The paper aims to study the impact of ozone (O3) treatment on the microstructural changes of the tooth enamel after the treatment at different time intervals. The ozonation was performed with gaseous O3 produced by HealOzone X4, the demineralization level was measured with the DiagnoDent Pen 2190 device, and the microstructure changes of enamel surface were observed using scanning electron microscopy (SEM) analysis. The results showed the exposure to O3 for 40-50 seconds enhanced enamel micro-hardness and ensures a rate of remineralization between 96.82-97.38%. In conclusion, in search of new minimally invasive solutions in the treatment of caries and to offer antimicrobial support of the oral cavity, the use of O3 as an alternative therapy to classical solutions may be a viable solution in dentistry.