There has been an increased demand for dental bleaching globally irrespective of age and gender. Main drawbacks associated with conventional tooth bleaching agents have been compromised strength and mineral-content of tooth enamel which results in sensitivity, discomfort, roughness, and structure loss of human teeth. Currently, nanoparticles synthesized by green synthesis have gained popularity especially in medical and dental applications because of their versatile and beneficial nano-scaled features. Titanium dioxide nanoparticles (TiO2Nps) in this study were prepared from green ecofriendly source using the aloe vera plant extract and were then characterized via dynamic light scattering (DLS), scanning electron microscope (SEM), X-ray diffraction spectroscopy (XRD), and energy dispersive X-ray (EDX), for size, shape, composition and true-phase. These TiO2 Nps were incorporated in commercial bleaching gel containing hydrogen peroxide to form a novel TiO2-bleaching gel which was used to bleach extracted anterior teeth belonging to four different age groups: 20–29 years, 30–39 years, 40–49 years and ≥50 years. These teeth were investigated for micro-hardness (Vickers microhardness tester) and mineral-content (EDX spectroscopy) including sodium, magnesium, phosphorus, calcium in an in-vitro environment both before and after bleaching. Results revealed that TiO2 Nps prepared by aloe vera plant were nanos-sized of about 37.91–49 nm, spherical shape, true anatase phase with pure titanium and oxygen in their composition. The values of Vickers micro-hardness and mineral-content (Na, Mg, P, Ca) of enamel specimens belonging to different age groups enhanced in a linear pattern before bleaching with the increase in age (p value < 0.05). There was negligible reduction observed in Vickers micro-hardness and mineral-content elements (Na, Mg, P, Ca) of all enamel specimens belonging to different ages after the bleaching (p value > 0.05). The novel TiO2-bleaching gel prepared was effective enough in preventing the declination in Vickers micro-hardness strength and mineral-content of all the enamel specimens belonging to different age groups even after the bleaching procedure which makes it a promising biomaterial.