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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

Effect of Lemon Juice, Lime Juice, Vinegar, Nonoxynol-9 (N-9), and Triton X-100 on Viability of Cervical Explant Tissues. Effects of lemon juice, lime juice, vinegar, Triton X-100, nonoxynol-9 (N-9), and UC781 on viability of cervical explant tissues. Presented are the percent viability for tissues treated with lemon juice (1-20%), lime juice (0.32-20%), vinegar (6%), Triton X-100 (0.00005-5%), nonoxynol-9 (N-9; 100 μg/mL), and UC781 (100 μM), compared to donor-matched, untreated controls (defined as 100%). Tissues were exposed from 2 hours to overnight. Each bar represents data from 1 to 6 donors. Bars indicate mean ± SD for each product/concentration. The concentration of juice or vinegar is expressed as percent (%) solution (v/v).
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Figure 4: Effect of Lemon Juice, Lime Juice, Vinegar, Nonoxynol-9 (N-9), and Triton X-100 on Viability of Cervical Explant Tissues. Effects of lemon juice, lime juice, vinegar, Triton X-100, nonoxynol-9 (N-9), and UC781 on viability of cervical explant tissues. Presented are the percent viability for tissues treated with lemon juice (1-20%), lime juice (0.32-20%), vinegar (6%), Triton X-100 (0.00005-5%), nonoxynol-9 (N-9; 100 μg/mL), and UC781 (100 μM), compared to donor-matched, untreated controls (defined as 100%). Tissues were exposed from 2 hours to overnight. Each bar represents data from 1 to 6 donors. Bars indicate mean ± SD for each product/concentration. The concentration of juice or vinegar is expressed as percent (%) solution (v/v).

Mentions: Since lemon and lime juice and vinegar were toxic to primary and transformed cell lines of various origin and vaginal Lactobacillus species, the next goal was to assess the cytotoxicity of these liquids in freshly obtained human cervical tissues (Figure 4). Human cervical explant tissues were exposed to the juices, vinegar, N-9, and Triton X-100, and the percent viability of the tissues is shown in Figure 4. The % viability of exposed tissues was higher for N-9 at 100 μg/mL than for the juices at ≥ 10%. Ten percent lemon juice reduced tissue mean viability by > 70%, and 10% lime juice reduced viability by > 80% as compared to tissue treated with culture medium only (Figure 4). Exposure of tissue to N-9 (100 μg/mL) or 0.3% acetic acid (6% household vinegar) reduced tissue viability by 50 and 30%, respectively. Five percent lemon or lime juice appeared less toxic than the 10% concentrations showing a clear dose-response effect on the tissues. Table 4 summarizes the anti-HIV-1 effect of lemon and lime juice compared to the untreated HIV-1 infected control in explant tissue. Virus replication was determined as a function of HIV-1 p24 in culture supernatants [21]. There was little or undetectable HIV-1 replication in the 5-20% lemon and lime juice treated samples compared to the untreated HIV-1 infected control, where an average of 3,090 pg/mL HIV-1 p24 Gag was measured. Treatment with 1% natural juice or neutralized juices up to 10% resulted in virus replication levels comparable to that of untreated virus controls.


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)

Effect of Lemon Juice, Lime Juice, Vinegar, Nonoxynol-9 (N-9), and Triton X-100 on Viability of Cervical Explant Tissues. Effects of lemon juice, lime juice, vinegar, Triton X-100, nonoxynol-9 (N-9), and UC781 on viability of cervical explant tissues. Presented are the percent viability for tissues treated with lemon juice (1-20%), lime juice (0.32-20%), vinegar (6%), Triton X-100 (0.00005-5%), nonoxynol-9 (N-9; 100 μg/mL), and UC781 (100 μM), compared to donor-matched, untreated controls (defined as 100%). Tissues were exposed from 2 hours to overnight. Each bar represents data from 1 to 6 donors. Bars indicate mean ± SD for each product/concentration. The concentration of juice or vinegar is expressed as percent (%) solution (v/v).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2913913&req=5

Figure 4: Effect of Lemon Juice, Lime Juice, Vinegar, Nonoxynol-9 (N-9), and Triton X-100 on Viability of Cervical Explant Tissues. Effects of lemon juice, lime juice, vinegar, Triton X-100, nonoxynol-9 (N-9), and UC781 on viability of cervical explant tissues. Presented are the percent viability for tissues treated with lemon juice (1-20%), lime juice (0.32-20%), vinegar (6%), Triton X-100 (0.00005-5%), nonoxynol-9 (N-9; 100 μg/mL), and UC781 (100 μM), compared to donor-matched, untreated controls (defined as 100%). Tissues were exposed from 2 hours to overnight. Each bar represents data from 1 to 6 donors. Bars indicate mean ± SD for each product/concentration. The concentration of juice or vinegar is expressed as percent (%) solution (v/v).
Mentions: Since lemon and lime juice and vinegar were toxic to primary and transformed cell lines of various origin and vaginal Lactobacillus species, the next goal was to assess the cytotoxicity of these liquids in freshly obtained human cervical tissues (Figure 4). Human cervical explant tissues were exposed to the juices, vinegar, N-9, and Triton X-100, and the percent viability of the tissues is shown in Figure 4. The % viability of exposed tissues was higher for N-9 at 100 μg/mL than for the juices at ≥ 10%. Ten percent lemon juice reduced tissue mean viability by > 70%, and 10% lime juice reduced viability by > 80% as compared to tissue treated with culture medium only (Figure 4). Exposure of tissue to N-9 (100 μg/mL) or 0.3% acetic acid (6% household vinegar) reduced tissue viability by 50 and 30%, respectively. Five percent lemon or lime juice appeared less toxic than the 10% concentrations showing a clear dose-response effect on the tissues. Table 4 summarizes the anti-HIV-1 effect of lemon and lime juice compared to the untreated HIV-1 infected control in explant tissue. Virus replication was determined as a function of HIV-1 p24 in culture supernatants [21]. There was little or undetectable HIV-1 replication in the 5-20% lemon and lime juice treated samples compared to the untreated HIV-1 infected control, where an average of 3,090 pg/mL HIV-1 p24 Gag was measured. Treatment with 1% natural juice or neutralized juices up to 10% resulted in virus replication levels comparable to that of untreated virus controls.

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