Research on photobiomodulation (PBM) has led to the development of various light-generating devices that can benefit a wide range of clinical indications. A novel approach of inducing PBM is through application of a Fluorescence Biomodulation (FB) System consisting of a blue light (peak wavelength between 440 and 460 nm) which activates topical photoconverter substrates containing specialized chromophores that generate fluorescence light energy (FLE). In clinical trials, FLE has been shown to modulate both healthy and disease-affected skin/soft tissue, providing a unique method for managing inflammatory skin conditions and accelerating healing. To better understand the biological impact of FB-induced FLE, we studied this system <i>in vitro</i> on dermal human fibroblasts (DHFs) and <i>in vivo</i> in canine deep pyoderma. In vitro data from stimulated DHFs exposed to an FB System showed a significant decrease in IL-6 production by 130.14% after 24 hr (p<0.001), compared to control groups. In canines with chronic deep pyoderma, the use of FB plus standard of care (SOC) treatment significantly reduced time to clinical resolution compared to controls that received SOC alone (p<0.001). Biopsies from lesional areas showed enhanced mitochondrial biogenesis in the FB lesions versus the SOC lesions, as supported by a significant increase in the number and size of mitochondria (89.31% and 90.15% respectively; p<0.0001). Significant modulation of inflammatory pathways, epithelialization, and angiogenesis were also demonstrated. These results support the use of FB Systems for skin conditions impacted by inflammation and offer a promising therapeutic solution to support its use in other medical conditions.