Adipose-Derived Stem Cells from Systemic Sclerosis Patients Maintain Pro-Angiogenic and Antifibrotic Paracrine Effects In Vitro

Round 1

Reviewer 1 Report

General Comments

This is a well referenced, well designed and thorough clinical study that addresses the properties of scleroderma patient ASC relative to healthy donors.  Complementary assays monitoring particular phenomena have been used appropriately.  The authors present their data clearly  and reach conclusions supported by the evidence. The findings have relevance to clinical decision making as autologous ASC and SVF cells become more frequently used as a therapetuic tool.  The specific comments below merit consideration.

Specific Comments

Line 50, Pg 2. The word "expect" should be "except".

Line 54.  In listing tissue sources of MSC, bone marrow should be included.

Line 71.  The word "month" should be replaced with "mouth".

Line 78.  The term 'ant-fibrotic" should  be replaced with "anti-fibrotic".

Line 179.  Section 2.9. Report cell plating density as cell number per square centimeter rather than cell number per well of a particular plate.

Line 206.  Rather than say "catch up" it may be more appropriate to say "capture".

Line 267, Table 2.  The number of HD patients responding regarding smoking history sums up to 6 not 7.  Was data on one subject lacking?

Liine 355, Table 4. I would recommend reporting values solely as the mean and SD for each paracrine factor and for each category (SSc or HD) rather than presenting the triplicate values.

Line 361. Change  "is" to "in".

Line 404.  This sentence needs to be re-written to begin with the word "the".

Line 414.  There is a need to expand on the findings of Virzi et al.  The single sentence here does not sufficiently relate Virzi et al's findings in a manner that explains how the current manuscript data differs from that previously published.

Line 457.  The phrase should read "allowed us  to address...".  Additionally, the  authors refer to their approach as a global assessment of the secretome.  This is an over-reach since only a handful of identified and predicted growth factors were evaluated using PCR and antibody based methods.  Neither transcriptomic or proteomic methods were employed.  Without those, it is inaccurate to call the approach global.  The authors  may wish to point out that broader surveys  of angiogenic and fibotic biomarkers may  be warranted in the future.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript by Velier et al. titled "Adipose-derived Stem Cells from Systemic Sclerosis patients maintain their pro-angiogenic and anti-fibrotic paracrine ehhects in vitro" reported that ASC cells derived from systemic sclerosis (SSc) patients manatined their angigenic and anti-fibrotic activity.

The manuscript is well written, the data are well presented and the experiments are well planned. However, some points shoud be addressed.

In the figure 3 the authors showed no differences between HD-ASC and SSc-ASC in terms of mRNA levels of anti-Col1A1, Col3, MMP-1, MMP-2, TIMP-1, TGF-B1, HGF and VEGF-a. However, often mRNA levels do not reflect amount of the total protein. Western blot analysis of the afore mentioned molecules should be performed in order to know their cytoplasmatic levels. In the figure legend there is also an typo mistake; please, modify "HD-SSc" with "HD-ASC". In Fig.2 additional senescence markers (i.e. p16INK4a, telomere associated foci - TAF) should provided. In the Table 2 the authors reported 7 HD, but only 6 (3/3) were reported as smokers or not smokers.What is the smoking status of the seventh patient? In the Table 3 was reported the percentage of positive cell for surface markers by HD-ASC and SSc-ASC. The authors should provide also MFI values for each markers. Also, a statistical comparison should be provide.

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

The authors were investigating adipose-derived stem cells from systemic sclerosis patients for their pro-angigenic and anti-fibrotic effects in vitro.

The paper is well written but needs here and there some changes in the formatting and rewriting for further clarification.

Tables should be labelled on top of the respective table. I would prefere Table 3 as graph, depicting the MFI ratio of the different antibodies to their respective isotype control. Figure 3: the legend should be under the actual figure. Figure 5a needs proper labelling. Each lane should be labelled, MW sould be indicated. Figure legend for Figure 5 needs reformatting. Figure legend for 5A needs rewording, it is unclear. The discussion is not very well written and needs some polishing.

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

The authors addressed most of the concerns raised.

Author Response

Thank you for your suggestion. The citation has been added (n°38) line 450.