Antimicrobial Activity of Ethanolic Propolis Extracts from Tame (Arauca) on Oral Biofilm Co-Cultures

Oral diseases such as dental caries, stomatitis, and periodontitis are closely associated with biofilms that are resistant to conventional therapeutic approaches. Streptococcus sanguinis and Streptococcus mutans play a key role as primary and secondary colonizers of oral surfaces, respectively, and interact synergistically with other species, including Candida albicans, to promote the establishment and progression of infection.

To evaluate the antimicrobial activity of ethanolic extracts of propolis from Tame (Arauca) on biofilms formed in co-cultures from reference strains and co-cultures with clinical isolates of oral pathogens.

Propolis was collected from Apis mellifera hives placed in rural Tame (Arauca), located in the foothills of the Eastern Andes (Colombia). Ethanolic extracts of propolis (EEP) were prepared in a 0.07 g/mL concentration and biological characterization was performed on single and complex co-cultures of S. mutans (serotype c), S. sanguinis, and C. albicans using disc diffusion test, determination of MIC and BMC, growth curves and biofilm formation. The cell viability and metabolic activity of primary cell cultures derived from a dental pulp explant were evaluated using the MTT assay.

EEP exhibited higher inhibition zones than chlorhexidine against S. mutans and C. albicans and lower efficacy against S. sanguinis. Among the microorganisms evaluated, S. mutans showed the lowest MIC and BCM values, followed by C. albicans and S. sanguinis. Growth curves and biofilm formation assays revealed higher inhibition in co-cultures of reference strains (S. mutans + C. albicans), while multi-species cultures (S. mutans + S. sanguinis + C. albicans), or clinical strains (S. mutans clinical isolated + S. sanguinis + C. albicans), showed higher resistance. Cell viability assays revealed low cytotoxicity (<30%) in primary cell cultures.

EEPs exhibited antimicrobial activity against relevant oral pathogens, especially in simple co-cultures, supporting their potential as natural therapeutic alternatives. However, their efficacy decreases in the presence of clinical strains and complex co-cultures, highlighting the importance of considering these variables in the development of oral treatments.