Fletcher1, P.S., Harman1, S.J., Boothe2, A.R., Doncel2, G.F. and Shattock1, R.J. 1St. George's University of London, UK and 2CONRAD, Eastern Virginia Medical School, USA

This study by researchers in the CONRAD (Contraceptive Research and Development) Program at Eastern Virginia Medical School and at St. George’s University/London demonstrated how MatTek’s EpiVaginal in vitro human cervico-vaginal tissue equivalent can be used pre-clinically to test potentially cytotoxic microbicide formulations in vitro. Background: The continued growth of the global HIV epidemic highlights the urgent need to develop novel prevention strategies to reduce HIV transmission. The development of topical microbicides is likely to take a number of years before such a product would be widely available. This has resulted in a call for the rapid introduction of simpler vaginal intervention strategies in the interim period. One suggested practice would be vaginal douching with natural products including lime or lemon juice. Here we present a comprehensive preclinical evaluation of lime juice (LiJ) as a potential intervention strategy against HIV. Results: Pre-treatment of HIV with LiJ demonstrated direct virucidal activity, with 10% juice inactivating the virus within 5 minutes. However, this activity was significantly reduced in the presence of seminal plasma, where inactivation required maintaining a 1:1 mixture of neat LiJ and seminal plasma for more than 5 minutes. Additionally, LiJ demonstrated both time and dose dependent toxicity towards cervicovaginal epithelium, where exposure to 50% juice caused 75– 90% toxicity within 5 minutes increasing to 95% by 30 minutes. Cervicovaginal epithelial cell monolayers were more susceptible to the effects of LiJ with 8.8% juice causing 50% toxicity after 5 minutes. Reconstructed stratified cervicovaginal epithelium (MatTek’s EpiVaginal tissues) appeared more resilient to LiJ toxicity with 30 minutes exposure to 50% LiJ having little effect on viability. However viability was reduced by 75% and 90% following 60 and 120 minutes exposure. Furthermore, repeat application (several times daily) of 25% LiJ caused 80–90% reduction in viability. Conclusion: These data demonstrate that the virucidal activity of LiJ is severely compromised in the presence of seminal plasma. Potentially, to be effective against HIV in vivo, women would need to apply a volume of neat LiJ equal to that of an ejaculate, and maintain this ratio vaginally for 5–30 minutes after ejaculation. Data presented here suggest that this would have significant adverse effects on the genital mucosa. These data raise serious questions about the plausibility and safety of such a prevention approach.


EpiVaginal, HIV, MTT, Microbicide, Microbicides, Repeat application, Repeat topical exposure, Toxicity, VEC-100

Materials Tested

Lime juice (LiJ)

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