Table 1.

In-vivo evaluation of 5-hydroxytyrosol toxicity. 5-Hydroxytyrosol was administered at three different doses for 14 consecutive days by topical route to the vaginal mucosa. All animals were sacrificed at the end of the treatment period and mortality/viability, clinical signs, microbiological assessment, vaginal pH and macroscopic and microscopic examination of the genital apparatus of each animal were performed.

			Vagina
						Microbiological assessment
	Clinical signs	Mortality/viability	Macroscopic examinationa	Microscopic examinationb	pH (mean ± SD)	Vaginal exudate before treatments	Vaginal mucosa after treatments
Group 1: blank control (n = 6)	Swelling in the genital area for several days (n = 1)	0	n = 3	LI: Grade 1 or 2 in proximal, medial and/or distal parts (n = 4)	7.46 ± 0.30	Streptococcus sp.: n = 6	Streptococcus sp.: n = 0
				VC: Grade 1 (n = 4)		Klebsiella pneumoniae: n = 0	Klebsiella pneumoniae: n = 4
				E: Grade 1 (n = 2)
Group 2: placebo HEC gel (n = 6)	–	0	n = 3	LI: Grade 1 or 2 in proximal or medial parts (n = 2)	7.80 ± 0.48	Streptococcus sp.: n = 6	Streptococcus sp.: n = 0
				VC: Grade 1 (n = 3)		Klebsiella pneumoniae: n = 0	Klebsiella pneumoniae: n = 1
				E: Grade 1 (n = 2)
Group 3: 5-hydroxytyrosol gel 30 mmol/l (n = 6)	–	0	n = 4	LI: Grade 1 or 2 in proximal, medial and/or distal parts (n = 4)	7.73 ± 0.27	Streptococcus sp.: n = 6	Streptococcus sp.: n = 0
				VC: Grade 1 (n = 4)		Klebsiella pneumoniae: n = 0	Klebsiella pneumoniae: n = 2
				E: Grade 1 (n = 2)
Group 4: 5-hydroxytyrosol gel 100 mmol/l (n = 6)	–	0	n = 6	VC: Grade 1 (n = 6)	7.66 ± 0.39	Streptococcus sp.: n = 5	Streptococcus sp.: n = 1
				E: Grade 1 (n = 3)		Klebsiella pneumoniae: n = 1	Klebsiella pneumoniae: n = 0
Group 5: 5-hydroxytyrosol gel 200 mmol/l (n = 6)	–	0	n = 5	LI: Grade 1 or 2 in proximal and medial parts (n = 5)	7.55 ± 0.35	Streptococcus sp.: n = 5	Streptococcus sp.: n = 1
				VC: Grade 1 (n = 5)		Klebsiella pneumoniae: n = 0	Klebsiella pneumoniae: n = 3
				E: Grade 1 (n = 3)

^aMacroscopic examination: n means number of animals with signs of irritation in the vaginal mucosa.

^bMicroscopic examination: leukocyte infiltration (LI) in vaginal mucosa. Vascular congestion (VC) on submucosa (proximal, medial and/or distal parts) and oedema (E) in mucosa (proximal, medial and/or distal parts): absent (grade 0), minimal (grade 1), mild (grade 2), moderate (grade 3) and marked (grade 4).