Introduction: The purpose of this study is to investigate and compare the effects of the antimicrobial agents Moringa oleifera and bioactive glass nanoparticles activated by femtosecond laser light on the biomimetic dentin remineralization using teeth having carious dentin ICDAS code 3. Methods and Materials: A total of 27 dentin surface samples were divided into three groups: the first group was treated with a Moringa oleifera extract, while the second group was treated with bioactive glass nanoparticles, and as for the control group, the third group received no additional agent. All groups were subjected to femtosecond laser light at three different wavelengths: 390 nm, 445 nm, and 780 nm. The photoactivation of each sample was achieved using the femtosecond laser light for 5 min with an average power rating of 300 mW, a pulse duration of 100 fs, and a pulse repetition rate of 80 Hz. The mineral content of the samples was obtained and analyzed using the laser-induced breakdown spectroscopy (LIBS). The LIBS analysis was conducted with the following laser light parameters: average power of ~215 mW, wavelength of 532 nm, pulse duration of 10 ns, and a pulse repetition rate of 10 Hz. Results: Most studied samples exhibited a relative increase in the mineral content that may enhance biomimetic remineralization. Moringa oleifera photoactivated by femtosecond laser light at 445 nm achieved a significant increase in mineral content. Conclusion: Using the femtosecond laser light to activate the relatively cheap and commercially available antimicrobial agent Moringa oleifera supports the strategy of minimal invasive approaches for the treatment and biomimetic remineralization of carious dentin ICDAS code 3.