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Objectives: The aim of this study was to evaluate the effect of in-vivo produced Nisin which is an antimicrobial peptide (AMP) added to adhesive resin on shear bond strength (SBS) and the adhesive remnant index (ARI) of orthodontic brackets., Methods: Bacterial AMP was produced by fermentation and the ideal AMP/Bond concentration and antimicrobial efficacy of the mixture were tested. To evaluate the SBS and ARI scores of AMP-added adhesive resins, 80 maxillary premolar teeth extracted for orthodontic purposes were used and randomly assigned into 2 groups (n = 40). Group 1: Control Group (teeth bonded with standard adhesive resin); Group 2: Experimental Group (teeth bonded with AMP-added adhesive resin). Statistical analysis was performed using the SPSS package program and applying the Mann-Whitney U and Fisher's exact tests. P < 0.05 was considered as statistically significant., Results: Nisin synthesized in-vivo from Lactococcus lactis (L. lactis) (ATCC 7962) bacteria was provided to form a homogenous solution at an ideal concentration To find the minimum AMP/Bond mixture ratio that showed maximum antimicrobial activity, AMP and Bond mixtures were tested at various concentration levels between 1/160 and 1/2 (AMP/Bond). As a result, the optimum ratio was determined as 1/40. The antimicrobial efficacy of Nisin-added adhesive resin was tested against Streptococcus mutans (S. mutans) (ATCC 35,688) and Lactobacillus strains (cariogenic microorganisms). AMP formed a 2.7 cm diameter zone alone, while 1/40 AMP-bond mixture formed a 1.2 cm diameter zone. SBS values of the teeth bonded with Nisin added adhesive (17.49 ± 5.31) were significantly higher than the control group (14.54 ± 4.96) (P = 0.004). According to the four point scale, Nisin added adhesive provided a higher ARI score in favour of the adhesive and tooth compared to the control group (ARI = 3, n = 20)., Conclusions: Nisin produced from L. lactis (ATCC 7962) had greater antimicrobial effects after mixing with adhesive bond against cariogenic microorganisms S. mutans (ATCC 35,688) and Lactobacillus strains. Nisin added adhesive increased shear bond strength (SBS) of orthodontic brackets and ARI scores in favor of adhesive & teeth., Clinical Relevance: Clinicians should take into account that using Nisin-added adhesive resin in orthodontic treatments can provide prophylaxis against tooth decay, especially in patients with poor oral hygiene., (© 2024. The Author(s).)