Methods: The experiment was carried out in Azra Naheed Center for Research and Development (ANCRD), Superior University, Lahore, Pakistan from September 2018 till May 2019. Biofilms and planktonic cells of C. albicans alone and in combination with streptococci were subjected to chlorhexidine, allium sativum and bakuchiol individually and to allium-bakuchiol combination. Kirby-Bauer test, antifungal susceptibility testing, CFU count and drug synergy assessment was done on planktonic cells. Dynamic biofilms were formed to mimic conditions similar to oral cavity and CFU was determined.
Results: MIC of all three agents was higher against mixed species when compared to single species planktonic cells and biofilm. Allium sativum and bakuchiol demonstrated synergistic effects. The decrease in CFU count and minimum biofilm reduction to salivary pellicle caused by allium sativum-bakuchiol was comparable to that of chlorhexidine.
Conclusion: Thus, allium sativum-bakuchiol combination demonstrated antimicrobial effects similar to chlorhexidine against planktonic cells and dynamic biofilm. It could serve as a possible natural, economical alternative to chlorhexidine mouthrinses usually recommended in dental clinics. However, in vivo studies are required to determine the correct dosage of these agents.
OBJECTIVES: Here, the efficacy of graphene oxide (GO), a carbon-based nanomaterial, was tested against the biofilms and intracellular S. aureus invitro. Following that, the mechanism for the intracellular antimicrobial activities and GO toxicities was elucidated.
METHODS: GO antibiofilm properties were evaluated based on the disruption of biofilm structure, and the intracellular antimicrobial activities were determined by the survival of S. aureus in infected bovine mammary cells following GO exposure. The mechanism for GO intracellular antimicrobial activities was investigated using endocytosis inhibitors. GO toxicity towards the host cells was assessed using a resazurin assay.
RESULTS: At 100 ug/mL, GO reduced between 30 and 70% of S. aureus biofilm mass, suggesting GO's ability to disrupt the biofilm structure. At 200 ug/mL, GO killed almost 80% of intracellular S. aureus, and the antimicrobial activities were inhibited when cells were pre-treated with cytochalasin D, suggesting GO intracellular antimicrobial activities were dependent on the actin-polymerization of the cell membrane. At
METHODOLOGY: Dentin blocks were sterilized and E. faecalis and C. albicans microbial colonies were counted for colony-forming-units against 2%k21, 2%CHX and Ca(OH)2 medicaments. Biofilm colonies after 7 days on dentin were analysed using confocal laser scanning microscopy with live/dead bacterial viability staining. TEM was done to study dentin collagen matrix. Dentin discs from 3rd day and 7th day well plate was used for Raman spectra and observed under fluorescent-microscope. Docking studies were carried out on MMP-2 S1 binding-domain with k21.
RESULTS: There was reduction of E. faecalis/C. albicans when k21, chlorhexidine and calcium hydroxide were used with highest percentage in 2%k21 treated specimens. 2%k21 showed dense and regular collagen network with intact cross-banding and decreased Raman intensity for 2%k21 on 3rd day. NaOCl + k21 showed least adherence, whereas saline groups showed highest adherence of E. faecalis and C. albicans to root-canal dentin. Alizarin red staining of hDPSCs revealed calcium deposition in all groups with significant difference seen amongst 2%k21 groups. MMP-2 ligand binding was seen accurately indicating possible target sites for k21 intervention.
CONCLUSION: 2%k21 can be considered as alternative intracanal medicament.