<p class="Abstract">Leaf juices as well as leaves of <em>Barleria prionitis</em> and <em>Barleria grandiflora</em> are being used by rural people across various regions of India in the treatment of oral ailments such as dental troubles, gum ailments, pyorrhoea, dental carries and mouth ulcers. Zone of inhibition and MIC values obtained for all the extracts suggest ethanolic extract of the herbs were more antimicrobial when compared to the aqueous extract. Results of biofilm suppression were found statistically significant (p&lt;0.05) when compared to control. Cytotoxicity was evaluated by MTT assay on human gingival fibroblast and human dermal fibroblast cell lines for ethanolic extract of the herbs. CTC<sub>50</sub> value was found to be more than 1,000 µg/mL for ethanolic extracts of both herbs. Chlorhexidine was found to be more cytotoxic with CTC<sub>50 </sub>value of 12.5–25 µg/mL. Ethanolic extract of <em>B. prionitis</em> and <em>B. grandiflora</em> found significantly cytotoxic (p&lt;0.05) in comparison with control.</p><p class="Abstract"><strong>Video Clip:</strong></p><p class="Abstract"><a href="https://youtube.com/v/DNByJJQT3RY">Cytotoxicity assay:</a> 18 min 45 sec</p><p> </p>