Gene expression analysis of vastus medialis cells after tourniquet-induced ischemia during Total Knee Arthroplasty: A randomized clinical trial
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PROVIDER: PRJNA512618 | ENA |
REPOSITORIES: ENA
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Project description:Purpose The effects of the commonly utilized intraoperative tourniquet on gene expression within the skeletal muscle cells are barely examined. Our group already showed that tourniquet application results in higher proteolytic activity within vastus medialis cells (KSSTA 2016), without influence on the amount and function of mitochondria (EJOT 2016). The purpose of the present study was to examine the gene expression within the skeletal muscle cells after tourniquet-induced ischemia and to identify differential expressed genes (DEGs). Methods As part of a randomized, controlled, monocentric trial (Clinical-Trials.gov NCT02475603) we included 20 patients scheduled to undergo primary total knee arthroplasty (TKA). The patients received a written consent and were randomly assigned to Group A (TKA with tourniquet) (n=10) and Group B (TKA without tourniquet) (n=10). Muscle biopsies of (5×5×5 mm) were obtained from the vastus medialis immediately after performing the surgical approach and exactly 60 min later. After preparation of a muscle homogenate, RNA extraction was performed (RNeasy Plus Universal Mini Kit Qiagen) and RNA integrity (RIN) was determined (Agilent 2100 Bioanalyzer,RNA 6000 Pico Kit). Gene expression profiling was performed methodological validated (GeneChip™ Human Transcriptome Array 2.0;Affymetrix). Statistical analysis include the number of significant DEGs with p-value < 0,05, number of DEGs with relative difference >25% and pathway analysis (SPSS-Version 24; SAS JMP10 Genomics, Version 6). The power analysis considered an effect size of 0,4 and a statistical power > 0,8, sample size n=10 for each group. The protocol was approved by our Institutional Ethics Committee (File reference 2012-334N-MA). Results and conclusion Numerous genes were differencially expressed during tourniquet induced ischemia. Further investigations and statistical analysis are ongoning.
2021-01-01 | GSE124595 | GEO
Project description:Ischaemic preconditioning is a method of protecting tissue against ischaemia-reperfusion injury. It is an innate protective mechanism that increases a tissue's tolerance to prolonged ischaemia when it is first subjected to short burst of ischaemia and reperfusion. It is thought to provide this protection by increasing the tissue's tolerance to ischaemia, therby reducing oxidative stress, inflammation and apoptosis in the preconditioned tissue. We used microarrays to investigate the genomic response induced by ischaemic preconditioning in muscle biopsies taken from the operative leg of total knee arthroplasty patients in order to gain insight into the ischaemic preconditioning mechanism. Patients undergoing primary knee arthroplasty were randomised to control and treatment (ischaemic preconditioning) groups. Patients in the treatment group received a preconditioning stimulus immediately prior to surgery. The ischaemic preconditioning stimulus consisted of three five-minute periods of tourniquet insufflation on the lower operative limb, interrupted by five minute periods of reperfusion. All patients had a tourniquet applied to the lower limb after the administration of spinal anaesthesia, as per normal protocol for knee arthroplasty surgery. Muscle biopsies were taken from the quadriceps muscle of the operative knee at the immediate onset of surgery (T0) and at 1 hour into surgery (T1). Total RNA was extracted from biospies of four control and four treatment patients and hybridised to the Affymetrix Human U133 2.0 chip.
2010-04-12 | E-GEOD-21164 | biostudies-arrayexpress
Project description:Transcriptome profiling was performed on muscle biopsies from patients immediately before Total Knee Arthroplasty and two hours after TKA and tourniquet application.
2016-02-01 | GSE75432 | GEO
Project description:Ischaemic preconditioning is a method of protecting tissue against ischaemia-reperfusion injury. It is an innate protective mechanism that increases a tissue's tolerance to prolonged ischaemia when it is first subjected to short burst of ischaemia and reperfusion. It is thought to provide this protection by increasing the tissue's tolerance to ischaemia, therby reducing oxidative stress, inflammation and apoptosis in the preconditioned tissue. We used microarrays to investigate the genomic response induced by ischaemic preconditioning in muscle biopsies taken from the operative leg of total knee arthroplasty patients in order to gain insight into the ischaemic preconditioning mechanism.
2010-04-02 | GSE21164 | GEO
Project description:These data are from a preclinical model of total knee arthroplasty (TKA) performed in healthy rats. One day after surgery, ipsilateral dorsal root ganglia (DRG) from L3 and L4 were collected for RNA extraction and sequencing.
2022-03-23 | GSE195833 | GEO
Project description:Arthrofibrosis is characterized by excessive extracellular matrix (ECM) deposition that results in restricted joint motion after total knee arthroplasty (TKA). Current surgical and pharmacologic treatment options are limited. Therefore, an in vitro model for joint myofibroblastogenesis is valuable to investigate the arthrofibrotic process and identify diagnostic biomarkers and treatment options. In this study, we obtained intraoperative posterior capsule (PC), quadriceps tendon (QT), and suprapatellar pouch (SP) tissue from knees of four patients undergoing primary TKA for osteoarthritis and characterized primary outgrowth cells from these tissues in the absence and presence of transforming growth factor beta 1 (TGFβ1), a pro-myofibroblastic cytokine. Light microscopy of knee outgrowth cells revealed spindle-shaped cells while immunofluorescence (IF) established staining for the fibroblast-specific antigen TE-7 and Vimentin, which are characteristics of fibroblasts. These fibroblasts differentiate readily into myofibroblasts as highlighted by enhanced alpha smooth muscle actin (ACTA2) mRNA and protein expression and increased collagen mRNA (i.e., collagen type 1 (COL1A1) and collagen type 3 (COL3A1)) expression and collagenous matrix deposition in the presence of TGFβ1. Of note, these studies also revealed that knee-derived fibroblasts are more sensitive to TGFβ1-mediated myofibroblastogenesis than adipose-derived mesenchymal stem cells. Importantly, while outgrowth fibroblasts isolated from four patients and three anatomical regions exhibit similar gene expression profiles, these knee fibroblasts form a unique gene expression cluster within the fibroblast niche as revealed by RNA-sequencing analysis. In conclusion, our study provides a fibroblast/myofibroblast model of outgrowth knee cells derived from patients undergoing primary TKA that can be employed to assess myofibroblast-related processes and test novel pharmacological strategies in vitro for arthrofibrosis.
2022-03-01 | GSE185333 | GEO
Project description:Gene Expression of Fibroblasts Derived from Knee Tissue of Patients Undergoing Primary Total Knee Arthroplasty for Osteoarthritis
| PRJNA768776 | ENA
Project description:Background: Endoprosthesis loosening is the major cause of arthroplasty failure. Currently, radiography and histo-pathological classification of the periprosthetic membrane are used retrospectively for diagnosis. Prospective options for early diagnosis and prevention of loosening are not available. In order to approach this challenge, the study presented here aimed to identify molecular biomarkers in single-centre prospectively collected cell and plasma samples from patients. Methods: Four patient cohorts (primary and revision arthroplasty of hip or knee) were defined. Aseptic loosening of implants was assessed by the standardised approach of the Knee Society Total Knee Arthroplasty Roentgenographic Evaluation and Scoring System (KSRESS) which is based on radiolucent lines. Synovial fluid, bone marrow, and blood were collected from a total of 96 patients. Bulk RNA-sequencing (RNA-seq) of mesenchymal stem cells (MSCs) isolated from the bone marrow of patients was performed for 28 samples. The data was analysed by bioinformatic tools. Quantitative real-time PCR served to validate and extend RNA-seq data. Western blots were performed for molecular differentiation. ELISAs were used to quantitate selected protein targets. Results: RNA-seq of MSCs identified glycoprotein non-metastatic melanoma protein B (GPNMB) as significantly upregulated in the cells from revision patients compared to patients with primary implantations. The protein encoded by this gene is plasma membrane-standing; its extracellular domain may be cleaved and secreted. In synovial fluid plasma, the soluble GPNMB protein exhibited increased levels in patients undergoing revision surgery compared to patients subjected to primary arthroplasty. No significant differences were observed in bone marrow plasma and blood plasma. The molecular species detected by Western blotting were similar in the plasma fractions of synovial fluid, bone marrow and blood. Conclusions: The use of GPNMB as a biomarker may be of prime importance for early detection of implant loosening, ahead of radiographic failure demonstration. Translational potential of this article: GPNMB has the potential to act as a prospective and quantitative parameter to assess different stages of implant loosening. Radiography and histo-pathological classification of the periprosthetic membrane, on the contrary, are used retrospectively for diagnosis and are subjective measures, performed mainly at the end of a full loosening process.
2025-09-17 | GSE276597 | GEO
Project description:To investigate the physiologic responses of whole osteoarthritic synovium to cycle tensile strain. We obtained 3 matched synovial tissue samples from 6 patients undergoing total knee arthroplasty, subjected them to cyclic tensile strain, and then performed gene expression profiling analysis using RNA seq from each cyclic tensile strain protocol.
2022-09-30 | GSE205196 | GEO