β-Estradiol is widely used in hormone replacement therapy (HRT) to alleviate symptoms of menopause, such as hot flashes, vaginal dryness, night sweats, and osteoporosis. It helps restore estrogen levels in postmenopausal women, improving overall quality of life. In transgender hormone therapy, it is administered to feminize secondary sexual characteristics in transgender women, such as breast development and fat redistribution. Additionally, β-Estradiol is utilized in the treatment of certain hormonal disorders, such as hypogonadism, primary ovarian failure, and estrogen deficiency following oophorectomy. It also plays a role in managing some types of breast and prostate cancer by regulating hormone-dependent tumor growth. In research, it is used to study estrogen signaling pathways and their effects on various physiological processes, including bone health where it helps prevent and treat osteoporosis by increasing bone density and reducing fracture risk.

However, the use of β-Estradiol should be under close medical supervision due to potential side effects and health risks, such as increased risk of blood clots and breast cancer in some cases.