Objectives:
Padma hepaten (PH) is a polyphenolic formula derived from traditional Tibetan medicine. It is composed of Chebulic myrobalan, amla fruit and belleric myrobalan.
Natural polyphenols were found to have the capacity of biofilm inhibition.
Here we examine whether PH can inhibit the caries-inducing properties of S. mutans and to explain its mechanism of action.
Methods:
Dental biofilm of S. mutans UA159 was grown in the presence of different concentrations of PH, three different wells for each concentration. Formation of biofilms was tested spectrophotometrically after crystal violet staining and by CSLM.
Biofilms morphology was demonstrated using a high resolution SEM magnification. Exopolysaccharides (EPS) secretion was verified and quantified using a computerized analysis of CSLM. Gene expression of selected caries-related genes was tested using qRT-PCR. The biofilm was also tested on orthodontic PVC surface (Essix) to assess biofilm inhibition ability on orthodontic appliances.
Results:
PH presentation had a dose-dependent inhibition on biofilm formation, with no inhibition on planktonic bacterial growth. The minimal biofilm inhibitory concentration needed to inhibit the formation by 50% (MBIC50) was found to be 0.16 mg/ml. EPS secretion of S. mutans grown in the presence of 0.16 mg/ml PH was inhibited by 88%.
This reduction in the bacterial count and EPS secretion was also demonstrated using SEM.
A strong reduction was found in the expression of the gtfB, gtfC, ftf and brpA caries associated genes.
Conclusions:
Natural PH polyphenolic extraction has a significant inhibitory effect on S. mutans biofilm formation. In an era of an ongoing-increase in bacterial resistance to antibiotics, the use of natural substances, which inhibit biofilm development in a specific manner, without killing of cells, is a novel therapeutic concept. This application is most suitable for children as orthodontic patients, it can slow the ever growing bacterial resistance to antibiotics and can shed light on new biofilm inhibition mechanisms.