Repetitive Bathing and Skin Poultice with Hydrogen-Rich Water Improve Wrinkles and Blotches Together with Modulation of Skin Oiliness and Moisture
Round 1
Reviewer 1 Report
The authors have presented a very interesting study in this paper. The results will be an important guideline for hydrogen usage in cosmetic field in the future. The questions for the authors are the following.
1 Photography conditions in some cases before and after treatment were not unified, for example in wrinkle study part from A to H, in skin blotch study part A, C, D.
2 Observing time were not the same in different cases.
3 Were the pathological diagnosis of the blotch in different cases the same?
Author Response
1 Photography conditions in some cases before and after treatment were not unified, for example in wrinkle study part from A to H, in skin blotch study part A, C, D.
→ The intensity of sunlight in the room and the of sunburn of the subject often differ between before and after using warm hydrogen water, so the skin background before and after was made as uniform as possible by Image J, then the degree of wrinkles and blotches were determined.
2 Observing time were not the same in different cases.
→ The observation period was different because we had regular interviews with the subject during the period of using hydrogen warm water, and took pictures and measured when the subject became aware of the improvement of wrinkles and blotches.
3 Were the pathological diagnosis of the blotch in different cases the same?
→ The criteria are the same because the concerned area was identified as a blotch where both the subject and the inspector recognized as a blotch without identification for pathological symptoms. The concerned sentences have been additively described in Materials & Methods Lines 98-99 of the new version.
Reviewer 2 Report
This manuscript evaluates the effect of repetitive bathing with hydrogen-rich in 5 participants. The idea is interesting, but the material and methods are not clear, and the results are reported in a confusing way.
The abstract should be re-structured and reorganized in a proper and understandable manner. You should not use etc in the abstract. You should include the most important information. You should include the population at the beginning and there is no need to mention it in every comparison (N=23).
Material and methods. It is not clear how many participants you included in the study. In the Treatments with HW bath and HW poultice section you mention that only 5 participants were included “The subjects were two 85 men and three women, aged from 49 to 66 years with no disease nor medical treatment 86 and no intake of hydrogen-related drinking/supplement for the latest 60 days”. The you say “The HW-application with bath and poultice for the prearranged periods were evalu-107 ated for effects on skin oiliness and moisture of the concerned face-parts (N=22) of five subjects”. I assume that only 5 participants were included, and you consider N each part, but this should be specified. Moreover, it is not common to consider each face part as a N, the N is usually the number of participants included in the study. You should extend the statistical analysis
Results. Why is so wide the range of days for bathing in hydrogen-rich warm water (11-61) with only 5 participants?
There is information included in the results that belongs to the material and methods “The prominent wrinkle regions on face were photographed and analyzed to be 123 classified to the degree of wrinkles into seven hierarchies (－, ±, +, ++, +++, ++++, +++++) ”.
In the figure 1 there are blue lines under the text that should be deleted. Figure D and H are pixeled. Figure E y F have different light. The footnotes should be consistent in all the figures.
“For every subject, several regions were found where the wrinkles became narrow, shal-125 low, faint or disappeared after HW bathing and face-washing (Figures 1A-H)”. This sentence should be explained widely
You mention that there are two types of treatment 10-minute bathing or repetitive three-time 3-minute face-poultice with hydrogen-rich warm water. You should include which participants receive each
“To investigate the statistically significant difference between before and after hydro-159 gen-rich warm bath, data of seven degrees of wrinkles were described as graphics” This sentence belongs to material and methods. You should include more information about the figure 2 in the text
“Twenty-eight out of 29-examined wrinkles (eye corner, forehead, cheek, middle 163 of eyebrows, nasolabial fold, etc.)” Etc is a scientific expression. Why do you say here that there are 28 regions and in material and method you say that there are 22?
Figure 2 headline have information in red.
Figure 3 also have pixeled images. T
The text regarding all the figures should include more information about them. The part corresponding to methos should be included in the corresponding section.
There are too many references together in the discussion
Author Response
The abstract should be re-structured and reorganized in a proper and understandable manner. You should not use etc in the abstract. You should include the most important information. You should include the population at the beginning and there is no need to mention it in every comparison (N=23).
→ First, the number of subjects (5) was inserted. The overall structure of Abstract has been rewritten to make it easier to understand.
Material and methods. It is not clear how many participants you included in the study. In the Treatments with HW bath and HW poultice section you mention that only 5 participants were included “The subjects were two 85 men and three women, aged from 49 to 66 years with no disease nor medical treatment 86 and no intake of hydrogen-related drinking/supplement for the latest 60 days”. The you say “The HW-application with bath and poultice for the prearranged periods were evalu-107 ated for effects on skin oiliness and moisture of the concerned face-parts (N=22) of five subjects”. I assume that only 5 participants were included, and you consider N each part, but this should be specified. Moreover, it is not common to consider each face part as a N, the N is usually the number of participants included in the study. You should extend the statistical analysis
→ Used n instead of N for the number of measurement points.
Results. Why is so wide the range of days for bathing in hydrogen-rich warm water (11-61) with only 5 participants?
→ The observation period was different because we had regular interviews with the subject during the period of using hydrogen warm water, and took pictures and measured at the time when the subject became aware of the improvement of wrinkles and stains.
There is information included in the results that belongs to the material and methods “The prominent wrinkle regions on face were photographed and analyzed to be 123 classified to the degree of wrinkles into seven hierarchies (－, ±, +, ++, +++, ++++, +++++) ”.
→ Deleted because there is a description similar to “materials and methods”
In the figure 1 there are blue lines under the text that should be deleted. Figure D and H are pixeled. Figure E y F have different light. The footnotes should be consistent in all the figures.
→ Fixed by Photoshop so that the pixel and the brightness are as similar as possible, between before and after examination.
“For every subject, several regions were found where the wrinkles became narrow, shal-125 low, faint or disappeared after HW bathing and face-washing (Figures 1A-H)”. This sentence should be explained widely
→ Rewritten
You mention that there are two types of treatment 10-minute bathing or repetitive three-time 3-minute face-poultice with hydrogen-rich warm water. You should include which participants receive each
→ Rewritten so that it can be understood that there are 5 people
“To investigate the statistically significant difference between before and after hydro-159 gen-rich warm bath, data of seven degrees of wrinkles were described as graphics” This sentence belongs to material and methods. You should include more information about the figure 2 in the text
→ The sentence has migrated to “material and methods”. In addition, the explanation of figure 2 has been conducted more in detail.
“Twenty-eight out of 29-examined wrinkles (eye corner, forehead, cheek, middle 163 of eyebrows, nasolabial fold, etc.)” Etc is a scientific expression. Why do you say here that there are 28 regions and in material and method you say that there are 22?
→ “22” is the measurement score of oiliness / moisture, which is different from the measurement score of wrinkles.
Figure 2 headline have information in red.
→ corrected
Figure 3 also have pixeled images. T
→ Fixed by Photoshop so that the brightness and the pixel are as close as possible before and after the examination in Figures 3B and C.
The text regarding all the figures should include more information about them. The part corresponding to methos should be included in the corresponding section.
→ Detailed description of figures.
There are too many references together in the discussion
→ Deleted 5 old papers preferentially.
Reviewer 3 Report
I would like to thank the authors for this article. The topic is very important and interesting. Using hydrogen water as a noninvasive method of skin rejuvenation is very promising.
I am missing the chapter: Study limitation. Please add it to the text.
Author Response
I am missing the chapter: Study limitation. Please add it to the text.
→ The indicated phrases have been added to the beginning of Discussion in the new version.
Round 2
Reviewer 2 Report
Changes proposed in the first review have not been made, the author still use etc, they have not ommited the N/n ang go on using them as participants. Abstract and matherial and methods have not been reorganised. There are still green lines in the images. Revise and make all the changes commented in the first review for further consideration.
Author Response
１ Changes proposed in the first review have not been made, the author still use etc.,
→ We have deleted “etc.” and the description of regions analyzed were moved to “Subjects and Methods” (Lines 16, 17, 22, 23 in the revised version)
２ they have not omitted the N/n ang go on using them as participants.
→ We have quitted to use N or n for the number of regions analyzed, instead used “loci” (Lines 17, 22, 23, 48, 112, 127, 163, 165, 172, 180, 196, 207, 220, 222, 228, 233, 248, 254, 259, 275, 282, 283 in the revised version)
.
３ Abstract and material and methods have not been reorganized.
→ We have re-written both of “Abstract” and “Subjects and methods” (Lines 16, 17, 22, 23, 96-114 in the revised version)
４ There are still green lines in the images.
→ We cannot see any green line in all figures, however, we found a bottom line of Fig.3C,thus replaced into a new one.
５ Revise and make all the changes commented in the first review for further consideration.
→ We have re-written the sentences as many as possible where you pointed out. (Lines 16, 17, 22, 23, 48, 112-113, 248, 275, 284 in the revised version)
Author Response File: 
Author Response.docx
Round 3
Reviewer 2 Report
Changes have been made
