Proteomics of callus tissue during fracture healing with and without shear stress
Ontology highlight
ABSTRACT:
ORGANISM(S): Oryctolagus Cuniculus
SUBMITTER: 
Weiyong Wu
PROVIDER: PXD045014 | iProX | Thu Aug 31 00:00:00 BST 2023
REPOSITORIES: iProX
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Project description:To identify the changes in gene expression profile caused by mechanical stress in the ligamentum flavum, we performed resection of the L3-4 supraspinal muscle and L2-3, L4-5 posterolateral fusion with instrumentation to concentrate the mechanical stress with segmental instability at the L3-4 level. The control group underwent only surgical exposure as a sham operation. Both groups of rabbits were sacrificed at 16 weeks after surgery and total RNA were extracted from ligamentum flavum.
2022-05-26 | GSE204798 | GEO
Project description:To identify the changes in gene expression profile caused by mechanical stress in the ligamentum flavum, we performed resection of the L3-4 supraspinal muscle and L2-3, L4-5 posterolateral fusion with instrumentation to concentrate the mechanical stress with segmental instability at the L3-4 level. The control group underwent only surgical exposure as a sham operation. Both groups of rabbits were sacrificed at 1 year after surgery and total RNA were extracted from ligamentum flavum.
2023-01-07 | GSE205215 | GEO
Project description:This is a study of femoral fracture healing in female rats 16 weeks old at fracture to compare intramedullary nailing, screw and plate fixation, and sham surgery. The sham surgery group received a surgical exposure of the femur, but no fracture, no plate, and no nail. Samples were collected at 1 day, 3 days, 1 week, 2 weeks, 4 weeks, and 6 weeks after surgery. Each sample is a pool of RNA from three rats from the same surgery group at the same time point after fracture. The middle third of the femur was collected with the cortical bone, fracture callus, and marrow elements. Mid-diaphyseal, simple, transverse fractures were induced by a Gigli saw. The no fracture sample was a time 0 control collected on the day of surgery from intact rats.
2007-11-03 | E-GEOD-1685 | biostudies-arrayexpress
Project description:Seventy-two female Sprague-Dawley rats underwent a 5 mm critical-sized femoral segmental defect followed by stabilization with a custom-made unilateral external fixator. Following femoral defect generation, absorbable type I bovine collagen sponges were loaded either with 50 ug GDF5wt or 50 ug GDF5N445T, dissolved in dilution buffer (10 mM HCl), and placed into the defect. The control group was treated with collagen sponges soaked with dilution buffer. For analysis of the early healing phase, 48 rats (n = 4 rats/group & time point) were sacrificed at day 3, 5, 7 and 14 post-operation and the callus tissue between the two inner pins was harvested and immediately flash frozen in liquid nitrogen for RNA extraction. Tissue was subsequently pulverized and total RNA was isolated using Trifast reagent and RNeasy Mini Kit. The total RNA from 3 animals per group and time point were pooled prior to hybridization on an Affymetrix GeneChip Rat Gene 1.0 ST Array.
2015-01-15 | E-MTAB-2249 | biostudies-arrayexpress
Project description:This is a study of femoral fracture healing in female rats 16 weeks old at fracture to compare intramedullary nailing, screw and plate fixation, and sham surgery. The sham surgery group received a surgical exposure of the femur, but no fracture, no plate, and no nail. Samples were collected at 1 day, 3 days, 1 week, 2 weeks, 4 weeks, and 6 weeks after surgery. Each sample is a pool of RNA from three rats from the same surgery group at the same time point after fracture. The middle third of the femur was collected with the cortical bone, fracture callus, and marrow elements. Mid-diaphyseal, simple, transverse fractures were induced by a Gigli saw. The no fracture sample was a time 0 control collected on the day of surgery from intact rats. Keywords = rat Keywords = fracture Keywords = plate Keywords = nail Keywords = time Keywords = femur Keywords: time-course
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Project description:Genome-wide comparative gene expression analysis of callus tissue of osteoporotic mice (Col1a1-Krm2 and Lrp5-/-) and wild-type were performed to identify candidate genes that might be responsible for the impaired fracture healing observed in Col1a1-Krm2 and Lrp5-/- mice. To investigate bone healing in osteoporosis, we performed fracture healing studies in wild-type mice (C57BL/6 genetic background) and the low bone mass strains Col1a1-Krm2 and Lrp5-/- (Schulze et al., 2010; Kato et al., 2002). Osteotomy was set in femora of female mice and stabilized by a semi-rigid fixator to allow fast bone healing (RM-CM-6ntgen et al., 2010). 21 days post surgery we analyzed the fracture calli by biochemical/histological methods, as well as micro-computed tomography, and observed impaired fracture healing in Col1a1-Krm2 and Lrp5-/- mice in comparison to wild-type. To identify genes that may be responsible for the impaired healing in osteoporotic mice, we performed microarray analysis of three independent callus samples of each genotype. The callus tissue was taken 10 days after surgery, because extensive bone formation took place at this point.
2013-10-25 | E-GEOD-51686 | biostudies-arrayexpress
Project description:Fracture healing is a process that involves many cell populations. In this study we characterized gene expression in a subset of cells involved in fracture healing. αSMACreERT2 mice crossed with Ai9 reporter mice that express tdTomato fluorescent protein after Cre-mediated activation were used as an experimental model. αSMA-expressing cells were labeled by tamoxifen administration, then periosteal cells from the tibia were isolated two days later (controls), or tibial fractures were performed and periosteum/soft callus tissue was collected after 2 and 6 days. The tdTomato positive cell population was isolated by flow cytometry, and subjected to microarray analysis. Histology and cell surface marker analysis indicates that αSMACreERT2 labels a mainly mesenchymal population in the periosteum that expands after fracture, and contributes to both osteogenic and chondrogenic elements of the fracture callus. We were therefore able to examine gene expression in a defined population during the early stages of fracture healing. Total RNA was obtained from the tomato positive cells within the periosteal compartment of fractures from αSMACreERT2/Ai9 mice. Control animals were given 2 doses of tamoxifen, and periosteum was collected and labeled cells sorted (8-9 sex-matched mice per group). Fractures were performed after the second dose of tamoxifen, and tomato positive cells from periosteum/callus tissue were isolated 2 and 6 days after fracture (4-8 animals per sample pooled). 3 replicates for each sample are included.
2013-11-27 | E-GEOD-45156 | biostudies-arrayexpress
Project description:Bone fractures, the most common musculoskeletal injuries, heal through three main phases: inflammatory, repair, and remodeling. Around 10% of fracture patients suffer from impaired healing that requires surgical intervention, a huge burden on the healthcare system. The rate of impaired healing increases with metabolic diseases such as obesity-associated hyperglycemia/type 2 diabetes (T2D), an increasing concern given the growing incidence of obesity/T2D. Immune cells play pivotal roles in fracture healing, and obesity/T2D is associated with defective immune-cell functions. However, there is a gap in knowledge regarding the stoichiometry of immune cells that populate the callus and how that population changes during different phases of healing. Here, we used complementary global and single-cell techniques to characterize the repertoire of immune cells in the fracture callus and to identify populations specifically enriched in the fracture callus relative to the unfractured bone or bone marrow. Our analyses identified two clear waves of immune-cell infiltration into the callus: the first wave occurs during the early inflammatory phase of fracture healing, while the second takes place during the late repair/early remodeling phase. Innate immune cells were activated during the early inflammatory phase, but in later phases they returned to homeostatic numbers and activation levels. Of the innate immune cells, distinct subsets of activated dendritic cells were particularly enriched in the inflammatory healing hematoma. In contrast to innate cells, lymphocytes, including B and T cells, were enriched and activated in the callus primarily during the late repair phase. The Diet-Induced Obesity (DIO) mouse, an established model of obesity-associated hyperglycemia and insulin resistance, suffers from multiple healing defects. Our data demonstrate that DIO mice exhibit dysregulated innate immune responses during the inflammatory phase, and defects in all lymphocyte compartments during the late repair phase. Taken together, our data characterize, for the first time, immune populations that are enriched/activated in the callus during two distinct phases of fracture healing and identify defects in the healing-associated immune response in DIO mice, which will facilitate future development of immunomodulatory therapeutics for impaired fracture healing.
2023-09-01 | GSE240390 | GEO
Project description:Immune cells are essential for proper fracture healing. The goal of this study was to compare the transcriptional profiles of the CD45+ immune cells isolated from the healing fracture callus of old (24 month) and young (3 month) mice (C57BL/6). Analysis of all CD45+ cells was completed via single cell RNAseq. Additional analysis of the macrophages involved in fracture healing in old and young mice was performed via bulk RNAseq. We show that there were multiple subpopulations of macrophages that were transcriptionally heterogenous and present simultaneously within the fracture callus tissue. Attenuated healing in the old mice was associated with a transcriptional profile that shifted towards more pro-inflammatory phenotypes.
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Project description:Loss or damage to the mandible due to trauma, treatment of oral malignancies, and other diseases is currently treated using bone grafting techniques that suffer from numerous shortcomings and contraindications. Zebrafish naturally heal large injuries to their mandibular bone, and thus offer an opportunity to understand how to boost intrinsic healing ability. Using a novel her6:mCherry Notch reporter, we show that canonical Notch signaling is induced during the initial stages of cartilage callus formation in both mesenchymal cells and chondrocytes. We also show that modulation of Notch signaling during the initial postoperative period results in lasting changes to regenerate bone quantity one month later. Notch signaling is required for mandibular bone healing, as pharmacological inhibition of Notch signaling blocks cartilage callus formation and results in non-union. Conversely, conditional transgenic activation of Notch signaling accelerates regenerative ossification. Mechanistically, we report that postoperative Notch signaling regulates multiple phases of chondroid regeneration and patterns callus metabolic landscape. Given conserved functions of Notch signaling in bone repair across vertebrates, we propose that targeted activation of Notch signaling during the early phases of bone healing may have therapeutic value.
2022-03-07 | PXD027280 | Pride