Ovahormon injection

Estradiol benzoate (OVAHORMON^®INJECTION; ASKA Pharmaceutical Co. Ltd., Tokyo, Japan) was applied to wounds at 0.75 μg/g/day every day following wounding procedures under inhalational anesthesia [21 (link)]. In brief, estradiol benzoate was applied to cover the wound surface after cleaning the wounds with normal saline. Estradiol benzoate was diluted at 0.75 μg/g in sesame oil (Wako Pure Chemical Industries Ltd., Tokyo, Japan). The sesame oil vehicle was applied at the same dose as estradiol benzoate to wounds every day following wounding.

Estradiol benzoate (estra-1,3,5(10)-triene-3,17β-diol 3-benzoate) (OVAHORMON^®INJECTION; ASKA Pharmaceutical Co. Ltd., Tokyo, Japan) at a dose of 0.75 μg/g/day was applied to the wounds daily according to our previous study [26 (link)]. Estradiol benzoate was diluted at 0.75 μg/g in sesame oil (Wako). The dose of sesame oil, which was used as a vehicle, was similar to that of estradiol benzoate, and it was applied on the wound daily.

In the topical EB wound treatment group, EB (Estra-1,3,5(10)-triene-3,17β-diol 3-benzoate) (OVAHORMON^®INJECTION; ASKA Pharmaceutical Co. Ltd., Tokyo, Japan) was applied at 0.75 μg/g/day to the wounds every day after wounding. EB was diluted at 0.75 μg/g in sesame oil (Wako Pure Chemical Industries Ltd., Tokyo, Japan). In the subcutaneous E2 (Estra-1,3,5(10)-triene-3,17β-diol) pellet group, a 0.05-mg, 21-d, slow-release E2 pellet (Innovative Research of a America, Sarasota, FL) was administrated at the time of wounding by s.c. implantation according to previous studies [8 (link)–13 (link), 17 (link)–21 (link)]. In the topical E2 skin application group, E2 gel (L’estrogel 0.06%; Bayer Yakuhin, Osaka, Japan) was applied at 0.01 g/day to the skin on the back, avoiding the wounds, every day after wounding according to our previous studies [14 (link)–16 (link)]. The dose administered was selected with successful estrogen replacement confirmed by an enzyme immunoassay on plasma samples, vaginal smears, or uterine weights.