Applying RNA sequencing technology to explore repair mechanism of Tuina on gastrocnemius muscle in sciatic nerve injury rats.
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ABSTRACT:
SUBMITTER: Lyu T
PROVIDER: S-EPMC9771165 | biostudies-literature | 2022 Oct
REPOSITORIES: biostudies-literature
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Applying RNA sequencing technology to explore repair mechanism of Tuina on gastrocnemius muscle in sciatic nerve injury rats.
Lyu Taotao T Liu Zhifeng Z Yu Tianyuan T Lu Mengqian M Zhang Yingqi Y Jiao Yi Y Wang Hourong H Xu Yajing Y Guan Qian Q
Chinese medical journal 20221005 19
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Project description:ObjectiveTo study the effects of the three methods and three-acupoint technique on DRG gene expression in SNI model rats and to elucidate the molecular mechanism of the three methods and three-acupoint technique on promoting recovery in peripheral nerve injury.Methods27 male SD rats were randomly divided into three groups: a Sham group, the SNI group, and the Tuina group. The Tuina group was treated with a tuina manipulation simulator to simulate massage on points, controlling for both quality and quantity. Point-pressing, plucking, and kneading methods were administered quantitatively at Yinmen (BL37), Chengshan (BL57), and Yanglingquan (GB34) points on the affected side once a day, beginning 7 days after modeling. Intervention was applied once a day for 10 days, then 1 day of rest, followed by 10 more days of intervention, totally equaling 20 times of intervention. The effect of the three methods and three-point technique on the recovery of injured rats was evaluated using behavior analysis. RNA sequencing (RNA-Seq) analysis of differentially expressed genes in DRGs of the three groups of rats was also performed. GO and KEGG enrichment was analyzed and verified using real-time PCR.ResultsRNA-Seq combined with database information showed that the number of differentially expressed genes in DRG was the largest in the Tuina group compared with the SNI group, totaling 226. GO function is enriched in the positive regulation of cell processes, ion binding, protein binding, neuron, response to pressure, response to metal ions, neuron projection, and other biological processes. GO function is also enriched in the Wnt, IL-17, and MAPK signaling pathways in the KEGG database. PCR results were consistent with those of RNA sequencing, suggesting that the results of transcriptome sequencing were reliable.ConclusionThe three methods and three-acupoint technique can promote the recovery of SNI model rats by altering the gene sequence in DRGs.
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Project description:Compared to conventional artificial nerve guide conduits (NGCs) prepared using natural polymers or synthetic polymers, acellular nerve grafts (ACNGs) derived from natural nerves with eliminated immune components have natural bionic advantages in composition and structure that polymer materials do not have. To further optimize the repair effect of ACNGs, in this study, we used a composite technology based on supercritical carbon dioxide (scCO2) extraction to process the peripheral nerve of a large mammal, the Yorkshire pig, and obtained an innovative Acellular nerve xenografts (ANXs, namely, CD + scCO2 NG). After scCO2 extraction, the fat and DNA content in CD + scCO2 NG has been removed to the greatest extent, which can better supported cell adhesion and proliferation, inducing an extremely weak inflammatory response. Interestingly, the protein in the CD + scCO2 NG was primarily involved in signaling pathways related to axon guidance. Moreover, compared with the pure chemical decellularized nerve graft (CD NG), the DRG axons grew naturally on the CD + scCO2 NG membrane and extended long distances. In vivo studies further revealed that the regenerated nerve axons had basically crossed the CD + scCO2 NG 3 weeks after surgery. 12 weeks after surgery, CD + scCO2 NG was similar to autologous nerves in improving the quality of nerve regeneration, target muscle morphology and motor function recovery and was significantly better than hollow NGCs and CD NG. Therefore, we believe that the fully decellularized and fat-free porcine ACNGs may be the most promising "bridge" for repairing human nerve defects at this stage and for some time to come.
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Project description:Background and purposeAfter peripheral nerve lesions, surgical reconstruction facilitates axonal regeneration and motor reinnervation. However, functional recovery is impaired by aberrant reinnervation.Materials and methodsWe tested whether training therapy by treadmill exercise (9 × 250 m/week) before (run-idle), after (idle-run), or both before and after (run-run) sciatic nerve graft improves the accuracy of reinnervation in rats. Female Lewis rats (LEW/SsNHsd) were either trained for 12 weeks (run) or not trained (kept under control conditions, idle). The right sciatic nerves were then excised and reconstructed with 5 mm of a congenic allograft. One week later, training started in the run-run and idle-run groups for another 12 weeks. No further training was conducted in the run-idle and idle-idle groups. Reinnervation was measured using the following parameters: counting of retrogradely labeled motoneurons, walking track analysis, and compound muscle action potential (CMAP) recordings.ResultsIn intact rats, the common fibular (peroneal) and the soleus nerve received axons from 549 ± 83 motoneurons. In the run-idle group, 94% of these motoneurons had regenerated 13 weeks after the nerve graft. In the idle-run group, 81% of the normal number of motoneurons had regenerated into the denervated musculature and 87% in both run-run and idle-idle groups. Despite reinnervation, functional outcome was poor: walking tracks indicated no functional improvement of motion in any group. However, in the operated hindlimb of run-idle rats, the CMAP of the soleus muscle reached 11.9 mV (normal 16.3 mV), yet only 6.3-8.1 mV in the other groups.ConclusionTreadmill training neither altered the accuracy of reinnervation nor the functional recovery, and pre-operative training (run-idle) led to a higher motor unit activation after regeneration.
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Project description:BackgroundThe aim of the present study was to investigate the effect of acellular nerve scaffold (ANS) containing human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) on nerve repair and regeneration in rats with sciatic nerve defect.MethodsSciatic nerve trunks were removed from 6 female Sprague-Dawley (SD) rats, and ANS was prepared by lyophilization + enzymatic method and divided into A, B, C, D and E groups according to different treatment times. hUC-MSCs were isolated from the collected umbilical cords and cultured, and then ANS-hUC-MSCs complexes were made. The other 24 adult female SD rats were randomly divided into the control, autograft, ANS, and ANS-hUC-MSCs groups, and a rat model of sciatic nerve defect was established. Hematoxylin-eosin (HE) staining, Luxol fast blue (LFB) staining, Masson staining, and scanning electron microscopy were used to observe the morphology and tissue structure of ANS. The performance of ANS was evaluated by mechanical detection, and hydroxyproline (HYP) content was evaluated using a biochemical kit. Flow cytometry was adopted to detect the levels of hUC-MSCs surface antigens CD29, CD44, and CD34, as well as electrophysiological detection and muscle wet weight recovery rate for measuring rat muscle performance.ResultsANS was prepared according to group A method and had good mechanical properties, with less residues of cells and myelin, and higher HYP content, indicating that this scaffold had the best performance. ANS-hUC-MSCs significantly reduced myelin injury in the sciatic nerve, and increased axonal regeneration, effectively improving sciatic nerve injury in rats. In addition, ANS-hUC-MSCs significantly increased compound muscle action potential (CMAP), nerve conduction velocity (NCV), and muscle wet weight, and reduced muscle atrophy.ConclusionsANS containing hUC-MSCs can promote nerve repair and regeneration in rats with sciatic nerve defects.
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Project description:[This corrects the article DOI: 10.21037/atm-22-1578.].
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Project description:Open surgical repair of proximal hamstring avulsions has been the standard of care for a long time, when surgery is needed. Endoscopic repair is a relatively new surgery, and its popularity increased in the last 10 years. This technique allows the surgeon an anatomic repair and a safe sciatic nerve exploration with small incisions and dissection. As a new technique, it has its limitations, mostly in chronic retracted tears, and long follow-up series are needed to assess long-term outcomes. We present an endoscopic repair of a right proximal hamstring avulsion performed along with a sciatic nerve neurolysis.
| S-EPMC9134099 | biostudies-literature
Project description:Peripheral nerve injuries (PNIs) are one of the common causes of morbidity and disability worldwide. Autograft is considered the gold standard treatment for PNIs. However, due to the complications associated with autografts, other sources are considered as alternatives. Recently, electrospun nanofibrous scaffolds have received wide attention in the field of tissue engineering. Exogenous tubular constructs with uniaxially aligned topographical cues to enhance the axonal re-growth are needed to bridge large nerve gaps between proximal and distal ends. Although several studies have used PLGA/PCL, but few studies have been conducted on developing a two-layer scaffold with aligned fibers properly orientated along the axis direction of the sciatic nerve to meet the physical properties required for suturing, transplantation, and nerve regeneration. In this study, we sought to design and develop PLGA-PCL-aligned nanofibers. Following the conventional examinations, we implanted the scaffolds into 7-mm sciatic nerve gaps in a rat model of nerve injury. Our in vivo evaluations did not show any adverse effects, and after eight weeks, an acceptable improvement was noted in the electrophysiological, functional, and histological analyses. Thus, it can be concluded that nanofiber scaffolds can be used as a reliable approach for repairing PNIs. However, further research is warranted.
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Project description:ObjectiveThe disordered growth of nerve stumps after amputation leading to the formation of neuromas is an important cause of postoperative pain in amputees. This severely affects the patients' quality of life. Regenerative peripheral nerve interfaces (RPNIs) are an emerging method for neuroma prevention, but its postoperative nerve growth and pathological changes are yet to be studied.MethodsThe rat sciatic nerve transection model was used to study the effectiveness of RPNI in this experiment. The RPNI (experimental) group (n = 11) underwent RPNI implantation after sciatic nerve transection, while the control group (n = 11) only underwent sciatic nerve transection. Autotomy behavior, ultrasonography, and histopathology were observed for 2 months postoperatively.ResultsCompared to the control group, the incidence and size of the neuromas formed and the incidence and extent of autotomy were significantly reduced in the RPNI group. The axon density in the stump and degree of stump fibrosis were also significantly reduced in the RPNI group.ConclusionRPNI effectively prevented the formation of neuromas.
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Project description:Nerve injuries requiring surgery are a significant problem without good clinical alternatives to the autograft. Tissue engineering strategies are critically needed to provide an alternative. In this study, we utilized aligned nanofibers that were click-modified with the bioactive peptide RGD for rat sciatic nerve repair. Empty conduits or conduits filled with either non-functionalized aligned nanofibers or RGD-functionalized aligned nanofibers were used to repair a 13 mm gap in the rat sciatic nerve of animals for six weeks. The aligned nanofibers encouraged cell infiltration and nerve repair as shown by histological analysis. RGD-functionalized nanofibers reduced muscle atrophy. During the six weeks of recovery, the animals were subjected to motor and sensory tests. Sensory recovery was improved in the RGD-functionalized nanofiber group by week 4, while other groups needed six weeks to show improvement after injury. Thus, the use of functionalized nanofibers provides cues that aid in in vivo nerve repair and should be considered as a future repair strategy.
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