Assessment of oral toxicity and vaginal irritation test of novel antimicrobial mucoadhesive herbal vaginal tablet for the treatment of vaginitis

Abstract

Introduction: The preclinical in vivo toxicity study aims to standardize new vaginal herbal formulation consisting of stem barks of Ficus glomerata Roxb. and Symplocos racemosa Roxb. which have been mentioned in Ayurvedic system of medicine as effective in the treatment of vaginitis. Acute and subacute toxicities of its aqueous extract have been evaluated using the oral route of administration through hematological and histopathological studies of the stomach, liver, and kidney. Vaginal irritation test has been also done to assess the redness, ulceration, bleeding, etc., during drug administration.Materials and Methods: Acute toxicity study was carried out on Swiss albino female mice weighing about 20–30 g following OECD guidelines. A single dose of aqueous extract of the research drug was administered orally at the level of 300, 600, 900, 1200, or 1500 mg/kg to different groups and animals were observed for the appearance of toxic symptoms up to 24 h. During subacute toxicity study, female albino mice divided into five groups of 6 animals were initially orally administered 500 mg/kg dose which was increased by 50 mg/kg every 3rd day to a maximum of 1200 mg/kg dose for a period of 4 weeks. After recording changes in body weight, food, and water intake and clinical signs, animals were sacrificed on the 30th day, and their hematological parameters and liver function test (LFT) were done. Histopathological studies were done on slides of dissected stomach, liver, and kidney and internal structures of cells, tissue, and mucous membrane, etc., were observed.Vaginal toxicity testing was performed on 18 female Wistar rats divided into 3 test groups, namely, control, drug-treated (500 mg/kg), and placebo-administered, using vaginal route of drug administration. Animals were necropsied 24 h after final vaginal dose and heart, kidney, liver, lung, ovary, pancreas, uterus, and vagina were excised, and histopathological examination of the internal structure of their cells, tissues, and glands was done. Results and Discussions: The acute toxicity tests performed on the research formulation revealed no signs of behavioral changes up to the dose of 1500 mg/kg and no mortality was reported up to 24 h. During the subacute toxicity (long-term toxicity) testing, no animal died when daily dose of the drug was gradually increased to 1200 mg/kg b.w. over 29 days, and no significant toxic effects were observed over a prolonged period. At the same time, the hematological studies, LFT and hormonal parameters of blood did not show any noticeable differences, and no significant damage of cells, tissues, epithelium lining, or other structures of stomach, liver, and kidney was observed during histopathological studies. The vaginal irritation (subacute toxicity) studies indicated that no differences were observed among treatment groups in respect of hematological parameters as well as weight and internal structures of all vital organs during the histopathological examination.Conclusion: The results clearly indicate the non-toxic attribute of research drug formulation on the basis of analysis of blood parameters and reproductive hormones using oral and intravaginal administration routes.