for lemon and lime juice in the CCR5-tropic assay, but not in the CXCR4-tropic assay. Neutralization of vinegar abolished both efficacy and toxicity in the CCR5-and CXCR4-tropic HIV-1 entry assays. For the CCR5-tropic cell-associated HIV-1 transmission assay, the CXCR4-tropic fusion assay, and the HIV-1 PBMC assays the therapeutic indices remained low (≤ 7.0) whether the juices and vinegar were neutralized or not. The therapeutic index was especially low in the HIV-1 fusion assays, where HeLa cells were exposed to the juices and vinegar for 48 hours.


Safety and anti-HIV assessments of natural vaginal cleansing products in an established topical microbicides in vitro testing algorithm.

Lackman-Smith CS, Snyder BA, Marotte KM, Osterling MC, Mankowski MK, Jones M, Nieves-Duran L, Richardson-Harman N, Cummins JE, Sanders-Beer BE - AIDS Res Ther (2010)

HIV-1 Replication Inhibition and Cytotoxicity with Increasing Concentrations of Lemon Juice, Lime Juice, and Vinegar in Cell-based Assays. CCR5-tropic cell-free HIV-1 entry assay (A), CCR5-tropic cell-associated HIV-1 transmission assay (B), CXCR4-tropic cell-free HIV-1 entry assay (C), CXCR4-tropic fusion assay (D), and PBMC antiviral assay (E). Virus growth (shown as % of virus control (VC), solid lines) and cell viability (shown as % viability of untreated cell control (CC), dashed lines) are presented for lemon juice, lime juice, and vinegar. Results are shown for both pH neutral (blue) and natural (red) formulations. Means ± standard deviations (SD) of replicate experiments are presented. The black, horizontal line indicates the level for 50% cell death (i.e., TC50) or 50% virus inhibition (i.e., IC50), respectively, for each assay. The number of experiments performed (n) is indicated within each figure for the pH neutralized (*) and the natural products (#). The concentration of juice or vinegar is expressed as percent (%) solution (v/v).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2913913&req=5

Figure 1: HIV-1 Replication Inhibition and Cytotoxicity with Increasing Concentrations of Lemon Juice, Lime Juice, and Vinegar in Cell-based Assays. CCR5-tropic cell-free HIV-1 entry assay (A), CCR5-tropic cell-associated HIV-1 transmission assay (B), CXCR4-tropic cell-free HIV-1 entry assay (C), CXCR4-tropic fusion assay (D), and PBMC antiviral assay (E). Virus growth (shown as % of virus control (VC), solid lines) and cell viability (shown as % viability of untreated cell control (CC), dashed lines) are presented for lemon juice, lime juice, and vinegar. Results are shown for both pH neutral (blue) and natural (red) formulations. Means ± standard deviations (SD) of replicate experiments are presented. The black, horizontal line indicates the level for 50% cell death (i.e., TC50) or 50% virus inhibition (i.e., IC50), respectively, for each assay. The number of experiments performed (n) is indicated within each figure for the pH neutralized (*) and the natural products (#). The concentration of juice or vinegar is expressed as percent (%) solution (v/v).
Mentions: The antiviral and cytotoxic effects of natural and pH neutralized lemon and lime juices and house-hold vinegar in cell-free and cell-associated HIV-1 transmission inhibition assays are presented in Figure 1a-e. In the CCR5-tropic and CXCR4-tropic cell-free HIV-1 entry assays (Figure 1a and Table 1), the IC50s and TC50s of natural lemon and lime juices and vinegar ranged from 1.7 to 4.1% solution (v/v), resulting in a very low therapeutic index (0.9-2.3). Neutralization of the juices increased the therapeutic index for lemon and lime juice in the CCR5-tropic assay, but not in the CXCR4-tropic assay. Neutralization of vinegar abolished both efficacy and toxicity in the CCR5-and CXCR4-tropic HIV-1 entry assays. For the CCR5-tropic cell-associated HIV-1 transmission assay, the CXCR4-tropic fusion assay, and the HIV-1 PBMC assays the therapeutic indices remained low (≤ 7.0) whether the juices and vinegar were neutralized or not. The therapeutic index was especially low in the HIV-1 fusion assays, where HeLa cells were exposed to the juices and vinegar for 48 hours.

Bottom Line: These products were also tested for their effect on viability of cervico-vaginal cell lines, human cervical explant tissues, and beneficial Lactobacillus species.All three liquid products inhibited viability of beneficial Lactobacillus species associated with vaginal health.Lemon and lime juice and household vinegar do not fulfill the safety criteria mandated for a topical microbicide.

View Article: PubMed Central - HTML - PubMed

Affiliation: Southern Research Institute, Frederick, MD, USA. lackmansmith@southernresearch.org.

ABSTRACT

Background: At present, there is no effective vaccine or other approved product for the prevention of sexually transmitted human immunodeficiency virus type 1 (HIV-1) infection. It has been reported that women in resource-poor communities use vaginally applied citrus juices as topical microbicides. These easily accessible food products have historically been applied to prevent pregnancy and sexually transmitted diseases. The aim of this study was to evaluate the efficacy and cytotoxicity of these substances using an established topical microbicide testing algorithm. Freshly squeezed lemon and lime juice and household vinegar were tested in their original state or in pH neutralized form for efficacy and cytotoxicity in the CCR5-tropic cell-free entry and cell-associated transmission assays, CXCR4-tropic entry and fusion assays, and in a human PBMC-based anti-HIV-1 assay. These products were also tested for their effect on viability of cervico-vaginal cell lines, human cervical explant tissues, and beneficial Lactobacillus species.

Results: Natural lime and lemon juice and household vinegar demonstrated anti-HIV-1 activity and cytotoxicity in transformed cell lines. Neutralization of the products reduced both anti-HIV-1 activity and cytotoxicity, resulting in a low therapeutic window for both acidic and neutralized formulations. For the natural juices and vinegar, the IC50 was

Conclusions: Lemon and lime juice and household vinegar do not fulfill the safety criteria mandated for a topical microbicide. As a result of their unphysiological formulation for the vaginal tract, they exhibit cytotoxicity to human cell lines, human vaginal tissues, and beneficial vaginal Lactobacillus species.

No MeSH data available.


Related in: MedlinePlus