Bifidobacterium longum BORI inhibits pain behavior and chondrocyte death, and attenuates osteoarthritis progression

Dong Keon Oh; Hyun Sik Na; Joo Yeon Jhun; Jeong Su Lee; In Gyu Um; Seung Yoon Lee; Myeong Soo Park; Mi-La Cho; Sung-Hwan Park

doi:10.1371/journal.pone.0286456

Peer Review History

Original SubmissionJanuary 20, 2023
3 Apr 2023 Decision Letter - Win Min Oo, Editor PONE-D-23-01788Bifidobacterium longum BORI reduces pain behavior and attenuates the progression of monosodium iodoacetate-induced osteoarthritisPLOS ONE Dear Dr. Cho, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== ACADEMIC EDITOR:This is a well written manuscript but some major improvements are required as indicated by the reviewer 1. The authors are suggested to correct the typo errors in the revised version. ============================== Please submit your revised manuscript by May 18 2023 11:59PM. 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Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Bifidobacterium longum BORI reduces pain behavior and attenuates the progression of monosodium iodoacetate-induced osteoarthritis Reviewer comments This paper submitted by the authors investigated whether B. longum BORI modulates arthritic symptoms and accompanying pain symptoms in an osteoarthritis model. Probiotics such as B. longum are currently being investigated and published for their disease-modulating effects in various inflammatory diseases. The results presented in this paper will add to our knowledge of probiotic therapies that can inhibit articular cartilage damage and pain in osteoarthritis models. I have several questions regarding this study. 1) In Background section authors present the topic well and with appropriate reference however little is said about the rational of their study. My suggestion is to expand on the relevance of in vitro models used, the bacteria strains they used, why they used that strains and the relevance these strains might have. 2) The author seems to have partially mentioned the number of experiments in the experiment method and results. It is desirable that the total number of experiments performed in this part be accurately reflected. 3) Why are some experiments used after killing B. longum BORI? An explanation is needed as to why dead bacteria were used. 4) In methods, "Preparation of bacteria" it is not mentioned what sample was used for bacteria isolation 5)In the same section, "Immunohistochemical staining of joint synovium showed that administration of B. longum BORI reduced the levels of IL-1b, LTB4r, PGE2, MMP9 and MMP13 (Figures 3 and 4)." . Histology in the method of Figures 3, 4 and 4 is not clear how to observe the reduction of the listed cytokines. It is necessary to add a description of how the cytokine reduction is observed. 6) The title refers only to experiments with rats. The experiment also includes the contents of the experiment in the patient's chondrocyte culture, so it would be good to mention these parts in the title. 7) In figure, B. longum BORI was not mentioned in Italics. It should be corrected throughout the Figure. 8) The authors should better check the manuscripts for any typographical errors. For example, LTB4 must be modified to LTB4r in figure 3. ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0286456.r001
Revision 1
6 Apr 2023 Author Response Reviewer comments This paper submitted by the authors investigated whether B. longum BORI modulates arthritic symptoms and accompanying pain symptoms in an osteoarthritis model. Probiotics such as B. longum are currently being investigated and published for their disease-modulating effects in various inflammatory diseases. The results presented in this paper will add to our knowledge of probiotic therapies that can inhibit articular cartilage damage and pain in osteoarthritis models. I have several questions regarding this study. 1) In Background section authors present the topic well and with appropriate reference however little is said about the rational of their study. My suggestion is to expand on the relevance of in vitro models used, the bacteria strains they used, why they used that strains and the relevance these strains might have. Response: We agree with referee’s comment. We revised the introduction section (marker by red color) Monosodium iodoacetate (MIA)-induced OA rats are widely used as a human osteoarthritis model, by destroying cartilage cells, causing degeneration and inflammation of the cartilage[14]. This causes symptoms similar to those of thoracic osteoarthritis. The administration of B. BORI is safe and has demonstrated positive effects in terms of inhibiting inflammation in cranial nerve cells, as well as hepatic and intestinal conditions [10, 15]. Furthermore, B. BORI demonstrated the maintenance of gut microbiota balance and stability by showing effectiveness in preventing and treating rotavirus infection in infants[16]. 2) The author seems to have partially mentioned the number of experiments in the experiment method and results. It is desirable that the total number of experiments performed in this part be accurately reflected. Response: Thank you for your comments. Osteoarthritis was induced in Wistar rat (n=6). The experiment was performed two times. We revised the Material and method section (marker by red color) 3) Why are some experiments used after killing B. longum BORI? An explanation is needed as to why dead bacteria were used. Response: Heat-killed probiotics are reportedly safer than live probiotics for eliminating antibiotic-resistance genes, preventing the production of recombinant strains, and controlling the microbial load during probiotic supplementation (Ishibashi, N. & Yamazaki, S. Probiotics and safety. Am. J. Clin. Nutr. 73,465S–470S (2001); Vintini, E.O. & Medina, M.S. Host immunity in the protective response to nasal immunization with a pneumococcal antigen associated with live and heat-killed Lactobacillus casei. BMC Immunol. 12, 46 (2011)). Thus, we elected to use heat killed B. longum BORI in this study. 4) In methods, "Preparation of bacteria" it is not mentioned what sample was used for bacteria isolation The authors would like to thank the comment of the reviewer. Following the reviewer’s comment, the authors revised the reference as follows: (marker by red color) B. longum BORI was isolated from human feces of healthy infant who lived in Chuncheon, South Korea between 1995 and 1998, identified with 16S rRNA sequencing and cultured with MRS (De Man, Rogosa and Sharpe, Becton Dickson, Franklin Lakes, NJ, USA) broth medium at 37 ◦C. For the initial screening, lyophilized powder of each strain was used. Freeze-dried powder of B. longum BORI was provided by BIFIDO Co. Ltd (Hongchun, Korea) 5)In the same section, "Immunohistochemical staining of joint synovium showed that administration of B. longum BORI reduced the levels of IL-1b, LTB4r, PGE2, MMP9 and MMP13 (Figures 3 and 4)." . Histology in the method of Figures 3, 4 and 4 is not clear how to observe the reduction of the listed cytokines. It is necessary to add a description of how the cytokine reduction is observed. Response: We agree and added the following text in method section. (marker by red color) The all IHC images were obtained each rat, and showing representative images. The percentage of positive cells showing in each image was measured by Image J. 6) The title refers only to experiments with rats. The experiment also includes the contents of the experiment in the patient's chondrocyte culture, so it would be good to mention these parts in the title. Response: Thank you for your comment. We have revised the title to “ Bifidobacterium longum BORI reduces pain behavior and attenuates the progression of monosodium iodoacetate-induced osteoarthritis and � Bifidobacterium longum BORI inhibits Pain behavior and chondrocyte death, and attenuates osteoarthritis progression. 7) In figure, B. longum BORI was not mentioned in Italics. It should be corrected throughout the Figure. Response: Thank you for your comment. We revised the manuscript. 8) The authors should better check the manuscripts for any typographical errors. For example, LTB4 must be modified to LTB4r in figure 3. Response: We apologize for this error, which has been corrected. Attachments Attachment Submitted filename: Rebuttal leter_BORI.docx https://doi.org/10.1371/journal.pone.0286456.r002
17 May 2023 Decision Letter - Win Min Oo, Editor Bifidobacterium longum BORI inhibits Pain behavior and chondrocyte death, and attenuates osteoarthritis progression. PONE-D-23-01788R1 Dear Dr. Cho, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Win Min Oo Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The authors submitted revised manuscript. After review, previous recommandations were well responded and corrected. I think this paper would be acceptable for publication. With regards, ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No ******** https://doi.org/10.1371/journal.pone.0286456.r003
Formally Accepted
13 Jun 2023 Acceptance Letter - Win Min Oo, Editor PONE-D-23-01788R1 Bifidobacterium longum BORI inhibits Pain behavior and chondrocyte death, and attenuates osteoarthritis progression. Dear Dr. Cho: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Win Min Oo Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0286456.r004

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