Introduction: Nigella sativa, also known as black seed, black cumin, and/or Habbatul Barakah, is a medicinal herbal plant. Thymoquinone (TQ), one of Nigella sativa's active ingredients, has been shown to be anticarcinogenic, anti-inflammatory, analgesic, and antimicrobial. Hydroxyapatite (HA) is an inorganic mineral with the apatite lattice structure (A10(BO4)6C2). The aim of the current study is to incorporate thymoquinone with blackcumin hydroxyapatite crystals to form a new nanoformulation and to find out its cytotoxicity and antimicrobial properties against dental pathogens. Materials and method: Streptococcus mutans, Staphylococcus aureus, Enterococcus faecalis, and Candida albicans were the species used. The antimicrobial efficacy of the nanoparticles was evaluated using the agar well diffusion method at different concentrations ranging from 25 micron litres to 100 micron litres. UV double beam spectrophotometers in the wavelength range of 250-750nm were used to characterise the synthesised nanoparticles. Results: The zone of inhibition against Staphylococcus mutans and Candida albicans was found to be highest at 100 L concentration. As the concentrations of new nano formulation solution increased, the mean zone of inhibition grew. Conclusion: The study showed that there is potential antimicrobial property of the new nano formulation against Streptococcus mutans and Staphylococcus aureus also there is antifungal activity against Candida albicans