Bone Healing

This study is about fracture healing and the development of new diagnostic methods essential for
diagnosing and treating chronic nonunion. Chronic Union is a fracture that persists for a minimum of
nine months without signs of healing for three months. This study explores the potential of using
Dipsaci Radix (DR), a traditional Chinese medicine, for treating chronic nonunion and improving bone
health. It involves bioinformatics analysis, microarray data, and systems pharmacology to identify
biomarkers for chronic nonunion and understand DR's mechanismsl.
The research first identifies differentially expressed genes in peripheral blood

mononuclear cells (PBMCs) related to chronic nonunion. Then, it screens for active ingredients in DR and
their target genes, finding potential biomarkers that overlap with DR's targets. Finally, the predicted
biomarkers are validated using experimental methods like ELISA and qPCR with patient blood samples.
This study offers insights into blood biomarkers for chronic nonunion and the mechanisms behind DR's
treatment potential.
The study utilized :
1. Microarray datasets (GSE93138, GSE93213, GSE93215) to identify differentially expressed genes

(DEGs) related to fracture healing, applying limma analysis with specific criteria.
Bioactive ingredients of Dipsaci Radix (DR) were sourced from the TCMSP database based on drug-
likeness and oral bioavailability thresholds. Corresponding target genes for these compounds were
identified from the PharmMapper and Swiss Target Prediction databases.
Protein-protein interactions among DEGs of healed and nonunion patients were analyzed using the
STRING database, and hub genes were determined. The study also established interactions between
DEGs and DR target genes. A drug-disease network was constructed using Cytoscape software, and the
GeneCards database was used to assess potential biomarker functions. Additionally, Gene Ontology
enrichment analysis was performed using DAVID to identify the biological aspects involving the
predicted target genes.
Blood samples were collected from participants and processed to obtain serum, which was stored at -
80°C. The serum biomarker concentrations were determined using ELISA kits. Total RNA was extracted
from PBMCs, reverse transcribed and mRNA expression was analyzed. Specific primers were used for
qPCR analysis. Statistical analysis was conducted with GraphPad Prism software version 7.0. Data are
presented as mean ± SEM. Comparisons between groups were assessed using two-tailed unpaired
Student's t-tests or one-way/two-way ANOVA as appropriate. Categorical data were analyzed using the
chi-squared (χ2) test, with significance set at p < 0.05.
In the study, three PBMC gene expression datasets were obtained from the GEO database, comparing
healed and nonunion patients. A total of 258 differentially expressed genes were identified between
these groups. To investigate the relationships among these genes, protein-protein interaction (PPI)
analysis was conducted using the STRING database, leading to the creation of a network. Within this
network, hub genes were identified using Cytoscape software, with the top 20 hub genes highlighted in
red and yellow, indicating their central importance. Notably, hub genes like RPS27A, RPS17, and RBX1
were found to potentially have a critical role in the biological activity of chronic nonunion.
In this study, Dipsaci Radix (DR), a traditional medicine for bone fracture treatment, was used to predict
biomarkers for nonunion patients. Ten potential active compounds from DR were identified from the
TCMSP database, and 443 target genes were obtained from the PharmMapper and Swiss Target
Prediction databases. By comparing drug target genes with nonunion-related genes, six interacting
genes were identified. Five active DR ingredients (Japonine, Gentisin, Gauloside A, Sylvestroside III, and
3,5-Di-O-caffeoylquinic acid) and six target genes (QPCT, CA1, LDHB, MMP9, UGCG, and HCAR2) were
found to be related to nonunion.
ELISA tests were conducted to assess six potential biomarkers in serum samples from healthy control
(HC) and acute injury (AI) patients. The serum concentrations of all six biomarkers significantly increased
in the AI group (p < 0.05), suggesting their association with the fracture healing process. Notably, there
were no significant differences in these biomarker concentrations between fracture-only and multi-
trauma patients.
To further validate the potential biomarkers CA1, MMP9, and QPCT in nonunion patients, qPCR analysis
was conducted on PBMCs from different patient groups. The results demonstrated significant changes in
PBMC mRNA expression levels of CA1, MMP9, and QPCT in the nonunion groups (p < 0.05). Additionally,
DR treatment significantly increased the PBMC mRNA expression levels of CA1, MMP9, and QPCT in
both healed and nonunion groups (p < 0.05), consistent with the ELISA findings. These results confirm
that CA1, MMP9, and QPCT can indeed serve as biomarkers for identifying nonunion patients, and DR
treatment appears to target these biomarkers.

Bone Healing

Uploaded by

Copyright:

Available Formats

Bone Healing

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Bone Healing

Uploaded by

Copyright:

Available Formats

This study is about fracture healing and the development of new diagnostic methods essential for

The research first identifies differentially expressed genes in peripheral blood

The study utilized :

1. Microarray datasets (GSE93138, GSE93213, GSE93215) to identify differentially expressed genes

You might also like