Ultraviolet rays from sunlight are involved in vitamin D synthesis, which is essential in maintaining health. Most in vivo vitamin D synthesis occurs via exposure to sunlight, whereas a small portion is obtained through diet. Therefore, appropriate exposure to sunlight is recommended to satisfy vitamin D requirement. However, vitamin D deficiency still poses a serious issue to numerous people. To resolve this problem, artificial lighting has been studied to provide rays of UV light with characteristics similar to that of sunlight. The present study introduced vitamin D synthesis stimulating UVB-LED light via the general diffusion lighting type to indoor dwellers. However, despite the strides of advancement made in this type of lighting, studies on UVB with relation to vitamin D synthesis are limited. This study aimed to suggest UVB-LED lighting for indoor dwellers and confirm the effectiveness of vitamin D stimulation through animal testing. For this, a UVB-LED providing safe UVB dosage was developed based on the photobiological safety (IEC 62471) of lighting. Next, a comparison study of the suggested lighting and general lighting was executed in an isolated system. The study continued for two weeks and the vitamin D levels within the animal test subject (Rat, Wistar, male) were measured. At this time, the lighting was turned on for 06:00~22:00 considering the sunrise timing and average lighting time for modern people, and blood was collected from the caudal veins of the rats and analyzed for the amount of 25-hydroxyvitamin D [25(OH)D]. Analysis results showed that the amount of 25-hydroxyvitamin D [25(OH)D] in blood was higher, on average, by 6.8% (0.97 ng/mL) and 7.3% (0.9 ng/ml) in rats reared under UVB-LED lighting on the seventh and fourteenth experimental days, respectively, than those under general lighting. It was, therefore, confirmed that vitamin D synthesis in the human body can be supported by the proposed lighting in this study.