Amoxicillin resistance of bacteria isolated from dental implants with peri-implant diseases
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Background Peri-implant mucositis is a reversible inflammatory lesion of the mucosa surrounding a dental implant, caused by the accumulation of bacterial plaque and biofilm formation, without bone loss. If peri-implant mucositis is not addressed, it can progress to peri-implantitis, characterized by significant inflammation of the peri-implant mucosa accompanied by the loss of supporting bone. Clinical evidence suggests that the management of peri-implant infections consist of mechanical debridement of implant, surgical intervention, and the administration of antibiotics. However, limited information is available regarding antibiotic resistance in bacteria causing peri-implant diseases. Gap statement Peri-implant bacteria are resistant to amoxicillin, a β-lactam antibiotic, which is the first choice medicine to treat peri-implant infections. Aim Evaluate the resistance of bacteria isolated from dental implants with peri-implant diseases, to amoxicillin and clindamycin. Methodology Biofilms were recovered from implants with peri-implant mucositis (n=4) and peri-implantitis (n=8). Microorganisms were cultured under aerobic and anaerobic conditions. Antibiotic sensitivity was assessed using the disk diffusion susceptibility assay using amoxicillin at a concentration of 8 μg per disk and clindamycin at a concentration of 30 μg per disk. Results Cultivation of microorganisms revealed predominant facultative anaerobic bacteria. Isolates recovered from implants with peri-implant mucosistis showed resistance to amoxicillin (100%; 4/4). In comparison, 75% of bacteria isolated from implants with peri-implantitis were resistant to amoxicillin (6/8). All isolates from implants with peri-implant diseases (n=12) were sensitive to clindamycin. Conclusion These findings suggest that bacteria isolated from dental implants with peri-implant infections are resistant to amoxicillin, a β-lactam antibiotic, raising concerns about the potential dissemination of this resistance to other oral bacteria.
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The reviewers have highlighted minor concerns with the work presented. Please ensure that you address their comments.
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Comments to Author
In the manuscript entitled: "Amoxicillin resistance of bacteria isolated from dental implants with peri-implant diseases" the author aimed to assess whether the resistance of bacteria isolated from dental implants with peri-implant diseases, to amoxicillin and clindamycin. The author found that Cultivation of microorganisms revealed predominant 36 facultative anaerobic bacteria. Isolates recovered from implants with peri-implant mucosistis showed resistance to amoxicillin (100%; 4/4). In comparison, 75% of bacteria isolated from implants with peri-implantitis were resistant to amoxicillin (6/8). All isolates from implants with peri-implant diseases (n=12) were sensitive to clindamycin. These findings suggest that bacteria isolated from dental implants with peri-implant infections are resistant to …
Comments to Author
In the manuscript entitled: "Amoxicillin resistance of bacteria isolated from dental implants with peri-implant diseases" the author aimed to assess whether the resistance of bacteria isolated from dental implants with peri-implant diseases, to amoxicillin and clindamycin. The author found that Cultivation of microorganisms revealed predominant 36 facultative anaerobic bacteria. Isolates recovered from implants with peri-implant mucosistis showed resistance to amoxicillin (100%; 4/4). In comparison, 75% of bacteria isolated from implants with peri-implantitis were resistant to amoxicillin (6/8). All isolates from implants with peri-implant diseases (n=12) were sensitive to clindamycin. These findings suggest that bacteria isolated from dental implants with peri-implant infections are resistant to amoxicillin, a β-lactam antibiotic, raising concerns about the potential dissemination of this resistance to other oral bacteria. Major comments: In general, the idea and innovation of this study regards the analysis of the link between periodontal tissues in health and disease is interesting and novel because the role these aspects in medicine are validated but further studies on this topic could be an innovative issue in this field could be open a creative matter of debate in literature by adding new information. Moreover, there are few reports in the literature that studied this interesting topic with this kind of study design. The study was well conducted by the authors; However, there are some concerns to revise that are described below. The introduction section resumes the existing knowledge regarding the important factor linked with the relationship between peri implant tissues and inflammation. However, as the importance of the topic, the reviewer strongly recommends, before a further re-evaluation of the manuscript, to update the literature through read, discuss and cites in the references with great attention all of those recent interesting articles, that helps the authors to better introduce and discuss the relationship and impact of chlorexidine and oxidative stress on periodontal and peri implant tissues mediators: 1) doi: 10.1186/s12967-024-05933-x. PMID: 39736760; 2) doi: 10.1002/JPER.24-0422. PMID: 39549247 The authors should be better specified, at the end of the introduction section, the rationale of the study and the aim of the study. In the central section, should better clarify inclusions and exclusions criteria of the selected sample. Please better state the results obtained in the abstract. The discussion section appears well organized with the relevant paper that support the conclusions, even if the authors should better discuss the relationship regarding the by periodontitis in and risk of oxidative stress evolution in periodontitis patients linked with inflammation. The conclusion should reinforce in light of the discussions. In conclusion, I am sure that the authors are fine clinicians who achieve very nice results with their adopted protocol. However, this study, in my view does not in its current form satisfy a very high scientific requirement for publication in this journal and requests a revision before a futher re-evaluation of the manuscript. Minor Comments: Abstract: - Better formulate the abstract section by better describing the aim of the study Introduction: - Please refer to major comments Discussion - Please add a specific sentence that clarifies the results obtained in the first part of the discussion
Please rate the manuscript for methodological rigour
Satisfactory
Please rate the quality of the presentation and structure of the manuscript
Poor
To what extent are the conclusions supported by the data?
Partially support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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Comments to Author
Aspects to be Improved: Title: Include the type of study. The objective also indicates that clindamycin resistance was evaluated. Abstract: The abstract lacks cohesion in presenting the background, gap statement, and aim. Ensure smooth transitions between these sections. The methodology section in the abstract could benefit from greater clarity and conciseness, especially regarding sample size and specific experimental conditions. The results presentation in the abstract may need to explicitly state statistical relevance or comparisons where applicable. The conclusion should be more concise and directly tied to the study's objectives, avoiding overgeneralization or repetition. Keywords should accurately reflect the study's focus, including terms specific to antibiotic resistance and peri-implant …
Comments to Author
Aspects to be Improved: Title: Include the type of study. The objective also indicates that clindamycin resistance was evaluated. Abstract: The abstract lacks cohesion in presenting the background, gap statement, and aim. Ensure smooth transitions between these sections. The methodology section in the abstract could benefit from greater clarity and conciseness, especially regarding sample size and specific experimental conditions. The results presentation in the abstract may need to explicitly state statistical relevance or comparisons where applicable. The conclusion should be more concise and directly tied to the study's objectives, avoiding overgeneralization or repetition. Keywords should accurately reflect the study's focus, including terms specific to antibiotic resistance and peri-implant diseases. Make sure to include MeSH terms Introduction - The introduction does not provide substantial background information. There is a very important literature with a high level of evidence on this subject that was not considered. The information presented in these studies limits the novelty of the current study (10.3390/ijerph192315609; 10.3390/antibiotics10060698). MAJOR CONCERN - It could better establish the novelty and clinical relevance of the study. Consider re-evaluating how the research gap is framed to make it more compelling. - Ensure that all references cited are current and relevant, and avoid overloading the introduction with excessive historical data. - Clarify the link between the observed prevalence of antibiotic resistance and its clinical implications, emphasizing why this study is critical for advancing knowledge. - Ensure that technical terms are consistently defined and that the introduction remains accessible to a broad audience. - Consider reorganizing the introduction to avoid redundancy and ensure that the study's objectives are clearly stated at the end of the section. Methods Patient Selection - Specify inclusion/exclusion criteria beyond location and condition to improve transparency. Peri-implant Disease Diagnosis - Ensure diagnostic criteria are explicitly stated and consistent with referenced sources. - Clarify if these diagnostic criteria align with international guidelines such as the EFP/AAP consensus. Collection of Extracted Dental Implants and Peri-implant Bacteria Cultivation - Countertorque ratchet technique (CTRT): Briefly explain the procedure or include a reference for readers unfamiliar with this technique. - Clarify if the chlorhexidine gel and povidone-iodine disinfection steps were randomized or standardized across samples. - Confirm if anaerobic conditions were continuously monitored during sample recovery to ensure bacterial integrity. Antimicrobial Susceptibility Testing - Specify the size of the inhibition zones used to determine susceptibility or resistance. - Verify whether controls (positive and negative) were included to ensure assay validity. - Include details on replicates or sample sizes to account for variability in the disk diffusion method. Molecular Analysis - For better clarity, mention whether a specific protocol (e.g., modified Aljanabi's protocol) was used and elaborate on the PCR thermal cycling conditions if applicable. - Add a brief rationale for selecting the V1-V3 region for amplification—highlight its relevance in differentiating peri-implant pathogens. - Clarify how sequencing errors were minimized (e.g., repeated sequencing or quality control measures). - Include references for software tools (e.g., Geneious and GraphPad) and describe their role in sequence alignment or data analysis. - Line 107: Change "bleeding within 30 seconds after probing (BOP)" to "bleeding on probing (BOP) observed within 30 seconds" for precision. - Sample Size Justification: Explain how patients were determined as an adequate sample size, including any power analysis if performed. - Statistical Analysis: Mention statistical methods used to analyze antimicrobial susceptibility or molecular results. Results/Discussion - The study included only 12 biofilm samples, which is not sufficient to make generalized conclusions about antibiotic resistance patterns in peri-implant diseases. - Incomplete Patient Information: Except for the patient from whom M29 and P30 samples were collected, no clinical or demographic data were collected for other patients. This prevents the exploration of correlations between antibiotic resistance and variables such as age, gender, or comorbidities. - Few Antibiotics Tested: The study only tested the susceptibility of peri-implant bacteria to amoxicillin and clindamycin. A broader panel of antibiotics could have provided a more comprehensive view of resistance patterns. -Single Dose-Dependent Susceptibility: The study used fixed concentrations for susceptibility testing. Exploring a range of concentrations would offer a clearer understanding of the minimum inhibitory concentration (MIC) and resistance thresholds. - Cultivation Bias: The cultivation methods used may have favored the growth of certain bacterial species while excluding others, potentially overlooking important pathogens involved in peri-implant diseases. - Limited Identification Techniques: The study identified bacterial species using 16S rRNA sequencing, but this method alone may not capture strain-level differences or virulence factors that could influence disease progression and treatment response. - The study does not investigate the molecular mechanisms of amoxicillin resistance, such as the presence of β-lactamase genes or efflux pumps. Understanding these mechanisms would provide insights into the drivers of resistance and inform therapeutic strategies. - Questionable Relevance of In Vitro Findings: While in vitro susceptibility testing is useful, its direct applicability to clinical outcomes is uncertain. The study does not address how the resistance observed impacts the success rates of actual treatments for peri-implant diseases.
Please rate the manuscript for methodological rigour
Poor
Please rate the quality of the presentation and structure of the manuscript
Poor
To what extent are the conclusions supported by the data?
Partially support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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