Recombinant Type XVII Collagen Inhibits EGFR/MAPK/AP-1 and Activates TGF-β/Smad Signaling to Enhance Collagen Secretion and Reduce Photoaging
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis manuscript clearly demonstrates the enhancement of collagen secretion by recombinant type VIII collagen and thoroughly describes its mechanism of action through both the inhibition of the EGFR/MAPK/AP-1 pathway and the stimulation of the TGF-β/Smad signaling pathway. There is just one minor comment regarding the methodology of the UVA irradiation experiment. In the UVA-irradiated group, the collection time after UV irradiation should be specified. For example, please clarify whether cells were harvested immediately after the 25-minute UVA irradiation or left for 24 hours after UV irradiation. Leaving the cells for 24 hours after irradiation would provide a similar condition to the collagen-treated UV-irradiated cell group.
Author Response
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Reviewer 2 Report
Comments and Suggestions for AuthorsThis paper is a thorough exploration of the effects of a col-17 peptide on a transformed keratinocyte cell line. The results are interesting and surprising. The authors could improve the paper by commenting on the mechanism by which a collagen peptide can exert so many effects on a keratinocyte line. This is particularly important since very large concentrations (1 mg/ml or 0.1%) of peptide are needed to evoke the response.
The paper should contain a paragraph on the limitations of the results. Foremost, HaCat is a transformed keratinocyte cell line. This cell type is not responsible for the vast majority of collagen production in skin, and is not the primary source of MMPs that degrade the collagen in the dermis. Further, the cell line is transformed so that it does not respond to growth controls as do resident keratinocytes, so gene expression profiles may have little relationship to the response of these cells in skin.
The description of the peptide in the Materials and Methods references a Chinese patent. This is an inadequate description, since the patent contains many variations of source cells, induction methods, etc. The research could not be repeated by another just by looking at the patent. A paragraph describing the precise method of preparing the peptide is needed.
Author Response
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Reviewer 3 Report
Comments and Suggestions for AuthorsThe present study therefore investigated the effects of recombinant COL17 on UVA- induced photoaging mechanisms, with a focus on classical signaling pathways and basement membrane proteins, in a HaCaT cell model. Overall, authors have a good job but I have a few suggestions and questions regarding this research.
1. In discussion, please mention figures while discussing about the result. It is really difficult to understand the discussion without mentioning figure numbers.
2. Figure 5, No. is duplicated,
i. Figure 5. Schematic diagram of the molecular mechanisms underlying UVA-induced skin photoaging.
ii. Figure 5. Effects of recombinant COL17 on collagen production in UVA-irradiated (20 ml/cm2) HaCaT cells. Images of HaCaT cells were obtained after staining with fluorescein isothiocyanate conjugated antibody (red) and 4',6-diamidino-2-phenylindole (DAPI; blue) to highlight the nuclei. (a) COL4. (b) COL17. (c) Recombinant anti-LAMA5. (d) Recombinant anti-integrin β1.
3. Figure 5-line no. 256, is it mJ/mc2, mj/mc2 or MJ/cm2? please correct
4. Why these three levels 0.25, 0.5, or 1.0 mg/mL of recombinant COL17 are specifically used?
5. Literature review should be added with more recent works.
Author Response
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Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for AuthorsThe research background is well-established. However, the background could be further expanded, especially regarding the specific mechanisms of type XVII collagen in skin aging. Current research in this area is limited, and the authors could cite more relevant literature to support the novelty and necessity of their study.
The experimental design of the study is reasonable, employing a HaCaT cell model to simulate UVA-induced photoaging and utilizing various molecular biology techniques (e.g., Western blot, RT-qPCR, immunofluorescence staining) to evaluate the anti-photoaging effects of recombinant type XVII collagen. The study design covers multiple signaling pathways (e.g., TGF-β/Smad, EGFR/MAPK/AP-1) and provides a comprehensive analysis. However, the study lacks in vivo experimental data, which limits the ability to fully simulate the photoaging process in human skin. Additionally, the study does not include gradient experiments with different UVA radiation doses, making it difficult to determine the dose-response relationship between UVA exposure and photoaging effects. It is recommended that future studies incorporate animal models and further optimize UVA radiation doses.
The language of the paper is generally clear, but there are some issues with grammar and word choice, particularly in the methods and results sections, where sentence structures can be complex and potentially confusing. It is recommended that the authors seek professional language editing before submission to improve readability.
The discussion section provides a detailed analysis of the study results, particularly in explaining the mechanisms of recombinant type XVII collagen and comparing the findings with other studies. However, the discussion does not sufficiently address the limitations of the study, particularly the lack of long-term efficacy data. Furthermore, the discussion does not include direct comparisons with other anti-photoaging ingredients (e.g., vitamin C, retinol), which limits the breadth of the discussion.
Overall, the study provides valuable molecular mechanism data on the anti-photoaging effects of recombinant type XVII collagen, but there is room for improvement in the experimental design and depth of discussion. It is recommended that the authors add in vivo experimental data and optimize UVA radiation doses in future revisions. Additionally, the discussion should include a more in-depth analysis of the study's limitations and broader comparisons with other anti-photoaging ingredients.
The research background is well-established. However, the background could be further expanded, especially regarding the specific mechanisms of type XVII collagen in skin aging. Current research in this area is limited, and the authors could cite more relevant literature to support the novelty and necessity of their study.
The experimental design of the study is reasonable, employing a HaCaT cell model to simulate UVA-induced photoaging and utilizing various molecular biology techniques (e.g., Western blot, RT-qPCR, immunofluorescence staining) to evaluate the anti-photoaging effects of recombinant type XVII collagen. The study design covers multiple signaling pathways (e.g., TGF-β/Smad, EGFR/MAPK/AP-1) and provides a comprehensive analysis. However, the study lacks in vivo experimental data, which limits the ability to fully simulate the photoaging process in human skin. Additionally, the study does not include gradient experiments with different UVA radiation doses, making it difficult to determine the dose-response relationship between UVA exposure and photoaging effects. It is recommended that future studies incorporate animal models and further optimize UVA radiation doses.
The language of the paper is generally clear, but there are some issues with grammar and word choice, particularly in the methods and results sections, where sentence structures can be complex and potentially confusing. It is recommended that the authors seek professional language editing before submission to improve readability.
The discussion section provides a detailed analysis of the study results, particularly in explaining the mechanisms of recombinant type XVII collagen and comparing the findings with other studies. However, the discussion does not sufficiently address the limitations of the study, particularly the lack of long-term efficacy data. Furthermore, the discussion does not include direct comparisons with other anti-photoaging ingredients (e.g., vitamin C, retinol), which limits the breadth of the discussion.
Overall, the study provides valuable molecular mechanism data on the anti-photoaging effects of recombinant type XVII collagen, but there is room for improvement in the experimental design and depth of discussion. It is recommended that the authors add in vivo experimental data and optimize UVA radiation doses in future revisions. Additionally, the discussion should include a more in-depth analysis of the study's limitations and broader comparisons with other anti-photoaging ingredients.
Author Response
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Author Response File: Author Response.pdf
Round 2
Reviewer 4 Report
Comments and Suggestions for AuthorsThe authors have provided a detailed response to the reviewer's comments, addressing each point thoroughly. They have made significant revisions to the manuscript, including supplementing the background research, modifying experimental methods, and improving the language quality. Additionally, they have engaged a professional language editing service to enhance readability and clarity.
The authors have revised the discussion section to include more details on the limitations of the study and future research directions. However, direct comparisons with other anti-photoaging ingredients (e.g., vitamin C, retinol) are still lacking.
Authors should consider including preliminary long-term efficacy data or at least a detailed plan for future long-term studies. This would address a significant limitation of the current study.
Author Response
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Author Response File: Author Response.pdf
