Open Access Research article

Quantitative assessment of microbicide-induced injury in the ovine vaginal epithelium using confocal microendoscopy

Gracie Vargas12*, Igor Patrikeev1, Jingna Wei1, Brent Bell1, Kathleen Vincent34, Nigel Bourne3 and Massoud Motamedi15

Author Affiliations

1 Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, TX, USA

2 Department of Neuroscience and Cell Biology, The University of Texas Medical Branch, Galveston, TX, USA

3 Department of Pediatrics, The University of Texas Medical Branch, Galveston, TX, USA

4 Department of Obstetrics and Gynecology, The University of Texas Medical Branch, Galveston, TX, USA

5 Department of Opthalmology, The University of Texas Medical Branch, Galveston, TX, USA

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BMC Infectious Diseases 2012, 12:48  doi:10.1186/1471-2334-12-48

Published: 29 February 2012



The development of safe topical microbicides that can preserve the integrity of cervicovaginal tract epithelial barrier is of great interest as this may minimize the potential for increased susceptibility to STI infections. High resolution imaging to assess epithelial integrity in a noninvasive manner could be a valuable tool for preclinical testing of candidate topical agents.


A quantitative approach using confocal fluorescence microendoscopy (CFM) for assessment of microbicide-induced injury to the vaginal epithelium was developed. Sheep were treated intravaginally with one of five agents in solution (PBS; 0.02% benzalkonium chloride (BZK); 0.2% BZK) or gel formulation (hydroxyethyl cellulose (HEC); Gynol II nonoxynol-9 gel (N-9)). After 24 hours the vaginal tract was removed, labeled with propidium iodide (PI), imaged, then fixed for histology. An automated image scoring algorithm was developed for quantitative assessment of injury and applied to the data set. Image-based findings were validated with histological visual gradings that describe degree of injury and measurement of epithelial thickness.


Distinct differences in PI staining were detected following BZK and N-9 treatment. Images from controls had uniformly distributed nuclei with defined borders, while those after BZK or N-9 showed heavily stained and disrupted nuclei, which increased in proportion to injury detected on histology. The confocal scoring system revealed statistically significant scores for each agent versus PBS controls with the exception of HEC and were consistent with histology scores of injury.


Confocal microendoscopy provides a sensitive, objective, and quantitative approach for non-invasive assessment of vaginal epithelial integrity and could serve as a tool for real-time safety evaluation of emerging intravaginal topical agents.

Benzalkonium chloride; Confocal endomicroscopy; Epithelial disruption; Imaging; Microbicides; Nonoxynol-9