We aimed to evaluate the degradation and antimicrobial activity of novel chlorhexidine-containing gelatin nanofabric devices and their cytotoxicity in animals. The electro-spun device with a size of 3×4×0.4 mm 3 was prepared by entrapping chlorhexidine gluconate (CHX) in a gelatin matrix. The devices were divided into three groups based on the CHX percentage (G1: 46%, G2: 50%, and G3: 54%), and the commercial product, PerioChip (CP), was used for the control group. We used an in vitro test for the degradation and antibacterial activity and investigated cytotoxicity using an in vivo test. Artificial saliva was used for the degradation test of chips and blood agar plates seeded with the oral bacteria Streptococcus sanguinis to address antibacterial activity. Furthermore, the devices were inserted between the skin and muscle of rat abdomens to evaluate infection and inflammatory reactions. We detected no obvious differences in the antibacterial or degradation test results between the test and control groups. In the histological analysis, the scaffold without CHX showed no signs of infection or inflammatory reaction. Both CHX-containing groups exhibited inflammation and necrosis in the muscle and skin, although the reaction in the test group was milder. In the degradation tests, antibacterial tests, and cytotoxic reactions in animal experiments, the electro-spun gelatin-based device showed similar results to those of the commercial device in the control group. Further studies on the quantitative analysis and clinical outcome evaluation of the electro-spun gelatin-based device in humans are necessary.