Spider Silk-Inspired SVX Biopolymer: A Novel Haircare Technology for Superior Color Retention, Heat Protection, and Hydration

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The author has performed a novel study in terms of hair related projects “Spider Silk-Inspired SVX Biopolymer: A Novel Haircare Technology for Superior Color Retention, Heat Protection, and Hydration”. This study could be beneficial for many researchers. However, further suggestion may enhance this works more and strengthen the paper:

1. Consider specifying quantitative improvements (e.g., “Eab reduced from X to Y” for color retention) in the abstract for higher impact.
2. The introduction could briefly reference known limitations or open questions in the current literature about protein-based hair protectants for greater balance.
3. The novelty compared with other recombinant silk fibroins or established bond-building agents (such as bis-aminopropyl diglycol dimaleate) should be more clearly delineated.
4. Details regarding the nature, provenance, and batch uniformity of hair tresses require elaboration, including donor variability and storage conditions.
5. Blind or randomized assignment of treatments to hair bundles is not stated and could inadvertently introduce bias.
6. Statistical significance testing is mentioned, but precise p-value reporting is inconsistent, and there is limited discussion of effect sizes or biological relevance alongside statistical significance.
7. Clearly state whether tests were one or two-tailed, confirm normality assumptions, and detail any corrections for multiple comparisons.
8. Most endpoints are in vitro or ex vivo; interpretation should be cautious regarding in-use, real-world consumer outcomes.
9. Claims about sensory properties (“silky,” “touchably smooth”) should be toned down or supported with formal consumer/user studies.
10. Consider including raw data (e.g., stress–strain curves, full colorimetric datasets) as supplementary material for reproducibility and evaluation.
11. The benchmark products are not clearly identified (brand, active ingredients). Without transparency on comparator choice, it is difficult to assess the relative performance of SVX.
12. The “serum without SVX” serves as a useful control, but additional relevant comparators (e.g., hydrolyzed keratin, collagen peptides) would place SVX in a broader performance context.
13. SEM images are described qualitatively, but no quantitative image analysis (e.g., porosity, cuticle thickness measurements) is provided. Reviewer suggests including such metrics to substantiate the visual claims.
14. The proposed mechanisms (film formation, hydrogen bonding, controlled release of lactic acid) are plausible but remain speculative. Direct biochemical or spectroscopic evidence (beyond FTIR confirmation of formulation) should be provided to support these mechanistic assertions. For example, surface binding affinity assays, protein–keratin interaction studies, or thermal stability curves of SVX under hair-relevant conditions would strengthen the mechanistic discussion.
15. Discuss the potential for long-term SVX buildup, interaction with common conditioners, or removal by repeated washing.
16. Outline next steps for in vivo validation, long-term consumer studies, and safety assessments in real-use scenarios.
17. The discussion could benefit from a schematic figure summarizing the proposed SVX mechanism and protection model.
18. Novelty is adequately established, but further clarity on what SVX uniquely offers compared to other protein polymers (e.g., silk fibroin, hydrolyzed keratin) would strengthen claims of innovation.
19. Clearly specify which datasets/methods are proprietary and which are available, and detail the avenue for requesting data if applicable.

Comments on the Quality of English Language

Please check word selection and connectivity between the sentences.

Author Response

Dear Reviewer,
We sincerely thank you for your valuable time, constructive comments, and professional insights. Your detailed observations and suggestions were extremely helpful in improving the quality, clarity, and scientific depth of our manuscript. We carefully addressed all the points raised, and we truly appreciate your contribution to strengthening this work.

1. Consider specifying quantitative improvements (e.g., “Eab reduced from X to Y” for color retention) in the abstract for higher impact. We completely agree. Numerical data were added to the abstract.

2 . The introduction could briefly reference known limitations or open questions in the current literature about protein-based hair protectants for greater balance. Absolutely, additional information was added to the introduction (lines 67–72).

3. The novelty compared with other recombinant silk fibroins or established bond-building agents (such as bis-aminopropyl diglycol dimaleate) should be more clearly delineated. Important point. Additional clarification was added to the Methods section specifying that SVX is a recombinant spider silk–inspired protein, distinct from fibroin or sericin.
4. Details regarding the nature, provenance, and batch uniformity of hair tresses require elaboration, including donor variability and storage conditions. Yes, we agree. A full paragraph was added to the Hair Samples section.
5. Blind or randomized assignment of treatments to hair bundles is not stated and could inadvertently introduce bias. An additional explanation was added to the Methods section.
6. Statistical significance testing is mentioned, but precise p-value reporting is inconsistent, and there is limited discussion of effect sizes or biological relevance alongside statistical significance. Addressed together with point 7.
7. Clearly state whether tests were one or two-tailed, confirm normality assumptions, and detail any corrections for multiple comparisons. We agree. Additional details were added to the Statistical Analysis section of Methods and further discussed in the Results and Discussion section..
8. Most endpoints are in vitro or ex vivo; interpretation should be cautious regarding in-use, real-world consumer outcomes. We expanded the Discussion to address this limitation.
9. Claims about sensory properties (“silky,” “touchably smooth”) should be toned down or supported with formal consumer/user studies. Well noted. We removed claims that are not scientifically substantiated.
10. Consider including raw data (e.g., stress–strain curves, full colorimetric datasets) as supplementary material for reproducibility and evaluation. Since all relevant data are already presented within the manuscript, we prefer not to include supplementary files.
11. The benchmark products are not clearly identified (brand, active ingredients). Without transparency on comparator choice, it is difficult to assess the relative performance of SVX. We understand this point. The comparison was performed using a multifunctional serum (Kerastase Chronologiste Thermique Régénérant), though we prefer not to disclose the brand name to avoid reputational concerns.
12. The “serum without SVX” serves as a useful control, but additional relevant comparators (e.g., hydrolyzed keratin, collagen peptides) would place SVX in a broader performance context. Same clarification as above
13. SEM images are described qualitatively, but no quantitative image analysis (e.g., porosity, cuticle thickness measurements) is provided. Reviewer suggests including such metrics to substantiate the visual claims. We understand this point. In this manuscript, SEM images are presented only for visual interpretation. To obtain statistically robust results, a much larger dataset would be required.
14. The proposed mechanisms (film formation, hydrogen bonding, controlled release of lactic acid) are plausible but remain speculative. Direct biochemical or spectroscopic evidence (beyond FTIR confirmation of formulation) should be provided to support these mechanistic assertions. For example, surface binding affinity assays, protein–keratin interaction studies, or thermal stability curves of SVX under hair-relevant conditions would strengthen the mechanistic discussion. Very important point. We have recently published a manuscript dedicated to the complex formation, hydrogen bonding interactions, and controlled release mechanisms of SVX. This reference was added to the current manuscript.
15. Discuss the potential for long-term SVX buildup, interaction with common conditioners, or removal by repeated washing. Well noted. This point was added to the Discussion section.
16. Outline next steps for in vivo validation, long-term consumer studies, and safety assessments in real-use scenarios. Additional information was added to the Discussion
17. The discussion could benefit from a schematic figure summarizing the proposed SVX mechanism and protection model. Excellent suggestion. Figure 11 was added to illustrate the proposed SVX mechanism and protection model.
18. Novelty is adequately established, but further clarity on what SVX uniquely offers compared to other protein polymers (e.g., silk fibroin, hydrolyzed keratin) would strengthen claims of innovation. SVX is a structural protein with a unique combination of strength and elasticity. Additional details were added to the Methods, Introduction, and Discussion
19. Clearly specify which datasets/methods are proprietary and which are available, and detail the avenue for requesting data if applicable. As stated in Section 6, all data are proprietary to Seevix.

Author Response File: Author Response.docx

Reviewer 2 Report

Comments and Suggestions for Authors

In the article “Spider Silk-Inspired SVX Biopolymer: A Novel Haircare Technology for Superior Color Retention, Heat Protection, and Hydration,” the authors presented the results of experimental studies on the effect of innovative products based on recombinant silk protein on the physicochemical parameters of hair after thermal and chemical treatments. The article is interesting and has practical potential. However, I have a few comments that will help the authors improve the article.

 

1. The list of keywords includes the word “vegan,” but this word appears only twice in the article and has no significant meaning (lane 23).
2. The authors emphasize that a biopolymer based on silk proteins has been created. It should be clarified what exactly the authors mean. The authors should specify this information, which proteins exactly—fibroin, sericin. Is it a complex of proteins? This is unclear from the text of the article.
3. The authors state that the effect of the created products was compared with commercial products, but it is unclear which ones (lane...88).
4. Section 2, “Materials and Methods,” states that the authors used benchmark commercial hair care products for comparison (lane 119). What are these products?
5. The authors claim that the SVX biopolymer was obtained in the form of particles with a highly porous and impermeable structure (lane 100). How then did they incorporate acids into the SVX complex?
6. Why do the authors consider the peaks at 1010 cm^-1 and 1730 cm^-1 to be analytical markers of hyaluronic acid and lactic acid, respectively? FTIR only provides identification of chemical groups. Which chemical groups specific to these acids did the authors mean?
7. The authors write that all data are presented as mean ± SEM. This is obviously a mistake (lane 291). SEM stands for scanning electron microscopy.
8. The authors indicate that the antioxidant properties of SVX can mitigate oxidative reactions….. (lane 299). It is unclear whether the authors studied the antioxidant properties of the complex or whether this is only an assumption.
9. Figures 3-4, 5, 6, and 8 show the results of experiments, but do not indicate whether there are statistically significant differences between the studied parameters. Conclusions about the effectiveness of innovative drugs can only be made on the basis of statistically significant differences.
10. Is 5-7 fibers per group sufficient in experimental research (lane 280)?
11. Fig. 3 should indicate the measurement units for water content (% ?).
12. The authors note that the use of SVX-based serum demonstrated almost complete preservation of the internal structure of the hair under thermal exposure compared to the reference product. However, visually (Image 3 and Image 5), this statement is unconvincing.

 

 

 

 

 

 

 

 

 

Author Response

Dear Reviewer 2,

We are very grateful for your thoughtful review, helpful remarks, and professional feedback. Your comments allowed us to clarify important aspects of the manuscript and improve both its structure and scientific accuracy. We highly appreciate the time and effort you dedicated to evaluating our work.

 

With sincere thanks,

Kosta Press and co-authors

 

1. The list of keywords includes the word “vegan,” but this word appears only twice in the article and has no significant meaning (lane 23). Completely agree. The word was removed from the keywords.
2. The authors emphasize that a biopolymer based on silk proteins has been created. It should be clarified what exactly the authors mean. The authors should specify this information, which proteins exactly—fibroin, sericin. Is it a complex of proteins? This is unclear from the text of the article. Important point. Additional clarification was added to the Methods section specifying that SVX is a recombinant spider silk–inspired protein, distinct from fibroin or sericin.
3. The authors state that the effect of the created products was compared with commercial products, but it is unclear which ones (lane...88). We understand this point. The comparison was performed using a multifunctional serum (Kerastase Chronologiste Thermique Régénérant), though we prefer not to disclose the brand name to avoid reputational concerns.
4. Section 2, “Materials and Methods,” states that the authors used benchmark commercial hair care products for comparison (lane 119). What are these products? Same clarification as above.
5. The authors claim that the SVX biopolymer was obtained in the form of particles with a highly porous and impermeable structure (lane 100). How then did they incorporate acids into the SVX complex? Excellent observation. We have recently published a manuscript describing the SVX complex formation, loading, and release of acids. This reference was added to the Methods and References
6. Why do the authors consider the peaks at 1010 cm^-1and 1730 cm^-1 to be analytical markers of hyaluronic acid and lactic acid, respectively? FTIR only provides identification of chemical groups. Which chemical groups specific to these acids did the authors mean? Section 2.2 (SVX Complex Preparation) was revised to include full details of the FTIR characterization and relevant chemical group assignments.
7. The authors write that all data are presented as mean ± SEM. This is obviously a mistake (lane 291). SEM stands for scanning electron microscopy. In this context, SEM refers to standard error of the mean. To avoid confusion with scanning electron microscopy, we removed “SEM” from the statistical section.
8. The authors indicate that the antioxidant properties of SVX can mitigate oxidative reactions….. (lane 299). It is unclear whether the authors studied the antioxidant properties of the complex or whether this is only an assumption. Excellent point. The antioxidant properties were evaluated by the DPPH assay, as described in our previous publication in Cosmetics. The reference has been added to the text and reference list.
9. Figures 3-4, 5, 6, and 8 show the results of experiments, but do not indicate whether there are statistically significant differences between the studied parameters. Conclusions about the effectiveness of innovative drugs can only be made on the basis of statistically significant differences. More detailed statistical information was added to the Discussion section.
10. Is 5-7 fibers per group sufficient in experimental research (lane 280)? This was a textual mistake. The Statistical Methods section was revised for clarity.
11. 3 should indicate the measurement units for water content (% ?). Corrected (%).
12. The authors note that the use of SVX-based serum demonstrated almost complete preservation of the internal structure of the hair under thermal exposure compared to the reference product. However, visually (Image 3 and Image 5), this statement is unconvincing. We understand this point. In this manuscript, SEM images are presented only for visual interpretation. To obtain statistically robust results, a much larger dataset would be required.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

The article titled “Spider Silk-Inspired SVX Biopolymer: A Novel Haircare Technology for Superior Color Retention, Heat Protection, and Hydration” is a well designed study with compelling data. the paper is generally well written but there are several critical issuess, primarily related to data consiistency and statistical analysis which must be addressed before publication.

1. Statistical analysis: The use of t tests to compare several groups increases the risk of a type 1 error. the appropriate statistical method would be a one way ANOVA followed by a suitable post-hoc test (like Tukeys HSD) to determine which specific groups differ from one another. Please revise the statistical analysis accordingly, additionally you should state whether the data met the assumptions for these tests (e.g normality and homogeneity of variances).
2. Line 99: the authors mentioned that SVX was purified to a (high degree of consistency and purity). For academic rigor, it would be beneficial to quantify this, for example, by stating the purity percentage (e.g >95% pure as determined by SDS-PAGE).
3. Line 165; the authors mentioned n=7 per group in the study protocol for clarity please mention the number of groups you used.
4. Line 301: the authors stated that the SVX protein is (thermally stable up to approximately 230∘C). This is a key piece of information for the mechanism of action. Could you please cite a reference for this or briefly mention the method used to determine this stability (e.g Differential Scanning Calorimetry)?
5. Lines 310, 339, 386, 378, 399, 401 and 411: Regarding the ImageJ analysis, it would be good to clarify how the regions of interest were selected on the hair tresses (e.g middle of the tress, randomized locations) to ensure there was no sampling bias.
6. Table 1 (Line 343): for the "baseline" group, the values for ΔL and ΔE should be 0 by definition, as no change has occurred.please correct the table to reflect this perhaps by entering (0).
7. The figure numbering appears to reset midway through the manuscript. The figure on line 520 is incorrectly labelled as figure 5 when it should be figure 7. This error continues for the subsequent figures (the Tensile Strength graph on line 536 should be Figure 8, not 6, and so on). Please re-number all figures sequentially throughout the manuscript.
8. Line 606: The text mentions "Instron LS5 tensile testing". The instrument specified in the methods section (line 262) is a Lloyd Instruments LS5. Please correct the instrument name in the discussion for accuracy.
9. Line 654: There is another significant data discrepancy in the conclusion. The text here states the color change (ΔE) for the SVX-serum treated hair was 5.0±0.6 and the results section (line 336) and Table 1 both report this value as 2.1±0.6. can you please clarify ? please write the sentence to make it more clear in table 1 or the one within the text.
10. Line 668: The conclusion states that the booster made hair (almost 30% smoother than with serum alone) i cannot find any data in the Materials and Methods or Results sections that quantifies smoothness (e.g via friction measurements, goniometry, or sensory panel). This claim appears to be unsubstantiated by the presented evidence. please either provide the data that supports this statement or remove the claim.
11. Line 688 & 709: It is unclear if the "Serum Benchmark" used for the glass hair test (Figure 5) is the same product as the "Serum Benchmark Kerastase" used for the wavy hair test (figure 6). please clarify this for consistency.
12. In the reference section; reference 2 and 4 duplicated.

 

Author Response

Dear Reviewer,

Thank you very much for your thorough and insightful review. We deeply appreciate your expert comments, especially regarding data consistency and statistical analysis. Your feedback has significantly improved the rigor and clarity of our manuscript.

 

With best regards,

Kosta Press and co-authors

 

1. Statistical analysis: The use of t tests to compare several groups increases the risk of a type 1 error. the appropriate statistical method would be a one way ANOVA followed by a suitable post-hoc test (like Tukeys HSD) to determine which specific groups differ from one another. Please revise the statistical analysis accordingly, additionally you should state whether the data met the assumptions for these tests (e.g normality and homogeneity of variances). Thank you for this important comment. The Statistical Methods section was completely revised to include one-way ANOVA followed by Tukey’s HSD post-hoc test, with verification of normality and homogeneity of variances.
2. Line 99: the authors mentioned that SVX was purified to a (high degree of consistency and purity). For academic rigor, it would be beneficial to quantify this, for example, by stating the purity percentage (e.g >95% pure as determined by SDS-PAGE). Great point. The purity level (>95%, determined by SDS-PAGE) was added to the Methods section.
3. Line 165; the authors mentioned n=7 per group in the study protocol for clarity please mention the number of groups you used. The text was corrected, and full details were added to the Statistical Methods section.
4. Line 301: the authors stated that the SVX protein is (thermally stable up to approximately 230∘C). This is a key piece of information for the mechanism of action. Could you please cite a reference for this or briefly mention the method used to determine this stability (e.g Differential Scanning Calorimetry)? This information was determined by Differential Scanning Calorimetry (DSC) and is cited from our related patent, now referenced in the text.
5. Lines 310, 339, 386, 378, 399, 401 and 411: Regarding the ImageJ analysis, it would be good to clarify how the regions of interest were selected on the hair tresses (e.g middle of the tress, randomized locations) to ensure there was no sampling bias. Well noted. Additional information on sampling and image selection was added to the Hair Sampling section.
6. Table 1 (Line 343): for the "baseline" group, the values for ΔL and ΔE should be 0 by definition, as no change has occurred.please correct the table to reflect this perhaps by entering (0). Corrected (ΔL = 0; ΔE = 0).
7. The figure numbering appears to reset midway through the manuscript. The figure on line 520 is incorrectly labelled as figure 5 when it should be figure 7. This error continues for the subsequent figures (the Tensile Strength graph on line 536 should be Figure 8, not 6, and so on). Please re-number all figures sequentially throughout the manuscript. Corrected throughout the manuscript.
8. Line 606: The text mentions "Instron LS5 tensile testing". The instrument specified in the methods section (line 262) is a Lloyd Instruments LS5. Please correct the instrument name in the discussion for accuracy. Corrected to “Lloyd Instruments LS5.”
9. Line 654: There is another significant data discrepancy in the conclusion. The text here states the color change (ΔE) for the SVX-serum treated hair was 5.0±0.6 and the results section (line 336) and Table 1 both report this value as 2.1±0.6. can you please clarify ? please write the sentence to make it more clear in table 1 or the one within the text. Corrected for consistency. The correct value (2.1 ± 0.6) is now reflected both in Table 1 and the text.
10. Line 668: The conclusion states that the booster made hair (almost 30% smoother than with serum alone) i cannot find any data in the Materials and Methods or Results sections that quantifies smoothness (e.g via friction measurements, goniometry, or sensory panel). This claim appears to be unsubstantiated by the presented evidence. please either provide the data that supports this statement or remove the claim. Removed.
11. Line 688 & 709: It is unclear if the "Serum Benchmark" used for the glass hair test (Figure 5) is the same product as the "Serum Benchmark Kerastase" used for the wavy hair test (figure 6). please clarify this for consistency. Clarified that the same product (Kerastase Chronologiste Thermique Régénérant) was used for all benchmark comparisons, without specifying the brand name in the text.
12. In the reference section; reference 2 and 4 duplicated. Corrected.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Thank you for the revision as per suggestion, which has increased the impact of the study. However, i thought after the major revision the author will add up more experiments and data in their manuscripts. Most of the data are repetitive and found to be splits instead of same kind of results in multiple data. Thus, I would like to request that the author please increase the scope of your experiment and submit again. Next, if author wants to publish with the available data only, I think this paper will not be suitable for publication at a moment.

Author Response

Dear Reviewer,

Many thanks for your careful evaluation and valuable feedback on our revised manuscript. We greatly appreciate the expertise and time you have dedicated to improving the impact and clarity of our study.

We have carefully considered each of your comments from the first round of revision and fully agree with their importance. In response, we have enhanced the manuscript in several significant ways. Specifically, we have added new experimental data on hair porosity and fiber thickness, which detail the effects of SVX treatment on hair microstructure. In addition, we have clarified the SVX:Lactic Acid complex creation and its mechanistic benefits, directly addressing concerns regarding both methodology and interpretation.

While our manuscript presents results from diverse experimental angles, we believe the data, especially after our additions, are not repetitive; rather, each result provides a unique layer of evidence that supports the overall claims and completes the mechanistic picture. The experiments complement one another and demonstrate the comprehensive impact of SVX technology on multiple hair parameters.

In light of these substantial additions and improvements, we kindly ask you to review our manuscript again and advise on its suitability for publication. We are grateful for your constructive suggestions, which have directly resulted in these new changes. Thank you once again for your time and invaluable expertise.

With best regards,

Konstantin Press

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The authors have revised the manuscript and improved it; it is now suitable for publication.

Author Response

Dear Reviewer,

We highly appreciate your time and expertise, and thank you sincerely for accepting the publication of our manuscript. As requested by another reviewer, we added a short paragraph describing the analysis of hair porosity. Many thanks again for your thoughtful input and support.

Best regards,

Konstantin Press

Author Response File: Author Response.pdf