In this study, we developed Lava-Dermabiotics HDB1234 using Lactobacillus plantarum HDB1234 and lava seawater fermentation technology to enhance skin barrier. The use of lava seawater increased the purity and concentration of nucleotides in Lava-Dermabiotics HDB1234 by using nanodrop 2000 spectrophotometer. Also, when 5%(v/v) Lava-Dermabiotics HDB1234 was treated, cytotoxicity on normal human epidermal keratinocytes (NHEKs) and human fibroblast cells was not observed by using MTT assay. The mRNA expressions of claudin-1 (CLDN-1), serine palmitoyltransferase (SPT), and filaggrin (FLG) on Lava-Dermabiotics HDB1234 treated cells were dose-dependent higher than those of non-treated cells. Especially, the protein expression of FLG was also confirmed by using western blotting and immunocytochemistry staining. These results indicated that Lava-Dermabiotics HDB1234 could be used as a potential cosmetic ingredient for skin barrier enhancement through CLDN-1, SPT, and FLG expression related to the formation of skin barrier.