Calcium Phosphate Treatment Enhances Osteogenic Differentiation of Porcine Adipose-Derived Stem Cells on Fibrin Scaffolds

Calcium Phosphate Treatment Enhances Osteogenic Differentiation of Porcine Adipose-Derived Stem Cells on Fibrin Scaffolds

Review Data

Q: Is the topic relevant to the journal area of interest? Is it contemporary and interesting for researchers?

Comments: Yes, this study is indeed relevant to the field of regenerative medicine. The development and optimization of biological scaffolds like the fibrin scaffold discussed in this study are vital topics within this field. Such scaffolds are key in tissue engineering and regenerative medicine because they support the growth, proliferation, and differentiation of cells into functional tissues.


The use of porcine adipose-derived stem cells (ASCs) in this study and their osteogenic differentiation is also a pertinent topic in regenerative medicine, given the increasing recognition of the potential of these cells in tissue regeneration and repair.


In addition, the novel approach to improve the fibrin scaffold's characteristics by adding calcium hydrogen phosphate, resulting in enhanced osteogenic differentiation, is an exciting development. It could potentially impact various areas, including bone grafting procedures, healing of bone fractures, and bone tissue engineering.


Moreover, the study is contemporary, considering the growing interest and research in optimizing biomaterials for tissue engineering applications. The findings could spark further research to improve scaffold design and the understanding of cell-scaffold interactions in bone regeneration.


Given all these reasons, the study is likely to be of interest to researchers working in the field of regenerative medicine, biomaterials, and tissue engineering.

Abstract & Keywords

Q: Are all required components included in the abstract? Are the keywords appropriately chosen?

Comments: Yes, the abstract does include all the necessary components. These components are:


Background and Purpose: The study's background is briefly explained, along with the purpose of the investigation, which is to develop a fibrin scaffold for optimum in vitro osteogenic differentiation of porcine ASCs.


Methods: The methods are described, including the creation and testing of different types of fibrin scaffolds.


Results: The primary findings are presented, notably that fibrin supplemented with granules of calcium hydrogen phosphate was found to be the most suitable formulation.


Conclusion: The abstract concludes with the implications of the findings and a brief mention of their potential applications in surgical operations and bone tissue engineering.


As for the keywords, they appear to be appropriately chosen and represent the main concepts explored in the study. The chosen keywords, "ASC," "fibrin," "calcium phosphate," "porcine," and "osteogenesis," will help researchers in related fields locate this study during their literature searches. These keywords accurately reflect the study's focus on osteogenic differentiation of adipose-derived stem cells (ASCs) in porcine models using calcium phosphate-modified fibrin scaffolds.


Q: Is the goal explicitly stated in the Introduction? Is its formulation clear and unambiguous?

Comments: Yes, the goal of the study is explicitly stated in the introduction. The authors state: "This study was designed to develop a fibrin scaffold for optimum in vitro osteogenic differentiation of porcine ASCs as a model system for human bone regeneration."


This formulation is clear and unambiguous as it explicitly outlines what the study intends to achieve: developing an optimized fibrin scaffold to enhance osteogenic differentiation of porcine Adipose-derived Stem Cells (ASCs), and this work is positioned as a model for human bone regeneration.


The introduction lays out the reasons for this study, citing the need for modification to fibrin scaffolds to improve their suitability for bone repair and the lack of existing research into methods to increase the stiffness of autologous fibrin scaffolds prior to stem cell culture and differentiation. This clear positioning and contextualization further support the clarity of the study's goal.


Q: Is the paper's structure coherent? Is it in coherence with the goal of the paper?

Comments: Yes, the structure of the paper is coherent and in line with the goal of the research paper. The overall structure adheres to the standard format of scientific research papers, which consists of the Abstract, Introduction, Materials and Methods, Results, Discussion, and Conclusion sections.


In the Introduction, the authors effectively presented the background information about the role of fibrin in the bone fracture healing process and the potential limitations of fibrin-based treatments. The hypothesis was clearly stated, setting up the stage for the study and the experimental design.


The Materials and Methods section detailed the steps taken to achieve the research goal, including blood collection, fibrin modification, and various treatment methods. This section is crucial for reproducibility and validation of results by other researchers in the field.


The Results section provided a clear and precise account of the findings obtained from the different treatments. The authors included statistical analysis and discussed the significance of the results.


The Discussion section effectively interpreted the results in the context of the research question. The authors also highlighted the potential implications of their findings in improving ASC therapy for bone defects, and they critically analyzed their methods and findings, including potential limitations.


The Conclusion succinctly summarized the findings and offered suggestions for future research, particularly the need for in vivo testing of CaHPO4-treated fibrin scaffolds.


Overall, the paper’s structure is coherent, and its content aligns with the stated goal: to develop an optimal preparation of whole blood-derived fibrin scaffold that is practical, biocompatible, and osteoconductive. It also offered a clear and logical progression from background, through experimental design and results, to interpretation and conclusion, maintaining a clear focus on the research goal throughout.

Tools and Methods

Q: Are methods the author uses adequate and well used?

Comments: The methods used in this study seem well considered and adequately applied for the research question. The researcher uses multiple techniques and procedures that collectively answer the questions posed in the study. Here are the key aspects:


1.     Fibrin Modification: The method used to modify fibrin by treating it with different substances (calcium chloride, calcium hydrogen phosphate, RBC lysis buffer, vacuum, and mechanical compression) is well outlined. This modification process is crucial to the experiment, as it directly relates to the study's goal of enhancing osteogenic differentiation.


2.     Coagulation Time: The timing of coagulation under various treatments is critical data to consider, as the coagulation rate affects the quality of the fibrin scaffold. The method used to measure this, although simple, is quite effective.


3.     In Vitro Degradation & Microstructural Analysis: The study uses standard techniques such as scanning electron microscopy (SEM) for evaluating the microstructure of the fibrin scaffolds, which is quite appropriate for the analysis of the treatment effects.


4.     Mechanical Testing: The use of standard mechanical testing procedures to assess the scaffold's strength and stiffness is suitable for assessing the physical properties of the fibrin scaffolds.


5.     ASC Co-Culture and Osteogenic Differentiation: The culture and osteogenic differentiation procedure for ASCs is well-documented. The imaging and subsequent quantification methods used to evaluate osteogenic differentiation seem appropriate and precise.


6.     Statistical Analysis: The use of one-way ANOVA and Fisher’s exact test are appropriate statistical methods for this study. The number of scaffolds and nodules analyzed per treatment also seems adequate.


7.     Microscopic and Histomorphometric Analysis: These are well-suited methods for evaluating cellular attachment, migration, and formation of osteogenic nodules.


In summary, the methods used in this study are adequate and well-executed, providing a comprehensive approach to answer the research question. They collectively allow for the evaluation of multiple aspects of the modified fibrin scaffolds' properties and their influence on osteogenic differentiation. Furthermore, the methods are sufficiently detailed to allow for the replication of this study by other researchers.

Discussion & Conclusion

Q: Is it related to the results presented before? Do you consider them as coherent?

Comments: Based on the information provided in the discussion and conclusion, it appears that the authors have logically connected their results to the discussion and have drawn a coherent conclusion from their findings.


In the discussion, they explained the observed effects of the different treatments on fibrin scaffolds and compared these to the expected outcomes based on current scientific knowledge. They also discussed potential mechanisms behind the results and provided detailed explanations for the results obtained.


The conclusion also seems coherent and closely tied to the results. They concluded that treating fibrin scaffolds with calcium phosphate improved several characteristics, which was clearly demonstrated in their results.


Overall, the authors have presented a clear thread from their results to the discussion and conclusion, and they have stayed consistent with the aim of their study. This coherence helps strengthen the validity of their findings and their potential implications in the field of bone tissue engineering.


Q: Does the author utilize relevant literature?

Comments: Yes, the authors do seem to utilize relevant literature appropriately. They cite multiple sources when discussing the context of their study, such as existing research on the properties of fibrin and calcium hydrogen phosphate, the applications of these materials in tissue engineering, and the methods used for scaffold fabrication and characterization.


The authors also reference relevant literature when discussing the results of their experiments. For instance, they cite studies on the role of calcium ions in coagulation, the impacts of scaffold stiffness and roughness on cell behavior, and the effects of different materials on osteogenic differentiation. This not only demonstrates their thorough understanding of the field, but also allows readers to follow their line of reasoning and verify the sources of their information.


In the discussion and conclusion, the authors again reference key studies, comparing their results with those from other research and explaining their findings in light of existing knowledge in the field. This strengthens the validity of their interpretations and helps place their study within the larger body of scientific literature.


Q: Is the length of the paper adequate to the significance of the topic? Do you suggest shortening the paper without losing its value?

Comments: Given the complex nature of the topic, the current length of the paper appears to be adequate. Bone tissue engineering is a multidisciplinary field that requires in-depth discussion and explanation to properly convey the findings. The author successfully details the experimental design, the observed results, their interpretation, and their significance within the broader context of bone tissue engineering.


The paper is also broken down into clearly defined sections, each serving a specific purpose, which improves readability and comprehension:


Introduction: Provides background information, establishes the research context, and states the research objective.

Materials and Methods: Describes the experimental design in detail.

Results: Presents the data gathered in a clear and precise manner.

Discussion: Interprets the results and draws connections with existing literature.

Conclusion: Summarizes the findings and outlines potential future research directions.

Any substantial reduction in the length of the paper could risk leaving out essential details or context, which would lessen its overall value.


Writing style

Q: Is it clear and understandable?

A: Yes, the abstract, introduction, methods, results, discussion, and conclusion all provide a clear, well-explained, and comprehensive description of your study. The use of fibrin scaffold supplemented with calcium hydrogen phosphate (CaHPO4) for enhancing in vitro osteogenic differentiation of porcine adipose-derived stem cells (ASCs) is thoroughly explored.

Overall, the document is detailed, scientifically rigorous, and well-structured, making it easily understandable to those with a background in the field.

Further comments on the paper


The study appears to be well-conducted with a logical, systematic approach. Here are a few additional comments:

1.     Clarity and Depth: The research paper is thorough and detailed. The introduction provides enough background information about fibrin, its role in wound healing and the potential benefits of modifying it to enhance stem cell therapy. The methods are clearly described and the results are well presented.

2.     Hypothesis and Investigation: The hypothesis is clearly stated, and the methods used to investigate it are appropriate and well explained. The researchers have considered several different methods of modifying fibrin scaffolds and examined their effects on coagulation time, in vitro degradation, mechanical properties, and osteogenic differentiation.

3.     Statistical Analysis: The statistical analysis seems to be well conducted, with appropriate tests applied. A potential improvement could be to provide exact p-values rather than just noting when they were less than 0.05 or 0.1. This can give the reader a clearer sense of the strength of the findings.

4.     Interpretation: The interpretation of the results aligns well with the data presented. It appears that modifying fibrin scaffolds with calcium hydrogen phosphate can improve several properties relevant to their potential use in surgical procedures and bone tissue engineering.

5.     Conclusion: The conclusion is appropriate, reflecting the results and suggesting the direction for future research. It's good that the authors acknowledge the need for further testing, especially in vivo studies, to confirm the potential benefits of the modified fibrin scaffolds.

Overall, this is an intriguing piece of research that suggests a promising direction for future studies.

Q: Would you recommend this manuscript for further publication?

A: Yes - This manuscript is recommended for further publication.

If you have any questions and clarifications you can write to the journal.

Science Repository Team


Author Info

Corresponding Author
Matthew B. Wheeler
Department of Animal Sciences, University of Illinois, Urbana-Champaign, Urbana, Illinois, USA

Article Info

Article Type
Research Article
Publication history
Received: Thu 04, May 2023
Accepted: Thu 18, May 2023
Published: Tue 13, Jun 2023
© 2023 Matthew B. Wheeler. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Hosting by Science Repository.
DOI: 10.31487/j.RGM.2023.02.01