Ex vivo Antimicrobial Evaluation of Different Drugs Using RT-PCR Test

Abstract

Introduction: Various medicaments are used in conjugation with mechanical cleaning and shaping to render root canals free of microbes. The different medicaments used tend to react among themselves sometimes resulting in deleterious products. For this reason use of alternative medicine, therapy is advocated. Aim: To evaluate the antimicrobial extent of different homeopathic drugs in comparison to the Ayurvedic and Allopathic medicines. Materials and Methods: A biofilm model of 8 mm dentine discs was prepared from single-rooted teeth, which were inoculated with E. faecalis for 21 days. 100 samples were divided into six groups Group I: Saline (negative Control), Group II: 2% Chlorhexidine Gluconate, Group III: Propolis, Group IV: RHUS Glabra, Group V: Zincum Oxydatum. The infected biofilm was then treated with the medicaments for seven days followed by testing to obtain threshold cycle (Ct) values using the Real-time polymerase chain reaction technique. DNA isolation was carried out and amplification was done using 16S rRNA gene sequence-based nested species-specific primers. DNA denaturation was done using DNA Thermocycler and a threshold cycle was obtained using Real-time Polymerize chain reaction technique.  Results: The threshold cycle (Ct) values obtained were subjected to statistical analysis that revealed a significant difference between the groups and among all 2% Chlorhexidine Gluconate, being the highest (30.3460±0.02505) followed by Propolis (30.1365±0.02621) then RHUS Glabra (30.0865±0.02581) better than Zincum Oxydatum (29.8070±0.02319) and least being Saline (29.6380±0.02505). Conclusion: All the homeopathic medicines showed antimicrobial properties comparable to the allopathic and ayurvedic drugs. Among RHUS Glabra and Zincum Oxydatum, RHUS Glabra performed significantly better.