This study aims to validate the efficacy of a new device specifically developed for equine lamellar biopsy. Nine adult horses were used. Under sedation and digital nerve perineural anesthesia and after keratinized tissue thinning, a sample from the dorsal lamellar stratum was obtained using an instrument called Falcão-Faleiros' lamellotome. Hoof pain sensitivity was evaluated for 60 days, and horses were monitored for 6 months. Lateromedial radiographic images to analyze the spatial relationship between the distal phalanx and the hoof capsule were obtained before and 30 days after the biopsy. The effect of time on the variables was statistically analyzed (P < .05). On average (±SD), the biopsies produced samples that were 2.32 (±0.64) cm in length, 0.48 (±0.09) in width, and 0.51 (±0.11) cm in depth. A mean of 69 of intact primary epidermal lamellae was obtained per biopsy sample. Lameness and sensitivity to hoof testers were evident during the first 4 days postbiopsy but returned to basal levels after the fifth day. Minimal radiographic changes were observed, and the horses completely returned to their regular activities after 60 days. All biopsied hoofs grew normally during the 6-month period, making the dorsal wall defects reach the ground level. The use of the lamellotome for equine hoof lamellar biopsy produced an adequate quality and quantity of tissue for histology and allowed for full clinical recovery of the horses.