The authors present data in support of using SW-120, an intestinal cancer cell line, as an in vitro model system for irritable bowel disease (IBD). IBD is a rather prevalent ailment that is also not entirely understood mechanistically so further study is warranted.

I am not an expert in intestinal disease so I can not say with authority if the idea to use SW-120 is novel but a quick internet search did not turn up any obvious conflict with existing literature.

In my opinion, this paper has two major weaknesses. The first is that the introduction does not reference previous work in establishing model systems for related diseases. The process of establishing a model system usually involves meeting certain criteria for cell identity and function. The authors would have been better served describing some of these criteria to more accurately explain how their work fits into that context.

The second major weakness of the paper has to do with the volume of data. The authors effectively only present one piece of data about a single marker, IL-8. Normally, a single marker is not enough to establish a model system so I wonder if the authors might want to consider repackaging this article as a pilot report or a presentation of preliminary data to more accurately reflect the fact that there is still a lot of work to be done. In addition, the main figure in the paper (Fig. 1) comes to us without error bars or an indication of the number of experiment repeats. This severly compromises its interpretability.

The experiment is logistically simple so I did not see anything in lacking in the methods.

The authors did a good job including a viability control (Fig. 2) which is essential to our ability to interpret the main data since the main ouput (IL-8) is being reported as a concentration. However, the viability is recorded as a percent and it was unclear to me how this number is being calculated. Clarification is required.

As mentioned above, data for a single marker is insufficient to establish a model system. At the same time, I do think the data is suggestive and good enough for a pilot report or preliminary report so long as error bars in Fig. 1 and a clarification of Fig. 2 can be provided.

I would have to say not. The data is not insignificant but there is still a lot of work that needs to be done before this can be used seriously by others as an established model.

Although the tone of the language overall was good, there was some imbalance in the paragraph size. In the discussion section, for example, one paragraph was a single sentence long while two paragraphs were two sentences long. This creates imbalance in the flow of the reading and makes the reader wonder if more polishing could be done.

Fendos JE, Engelman DM. (2012) pHLIP and acidity as a universal biomarker for cancer. Yale Journal of Biology and Medicine.

Cell biology, cancer biology.

The authors developed an in vitro model for IBD using colon cancer cell line SW-620. They measured the IL-8 levels om HT-29 aftehr the treatment with LPS isolated from various bacterial strains. They further studied the effect of drug Ozagrel and found that it regulates of IL-8 expression.

The claims are novel. However, the number of the experiments they conducted was not clear, which weakens the reliability of the claims.

NA

Yes. But they need to show mutiple runs of the same experiments to show that the data is reproducible.

NA

Yes

Need to repeat the experiments

It's an OK paper.

1) What is MTT assay? I wound not find the full name of it.

2) The legend for the x axis in fig.1 is not clear. They also need to show error bars and If possible p-values for figure 1.

NA

NA

Authors claimed that they developed an in vitro model for IBD using colon cancer cell line SW-620. In this in-vitro model of IBD, IL-8 was playing a key role in inflammation cascade. Action of anti-inflammatory drug Ozagrel was studied and was found to block the up regulation of IL-8.

The claims are well known in literature.

Gastroenterology. 2003 Apr;124(4):1001-9.

TNF-alpha and IFN-gamma regulate the expression of the NOD2 (CARD15) gene in human intestinal epithelial cells.

Rosenstiel et. al.

Yes

Yes, result poorly written. Even results were not statistically verified (fig1. Contains no standard deviation/error bar).

No

Yes. But it should be updated with detailed information. Several information are missing as

“Specific cell line was selected based on the experiment and the cells were allowed to get confluent. Post confluent cells were seeded in 96 well plate as described above, with a concentration of 30,000-40,000 cells/well. Plate was then left at CO2 incubator for overnight incubation. As designed for the experiment 1) if the cells are to be pre-incubated with drug followed by LPS treatment, the cells in the plate were incubated with drug in different concentrations. Based on the solubility of the drug if soluble in water was directly diluted with medium and applied to the cells or if insoluble was the diluted in DMSO and the applied. 3 hours later of drug incubation cells were treated with LPS to get inflamed.”

Some Questions are-

What kind of cell line?

LPS treatment at what Conc.?

Final Conc. of DMSO ?

NA

No. The paper is not outstanding as it contains common data.

Poor article. I would like to advise the authors to be cautious, it is poorly written. there are several  mistakes, e.g.

It should be

CO₂ instead of CO2

Strain names should be in italics (as in fig1)

Drug development and drug mechanism of action

The manuscript analyzes  the effect of the anti-inflammatory drug Ozagrel on IL-8 production induced by LPS isolated from various bacterial. Authors utilized the colon cancer cell line SW-620 as experimental model of Inflammatory Bowel Disease (IBD)

No this paper is not novelty because other studyes have already demonstrated a role of thromboxane synthase inhibition such as NV52, E3040 and Ridogrel in IBD

No, the paper needs a major revision of current literature on the effects of thromboxane synthase inhibition in IBD

The data presented in the manuscript are not convincing, the article lacks an adequate statistical analysis of the results, making questionable the whole body of the study. Moreover , the authors have to explain how a colon cancer cell line could be a suitable model for IBD

No, this cannot be applied to this paper

No, the paper lacks a Result section and an adequate statistical analysis of the results. Moreover some methodology are not explained: no information on the isolation, quantification and purity control of LPS were reported

The manuscript could be considered for a publication only if:

1. A  major revision of the entire manuscript and a major revision of English grammar are undertaken;
2. If the authors explain their experimental model of IBD;
3. If this study will be extended to other citokines involved in inflammation, the manuscript could be considered for publication

At the moment this paper is not outstanding in its discipline since a cancer cell line cannot be a model for IBD and the effects of Ozagrel need to be extended to other citokines than only IL-8

The manuscript entitled “SW-620 Cells Evoke IL-8 Expression and Downregulation by Ozagrel: An Intervention in IBD”:by Bhargava V, Singh N, Bhargava R, analyzes  the effect of the anti-inflammatory drug Ozagrel on IL-8 production induced by LPS isolated from various bacterial. Authors utilized colon cancer cell line SW-620 as experimental model of Inflammatory Bowel Disease (IBD).

In my opinion, the manuscript is badly structured and lacks novelty .

The data presented in the manuscript are not convincing, the article lacks adequate description and statistical analysis of the results, making questionable the whole body of the study.

Authors refer to have developed an in vitro model for IBD.  Culture experiments have provided insights into the effects of individual cytokines and other inflammatory mediators on epithelial pathophysiology  that may be involved in IBD, not for IBD as such. In addition, the authors have to explain how a colon cancer cell line could be suitable model for  IBD, except than IL-8 production after LPS stimulation.

Author reported that “LPS treatment was optimized using different strains of bacteria and maximum efficient strain to cause inflammation was chosen for the experiment.” No information on the isolation, quantification and purity control of LPS were reported.

Moreover, as Ozagrel are popularly used to inhibit the upregulation of cytokine IL-8 (Bonner GB 1996), the paper of Bhargava at al. does not add very much to our knowledge on the effect of anti-inflammatory drugs for IBD pharmacological treatment.

Thus, as many points need to be deeply modified, the manuscript is not suitable for publication.

Specific comments:

1. The Material and Methods section is confusing and insufficient.

2. The Result section is completely lacks.

3. The Discussion section has to be deepened.

4. The case number must be indicated in Figure Legends, and statistical significance too.

5. The authors should have their paper revised by a mother tongue speaker with scientific knowledge.

Study of oxidative stress in normal and tumor cell lines Study of PARP in DNA damage/repair and gene regulation in normal and tumor cell lines