The purposes of this study were to verify the skin safety of the fermented broth of Lactobacillus rhamnosus by means of clinical trial, and to secure its applicability as an ingredient of cosmetics for recovering skin barrier as well as anti-aging. To clinical trial, the fermented broth of L. rhamnosus with different substrates, namely fermented MRS broth (MRS), fermented milk broth (FM), fermented milk-cream broth (FMC) and distilled water (C) were made into the water base of cosmetics for different experimental groups, And applied on the skin of middle-aged women for 7 weeks. Thereafter, a survey was conducted on actual skin irritation. Also, any other resulting changes in the neck skin were observed before and after clinical trial. The results showed that the fermented milk-cream broth of L. rhamnosus did not induce skin itchiness, dryness, redness, allergic reaction and eye-stimulation. Furthermore, the fermented broth of L. rhamnosus resulted in different changes on the neck skin, depending on their respective substrates. In particular, the FMC group obtained the following statistically significant results: increases in moisture contents (p<.001) and sebum contents (p<.05), decreases in TEWL (p<.05), and skin roughness (p<.01), increases in skin elasticity (p<.05), as well as a drastic drop (4.92%) in the melanin index compared to the control group. Thus, the fermented milk-cream broth of L. rhamnosus is expected to be safe as an ingredient of cosmetics for skin. In addition, the fermented milk-cream broth of L. rhamnosus was proven as an applicable ingredient of cosmetics that recovers damaged skin barriers and improves the neck skin of middle-aged women.