Wound healing is a multifaceted process that can be interrupted, resulting in chronic wounds and infections. It is essential to develop treatments that are not only effective and maintain cellular bioactivity but are also cost-effective and easy to store. The amniotic membrane (AM) stands out for its high biocompatibility and abundance of bioactive factors, which makes it a promising option in regenerative medicine. This study investigates the efficacy of fresh bovine AM in promoting the healing of full-thickness skin wounds in dogs. Bovine AM was collected, washed, and sterilized. In vivo, two full-thickness skin wounds were created on the backs of 8 dogs and treated with saline (control) or fresh AM. Wound healing was monitored over 3 and 5 weeks, and tissue samples were collected for histological examination. Grossly, wounds treated with fresh AM healed faster than those in control wounds. Histologically, the fresh AM group exhibited accelerated healing compared to the control, with signs of angiogenesis, collagen fiber remodeling, resolution of inflammation, and re-epithelialization. Additionally, hair follicles and their associated glands were observed only in the fresh AM-treated wounds. In conclusion, the application of fresh bovine amniotic membrane greatly promotes wound healing, making it a practical and beneficial choice for managing full-thickness skin wounds.