Clinical Study and Microbiological Analysis of Periodontopathogenic Microflora Analyzed among Children and Adolescents with Cardiovascular Diseases Compared to Group with Good General Status
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsIn the present study, 62 children and adolescents, of both sexes, who had gingival inflammation and poor oral hygiene (IP over 25%) were examined. The subjects were divided into two groups according to the presence or absence of cardiovascular diseases. Sulcus samples were collected for PCR determinations of selected periodontitis associated bacterial species (Aa, Pg, Td, Tf, Pi, Pm, Fn) before and three months after specialist treatment. An extensive periodontal examination was performed at both these timepoints.
General aspects:
The study consists of an interesting study population with the potential to provide valuable information about onset of periodontal disease. The focus on children with cardiovascular disease makes the study unique. However, below I´ll provide a number of suggestions that substantially can improve the present paper before publication. Suggest that “group A” and “group C”, as well as T1 and T2 should be replaced with full words, maybe before treatment, after treatment, coronary healthy, coronary healthy.
Specific points:
Line 13-36 Abstract needs to be extensively revised. Suggest that abbreviations (group A, C, T1, T2) and some general information like “. Subgingival microorganisms are influenced by local but also general factors: oral hygiene status, dental eruption, saliva, oral pH, immune system, hormonal changes, vicious habits, systemic conditions and possible medication” with more specific information. Which clinical parameters have been recorded and who are the bacterial species that been analyzed by PCR? Valuable information in an abstract that will be useful for complete understanding.
Line 51 Suggest that installation should be replaced with initiation.
Line 58 Move reference 3-6 up to line 52.
Line 61-65 Use the new diagnostic criteria for periodontal disease stage I-IV and grade 1-3 (https://aap.onlinelibrary.wiley.com/doi/full/10.1002/JPER.18-0006). Aggressive periodontitis is not a diagnose anymore!
Line 84-88 It should be of great interest to see the authors opinion about the importance of microorganisms for the onset of cardiovascular disease in children? Is the common link an impaired immune function? As an alternative, what is written today about this correlation by others.
Line 93 The study need a distinct aim at the end of the introduction.
Line 96-265 The materials and method section needs to be substantially improved.
Divide into sub-sections 1) study populations 2) clinical examinations 3) sampling for PCR-detection of selected bacterial species 4) Extraction of DNA 4)PCR- methods.
Moreover, a table of background demographics from the two study populations would be an important information in a revised manuscript. Sex, age (mean) and geographic background of the two cohorts could be of interest (in section 1 of Mat Met).
Many of the majority of the illustrations can be omitted or moved to the section for supplementary materials.
Line 282 Suggest that data from table 1-2 and fig 18-19 should be moved to the material and method section and presented together with other demographics.
Line 308 Suggest that the authors present the clinical parameters in table 4 in accordance to the diagnostic criteria that are used today (https://aap.onlinelibrary.wiley.com/doi/full/10.1002/JPER.18-0006)
Line 321-327 Figure 20 is fine, but need an improved design. Data in figure 21 and 22 could be presented in the same illustration.
Line 362 The table lack legend and the text in the table needs to be translated to English.
Line 365-369 Change th bacterial species name to italic.
Line 381 Figure legend 24 should be placed under the illustration. Remove heading inside the graph and write the complete words for NTG in the figure legend.
Line 382-385 Adjust y-axis scale in figure 22-26, and see comments above.
Line 402-403 Translate table head text to English. Here is also data for 9 bacterial species presented. Information provided in the materials and methods section say 7 different bacterial species. What is correct?
Line 404 Figure 27. What is represented for the brown and grey bars? Remove heading in graph and add missing information in the text. It should be nice to see this illustration for the two patient groups (A and C).
Line 428-429 Again, here is data for 9 bacterial species presented?
Line 267-429 In general, there are to many figures and tables in the result presentation. There is no doubt that there is an effect of treatment, and also a slight difference between A and C. However, data that look for correlations between prevalence of certain bacterial species and clinical parameters are missing.
Line 446 Write the bacterium in italic letters!
Line 441-466 Several statements in these sections needs addition of several references.
Author Response
Hello,
Thank you very much for your time for the article I sent you.
I have modified the article and taken into account your indications.
Reviewer1 I General aspects:
The study consists of an interesting study population with the potential to provide valuable information about onset of periodontal disease. The focus on children with cardiovascular disease makes the study unique. However, below I´ll provide a number of suggestions that substantially can improve the present paper before publication. Suggest that “group A” and “group C”, as well as T1 and T2 should be replaced with full words, maybe before treatment, after treatment, coronary healthy, coronary healthy.
We replaced the A and C group with the group with cardiovascular disease and the group without cardiovascular disease.
T1 and T2, we replaced at baseline and 3 months after treatment. However, these notations were kept for the statistics part, the tables and graphs are clearer with these abbreviations.
Reviewer1 Specific points:
Line 13-36 Abstract needs to be extensively revised. Suggest that abbreviations (group A, C, T1, T2) and some general information like “. Subgingival microorganisms are influenced by local but also general factors: oral hygiene status, dental eruption, saliva, oral pH, immune system, hormonal changes, vicious habits, systemic conditions and possible medication” with more specific information. Which clinical parameters have been recorded and who are the bacterial species that been analyzed by PCR? Valuable information in an abstract that will be useful for complete understanding.
I have completely rephrased the abstract and provided information related to clinical parameters and microorganisms analyzed.
Subjects were examined in the initial consultation, the state of hygiene and periodontal inflammation was assessed using plaque index(PI) and gingival index(GI), samples were taken from the gingival sulcus using sterile paper cones to determine nine subgingival microorganisms. Nine subgingival microorganisms were identified Aggregatibacter actinomycetemcomitans (Aa),Porphyromonas gingivalis (Pg),Treponema denticola (Td),Tannerella forsythias (Tf) Prevotella intermedia (Pi), Peptostreptococcus (Micromonas) micros (Pm), Fusobacterium nucleatum (Fn) ,Eubacterium nodatum(En), Capnocytophaga gingivalis( Cg).The patients were included in the specialist treatment programme which aimed to relieve the inflammatory condition, remove local irritative factors and train the patients to perform proper oral hygiene at home by using primary and secondary oral hygiene products. Subjects were re-evaluated 3 months after treatment, when measurements from PI and GI and microbiological determinations were repeated.
Reviewer1
Suggest that installation should be replaced with initiation.
I replaced
Reviewer1
Move reference 3-6 up to line 52.
I moved the references
Reviewer1
Use the new diagnostic criteria for periodontal disease stage I-IV and grade 1-3 (https://aap.onlinelibrary.wiley.com/doi/full/10.1002/JPER.18-0006). Aggressive periodontitis is not a diagnose anymore!
I have removed the diagnosis of aggressive periodontitis
Reviewer1
It should be of great interest to see the authors opinion about the importance of microorganisms for the onset of cardiovascular disease in children? Is the common link an impaired immune function? As an alternative, what is written today about this correlation by others.
There is some evidence for alterations in the oral microflora as a result of physiopathological and treatment-related factors in children with children with congenital heart disease, but additional research is required to validate these findings. [39,40].
Reviewer1
The study need a distinct aim at the end of the introduction.
The aim of this study was to detect the presence and quantify the nine periodontopathogenic bacterial strains using PCR assays. We performed the association of these microorganisms with local clinical parameters and the diagnosis of periodontal disease in a group of patients with cardiovascular disease compared to a control group of subjects without systemic pathology.
Reviewer1
The materials and method section needs to be substantially improved.
Divide into sub-sections 1) study populations 2) clinical examinations 3) sampling for PCR-detection of selected bacterial species 4) Extraction of DNA 4)PCR- methods.
I have modified as indicated
Reviewer1
Moreover, a table of background demographics from the two study populations would be an important information in a revised manuscript. Sex, age (mean) and geographic background of the two cohorts could be of interest (in section 1 of Mat Met).
Demographic data related to gender and age are presented.
Reviewer1
Many of the majority of the illustrations can be omitted or moved to the section for supplementary materials.
Pictures can be moved to the additional material section. If this makes it easier to follow the article
Reviewer1
Suggest that data from table 1-2 and fig 18-19 should be moved to the material and method section and presented together with other demographics.
The pictures are part of the clinical determinations and sample collection kit, we considered them part of the material and method
Reviewer1
Suggest that the authors present the clinical parameters in table 4 in accordance to the diagnostic criteria that are used today (https://aap.onlinelibrary.wiley.com/doi/full/10.1002/JPER.18-0006)
Patients who are children, show only gingival inflammation, damage to the superficial periodontium, not periodontal disease with damage to the deep periodontium, to be classified into stages and grades. We used the gingival index to diagnose localized or generalized gingival inflammation, mild, moderate, severe.
- Trombelli L, Farina R, Silva CO, Tatakis DN. Plaque-induced gingivitis: Case definition and diagnostic considerations. J. Clin. Periodontol. 2018;45:S44–S67. doi: 10.1111/jcpe.12939
Reviewer1
Figure 20 is fine, but need an improved design. Data in figure 21 and 22 could be presented in the same illustration.
I have modified
The table lack legend and the text in the table needs to be translated to English.
I have modified
Change th bacterial species name to italic.
I have modified
Figure legend 24 should be placed under the illustration. Remove heading inside the graph and write the complete words for NTG in the figure legend.
I have modified
Adjust y-axis scale in figure 22-26, and see comments above.
I have modified
Translate table head text to English. Here is also data for 9 bacterial species presented. Information provided in the materials and methods section say 7 different bacterial species. What is correct?
I have modified
Figure 27. What is represented for the brown and grey bars? Remove heading in graph and add missing information in the text. It should be nice to see this illustration for the two patient groups (A and C).
I have modified
Again, here is data for 9 bacterial species presented?
I have modified . There are nine microorganisms, two of which are also compatible with periodontal health, hence the confusion
In general, there are to many figures and tables in the result presentation. There is no doubt that there is an effect of treatment, and also a slight difference between A and C. However, data that look for correlations between prevalence of certain bacterial species and clinical parameters are missing.
I have modified
Write the bacterium in italic letters!
I have modified
Several statements in these sections needs addition of several references.
I have modified
Reviewer 2 Report
Comments and Suggestions for AuthorsThe manuscript has many flaws and needs significant modifications. Please see the enclosed PDF
Comments for author File: 
Comments.pdf
Should be checked by a specialist
Author Response
Hello,
Thank you very much for your time for the article I sent you.
I have modified the article and taken into account your indications.
We replaced the A and C group with the group with cardiovascular disease and the group without cardiovascular disease.
T1 and T2, we replaced at baseline and 3 months after treatment. However, these notations were kept for the statistics part, the tables and graphs are clearer with these abbreviations.
I have completely rephrased the abstract and provided information related to clinical parameters and microorganisms analyzed.
Subjects were examined in the initial consultation, the state of hygiene and periodontal inflammation was assessed using plaque index(PI) and gingival index(GI), samples were taken from the gingival sulcus using sterile paper cones to determine nine subgingival microorganisms. Nine subgingival microorganisms were identified Aggregatibacter actinomycetemcomitans (Aa),Porphyromonas gingivalis (Pg),Treponema denticola (Td),Tannerella forsythias (Tf) Prevotella intermedia (Pi), Peptostreptococcus (Micromonas) micros (Pm), Fusobacterium nucleatum (Fn) ,Eubacterium nodatum(En), Capnocytophaga gingivalis( Cg).The patients were included in the specialist treatment programme which aimed to relieve the inflammatory condition, remove local irritative factors and train the patients to perform proper oral hygiene at home by using primary and secondary oral hygiene products. Subjects were re-evaluated 3 months after treatment, when measurements from PI and GI and microbiological determinations were repeated.
We have made the recommended changes.
Thank you
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe revised version is substantially improved, however, I suggest that material and methods as well as reuslts should be devided with subheadings 2.1 2.2 2.3........... and 3.1 3.2 3.3........... in accordance to authors instructions. In addition, replace , with . as decimal in tables, and check out where the number of decimals could be reduced. Finally, suggest that A, A, T1 and T2 in legend for figures and tables should be replaced in accordance to the terminolgy used in the revised text.
Author Response
Hello,
Thank you for your thoughtful review
I have modified the article as indicated
The changes have been marked in red
Thank you
Reviewer 2 Report
Comments and Suggestions for AuthorsThe manuscript has been improved
Author Response
Hello,
Thank you for your thoughtful review
I have modified the article as indicated
The changes have been marked in red
Thank you
