Review for 10.1186/s12876-025-03923-x (2025)

Title: Effect of chitosan nanoparticles conjugated with the cell free supernatant of
Bifidobacterium bifidum on the expression of genes related to colorectal cancer in colon
adenocarcinoma (Caco-2) cell line
Version: 0 Date: 06 Nov 2024
Reviewer:  Dingpei Long
Reviewer's report:
This Manuscript demonstrates a certain degree of innovation by exploring the regulatory effects
of CTNP combined with B.b-sup on CRC signaling pathway genes. However, there is significant
room for improvement in experimental design, data analysis, and discussion. It is recommended
to provide more detailed descriptions of experimental methods, include necessary statistical
analyses, and improve the language and structure.
1. Abstract and Introduction
(1) Abstract: There are inaccuracies in grammar and wording. For example, "Following the
investigate of CTNP/B.b-sup toxicity on Caco-2 cells" should be corrected to "Following the
investigation of the toxicity of CTNP/B.b-sup on Caco-2 cells." It is advised to review and
improve the abstract for clearer and more fluent language.
(2) Introduction: It is recommended to divide the introduction into three paragraphs to make the
logical flow clearer:
>First Paragraph: Background and Problem Statement
Content: Describe the global prevalence of colorectal cancer (CRC) and its high mortality rate.
Briefly introduce current treatment options (e.g., endoscopy, surgery, chemotherapy) and their
side effects and limitations.
>Second Paragraph: Potential of Probiotics in Cancer Treatment
Content: Discuss the potential role of probiotics in cancer treatment, particularly the benefits
of probiotics such as Bifidobacterium bifidum in anti-inflammatory and tumor-suppressing
activities. Elaborate on their mechanisms of action and reference related studies that support
their efficacy against cancer.
>Third Paragraph: Research Objectives and Innovation
Content: Explain the background and potential of using chitosan nanoparticles combined
with B. bifidum culture supernatant (CTNP/B.b-sup). Emphasize the advantages of this new
combination in overcoming the limitations associated with probiotics when used alone in
gastrointestinal environments.
2. Materials and Methods
(1) The methods section lists specific experimental procedures in detail, but it lacks information
about experimental replicates. It is suggested to add the number of repetitions for each
experiment and the method for calculating the average values.
(2) Release Study: The term "48" should be corrected to "48 h" to indicate the time clearly.
3. Results and Discussion
(1) The results section contains too many subsections; it is suggested to consolidate some of the
headings and content for better coherence.
(2) There is insufficient clarity in some statistical analyses, such as in the protein release
experiments. For example, the data on protein release under different pH conditions (Figure 3)
lacks detailed discussion on the statistical significance between groups. It is recommended to
add more comprehensive statistical descriptions.
(3) The discussion regarding the effects of CTNP/B.b-sup on oncogenes and tumor suppressor
genes is too general. It is suggested to cite more relevant literature and discuss the potential
molecular mechanisms in depth. For instance, regarding the inhibition of TGF-# and Bcl2 genes,
further explanation of their roles in signaling pathways and the specific mechanisms of action
of CTNP/B.b-sup is recommended.
4. Scientific Rigor and Logical Coherence
(1) There are some logical issues in the experimental design. For instance, the paper states that
CTNP/B.b-sup significantly regulates CRC signaling pathway genes, but lacks discussion on
the interactions between different pathways. It is recommended to supplement information on
the complementary or synergistic effects of CTNP and B.b-sup within signaling pathways.
(2) The data in the tables and figures lack detailed information about the control groups,
particularly in the Caco-2 cell experiments. It is suggested to provide more explicit control group
settings and results discussion.
5. Language and Structure
There are some grammatical errors and improper word choices throughout the article, affecting
its professionalism and readability. It is suggested to have the entire manuscript professionally
edited for academic English, especially in the results and discussion sections.
6. Figures
The SEM images of the nanoparticles in Figure 2 are not clear enough. Please dilute the sample
to an appropriate concentration and retake the images, and consider using TEM for better
visualization.