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Study Protocol
Revised

A scoping review protocol to identify clinical signs, symptoms and biomarkers indicative of biofilm presence in chronic wounds

[version 2; peer review: 2 approved, 1 approved with reservations]
PUBLISHED 23 Nov 2021
Author details Author details
OPEN PEER REVIEW
REVIEWER STATUS

Abstract

Introduction: Wound healing is characterised by haemostatic, inflammatory, proliferative and remodelling phases. In the presence of comorbidities such as diabetes, healing can stall and chronic wounds may result. Infection is detrimental to these wounds and associated with poor outcomes. Wounds are contaminated with microbes and debris, and factors such as host resistance, bacterial virulence, species synergy and bioburden determine whether a wound will deteriorate to critically colonised/infected states. Biofilms are sessile microbial communities, exhibiting high-level antibiotic tolerance and resistance to host defences. Biofilm in critically colonised wounds can contribute to delayed healing. Little is known about clinical presentation and diagnosis of wound biofilms.
Objective: To identify from the literature clinical signs, symptoms and biomarkers that may indicate biofilm presence in chronic wounds.
Methods: This review will be guided by the Preferred Reporting Items for Systematic Reviews extension for Scoping Reviews (PRISMA-ScR), and the Joanna Briggs Institute Manual for Evidence Synthesis. Studies of any design in any language recruiting adult patients with  venous, diabetic, pressure or mixed arterial-venous ulcers and reporting data on clinical signs/symptoms of biofilm are eligible. Searches of Medline, Embase, CINAHL, Cochrane Central and BASE will be conducted from inception to present. Reference scanning and contact with content experts will be employed. Title/abstract screening and full text selection will be executed by two reviewers independently. Discrepancies will be resolved by discussion between reviewers or through third party intervention. Data will be extracted by a single reviewer and verified by a second. Clinical signs and symptoms data will be presented in terms of study design, setting and participant demographic data.
Discussion: Understanding biofilm impact on chronic wounds is inconsistent and based largely on in vitro research. This work will consolidate clinical signs, symptoms and biomarkers of biofilm in chronic wounds reported in the literature.

Keywords

Chronic wound, Wound healing, Infection, Biofilm, Clinical signs and symptoms

Revised Amendments from Version 1

One typographical error has been fixed in the inclusion criteria section:  "any cliinical any clinical signs, symptoms and/or biomarkers." has been changed to "any clinical signs, symptoms and/or biomarkers.

The sentence "Pay-per-view articles will not be included." has been removed from the exclusion criteria section.

The Medline search strategy was also updated.

See the authors' detailed response to the review by Karen Ousey
See the authors' detailed response to the review by Gregory Schultz

Introduction

Wound healing occurs via a complex sequence of events which, under normal circumstances, proceed in an orderly fashion through haemostatic, inflammatory, proliferative and remodelling/maturation phases to restore cutaneous integrity and barrier function. However, in the presence of complicating factors such as diabetes or chronic venous insufficiency, the healing process can break down and the wound becomes chronic, failing to heal in a timely manner1,2. There is a lack of consensus regarding the definition of chronic wounds and they have for example, been described as ‘wounds that have not proceeded through an orderly and timely reparation to produce anatomic and functional integrity after 3 months’, ‘wounds that lack a 20–40% reduction in size after 2–4 weeks of optimal treatment or when there is not complete healing after 6 weeks’, or simply as ‘wounds that fail to proceed through the normal phases of wound healing in an orderly and timely manner’2,3. Typically, these wounds include but are not limited to venous, diabetic and pressure aetiologies4. Chronic wounds of mixed aetiologies were estimated to have a pooled prevalence of 2.21 per 1000 population in a 2019 meta-analysis of three studies, while a second meta-analysis from the same year including nine studies estimated the pooled prevalence of chronic leg ulcers to be 1.5 per 1000 population5. Chronic wounds are burdensome to the individual in terms of finances and quality of life, and to healthcare systems. Unhealed diabetic foot ulcers (DFU), venous leg ulcers (VLU) and pressure ulcers (PU) cost the United Kingdom National Health Service (NHS) approximately £4 billion in 2017–2018, or an average of £6305 per patient6,7.

Infection commonly affects chronic wounds and is associated with poor clinical outcomes8. The risk of hospitalisation for patients with a DFU is increased by a factor of 50 while the risk of lower-extremity amputation is 150 times higher if their wounds become infected. Development of infection in chronic wounds is a complex process. Multiple factors including virulence of colonising organisms, synergy between multiple microbial species, bioburden and host resistance interact with each other and determine whether a wound will progress from a non-threatening, contaminated/colonized state through to critically colonized or infected states9.

Bacteria can manifest in wounds as a free-floating planktonic phenotype or as a sessile biofilm phenotype10. Biofilms occur when microbial cells organise themselves into aggregates or communities encased in a self-produced polymeric matrix which typically attach to surfaces. Biofilms exhibit high levels of tolerance to antimicrobial agents and host defences11. When they form in critically colonized chronic wounds, healing stalls and the wound remains stuck in the inflammatory phase1216.

In 2017, a panel of specialists, chosen for their expertise in chronic wounds and biofilms, for their scholarly activity and publication record, issued a consensus document17. The document aimed to clarify the role of biofilms in clinical practice, help clinicians to recognize biofilms in chronic non-healing wounds and optimise patient management. A modified Delphi process was used to achieve consensus on a series of statements formulated to address issues in ten areas relevant to management of non-healing chronic wounds. Five-point likert scales of agreement (1 = disagree strongly – 5 = agree strongly) and ranking (1 = not important – 5 = most important) were used to score the statements. There was strong agreement (mean 4.0, standard deviation [SD] 0.82) that specific clinical signs and symptoms should be used to confirm presence of biofilm in the absence of diagnostic bedside tests. Clinical features, such as a recurring gelatinous material on the wound edge, have been proposed as surrogate markers of wound biofilm but there was weak agreement (mean 3.6, SD 1.5) that the clinical signs and symptoms that could indicate the presence of biofilm. Little is known about presentation and diagnosis of wound biofilms and knowledge of their characteristics is limited18. Generally, biofilms are difficult to diagnose and currently no guidelines exist to help clinicians and microbiologists in diagnosis and treatment19.

In addition, little quantitative work has been done with respect to clinical signs and symptoms of biofilm in chronic wounds, especially in human patients, and existing published research is mainly observational rather than incorporating more rigorous study designs12,17,18,20,21.

For these reasons we suspect that a rigorous systematic review with a focused research question and strict criteria with respect to eligible study design may be too exclusive and fail to answer the research question.

A scoping review methodology will therefore be employed to identify any associated clinical signs and symptoms thought to determine the presence of biofilm in chronic wounds.

The research question for the study is: what clinical signs, symptoms and biomarkers are proposed within the literature to determine the presence of biofilm in chronic wounds?

Methods

The Preferred Reporting Items for Systematic Review and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) statement, and the Joanna Briggs Institute Manual for Evidence Synthesis will guide this work22,23.

Inclusion criteria

Eligible participants will be adults (18 years +). Eligible wound aetiologies will be venous leg ulcers (VLU), diabetic foot ulcers (DFU), pressure ulcers (PU) and/or mixed arterial/venous leg ulcers (MAVLU), treated in any setting.

Databases will be searched from inception to present without limits on language. Study designs including but not limited to systematic reviews, randomized controlled trials (RCTs), controlled clinical trials, cohort studies, case series, case reports, letters to the editor with relevant data and editorials will be included. These articles must report any clinical signs, symptoms and/or biomarkers, validated or otherwise, thought to be associated with the presence of biofilm in chronic wounds.

Exclusion criteria

Patients with wounds resulting from burns, malignant fungating wounds, wounds secondary to conditions such as rheumatoid arthritis or pyoderma gangrenosum are ineligible for this study.

Search strategy

A search strategy will be developed in Medline, reviewed according to Peer Review of Electronic Search Strategies (PRESS) guidelines24 and adapted for use in Embase, CINAHL, Cochrane Central and The Bielefeld Academic Search Engine (BASE).

Box 1. Search strategy for the Medline database

Medline

      1.      “wounds and injuries”/

      2.      Wound healing/

      3.      Skin ulcer/

      4.      Leg ulcer/

      5.      Varicose ulcer/

      6.      Foot ulcer/

      7.      Diabetic foot/

      8.      Diabetes Mellitus, Type2/co [Complications]

      9.      Diabetic neuropathies/

      10.      Peripheral nervous system diseases/

      11.      Peripheral arterial disease/

      12.      Pressure ulcer/

      13.      Wound infection/

      14.      Debridement/

      15.      Re-epithelialization/

      16.      (chronic adj3 ulcer*).tw.

      17.      (skin adj3 ulcer*).tw.

      18.      (vascular adj3 ulcer*).tw.

      19.      (varicose adj3 ulcer*).tw.

      20.      (venous adj3 ulcer*).tw.

      21.      (stasis adj3 ulcer*).tw.

      22.      (leg adj3 ulcer*).tw.

      23.      (foot adj3 ulcer*).tw.

      24.      (diabetic adj3 ulcer*).tw.

      25.      (neuropathic adj3 ulcer*).tw.

      26.      (isch?emic adj3 ulcer*).tw.

      27.      (neuro-isch?emic adj3 ulcer*).tw.

      28.      (neuroisch?emic adj3 ulcer*).tw.

      29.      (pressure adj3 ulcer*).tw.

      30.      (decubitus adj3 ulcer*).tw.

      31.      (arterial adj3 ulcer*).tw.

      32.      (mixed adj3 ulcer*).tw.

      33.      (care adj3 ulcer*).tw.

      34.      (heal* adj3 ulcer*).tw.

      35.      (nonheal* adj3 ulcer*).tw.

      36.      (non-heal* adj3 ulcer*).tw.

      37.      (“non heal*” adj3 ulcer*).tw.

      38.      (re-epitheliali* adj3 ulcer*).tw.

      39.      (reepitheliali* adj3 ulcer*).tw.

      40.      (surface adj3 ulcer*).tw.

      41.      (“lower extremit*” adj3 ulcer*).tw.

      42.      (lower-extremit* adj3 ulcer*).tw.

      43.      (debrid* adj3 ulcer*).tw.

      44.      (manag* adj3 ulcer*).tw.

      45.      (bed adj3 ulcer*).tw.

      46.      (“hard to heal” adj3 ulcer*).tw.

      47.      (hard-to-heal adj3 ulcer*).tw.

      48.      (infect* adj3 ulcer*).tw.

      49.      (chronic adj3 wound*).tw.

      50.      (skin adj3 wound*).tw.

      51.      (vascular adj3 wound*).tw.

      52.      (varicose adj3 wound*).tw.

      53.      (venous adj3 wound*).tw.

      54.      (stasis adj3 wound*).tw.

      55.      (leg adj3 wound*).tw.

      56.      (foot adj3 wound*).tw.

      57.      (diabetic adj3 wound*).tw.

      58.      (neuropathic adj3 wound*).tw.

      59.      (isch?emic adj3 wound*).tw.

      60.      (neuro-isch?emic adj3 wound*).tw.

      61.      (neuroisch?emic adj3 wound*).tw.

      62.      (pressure adj3 wound*).tw.

      63.      (decubitus adj3 wound*).tw.

      64.      (arterial adj3 wound*).tw.

      65.      (mixed adj3 wound*).tw.

      66.      (care adj3 wound*).tw.

      67.      (heal* adj3 wound*).tw.

      68.      (nonheal* adj3 wound*).tw.

      69.      (non-heal* adj3 wound*).tw.

      70.      (“non heal*” adj3 wound*).tw.

      71.      (re-epitheliali* adj3 wound*).tw.

      72.      (reepitheliali*adj3 wound*).tw.

      73.      (surface adj3 wound*).tw.

      74.      (“lower extremit*” adj3 wound*).tw.

      75.      (lower-extremit* adj3 wound*).tw.

      76.      (debrid* adj3 wound*).tw.

      77.      (manag* adj3 wound*).tw.

      78.      (bed* adj3 wound*).tw.

      79.      (“hard to heal” adj3 wound*).tw.

      80.      (hard-to-heal adj3 wound*).tw.

      81.      (infect* adj3 wound*).tw.

      82.      (diabetic adj3 foot).tw.

      83.      (diabetic adj3 feet).tw.

      84.      (pressure adj3 sore*).tw.

      85.      bedsore*.tw.

      86.      “bed sore*”.tw.

      87.      bed-sore*.tw.

      88.      or/1-87

      89.      (“mixed etiolog*” adj3 ulcer*).tw.

      90.      (“mixed aetiolog*” adj3 ulcer*).tw.

      91.      (“mixed etiolog*” adj3 wound*).tw.

      92.      (“mixed aetiolog*” adj3 wound*).tw.

      93.      or/89-92

      94.      88 or 93

      95.      Exp Biofilms/

      96.      biofilm*.tw.

      97.      “EPS matrix”.tw.

      98.      “EPS matrices”.tw.

      99.      “extracellular polymeric substance*”.tw.

      100.          or/95-99

      101.           94 AND 100

      102.           Exp Animals/ NOT (Humans/ and exp Animals/)

      103.           101 NOT 102

Reference scanning. Reference lists of included articles will be scanned to locate subsequent, potentially relevant articles.

Content experts and organisations. Content experts and relevant organisations will be consulted to obtain information about unpublished or ongoing studies and where applicable, to request access to known but unavailable sources of evidence.

Search results will be exported to EndNote X9TM for storage and to RAYYAN25 for screening against eligibility criteria.

Evidence screening and selection

Level 1 screening (title and abstract screening). Pairs of researchers will independently screen titles and abstracts for inclusion according to the pre-determined eligibility criteria. A single failed eligibility criterion will be considered sufficient to exclude a study from this review. Discrepancies will be resolved by discussion between researchers in a pair. In cases where disagreements cannot be resolved, a final decision on the discrepancy will be made by a third party.

The screening process will be pilot tested on a random sample of 50 titles and abstracts.

Level 2 screening (full text screening). Pairs of researchers will independently screen all located full text articles for inclusion into this review according to eligibility criteria. A single failed eligibility criterion will be considered sufficient to exclude a study from this review. Discrepancies will be resolved by discussion between researchers in a pair. In cases where disagreements cannot be resolved, a final decision on the discrepancy will be made by a third party.

This level of the screening process will be pilot tested on a random sample of 10 articles if available.

A chart of the screening and selection process detailing the flow of studies from the search to data extraction and including duplicate removal will be presented with the findings.

An appendix of excluded full -text articles will also be included along with reasons for exclusion.

Data extraction

A data extraction form will be developed a priori in Microsoft Excel (2016). The form will capture the following data:

  • - Study data including authorship, year of publication, article type/study design, country of origin, setting and study objective.

  • - Participant data including sample size, age, gender and wound aetiology.

  • - Study concept data i.e. reported clinical signs, symptoms and biomarkers of biofilm in chronic wounds.

Data will be extracted by a single researcher and verified by a second.

Critical appraisal/risk of bias assessment

The aim of this review is to collate a comprehensive list of signs, symptoms and biomarkers used to indicate presence of biofilm in chronic wounds regardless of their level of refinement. Relevant data may be found in articles that span the evidence hierarchy and range from systematic reviews to opinion/editorial articles21. For these reasons, risk of bias assessment of included articles will not be necessary.

Data analysis, summary and presentation

Extracted data will be tabulated. Concept data will be presented in terms of country of origin, setting, study design, and in terms of participant data i.e. sample size, wound aetiology, age and gender. Data will be analysed with SPSS statistical package version 26. Demographic data will be presented descriptively in terms of mean and standard deviation or median and range.

Discussion

Little work regarding biofilms’ impact on chronic wounds involving human subjects has been done and much of our clinical understanding is based on in vitro work. Clinician’s knowledge of research data and of the importance of biofilms in the management of non-healing chronic wounds is inconsistent17. This review will for the first time, consolidate those signs, symptoms and biomarkers of biofilm in chronic wounds reported in the literature into one document which may serve to open an avenue for future clinical research in this area.

Dissemination

The findings of this scoping review will be published in a peer-reviewed medical journal.

The research team will also regularly update and disseminate project findings to key stakeholders, research colleagues, patient representatives and knowledge users.

Study status

The review has not yet initiated.

Data availability

No data are associated with this article.

Comments on this article Comments (1)

Version 2
VERSION 2 PUBLISHED 23 Nov 2021
Revised
  • Author Response 29 Nov 2021
    John Ivory, School of Nursing & Midwifery, National University of Ireland, Galway, Galway, H91TK33, Ireland
    29 Nov 2021
    Author Response
    The literature search strives to maximize comprehensiveness of returns while simultaneously screening out irrelevant material. It provides the evidence base for a review thus making it a foundational aspect whose ... Continue reading
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how to cite this article
Ivory JD, Vellinga A, O'Gara J and Gethin G. A scoping review protocol to identify clinical signs, symptoms and biomarkers indicative of biofilm presence in chronic wounds [version 2; peer review: 2 approved, 1 approved with reservations]. HRB Open Res 2021, 4:71 (https://doi.org/10.12688/hrbopenres.13300.2)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Open Peer Review

Current Reviewer Status: ?
Key to Reviewer Statuses VIEW
ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions
Version 2
VERSION 2
PUBLISHED 23 Nov 2021
Revised
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31
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Reviewer Report 18 Feb 2022
Luc Teot, Department of Plastic and Reconstructive Surgery and Wound Healing, Montpellier University Hospital, Montpellier, France 
Approved with Reservations
VIEWS 31
The eligibility criteria no. 1 may avoid the inclusion of a group of articles where biofilm is not mentioned in the abstract, even if in the full text there is a part concerning biofilms. On the contrary "optimistic" colleagues pretending ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Teot L. Reviewer Report For: A scoping review protocol to identify clinical signs, symptoms and biomarkers indicative of biofilm presence in chronic wounds [version 2; peer review: 2 approved, 1 approved with reservations]. HRB Open Res 2021, 4:71 (https://doi.org/10.21956/hrbopenres.14586.r31438)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
Views
32
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Reviewer Report 15 Feb 2022
Gregory Schultz, Department of Obstetrics and Gynecology, College of Medicine, University of Florida, Gainesville, FL, USA 
Approved
VIEWS 32
The study proposal addresses an important issue for the field of chronic wound care, specifically, how can clinicians reliably assess if a chronic wound contains bacterial biofilm communities as part of the bacterial bioburden in the wound bed. The proposed ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Schultz G. Reviewer Report For: A scoping review protocol to identify clinical signs, symptoms and biomarkers indicative of biofilm presence in chronic wounds [version 2; peer review: 2 approved, 1 approved with reservations]. HRB Open Res 2021, 4:71 (https://doi.org/10.21956/hrbopenres.14586.r31436)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
  • Author Response 01 Mar 2022
    John Ivory, School of Nursing & Midwifery, National University of Ireland, Galway, Galway, H91TK33, Ireland
    01 Mar 2022
    Author Response
    Hi Dr. Schultz,

    Many thanks for taking the time to review this protocol and for providing such favourable feedback.

    Best regards,

    John.
    Competing Interests: N/A
COMMENTS ON THIS REPORT
  • Author Response 01 Mar 2022
    John Ivory, School of Nursing & Midwifery, National University of Ireland, Galway, Galway, H91TK33, Ireland
    01 Mar 2022
    Author Response
    Hi Dr. Schultz,

    Many thanks for taking the time to review this protocol and for providing such favourable feedback.

    Best regards,

    John.
    Competing Interests: N/A
Version 1
VERSION 1
PUBLISHED 08 Jul 2021
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Reviewer Report 14 Jul 2021
Karen Ousey, Institute of Skin Integrity and Infection Prevention, School of Human and Health Sciences, University of Huddersfield, Huddersfield, UK 
Approved
VIEWS 46
A clear well, written, and structured scoping review. The area of biofilms is relevant to clinical practice and will be of interest to the multi-disciplinary team. I am sure the outcome of the review will identify areas for future research. ... Continue reading
CITE
CITE
HOW TO CITE THIS REPORT
Ousey K. Reviewer Report For: A scoping review protocol to identify clinical signs, symptoms and biomarkers indicative of biofilm presence in chronic wounds [version 2; peer review: 2 approved, 1 approved with reservations]. HRB Open Res 2021, 4:71 (https://doi.org/10.21956/hrbopenres.14477.r29794)
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
  • Author Response 12 Aug 2021
    John Ivory, School of Nursing & Midwifery, National University of Ireland, Galway, Galway, H91TK33, Ireland
    12 Aug 2021
    Author Response
    Dear Professor Ousey,

    Many thanks for taking the time to review this protocol and for the valuable feedback.

    We will change our eligibility criteria to admit pay-per-view articles.
    Competing Interests: No competing interests were disclosed.
COMMENTS ON THIS REPORT
  • Author Response 12 Aug 2021
    John Ivory, School of Nursing & Midwifery, National University of Ireland, Galway, Galway, H91TK33, Ireland
    12 Aug 2021
    Author Response
    Dear Professor Ousey,

    Many thanks for taking the time to review this protocol and for the valuable feedback.

    We will change our eligibility criteria to admit pay-per-view articles.
    Competing Interests: No competing interests were disclosed.

Comments on this article Comments (1)

Version 2
VERSION 2 PUBLISHED 23 Nov 2021
Revised
  • Author Response 29 Nov 2021
    John Ivory, School of Nursing & Midwifery, National University of Ireland, Galway, Galway, H91TK33, Ireland
    29 Nov 2021
    Author Response
    The literature search strives to maximize comprehensiveness of returns while simultaneously screening out irrelevant material. It provides the evidence base for a review thus making it a foundational aspect whose ... Continue reading
Alongside their report, reviewers assign a status to the article:
Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested
Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.
Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions

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