Abstract
Background
Guided bone self-generation with periosteum-preserved has successfully regenerated mandibular, temporomandibular and interphalangeal joint. The aim of this study was to investigate the dynamic changes of gene expression of periosteum which was involved in the guided bone self-generation.
Methods
Rib defects of critical size were created in mature swine with periosteum-preserved. The periosteum was sutured into a sealed sheath that closed the bone defect. The periosteum of trauma and control sites were harvested at postoperative 9 time points, and total RNA was extracted. Microarray analysis was conducted to identify the differences in the transcriptome of different time points between two groups.
Results
The differentially expressed genes (DEGs) between control and trauma group were different at postoperative different time points. The dynamic changes of the number of DEGs fluctuated a lot. There were 3 volatility peaks, and we chose 3 time points of DEG number peak (1 week, 5 weeks and 6 months) to study the functions of DEGs. Oxidoreductase activity, oxidation–reduction process and mitochondrion are the most enriched terms of Go analysis. The major signaling pathways of DEGs enrichment include oxidative phosphorylation, PI3K-Akt signaling pathway, osteoclast differentiation pathway and Wnt signaling.
Conclusions
The oxidoreductase reaction was activated during this bone regeneration process. The oxidative phosphorylation, PI3K-Akt signaling pathway, osteoclast differentiation pathway and Wnt signaling may play important roles in the guided bone self-generation with periosteum-preserved. This study can provide a reference for how to improve the application of this concept of bone regeneration.
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Introduction
Periosteum, a highly vascularized connective tissue, plays a key role in the growth, development and regeneration of bone [39]. Fracture repair was considered to be a recapitulation of embryonic development, and members of the Wnt signaling pathway were activated [40]. Nan et al. found that Wnt signaling pathway was activated during bone regeneration [40]. The Wnt signaling pathway may play an important role in the regenerative process.
Several strategies, including gene therapy and tissue engineering together with mesenchymal stem cells (MSC), have been proposed to promote the healing of the musculoskeletal tissue. Moreover, a recent technology has revolutionized gene editing: Clustering regulatory interval short palindromic repeats (CRISPR) features simple target design, affordable, versatile and efficient, but requires more research to be the preferred platform for genome editing. Predictive genomics DNA analysis can understand which genetic advantages (if any) can be exploited and why specific rehabilitation programs are more effective in some people than others [41, 42]. Therefore, a better understanding of the genetic impact on musculoskeletal system function and disease healing is needed to plan and develop patient-specific management strategies. Currently, while some results are promising, all biological interventions are experimental and the cost/effectiveness has not been proven. In addition, the short follow-up time of most studies questioned the durability of treatment [42]. In this study, autologous periosteum was used to guide bone regeneration in vivo, and the regenerated bone was used for precise repair of the body. This technology has a promising clinical application prospect. The related differential genes and signaling pathways identified in this study can provide a rich theoretical basis for later gene and molecular intervention.
Though this in vivo study provides a better understanding in the molecular mechanisms involved in guided self-generation, it also has limitations. First, micro-CT can be conducted to evaluate the conditions of bone regeneration at different time points. The association of changes of molecules and bone regeneration can be analyzed to provide more precise explanations of the mechanisms. Besides, different parts of the regeneration may be at different stage of healing process. Immuno-histochemistry to localize the transcript and protein expression may provide deeper appreciation of the function of specific genes.
Conclusions
This study shows the guided bone regeneration involves DEGs associated with oxidation–reduction process, mitochondrion and oxidoreductase activity. The main signaling pathways includes oxidative phosphorylation, PI3K-Akt signaling pathway, osteoclast differentiation pathway and Wnt signaling. This study could deepen our understanding of the molecular mechanisms involved in the guided bone regeneration. With these findings of molecular changes at different time points, it gains the potential of regulating the specific mechanism to enhance the bone regeneration.
Availability of data and materials
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
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Funding
This research was supported by grants from the Shanghai Municipal Key Clinical Specialty (shslczdzk00901 to J.W) and the National Natural Science Foundation of China (No. 81871572).
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DCC and WJ designed the study and supported funding. YBF, WZ and CXX were major contributors in writing and reviewing the manuscript. YBF, WZ, CXX, ZQ and YN completed experiments and analyzed the data. All authors read and approved the final manuscript.
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Supplementary Information
Additional file 1: Figure S1
. Volcano maps of the DEGs between control and trauma group at postoperative different time points (A: 1 day, B: 3 days, C: 1 week, D: 2 weeks, E: 1 month, F: 5 weeks, G: 3 months, H: 6 months, I: 7 months).
Additional file 2: Figure S2
. Results of cluster analysis revealed unknown biological connections between genes through expression clustering.
Additional file 3: Figure S3
. Top 10 GO term entries with the smallest p-value. A, The top 10 GO term entries of postoperative 1 week. B, The top 10 GO term entries of postoperative 5 weeks. C, The top 10 GO term entries of postoperative 6 months.
Additional file 4: Figure S4
. PI3K-Akt signaling pathway.
Additional file 5: Figure S5
. Osteoclast differentiation pathway.
Additional file 6: Figure S6
. Wnt signaling pathway.
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Yu, BF., Wang, Z., Chen, XX. et al. Continuous dynamic identification of key genes and molecular signaling pathways of periosteum in guided bone self-generation in swine model. J Orthop Surg Res 18, 53 (2023). https://doi.org/10.1186/s13018-023-03524-y
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DOI: https://doi.org/10.1186/s13018-023-03524-y