Recent studies demonstrated that photoaging and skin damage were associated with longer wavelengths, in particular near infrared radiation. Studies have been conducted in order to determine the damage caused by infrared-A (IR-A) radiation, particularly the upregulation of MMP-1, using in vitro and/or in vivo investigation. For detection of MMP-1, authors used several IR simulators devices and determined increases in MMP-1 levels through gene expression, western blotting and immunohistochemistry techniques. Nevertheless, no information concern MMP-1 quantification using ELISA assay had been published. In this study, we validated the optimal incubation time for the release of MMP-1 in cultures supernatants by human dermal fibroblasts (HDF) stimulated with IR-A radiation. Our results demonstrated that irradiation with IR-A resulted in a significant upregulation of MMP-1 production 1.7-fold in relation to non-irradiated group only in incubation time of 72 hours after radiation. After 24 and 48 hour IR-A radiation no significant differences were observed when compared to control groups. In this work, we standardized the enough time of human fibroblasts incubation to produce MMP-1 after IR-A radiation. Conversely previous studies in the literature, where MMP-1 gene expression is observed after 24 hours from IR-A exposure, the production of this protease in supernatants in only detected after 72 hours incubation.