Periodontitis, one of the most common chronic inflammatory diseases, affects the quality of life. Osteogenesis makes an important role of the disease. There is a connection between hydrogen sulfide (H2S) and periodontitis, but according to the study has been published, the precise role of H2S in inflammation remains in doubt. The main reason of the lack of research is that H2S is an endogenous gasotransmitter, difficult to discern through testing. So, we synthesis a novel fluorescence probe which can detective H2S in vitro. By using the novel H2S fluorescence probe, we found that H2S changes in osteoblasts mainly by cystathionine-γ-lyase, and H2S increases under LPS stimulation. H2S could be a potential marker for diagnosis of inflammatory diseases of bone, and might help deeper studies of the changes of H2S level and promote the progression on the researches about pathogenesis of periodontitis.