Design and Evaluation of Complex Polypeptide-Loaded Dissolving Microneedles for Improving Facial Wrinkles in Different Areas
, Yuning Ma 2, Suohui Zhang 1,5,* and Yunhua Gao 1,3,5,*Round 1
Reviewer 1 Report
1. The structure of presenting both the methodology part as well the result and discussion part should be rearranged. First the preparation part and characterization of microneedles. Then the in vitro studies nd the in vivo studies and finally the clinical studies part.
2. In section 2.3. the authors mentioned that the preparation pf the molds as well as the microneedles were done according o previous studies. This should be also added as paragraph explaining in details the procedures in addition to the citations.
3. It is preferable for the authors to add an experiment showing the release profile or pattern of the loaded cargo from the developed microneedles.
Author Response
Response to Reviewer 1 Comments
Point 1: The structure of presenting both the methodology part as well the result and discussion part should be rearranged. First the preparation part and characterization of microneedles. Then the in vitro studies nd the in vivo studies and finally the clinical studies part.
Response 1: Thank you for your kind suggestion. We have adjusted the structure of the methodology part and result and discussion part to make it more readable and logical.
Point 2: In section 2.3. the authors mentioned that the preparation pf the molds as well as the microneedles were done according o previous studies. This should be also added as paragraph explaining in details the procedures in addition to the citations.
Response 2: We have made detailed supplements about preparation of molds and microneedles in section 2.3 to ensure the readability and coherence of the study. Thank you for your advice.
Point 3: It is preferable for the authors to add an experiment showing the release profile or pattern of the loaded cargo from the developed microneedles.
Response 3: As you suggested, we had tried to establish the quantitative analysis methods to measure the release profile of three active ingredients loaded in CP-DMNs. Nevertheless, the concentrations of three peptides in the microneedle solutions were extremely low: oligopeptide-1 (0.15%, w/v), acetyl hexapeptide-8 (0.03%, w/v), and palmitoyl pentapeptide-4 (0.03%, w/v), making it difficult to conduct in vitro percutaneous release experiments by HPLC method. Alternatively, the in vitro skin dissolution test of CP-DMNs has been performed to verify that the active ingredients loaded in DMNs can effectively enter the skin and exert their effectiveness, as shown in Figure 3. Thanks again for your professional comment. In the future, we may consider the UPLC-MS/MS method (more sensitive) for transcutaneous release studies of CP-DMNs. We look forward to your continued attention.
Reviewer 2 Report
In this manuscript, titled “Design and evaluation of complex polypeptide loaded dissolving microneedles for improving facial wrinkles in different areas”, the authors developed a complex polypeptide loaded dissolving microneedles (CP-DMNs) for reducing the facial wrinkles in different areas. The authors firstly analyzed the structural and morphological aspects of the microneedles, then they performed in vivo and in vitro studies in order to demonstrate the efficacy and safety of their system in reducing facial wrinkles.
The manuscript may be quite interesting and relevant to the pharmaceutics, medical, and bio-medical communities; anyway, several technical, formal, and scientific issues must be addressed.
1. From a formal point of view, the article must be revised. In my opinion, in the abstract and in the introduction there are some sensationalistic expressions such “it is an urgent need to develop a safe, effective, and multi-functional anti-wrinkle product to improve facial wrinkles in different areas (lines 18-20), or “wrinkled face is an intuitive manifestation that a person is no longer young, for those who pay great attention to their appearance, wrinkles will undoubtedly cause anxiety and frustration of self-esteem (lines 42-44)”. This should be avoided in a scientific paper and the authors should modify them.
2. The manuscript is rich in abbreviations and acronyms, some of which are commonly used in numerous scientific articles while others are more specific and less common. Usually, the use of acronyms should facilitate the reader but, not in this case. In my opinion, it is mandatory to insert a list of acronyms at the beginning of the manuscript.
3. In section 3.1: the authors should indicate how much pressure they applied to allow the microneedles to penetrate the skin. In addition, the authors state that the needles penetrate 100, 180, and 240 µm through the skin but the error is not reported on the measurements; how many measurements were performed? Also, there are no pictures of the pillars after removal from the skin. This would be useful to understand if the needles were broken or dissolved in the skin. Finally, the scale bars are not visible (too small) and the references in the text of Figure 1 a), b) and c) are missing.
4. In section 3.2: the authors should be clear on how they performed the cytotoxicity test: did they apply the microneedles pad on cell culture? The same goes for microbial limit tests: authors should provide a more accurate description of the procedure.
5. A technical issue: All the figures in the manuscript must be revised in depth. The authors must modify all the figures reported in the manuscript because the pictures are very difficult to read. For example:
Fig.1: the scale bars are too small.
Fig. 3 and 4: Please, increase the size of the picture, the size of the bars, the titles of the axes of the graphs, and the data reported in the graphs.
Fig. 5: the graphs on the right side of the figure are too small and illegible.
Fig. 4 and 6: In my opinion, the colours chosen by the authors are too brilliant and it is not possible to easily appreciate the differences between the bars, as well as the error bars. Please, modify the colors of the graphs.
Author Response
Response to Reviewer 2 Comments
Point 1: From a formal point of view, the article must be revised. In my opinion, in the abstract and in the introduction there are some sensationalistic expressions such “it is an urgent need to develop a safe, effective, and multi-functional anti-wrinkle product to improve facial wrinkles in different areas (lines 18-20), or “wrinkled face is an intuitive manifestation that a person is no longer young, for those who pay great attention to their appearance, wrinkles will undoubtedly cause anxiety and frustration of self-esteem (lines 42-44)”. This should be avoided in a scientific paper and the authors should modify them.
Response 1: Thanks to the reviewer for pointing out our mistakes in expressions. The improper descriptions in the abstract and introduction have been replaced by appropriate words. Further, we have also carefully checked the language expression of the full text to standardize the scientific and rigor of the manuscript.
Point 2: The manuscript is rich in abbreviations and acronyms, some of which are commonly used in numerous scientific articles while others are more specific and less common. Usually, the use of acronyms should facilitate the reader but, not in this case. In my opinion, it is mandatory to insert a list of acronyms at the beginning of the manuscript.
Response 2: According to the reviewer’ suggestions, a list of acronyms has been inserted at the beginning of the manuscript in red font to facilitate reviewers’ viewing and readers’ understanding.
Point 3: In section 3.1: the authors should indicate how much pressure they applied to allow the microneedles to penetrate the skin. In addition, the authors state that the needles penetrate 100, 180, and 240 µm through the skin but the error is not reported on the measurements; how many measurements were performed? Also, there are no pictures of the pillars after removal from the skin. This would be useful to understand if the needles were broken or dissolved in the skin. Finally, the scale bars are not visible (too small) and the references in the text of Figure 1 a), b) and c) are missing.
Response 3: In the in vitro skin insertion studies, all microneedles were pierced into the skin via a homemade applicator under the force of 20 N/cm2. Additionally, the experiments were performed three times in parallel, and we have supplemented the results in section 3.1 in the form of mean ± SD. Further, the intradermal dissolution test of CP-DMNs has been complemented to verify that the active ingredients loaded in the microneedles can effectively enter the skin. Finally, both the scale bars and references have been revised and added in the manuscript. Thank you for your meticulous review and advice.
Point 4: In section 3.2: the authors should be clear on how they performed the cytotoxicity test: did they apply the microneedles pad on cell culture? The same goes for microbial limit tests: authors should provide a more accurate description of the procedure.
Response 4: We are grateful for the kind reminder from the reviewer. We have supplemented all the necessary details and accurate procedures around the cytotoxicity test and the microbial limit tests in the manuscript in section 2.5.
Point 5: A technical issue: All the figures in the manuscript must be revised in depth. The authors must modify all the figures reported in the manuscript because the pictures are very difficult to read. For example:
Fig.1: the scale bars are too small.
Fig. 3 and 4: Please, increase the size of the picture, the size of the bars, the titles of the axes of the graphs, and the data reported in the graphs.
Fig. 5: the graphs on the right side of the figure are too small and illegible.
Fig. 4 and 6: In my opinion, the colours chosen by the authors are too brilliant and it is not possible to easily appreciate the differences between the bars, as well as the error bars. Please, modify the colors of the graphs.
Response 5: Thank you for pointing out the deficiency existed in the figures, and we have modified all figures in this manuscript to make it easier for the readers to see, according to the reviewer’s detailed suggestions.
Round 2
Reviewer 1 Report
The comments have been considerably addressed. Thak you.
Author Response
Thank you for taking the time to review the manuscript.
Reviewer 2 Report
I very much appreciated the changes you have made to the text and believe that the answers to the questions are adequate.
Author Response
Thank you again for your contributions in the review of this manuscript.
