This study aims to assess the antimicrobial efficacy and impact on color stability of Thymus (T.) vulgaris solution compared to conventional disinfectants on maxillofacial silicones. Various solutions were evaluated, including T. vulgaris solutions at 5 and 10%, saline (control), chlorhexidine (4%), and soap water. The substrates were MDX4-4210 silicone elastomers, and the microorganisms tested were Candida (C.) albicans and Staphylococcus (S.) aureus. The viability of microorganisms was determined through an 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) reduction assay, and color stability was measured using a spectrophotometer with X-Rite Europe software. Statistical analysis was performed using the Kruskal-Wallis test, Mann-Whitney U post hoc test, and Wilcoxon Signed Rank test. Soap water demonstrated superior disinfectant action against both microorganisms, while T. vulgaris solutions at 5 and 10% exhibited comparable antimicrobial efficacy. Chlorhexidine and 10% T. vulgaris solution showed minimal color changes in the silicone material. In contrast, soap water and the 5% T. vulgaris solution resulted in clinically unacceptable color alterations. This study underscores the potential of T. vulgaris as an herbal disinfectant for combating microbial biofilms on maxillofacial silicones, particularly at concentrations of 5 and 10%. The importance of maintaining color stability is emphasized, with Chlorhexidine and the 10% T. vulgaris solution demonstrating effective preservation of esthetics. These findings suggest the viability of considering T. vulgaris as an alternative disinfectant in clinical settings for maxillofacial silicone prostheses. Maxillofacial silicones are vital in restoring aesthetic features for individuals with facial trauma, congenital deformities, or post-surgical interventions. Yet, biofilm-related infections jeopardize their durability and visual integrity. Clinically, T. vulgaris signifies a potential advance in prosthodontic care, offering valuable insights for improving antimicrobial performance and aesthetic durability in maxillofacial prostheses. How to cite this article: Peter M, Kanathila H, Bembalagi M, et al. An In Vitro Comparative Evaluation of Conventional and Novel Thymus vulgaris Derived Herbal Disinfectant Solutions against Pathogenic Biofilm on Maxillofacial Silicones and Its Impact on Color Stability. J Contemp Dent Pract 2023;24(12):967-973.