Percutaneous Needle Electrolysis Accelerates Functional Muscle Regeneration in Mice
, Ramon Margalef 4, Marc Bosque 4, Francisco Minaya-Muñoz 1,2,5 and Manel M. Santafé 4,*
Round 1
Reviewer 1 Report
The authors have evaluated the effect of percutaneous needle electrolysis (PNE) in muscle regeneration. Following needs to be addressed before acceptance:
1. Introduction should be strengthen by describe the novelty and more details about rationality of the study.
2. The methodology section needs major modification. The section 2.2 muscles should be place after section 2.5 since once all the in vivo studies were completed, the muscles would be excised. Experimental groups should be put first, there after the experimental protocols, followed by endplate noise recordings and finally “muscles”. The “muscles” section should be renamed to “processing of muscles” or similar such title for better understanding.
3. Results: The figure 2 mentions magnification as 100X. Plz check as it seems to be more than 100X. Also the staining in the images A, B and C seem to be different in terms of intensity. Can authors explain reason behind this?
4. Figure 4 legends is not clear; please clearly state what is (a) and (b).
5. The discussion should include a section on limitations of current study for broader readership.
6. Grammatical and spelling errors need correction eg. Page 1, line 43 “a sex vivo” should be ‘as ex vivo’
Author Response
- INTRODUCTION SHOULD BE STRENGTHENED BY DESCRIBE THE NOVELTY AND MORE DETAILS ABOUT RATIONALITY OF THE STUDY.
O.R.: We have tried to be more specific by introducing the procedures that are used later in the paper.
- THE METHODOLOGY SECTION NEEDS MAJOR MODIFICATION. THE SECTION 2.2 MUSCLES SHOULD BE PLACE AFTER SECTION 2.5 SINCE ONCE ALL THE IN VIVO STUDIES WERE COMPLETED, THE MUSCLES WOULD BE EXCISED. EXPERIMENTAL GROUPS SHOULD BE PUT FIRST, THERE AFTER THE EXPERIMENTAL PROTOCOLS, FOLLOWED BY ENDPLATE NOISE RECORDINGS AND FINALLY “MUSCLES”. THE “MUSCLES” SECTION SHOULD BE RENAMED TO “PROCESSING OF MUSCLES” OR SIMILAR SUCH TITLE FOR BETTER UNDERSTANDING.
O.R: Suggested changes have been made.
- RESULTS:
3.A.- THE FIGURE 2 MENTIONS MAGNIFICATION AS 100X. PLZ CHECK AS IT SEEMS TO BE MORE THAN 100X.
O.R.: Microphotographs were initially taken at 100X, as shown in the legend of the figure. The size of the mast cells is the same in the 3 photomicrographs.
3.B.- ALSO THE STAINING IN THE IMAGES A, B AND C SEEM TO BE DIFFERENT IN TERMS OF INTENSITY. CAN AUTHORS EXPLAIN REASON BEHIND THIS?
O.R.: Lines 218-222: This paragraph has been modified to account for the different intensities of blue in case any other reader has this question.
- FIGURE 4 LEGENDS IS NOT CLEAR; PLEASE CLEARLY STATE WHAT IS (A) AND (B).
O.R.: The legend of figures 4 and 5 have been modified for clarity.
- THE DISCUSSION SHOULD INCLUDE A SECTION ON LIMITATIONS OF CURRENT STUDY FOR BROADER READERSHIP.
O.R: A section of limitations has been added.
- GRAMMATICAL AND SPELLING ERRORS NEED CORRECTION EG. PAGE 1, LINE 43 “A SEX VIVO” SHOULD BE ‘AS EX VIVO’
O.R.: The change has been made.
Author Response File: 
Author Response.docx
Reviewer 2 Report
General Comments to Authors:
The manuscript describes a neat study of accelerated muscle regeneration following the use of needle electrolysis. An important key finding of the study is that needle electrolysis could improve recovery of damaged muscle tissue.
The Introduction provides a good background of the subject matter of the study.
The Methods section could be improved, and specific recommendations regarding the re-ordering and editing of the Methods sections are made on this matter (see below).
The Results are presented clearly and the Discussion is organised well and deals with the findings of the study appropriately.
Specific Comments to Authors:
The Methods are not written clearly in respect of the use of animals. I presume the better order of the description of the methods is to move section 2.2 "Muscles" so that it appears between the current sections 2.5 and 2.6, and it should be re-named to "Muscle Histology". I would also move the final sentence of the current "Muscles" section (that being "The gastrocnemius muscles were used for electromyographic recordings.") to appear at the beginning the current section 2.3 "Experimental protocols: ...".
In the new "Muscle Histology" section (which would be 2.5) the first sentence should read as follows: "At the conclusion of the experiment animals were deeply anaesthetized with isoflurane before being euthanized by exsanguination."
I recommend placing the terms "in vivo" and "ex vivo" into italic font (which is a convention) throughout the manuscript.
In Line 160 the manuscript suggests that not all animals were anaesthetized during the recording sessions. Is that the case? If so, the authors should provide the breakdown of the numbers of recordings obtained from anaesthetized animals and unanaethetized animals. If all the animals were anaesthetized then the authors must remove the word "usually" in line 160.
Other Specific Comments:
Line 43 - the sentence should end with "... as well as ex vivo." (typographical error)
Line 74 - this should be "... by endplate noise recording, and functional rein nervation ..." (insert the word "and" before "functional).
Line 86 - The beginning of the sentence that begins on this line should be either "The animals' room was ..." if all animals were in one room, or "The animals' rooms were ..." is there was more than one room housing the animals.
Lines 116 and 119 - there should be spaces between the numbers and the words, for example "72 hours" and "and 10 days".
Line 139 - There is the number 1 at the end of the word "repetitions". The number should be removed.
Line 152 - a space should be introduced between "and" and "BPV" (eg. and BPV).
Line 157 - there should be spaces between the numbers and the words, for example "72 hours" and "and 10 days".
Line 161 - the temperature information should be written as "22 - 25.8o C" (the spacing is important).
Line 218 - the sentence starting on this line should start with "After treatment, this ..." (remove the word "An").
Line 267 - always place spaces between the numbers and the + sign (eg. 123.23 + 6.87).
Line 352 - the sentence would read better if is started as "The myotoxic effects of BPV have been known since ..." (remove the words "study of the").
Line 353 - place a space between "[23]" and "have".
END
Author Response
1.- THE METHODS ARE NOT WRITTEN CLEARLY IN RESPECT OF THE USE OF ANIMALS. I PRESUME THE BETTER ORDER OF THE DESCRIPTION OF THE METHODS IS TO MOVE SECTION 2.2 "MUSCLES" SO THAT IT APPEARS BETWEEN THE CURRENT SECTIONS 2.5 AND 2.6, AND IT SHOULD BE RE-NAMED TO "MUSCLE HISTOLOGY". I WOULD ALSO MOVE THE FINAL SENTENCE OF THE CURRENT "MUSCLES" SECTION (THAT BEING "THE GASTROCNEMIUS MUSCLES WERE USED FOR ELECTROMYOGRAPHIC RECORDINGS.") TO APPEAR AT THE BEGINNING THE CURRENT SECTION 2.3 "EXPERIMENTAL PROTOCOLS: ...". IN THE NEW "MUSCLE HISTOLOGY" SECTION (WHICH WOULD BE 2.5) THE FIRST SENTENCE SHOULD READ AS FOLLOWS: "AT THE CONCLUSION OF THE EXPERIMENT ANIMALS WERE DEEPLY ANAESTHETIZED WITH ISOFLURANE BEFORE BEING EUTHANIZED BY EXSANGUINATION."
O.R: Suggested changes have been made
2.- I RECOMMEND PLACING THE TERMS "IN VIVO" AND "EX VIVO" INTO ITALIC FONT (WHICH IS A CONVENTION) THROUGHOUT THE MANUSCRIPT.
O.R.: The change has been made
3.- IN LINE 160 THE MANUSCRIPT SUGGESTS THAT NOT ALL ANIMALS WERE ANAESTHETIZED DURING THE RECORDING SESSIONS. IS THAT THE CASE? IF SO, THE AUTHORS SHOULD PROVIDE THE BREAKDOWN OF THE NUMBERS OF RECORDINGS OBTAINED FROM ANAESTHETIZED ANIMALS AND UNANAETHETIZED ANIMALS. IF ALL THE ANIMALS WERE ANAESTHETIZED THEN THE AUTHORS MUST REMOVE THE WORD "USUALLY" IN LINE 160.
O.R.: The word "usually" has been removed
4.- OTHER SPECIFIC COMMENTS:
LINE 43 - THE SENTENCE SHOULD END WITH "... AS WELL AS EX VIVO." (TYPOGRAPHICAL ERROR)
LINE 74 - THIS SHOULD BE "... BY ENDPLATE NOISE RECORDING, AND FUNCTIONAL REINNERVATION ..." (INSERT THE WORD "AND" BEFORE "FUNCTIONAL).
O.R: Suggested changes have been made
LINE 86 - THE BEGINNING OF THE SENTENCE THAT BEGINS ON THIS LINE SHOULD BE EITHER "THE ANIMALS' ROOM WAS ..." IF ALL ANIMALS WERE IN ONE ROOM, OR "THE ANIMALS' ROOMS WERE ..." IS THERE WAS MORE THAN ONE ROOM HOUSING THE ANIMALS.
O.R: "The animals' room was ..." : It has been modified
LINES 116 AND 119 - THERE SHOULD BE SPACES BETWEEN THE NUMBERS AND THE WORDS, FOR EXAMPLE "72 HOURS" AND "AND 10 DAYS".
LINE 139 - THERE IS THE NUMBER 1 AT THE END OF THE WORD "REPETITIONS". THE NUMBER SHOULD BE REMOVED.
LINE 152 - A SPACE SHOULD BE INTRODUCED BETWEEN "AND" AND "BPV" (EG. AND BPV).
LINE 157 - THERE SHOULD BE SPACES BETWEEN THE NUMBERS AND THE WORDS, FOR EXAMPLE "72 HOURS" AND "AND 10 DAYS".
LINE 161 - THE TEMPERATURE INFORMATION SHOULD BE WRITTEN AS "22 - 25.8O C" (THE SPACING IS IMPORTANT).
LINE 218 - THE SENTENCE STARTING ON THIS LINE SHOULD START WITH "AFTER TREATMENT, THIS ..." (REMOVE THE WORD "AN").
LINE 267 - ALWAYS PLACE SPACES BETWEEN THE NUMBERS AND THE + SIGN (EG. 123.23 + 6.87).
LINE 352 - THE SENTENCE WOULD READ BETTER IF IS STARTED AS "THE MYOTOXIC EFFECTS OF BPV HAVE BEEN KNOWN SINCE ..." (REMOVE THE WORDS "STUDY OF THE").
LINE 353 - PLACE A SPACE BETWEEN "[23]" AND "HAVE".
O.R: Suggested changes have been made.
Other uncommented spaces have been reviewed. Probably inserting the text in the publisher's pro forma document randomly deleted some spaces.
Author Response File: Author Response.docx
