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Preclinical In‐vivo Safety Evaluation of Concanavalin A‐based Nanomicrobicide in Female Mouse Reproductive Tract
Author(s) -
Ezoulin Miezan JM,
Coulibaly Fohona,
Dim Daniel
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1195.6
Subject(s) - proinflammatory cytokine , concanavalin a , h&e stain , spleen , medicine , pathology , staining , andrology , chemistry , inflammation , immunology , in vitro , biochemistry
Objective Tenofovir (TFV) loaded Concanavalin A nanoparticles (ConA‐NPs) was recently synthesized as mannose sensitive nanomicrobicide template for the prevention of HIV/AIDS transmission. To advance into clinical trials, such nanomicrobicide must be evaluated in preclinical safety and efficacy models. This study aims at examining the mucosal safety of these specific ConA‐NPs in the female mouse reproductive tract. Method TFV loaded ConA‐NPs (~ 670 ± 234 nm, −15 ± 0.55 mV) were prepared via the “layer‐by‐layer” technology by coating a core template, calcium carbonate (CaCO3) crystal, with Con A crosslinked with glycogen. Pre‐clinical safety studies of intravaginally administered TFV loaded ConA‐NPs (100 mg/kg) were performed using a female mouse model C57Bl6. Mouse cervicovaginal lavage (CVL) and tissues (CVT) were collected 24h after a single application. Changes in the levels of inflammatory cytokines (IL‐1l, IL‐1β, IL‐6, IL‐7, IP‐10, MKC, and TNF‐l) were quantitated by a high‐sensitivity multiplexed bead‐based immunoassay. The histopathology of the genital tract (vagina, cervix, uterus, and ovary), rectum and other major organs (spleen, lung, liver, kidney, heart, brain) was studied using Hematoxylin & Eosin staining. The vaginal tissues were evaluated for localized immune cells (lymphocytes) infiltration by immunohistochemical staining for CD45. Results After 24h exposure to these NPs, the basal levels of proinflammatory cytokines (IL‐1l, IL‐1β, IL‐6, IL‐7, IP‐10, MKC, and TNF‐l) in mouse CVL and CVT increased 3 – 56 fold and 1.2 – 5.2 fold, respectively when compared to phosphate buffered saline (PBS) used as a negative control. The induced inflammation was associated with the infiltration of immune cells to the vaginal epithelium. The histological analysis of the reproductive tract revealed signs of epithelial disruption in the vagina. However, all other tissues were histologically similar to control tissue at 24 h post application. Conclusion These experiments showed that the present ConA‐NPs are cytotoxic to the genital tract epithelial cells at 100 mg/kg. These data suggest the need for further preclinical studies to determine the maximum tolerable dose of the nano formulation for vaginal application. Support or Funding Information The project was supported by Award Number RO1 AI087304 from the National Institutes of Allergy and Infectious Diseases (Bethesda, MD, USA). The content is the sole responsibility of the authors and does not necessarily represent the official views of the National Institutes of Allergy and Infectious Diseases or the National Institutes of Health.