Sonication-Assisted Decellularization of Waste Tilapia (Oreochromis niloticus) Heads for Extracellular Matrix Extraction

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The authors of this report have specifically looked at finding the most efficient method for decellularizing Tilapia fish heads as a means to enable this waste product's use in various biomass related applications. In order to do so, the authors explore methods using SDS and TX-100 with and without sonication.

While the methods portrayed, results generated and ensuing conclusions are valid, I would encourage the authors to engage the audience in greater detail about why only one concentration of decellularization agents were used (i.e., What about other decellularization agents? What about other concentrations of SDS and TX-100? What about other contact times)? While I understand that no study can be fully comprehensive, I would want to see why these specific concentrations were chosen in the first place, and what might happen when one uses different agents, ratios, or durations.

Simultaneously, in the discussion section, I would like to see acknowledgement that a full Design of Experiments would be required to accurately determine the best decellularization technique, and a listing of all the variables.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Title

Sonication-assisted decellularization of waste Tilapia (Oreochromis niloticus) heads for extracellular matrix extraction

 

Manuscript ID: biomass-3156607

After reviewing the manuscript critically, it is a manuscript that presents very important information for the utilization of tilapia waste.  In fact, the objective of the study was to evaluate the effects of sonication as a viable decellularization method for the extraction of ECM from tilapia heads, where, the findings obtained show the benefit of sonication in the decellularization of waste tilapia heads with the purpose that in the future the products obtained can be applied practically in the clinical, food and environmental fields.

This is a very well structured, organized and well thought out manuscript. It is an original, innovative and novel manuscript, with potential for publication in the prestigious journal Biomass, however, I consider that from my humble point of view that it is necessary to make some adjustments to the manuscript.

The abstract does not include a brief description of the materials and methods because the results are described immediately after the objective. The abstract does not explain what was done and how it was done. Also, it is necessary to include a conclusive statement related to the results obtained and not necessarily to their potential application.

The introduction could include statistical aspects of tilapia production worldwide and the amount of waste generated. This could contribute to emphasize the global importance of the study and not only in the Philippines. Also, it would be very interesting to include in the introduction the advantages of sonication in decellularization in general.

In the materials and methods section, the characteristics of the organisms used, especially those aspects related to their freshness, need to be more detailed. How would the degree of freshness of the organisms or their chemical composition affect the results of the present study? Were they live or dead organisms? If dead, how many days old were they and how were they stored?

It is necessary to indicate more details of the characteristics and conditions under which sonication was applied.

On line 138, what do you mean by room temperature?

Could table 1 be clarified or rearranged? Explain why the contact time was 5 or 10 min.

Could you explain in more detail the quantification of DNA?

Could you explain in more detail the differential scanning calorimetry determination.

In which analysis was the tukey test used? Superscripts to indicate significant differences between groups are not included.

The discussion of results section is based on few bibliographical references; I recommend increasing the number of references in order to obtain a more robust discussion based on available scientific information. The discussion of differential scanning calorimetry should be improved, and the changes in function of the protein should also be discussed.

It is necessary to reduce the conclusion based on the objective and results of your work. The conclusion itself represents half of the discussion. Please be more concrete in the conclusion.

In the bibliographic references, it is necessary to standardize the format in which it is reported, because there are some that are incomplete (10, 20, 23, 32) please check.

Comments on the Quality of English Language

The manuscript is well written in the English language. 

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The article presents an insightful and innovative approach to addressing the environmental concerns associated with tilapia farming in the Philippines. By exploring the potential of tilapia heads—a commonly discarded by-product—as a valuable source of extracellular matrix (ECM) and bioactive materials, the study effectively highlights the untapped potential of these waste materials.

Comments and questions for manuscript improvements follow below:

 40: Include enzymes derived from fish viscera as products that originate from fish by-products

https://doi.org/10.1016/j.bbrep.2016.08.003

https://doi.org/10.1016/j.bcab.2020.101584

 

1-What environmental concerns are associated with the underutilization of waste materials generated from tilapia farming in the Philippines?

2- How does the sonication method compare to non-sonicated methods in terms of DNA content reduction and protein retention during the decellularization process?

3-What were the key findings from the SDS-PAGE analysis regarding the structural integrity of collagen in sonicated versus non-sonicated samples?

4-What practical applications are suggested for the extracellular matrix (ECM) extracted from decellularized tilapia heads, and how might this contribute to sustainable environmental practices?

Comments on the Quality of English Language

The manuscript has appropriate language

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

Undoubtedly, the authors greatly improved the quality of the manuscript. In addition, all my suggestions indicated in the first revision were considered and included in the manuscript. Therefore, I don't have any additional comments or modifications for this manuscript.

Comments on the Quality of English Language

The manuscript is well written in the English language.