Project description:10X Genomics single cell RNA-sequencing of freshly isolated nonunion tissue from 63-year old male was conducted to interrogate non-hematopoietic tissue composition. The goal was to generate a dataset that can be compared with publicly available datasets of normal fracture tissue and cells associated with bone healing.
Project description:Background: Bone nonunion is a serious complication of fracture. This study explored the differentially expressed lncRNAs (DELs) and mRNAs (DEGs) and identified potential lncRNA-mRNA interactions in bone nonunion. Methods: We extracted total RNA from three bone nonunion and three bone union patient tissue samples. RNA sequencing was performed to detect DELs and DEGs between bone nonunion and union tissue samples. The lncRNAs and genes with absolute log2-fold change (log2FC) > 1 and adjusted p value < 0.05 were further chosen for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. lncRNA and targeted mRNA interaction networks were constructed. Results: We observed 179 DELs and 415 DEGs between the bone nonunion and union tissue samples. GO analysis indicated that DELs and DEGs were mainly enriched in chondroitin sulfate proteoglycan biosynthetic process. DELs and DEGs were enriched in 'ECM-receptor interaction' and 'Staphylococcus aureus infection' KEGG pathways. Several potential lncRNA-mRNA interactions were also predicted. Conclusions: This study identified bone nonunion-associated lncRNAs and mRNAs using deep sequencing that may be useful as potential biomarkers for bone nonunion.
Project description:20 miRNAs were identified as differentially expressed in patients with infected nonunion of long bones, which may facilitate the diagnosis of infected nonunion of long bones.
Project description:Profiling of miRNA expressions comparing standard fracture healing models with nonunion models in rats 12w, male, Sprague–Dawley rats were used in this study. Animals were randomized to receive either a surgical treatment that has been shown to produce a nonunion or to a standard stabilized closed femoral shaft fracture that is known to successfully heal. The details of these procedures have been previously described. Briefly, to produce standard healing models, a 1.2-mm diameter K-wire was inserted retrograde into the right femoral intramedullary canal and a closed transverse femoral shaft fracture was produced using a three-point bending apparatus with a drop weight . To produce the nonunion, the fractured site was additionally exposed, and the periosteum was cauterized circumferentially for a distance of 2 mm on each side of the fracture . Five animals from each group were euthanized on post-fracture day 14 for microarray analysis.
Project description:Fresh resected human lung tissue (parenchymal lung and distal airway specimens) was obtained via the CPC BioArchive at the Comprehensive Pneumology Center Munich (CPC-M, Munich, Germany) and profiled using single cell RNA sequencing technology (Drop-seq). In total, we analysed parenchymal tissue of uninvolved areas of tumour resection material from four patients.