In the present work, we report the novel natural transdermal otoliths/collagen/bacterial cellulose patch for osteoporosis treatment. This biomaterial is an osteoinductor, or be, stimulates the bone regeneration, enabling bigger migration of the cells for formation of the bone fabric. Otolith is a typical biomaterial that is composed of calcium carbonate and organic matrix. Otoliths are calcareous concrescences present in the inner ear of fishes. Since they are rich in minerals, they are considered essential to the bone mineralization process on a protein matrix (otolin). The objective in this study was to analyze the regeneration capacity of bone defects treated with otoliths network preparation. Collagen and nano-otoliths influences in bacterial cellulose was analyzed using transmission infrared spectroscopy (FTIR). In vivo analysis shows bone surface tissue with high regularity, higher osteoblast activity, and osteo-reabsorption activities areas. These results indicated that the transdermal permeation of otollith using this patch system was sufficient for the treatment of bone diseases. These findings indicate that our novel transdermal delivery system for otolith/collagen/bacterial cellulose is a promising approach to improve compliance and quality of life of patients in the treatment of bone diseases.