To investigate the dynamics of a disinfection regimen using 1% trypsin and 1% proteinase K

in combination with 2% chlorhexidine (with or without ultrasonics) using a nutrient-stressed

endodontic multispecies model biofilm.
Methodology
Nutrient-stressed biofilms (Propionibacterium acnes, Staphylococcus epidermidis,
Actinomyces radicidentis, Streptococcus mitis and Enterococcus faecalis OMGS 3202) were
grown in prepared root canals of single-rooted teeth. The treatment groups included 1%
trypsin and 2% chlorhexidine (CHX), 1% proteinase K and 2% CHX (with and without
ultrasonics). 2% CHX was the positive control and untreated group, and sterile saline (with
and without ultrasonics) was the negative control. The biofilms were investigated using
confocal laser scanning microscopy (CLSM) with live/dead staining and quantitative
microbial culture.
Results
The trypsin and CHX group with ultrasonics was significantly more effective in reducing
viable counts and the substratum coverage than those of all other groups (P < 0.05). The
viable counts of the proteinase K and CHX group used with (4.26 0.58 log10 cfu mL1) or
without ultrasonics (5.05 1.36 log10 cfu mL1) were significantly reduced (P < 0.05) as
compared with the untreated control (7.67 0.84 log10 cfu mL1) and saline groups used with
(6.57 0.73 log10 cfu mL1) and without ultrasonics (6.74 0.10 log10 cfu mL1). The CHX
group was less effective in biofilm disruption compared to when used in combination with
trypsin and proteinase K.
Conclusion
The trypsin and CHX group with ultrasonics was significantly more effective at reducing
bacterial viable counts and disrupting biofilm.