Cartilage Tissue in Forensic Science—State of the Art and Future Research Directions

Round 1

Reviewer 1 Report

Please see the attached file.

Comments for author File: Comments.docx

Author Response

1. First of all, the Authors claim it is a review of the current literature; yet, there is no reference of it in the title, nor there is a method paragraph describing how the choices were made. If the Authors want to do a really good job, they should focus on a systematic review; if they do not want to do that, they should refrain from stating their work is a review.

Thank you for this suggestion. We added the phrase - "state of the art" to the title of our review. We also added a Methods section that describes the search method for the articles under review.

2. Moreover, although I could agree that cartilage tissue should be studied for its potential use in Forensic Science, I cannot see a discussion section where the Authors should underline limitations, costs, actual difficulties, etc. They should comment these issues, even at the end of each paragraph.

Thank you for this comment. In each chapter, we added information on cartilage use limitations for the purposes described. Each chapter was designed to demonstrate cartilage advantages and potential in forensics first. At the end of the respective chapter we added a paragraph about cartilage disadvantages - costs, requirements for specialized equipment, and issues for future researchers’ consideration.

3. As far as English is concerned, there are many typos and banalities, like (I do not have the lines numbered):

• Page 2: occurrence in tpatients aged 15-30
• Page 3: Compared to CT, MRI minimizes the body’s or material’s exposure to radiation (You don’t say! And what about radiation danger in dead people)
• Page 5: The study studies showed that changes

Thank you very much for noticing these mistakes. They are the remnants of review mode but should not have taken place in the manuscript presented for review, as they are indeed a major shortcoming. We rechecked the work in terms of language and removed all typos and linguistic errors. Regarding the phrase "Compared to CT, MRI minimizes the body's or material's exposure to radiation," the forensic MRI imaging method described by Ekizoglu clearly demonstrates minimizing the risk of radiation in the living. By mentioning this work, we meant the rare, casuistic cases where, for forensic purposes, it is necessary to identify living people who lost their sense of their own personality and identity due to the injuries. Indeed, the original phrasing might have been interpreted as nonsense, so we modified the phrase in the revised version.

The study has good potential, but it has to be structured in a more suitable form. 

Reviewer 2 Report

Dear Authors, 

This is an interesting paper exploring the applications of cartilage analysis in forensic science. I have some comments: 

- Introduction, the last two paragraphs described two different aims of the review and should be combined. With the first paragraph seems like you are going to describe one thing, and then, the first description is related to age estimation. Combine these two aims and include them in the order of appearance in the text. 

- The subsequent sections described applications, however, you included one called "microscopic analysis". This is a methodology, no application, please rename it with the specific application (murder weapons, cause of death). 

- Include a discussion. Although it is a review, you should discuss the applications, advantages and disadvantages,... 

- Please, review the abstract, it is a little bit disorganize. 

Thank you very much. 

Author Response

Dear Authors,

This is an interesting paper exploring the applications of cartilage analysis in forensic science. I have some comments:

1. Introduction, the last two paragraphs described two different aims of the review and should be combined. With the first paragraph seems like you are going to describe one thing, and then, the first description is related to age estimation. Combine these two aims and include them in the order of appearance in the text.

Thank you for pointing this out. We have changed the order of the paragraphs in the Introduction. We also added a Methods section describing the methodology behind the preparation for the presented review.

2. The subsequent sections described applications, however, you included one called "microscopic analysis". This is a methodology, no application, please rename it with the specific application (murder weapons, cause of death).

Thank you for this valuable comment. We changed the title accordingly.

3. Include a discussion. Although it is a review, you should discuss the applications, advantages and disadvantages

We have decided to add a final paragraph to each chapter describing cartilage use’s limitations. Moreover, we proposed the areas that need to be explored in future research to start routinely using cartilage for forensic purposes.

4. Please, review the abstract, it is a little bit disorganize.

As suggested, we improved the abstract for its structure and accessibility.

Thank you very much.

Reviewer 3 Report

This review is a comprehensive appraisal of work done on cartilage and its applications to forensic science. 

The authors are advocating for the use of cartilage based workflows as well as future research in this understudied field. I am in agreement with this approach. However, the slow degradation rate is situational and highly variable given the reality of case work admitted for forensic analysis. Remains are evaluated in different conditions and at various stages of decomposition. In situations of advanced decomposition or remains found in water, cartilage will decompose quickly thereby hindering findings at autopsy or axillary testing. Therefore, cartilage isn't as resilent to degradation as dental and skeletal matter. Degradation effects molecular assays as abundance and integrity of proteins and nucleic acids is inversely proportional to increasing PMI. This needs to be highlighted and emphasized in the manuscript. 

With respects to age prediction, subadult and adult age prediction has been covered. However, there is a paucity of information on age estimation for paediatric decedents (<12 years). Is this due to a lack of data or a focus on forensic identification of older individuals? As a precursor for endochondral ossification, surely correlations can be made between the appearance of primary ossification sites/ patterns of ossification associated with a region of the body (eg skull) and biological age. 

Please delineate between ossification and calcification in text. Please address jargon use and minor rephrasing in sentence construction. e.g. page 2 - "including those for fixing murder weapons". This sentence is confusing. Terminology: The use of the word "cadaver" relates to medical training. The use of the word " decedent" or "individual" is more appropriate for forensic remains. 

The review has touched on NGS applications. However, what about proteomics or mass spectrometry based technologies. Please review multidisciplinary approaches about cartilage development and degradation. Example: Bielajew et al.,(2022)Proteomic, mechanical, and biochemical characterization of cartilage development.Acta Biomaterialia,143,52-62.

There is a deficit of forensic based work on cartilage. However, fundamental research provides insight into the reliablity and validity of this tissue to the medicolegal setting. Please consider this when editing your manuscript. 

 

Author Response

This review is a comprehensive appraisal of work done on cartilage and its applications to forensic science.

1. The authors are advocating for the use of cartilage based workflows as well as future research in this understudied field. I am in agreement with this approach. However, the slow degradation rate is situational and highly variable given the reality of case work admitted for forensic analysis. Remains are evaluated in different conditions and at various stages of decomposition. In situations of advanced decomposition or remains found in water, cartilage will decompose quickly thereby hindering findings at autopsy or axillary testing. Therefore, cartilage isn't as resilent to degradation as dental and skeletal matter. Degradation effects molecular assays as abundance and integrity of proteins and nucleic acids is inversely proportional to increasing PMI. This needs to be highlighted and emphasized in the manuscript.

Thank you for this comment. Indeed, the text fails to highlight the limitations of cartilage and the fact that cartilage may not always be available for analysis. We have mentioned this in the chapter on cartilage genetic postmortem analysis. We believe that due to corpse decomposition, genetic analysis is the most challenging analysis performed on cartilage. Additionally, inspired by your comment, we have added information on other cartilage limitations in other chapters.

2. With respects to age prediction, subadult and adult age prediction has been covered. However, there is a paucity of information on age estimation for paediatric decedents (<12 years). Is this due to a lack of data or a focus on forensic identification of older individuals? As a precursor for endochondral ossification, surely correlations can be made between the appearance of primary ossification sites/ patterns of ossification associated with a region of the body (eg skull) and biological age.

Indeed, during our research, we noticed a glaring lack of studies on children and infants. We are aware that in this age group, assessing the degree of skeletal cartilage ossification would be an opportunity to increase the accuracy of age prediction methods for deceased children. Inspired by the comment, we added a paragraph on this subject to the age prediction chapter and expressed the need to investigate this area in the future.

3. Please delineate between ossification and calcification in text. Please address jargon use and minor rephrasing in sentence construction. e.g. page 2 - "including those for fixing murder weapons". This sentence is confusing. Terminology: The use of the word "cadaver" relates to medical training. The use of the word " decedent" or "individual" is more appropriate for forensic remains.

Thank you for these comments. We checked the work again in terms of language and style and corrected any phrases and statements that may confuse the reader. Additionally, we have modified the work in terms of terminology, paying attention to the term "cadaver" in places where its actual use is inappropriate. We also added a sentence in the text that distinguishes calcification from ossification.

4. The review has touched on NGS applications. However, what about proteomics or mass spectrometry based technologies. Please review multidisciplinary approaches about cartilage development and degradation. Example: Bielajew et al.,(2022)Proteomic, mechanical, and biochemical characterization of cartilage development.Acta Biomaterialia,143,52-62.

Thank you for this suggestion. We decided to add a section on the possibilities and examples of molecular methods applications, along with suggested areas to explore in the future. We did not want our review to contain too much speculation about the use of different methods in forensics but to be a collection of existing research strictly on forensics. The proposals for new methods’ applications were only suggestions for future researchers.

There is a deficit of forensic based work on cartilage. However, fundamental research provides insight into the reliablity and validity of this tissue to the medicolegal setting. Please consider this when editing your manuscript.

Thank you

 

Round 2

Reviewer 2 Report

Dear Authors, 

Thank you very much for addressing my comments. I only have one additional suggestion, please, substitute "age prediction" for "age estimation", as it is the correct term. 

Thank you very much. 

Author Response

Dear Reviewer,

Thank you for pointing this out. We have substituted the phrase "age precticion" with the correct term "age estimation" as marked in yellow in the manuscript (please see the attachment). Also, according to Editor’s suggestion, we changed the title of last section to Conclusions.

Thank you very much,

Authors

Author Response File: Author Response.docx