UV-induced skin damage is involved in ROS overproduction and the overexpression of matrix metalloproteinases (MMPs), which are inhibited by TIMPs (tissue inhibitor of neural stem cells (NSCs)). These proteins may be associated with skin regeneration through the activation of TIMP proteins, but there have been no reports of treatment of skin photodamage using NSCs and their secreted proteins TIMP-1 and TIMP-2. Here we investigated the photoprotective role of NSCs and their TIMP proteins for the inhibition of UVB-irradiation damage in fibroblasts in SKH-1 mice. SKH-1 hairless mice were divided into three groups (n = 4 per group): normal, treatment, and control groups. The latter two groups were dorsally exposed to UVB irradiation for 12 weeks. After UVB irradiation, treatments with NSC-CM and its secreted factors TIMP-1 and TIMP-2, markedly ameliorated the photodamage triggered by the increase in MMP expression and activity through ROS production, and the subsequent activation of the NF-kappa B pathway in UVB-irradiated fibroblasts and the treatment mouse group. In addition, the topical application of NSC-CM to mice in the treatment group after irradiation clearly inhibited the expression of gamma-H2AX, a DNA damage marker, through the activation of the DNA repair enzyme Rad50. These results demonstrate that NSC-CM or TIMPs proteins can ameliorate skin photodamage induced by UVB-irradiation in in vitro and in vivo systems. (C) 2019 Elsevier Inc. All rights reserved.