title: Evaluation of safety margins of Chenopodium album seed decoction: 14-day subacute toxicity and microbicidal activity studies.
abstract: BACKGROUND
  Sperm immobilizing activity and plausible mechanism of action of Chenopodium album seed decoction (CAD) have been elucidated in our earlier studies. The present study has been carried out to explore the safety standards of CAD along with microbicidal properties as prerequisite for its use as a topically applicable vaginal contraceptive.
  
  
  METHODS
  The safety standards of CAD were assessed by a) Hemolytic index determination using rabbit erythrocytes, to set the doses of the other experiments, b) Dermal irritancy test using refined version of Draize scoring system on rabbits, c) Possible effect on local tissues and reproductive performance in female rats after fourteen daily single dose application, d) PCNA staining- to evaluate the effect of CAD on vaginal tissue proliferation, e) TUNEL assay- to examine its ability to induce in situ apoptosis in the vaginal tissue sections of the treated animals, and f) Microbicidal activity- to explore the effect of CAD on the growth of Lactobacillus acidophilus and Candida albicans.
  
  
  RESULTS
  In vitro irritation studies on rabbit erythrocytes revealed the hemolytic index of CAD to be 8.2 mg/ml. The dermal irritation test showed it to be a non-irritant even at higher doses. Intra vaginal application of CAD in rat vagina for 14 consecutive days caused slight reversible inflammation on vaginal epithelial cells at doses as high as 82 mg/ml. However, at this dose level it neither had any adverse effect on vaginal tissue proliferation nor did it cause in situ apoptosis as evident from PCNA staining and TUNEL assay. Fertility and fecundity were restored 4-15 days after withdrawal of CAD application. At dose level 10 times that of its spermicidal MEC (minimum effective concentration), CAD did not block the growth of Lactobacillus, although the size of individual colony was marginally reduced. However, growth of the pathogenic fungus Candida albicans was completely inhibited with 20 mg/ml of CAD.
  
  
  CONCLUSIONS
  The overall result evolved from the study strengthens the candidature of CAD as a safe microbicidal spermicide. It is almost non-irritant to rabbit skin and rat vaginal tissues at doses 10 fold higher than its hemolytic index. The effect of CAD on Lactobacillus culture was not highly encouraging but it prevented the growth of the fungal pathogen Candida albicans at 20 mg/ml of CAD.
authors.affiliation: Department of Chemistry, Indian Institute of Chemical Biology, Council of Scientific and Industrial research, 4 Raja S C Mullick Road, Jadavpur, Kolkata, India.
authors.forename: Shrabanti
authors.initials: S
authors.lastname: Kumar
authors.forename: Shampa
authors.initials: S
authors.lastname: Biswas
authors.forename: Sukdeb
authors.initials: S
authors.lastname: Banerjee
authors.forename: Nirup B
authors.initials: NB
authors.lastname: Mondal
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002176:Candida albicans / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D027463:Chenopodium album / Q000737:chemistry*; D064420:Drug-Related Side Effects and Adverse Reactions; D004912:Erythrocytes / Q000187:drug effects*; D005123:Eye / Q000187:drug effects; D005260:Female; D006461:Hemolysis / Q000187:drug effects; D020287:In Situ Nick-End Labeling; D007779:Lactobacillus acidophilus / Q000187:drug effects; D010936:Plant Extracts / Q000633:toxicity*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D012639:Seeds / Q000633:toxicity*; D013089:Spermatocidal Agents / Q000494:pharmacology*; D014621:Vagina / Q000187:drug effects / Q000473:pathology; D014627:Vaginitis / Q000139:chemically induced
chemical_list: D010936:Plant Extracts; D013089:Spermatocidal Agents