Comparison of Three Endodontic Irrigant Regimens against Dual-Species Interkingdom Biofilms: Considerations for Maintaining the Status Quo
, Saeed Alqahtani, Suror Shaban, Khawlah Albashaireh, William McLean and Gordon Ramage *Round 1
Reviewer 1 Report
This manuscript measures the effectiveness of three endodontic antimicrobial treatments against mono and dual species biofilms composed of Candida albicans and Enterococcus faecalis. The treatments consisted 3% bleach, 3% bleach followed by 17% EDTA, or 9% HEDP dissolved in 3% bleach. HEDP is a mild chelating agent that can be dissolved in 3% bleach, unlike EDTA, which must be used sequentially with bleach. Importantly, HEDP has never been evaluated in a dual species biofilm. Immediate kill and biofilm regrowth after 72 h was measured using qPCR.
All solutions containing bleach were immediately effective against mono- and co-cultured biofilms. However, only the sequential bleach/EDTA treatment suppressed 72 h regrowth to a significant extent. The use of a dual species fungal/bacterial biofilm makes this study relevant since these species are found together in teeth undergoing decay. This study supports the continued use of bleach/EDTA as a cleaning regimen in root canals.
The work is carefully performed, clearly reported and written. Statistics are used appropriately.
I have a few minor points requiring additional editing.
Line 27: State concentration of HEDP, i.e. “9% HEDP”, in abstract for consistency.
Line 28: “after 72 hr of regrowth”
Line 81: “Initially,”
line 82: define "PMIC" when first used. Also, “PMIC” It is sometimes spelled with a lower case "p" and sometimes an upper case "P". Be consistent.
Line 202: A good references for the sentence about EDTA disruption of OM is: https://www.jbc.org/content/243/9/2373.long
Figure 1, 3 and 4: In the key for each graph, the condition “NaOCl+HEDP” should be changed to “NaOCl:HEDP” to be consistent with terminology used in Table 3. Also, if possible, make the label fonts a bit larger in Figure 1.
Table 3 should be labeled Table 2 and vice versa to coincide with the order they appear in the paper.
Author Response
We thank the reviewer for their careful assessment of the manuscript. We have edited the manuscript accordingly as indicated below:
Line 27: “9% HEDP” was added instead of “HEDP”
Line 28: “after 72h of regrowth” was added instead of “after 72h”
Line 84: “Initially,” was added instead “initially”
Line 85: The full form “planktonic minimum inhibitory concentration” of PMIC was added as this line has the first use of this term. Also, all “pMIC” in the manuscript was changed to “PMIC” for consistency
Line 210: the reference (29) (https://www.jbc.org/content/243/9/2373.long) was added
In figures 1, 3 and 4, in the key for each graph, the condition NaOCl+HEDP was changed to NaOCl:HEDP for consistency
In Figure 1: the label fonts size was increased
Table 3 was labelled as Table 2 and vice versa to coincide with the order they appear in the paper.
Reviewer 2 Report
The paper is interesting and presented a comparison between irrigants for endodontic antimicrobial regimens.
The methods are clearly described and the article can be interesting for the readers
However, some clinical aspects of endodontic infections have to be described, in particular in the introduction, because they are linked to bone loss around teeth or bone defects, if treated in a wrong way, that can bring to the loss of teeth. In these cases biomaterials have to be applied and grafted. Moreover, a description on some anatomical features of roots have to be described, because sometimes this particular anatomy can lead to unsuccessfully therapy.
So I suggest to include the following papers in the introduction:
- PMID: 21841997
- PMID: 32905137
- PMID: 31215045
- PMID: 22010090
- PMID: 30833097
- PMID: 25069906
Author Response
We thank the reviewer for their comments and suggestions. As a group we have discussed the suggestions and looked at the papers in detail. While these are interesting, they are do not in our opinion fit directly within the narrative and logic of the introduction. However, we have taken on board the comments in general and added the following lines:
Line 65: A sentence explaining the complex anatomy of root canal system with appropriate reference was added in response to review report 2.
"The complex anatomy of root canal system has been well described by Vertucci (14). The presence of lateral canals, multiple foramina, deltas, isthmus and c-shaped canals are some forms of the complex anatomy that may renders root canal disinfection more challenging."
