Facial packs or masks are popular beauty treatments that are thought to improve skin quality. We formulated a yoghurt pack using natural ingredients (F-YOP), with consideration of skin affinity, safety, health, and beauty. Then, we performed an in vitro assessment of biological activity and in vivo assessments of moisture, TEWL, melanin content, and elasticity. Facial areas treated with F-YOP showed increased moisture compared to control regions: 89±6.26% (forehead), 140.72±10.19% (cheek), and 123.29±6.67% (chin). Transepidermal water loss (TEWL) values were decreased in the treated areas compared to control: 101.38±6.95% (forehead), 50.37±5.93% (cheek), and l57.81±10.88% (chin). Elasticity was decreased in the control region, whereas the treatment region did not change. The initial elasticity was maintained in the cheek. F-YOP exhibited activity on DPPH radical scavenging, SOD-like activity, and lipoxygenase activity. F-YOP treatment successfully improved the moisture, brightness, and elasticity of treated skin.