{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T01:51:18Z","timestamp":1791856278564,"version":"4.3.4"},"update-to":[{"DOI":"10.12688\/f1000research.152876.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T00:00:00Z","timestamp":1757980800000}}],"reference-count":30,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T00:00:00Z","timestamp":1757980800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012521","name":"Universitas Gadjah Mada","doi-asserted-by":"publisher","award":["5075\/UNI.P.II\/Dit-Lit\/PT.01.01\/2023"],"award-info":[{"award-number":["5075\/UNI.P.II\/Dit-Lit\/PT.01.01\/2023"]}],"id":[{"id":"10.13039\/501100012521","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns3:p>Background Pelvic organ prolapse (POP) is a prevalent condition caused by weakened pelvic floor support structures. Extracellular matrix alterations, including changes in collagen type I, collagen type III, and matrix metalloproteinase 1 (MMP-1), contribute to the pathogenesis of this condition. Human platelet-rich fibrin lysate (hPRF-L) is a novel regenerative treatment that has shown beneficial results in treating structural weaknesses related to various pelvic floor diseases, including POP. Methods This study protocol aims to investigate the effects of hPRF-L injection on collagen I, III, and MMP-1 in the vaginal mucosa of a rat POP model. POP will be induced in female Sprague-Dawley rats, which will be randomly assigned to control, sham, and hPRF-L treatment groups. The hPRF-L group will receive weekly injections of hPRF-L (25, 50, or 75 \u03bcL) into the vaginal mucosa for 4 weeks. Vaginal tissue samples will be collected, and collagen type I, collagen type III, and MMP-1 expression will be evaluated using quantitative reverse transcription polymerase chain reaction and immunohistochemical analyses. Data analysis will be performed with ANOVA and post-hoc tests. Discussion The findings from this study protocol are expected to provide valuable insights into the mechanisms by which hPRF-L impacts the structural integrity of the pelvic floor. By elucidating these mechanisms, this study aims to inform future POP treatment strategies. The anticipated results are an increase in collagen type I and III expression and a reduction in MMP-1 levels in the hPRF-L treatment group compared to the control and sham groups. These outcomes could support the use of hPRF-L as a regenerative therapy for managing POP, offering a potential alternative to more invasive surgical interventions. Conclusion The expected results will contribute to the development of less invasive treatments for POP, improving patient outcomes and quality of life.<\/ns3:p>","DOI":"10.12688\/f1000research.152876.2","type":"journal-article","created":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T03:00:07Z","timestamp":1759892407000},"page":"1056","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Effect of human platelet-rich fibrin lysate on collagen type I, collagen type III, and matrix metalloproteinase 1: a protocol study on rat models with pelvic organ prolapse"],"prefix":"10.12688","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8829-786X","authenticated-orcid":false,"given":"Akbar Novan Dwi","family":"Saputra","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4775-282X","authenticated-orcid":false,"given":"Dicky Moch","family":"Rizal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nandia","family":"Septiyorini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad Nurhadi","family":"Rahman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yohanes Widodo","family":"Wirohadidjojo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1126-4939","authenticated-orcid":false,"given":"Dwi cahyani Ratna","family":"Sari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raden Mas Sonny","family":"Sasotya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2025,9,16]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"3-11","DOI":"10.1089\/gyn.2022.0049","article-title":"Pelvic Organ Prolapse: Overview, Diagnosis and Management.","volume":"39","author":"S El-Nashar","year":"2023 Feb 1","journal-title":"J. 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Rationale for Protocol Study Publication  \n<b>Reviewer comment:<\/b> &quot;My major comment is related to the rationale of writing a protocol study. What was your point for writing a protocol study.&quot; \n<b>Author\u2019s Response\/ Revision:<\/b> We appreciate this fundamental question about our publication strategy. The rationale for publishing this protocol study is multifaceted and follows established best practices in biomedical research: \n<b>- Scientific Transparency and Reproducibility:<\/b> Protocol publication enhances research transparency by providing detailed methodological information before study completion, reducing the risk of selective reporting and post-hoc modifications to study design. This approach aligns with the CONSORT guidelines for protocol publications and strengthens the overall scientific integrity of our research program. \n<b>- Methodological Innovation:<\/b> Our study introduces novel methodological approaches, including the use of human platelet-rich fibrin lysate (hPRF-L) in a rat POP model and specific outcome measures combining molecular, histological, and functional assessments. Publishing the protocol allows the scientific community to evaluate, critique, and potentially adopt or modify our methodology before results are available. \n<b>- Contribution to Evidence Base:<\/b> This protocol addresses identified gaps in regenerative therapy research for POP, and early publication facilitates collaboration and prevents unnecessary duplication of research efforts.   2. Surgical Complications Statement  \n<b>Reviewer comment:&nbsp;<\/b> &quot;In the Introduction part in second chapter starting 'Pelvic floor reconstruction surgery...'. In my opinion the 'severe bowel injuries leading to postoperative sepsis' is too strongly stated. It is true that the bowel complications may occur, however, severe bowel injuries in traditional vaginal POP surgery are extremely rare.&quot; \n<b>Author\u2019s Response\/ Revision:&nbsp;<\/b> We acknowledge this valid concern and agree that our statement was overly dramatic. We have revised this section to reflect more accurate complication and severity. However, it is associated with several risks that must be carefully considered. Surgical complications can include damage to intestines, bladder, rectum or other organs, excessive bleeding due to damaging a vein or artery, and blood clots.  3. Recurrence Rate Statistics  \n<b>Reviewer comment:&nbsp;<\/b> &quot;Further, the recurrence rate 58% within the first year after surgery is too pessimistic. Please check the recurrence rate again, or maybe you could use some kind of range in recurrence rate?&quot; \n<b>Author\u2019s Response\/ Revision:&nbsp;<\/b> We appreciate this important correction. Our cited recurrence rate was indeed overly pessimistic and not representative of current surgical outcomes. We have revised this with more accurate, evidence-based statistics. The recurrence rate of POP after surgery was 37.7%.  4. Blood Donor Clarification  \n<b>Reviewer comment:&nbsp;<\/b> &quot;At page 6 (Preparation of human platelet-rich fibrin lysate): What do you mean by the participants, whose blood was used to make hPRF-L? Were they voluntary donors?&quot; \n<b>Author\u2019s Response\/ Revision:&nbsp;<\/b> We apologize for the confusion in our terminology. We have clarified this section to eliminate ambiguity about human involvement. The hPRF-L will be prepared from venous blood collected from &nbsp;one healthy female volunteer (age 25-35 years, nulliparous) following informed consent procedures approved by the Institutional Ethics Committee (Ref.No: KE\/FK\/0092\/EC\/2023). The following criteria will be used: \n<b>Inclusion Criteria<\/b>  \n<b>Healthy female, aged 25-35 years:<\/b> To ensure a consistent and healthy donor pool, we will only include healthy female between 25 and 35 years old. This age range is selected to minimize variability associated with age-related physiological changes that could affect blood composition. \n<b>Nulliparous status:<\/b> We will only include nulliparous donor to eliminate the potential influence of pregnancy and childbirth on blood factors, which could alter the composition of the platelet-rich fibrin lysate (hPRF-L). \n<b>No history of pelvic floor disorders:<\/b> To avoid any confounding variables related to pre-existing pelvic conditions, we will exclude donor with a history of pelvic floor disorders. \n<b>No use of hormonal contraceptives or medications affecting coagulation:<\/b> The use of hormonal contraceptives or other medications that could impact blood coagulation will be an exclusion criterion to ensure the quality and consistency of the hPRF-L preparation. \n<b>Normal complete blood count and coagulation parameters:<\/b> Donor must have normal complete blood count and coagulation parameters to ensure the safety of blood donation and the quality of the hPRF-L product.  \n<b>Exclusion Criteria<\/b>  \n<b>Pregnancy or breastfeeding:<\/b> Pregnancy and breastfeeding are known to alter hormonal and physiological states, which could affect blood composition. \n<b>History of autoimmune disorders:<\/b> Autoimmune disorders can impact blood and inflammatory markers, making these donors unsuitable for providing a consistent hPRF-L product. \n<b>Anticoagulant therapy:<\/b> Donor undergoing anticoagulant therapy will be excluded to prevent any interference with the natural coagulation process required for hPRF-L preparation. \n<b>Active infection or inflammatory conditions:<\/b> Active infections or inflammatory conditions can alter the donor's blood composition, including cytokine and growth factor profiles, which could affect the therapeutic properties of hPRF-L. \n<b>Smoking or excessive alcohol consumption:<\/b> These lifestyle factors are known to influence overall health and blood parameters, and thus will be used as an exclusion criterion to maintain the consistency of the donor pool.    5. Overstated Efficacy Claims  \n<b>Reviewer comment:&nbsp;<\/b> &quot;At page 9 in the discussion part, you are writing that the hPRF-L used in this study is more advanced and possibly more efficient method for regenerative therapy in POP. I think that is overstated based on this protocol study writing. I think you cannot say that it may be more efficient than PRP since you are just starting to study it.&quot; \n<b>Author\u2019s Response\/ Revision:&nbsp;<\/b> This is an excellent point, and we completely agree that our language was inappropriately definitive for a protocol study. We have revised this section to reflect the investigational nature of our work. Our study will investigate whether hPRF-L offers theoretical advantages over traditional PRP based on established principles of growth factor biology. The lysate preparation may potentially provide more consistent growth factor release compared to intact platelet preparations, as suggested by in vitro studies in other tissue types. Unlike PRP, which requires the use of anticoagulants and external thrombin to activate platelets, hPRF-L is produced using a more natural method that uses the patient\u2019s own blood coagulation. The intrinsic coagulation process enables the entrapment of a broader range of growth factors, cytokines, and other biologically active substances within the fibrin structure. Subsequently, these substances are gradually released over a period of time to facilitate tissue regeneration and restructuring. Furthermore, the absence of anticoagulants and the limited use of centrifugation processes in the hPRF-L preparation protocol aid in maintaining the integrity and functionality of the platelets and their associated biomolecules. \n<sup>14<\/sup>\n<sup> ,<\/sup> \n<sup>15<\/sup>\n<sup> ,<\/sup> \n<sup>24<\/sup>\n<sup> <\/sup>This can improve the therapeutic effectiveness of hPRF-L compared with PRP. Moreover, the fibrin-based composition of hPRF-L offers a more enduring and consistent release of bioactive substances. This characteristic can be beneficial in managing the persistent and advancing nature of POP. However, the clinical superiority of hPRF-L over conventional PRP remains hypothetical and requires empirical validation. The theoretical advantages of hPRF-L are based on extrapolation from growth factor kinetics studies rather than direct comparative evidence. This limitation underscores the importance of systematic investigation through well-designed preclinical studies such as the one described in this protocol. This protocol study is designed to provide initial mechanistic evidence for hPRF-L effects on collagen metabolism in a controlled animal model. The outcomes of this investigation will inform whether hPRF-L merits further development and clinical evaluation. We emphasize that any claims regarding therapeutic superiority must await completion of this study and subsequent clinical trials.","URL":"https:\/\/f1000research.com\/articles\/13-1056\/v2#referee-comment-14556","order":1,"name":"referee-comment-14556","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.188094.r414776, Kristina Allen-Brady, The University of Utah, Salt Lake City, Utah, 26 Sep 2025, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/13-1056\/v2#referee-response-414776","order":2,"name":"referee-response-414776","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.188094.r415257, Nur Anisah Rahmawati, Universitas Airlangga, Jawa Timur, 08 Oct 2025, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/13-1056\/v2#referee-response-415257","order":3,"name":"referee-response-415257","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"The authors  received funding from Universitas Gadjah Mada for the Final Project Recognition Grant, with reference number 5075\/UNI.P.II\/Dit-Lit\/PT.01.01\/2023. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":4,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}