Peripheral Nerve Crush Injury Research Articles

Neuromuscular Junction Remodeling: the Role of Sub-Synaptic Nuclei, Terminal Schwann Cells, and Trophic Signaling Following Neuromusculoskeletal Injuries

Peripheral nerve crush injury is a well-established model of neuromuscular junction (NMJ) denervation and subsequent re-innervation. Functionally, the muscle tissue follows a similar pattern as neural recovery, with immediate loss of force production that steadily improves in parallel with rates of re-innervation. On the other hand, traumatic injury to the muscle tissue itself, specifically volumetric muscle loss (VML), results in an irrecoverable loss of muscle function. Recent work has indicated significant impairments to the NMJ following VML that appear chronic in nature, alongside functional recovery. Thus, the goal of this study was to compare the effects of nerve and muscle injury on NMJ remodeling, with the hypothesis that VML will result in chronic alterations to the NMJ, including a higher degree of cellular localization to the NMJ. Even numbers of adult male and female mice were used with three experimental groups: injury naive, nerve crush, and VML-injury; and three terminal timepoints: 3-, 48-, and 112-days post injury. Data were analyzed by three-way ANOVA with Tukey’s HSD post-hoc. Confirming the assumed recoverability of the two injury models, we found in vivo maximal torque was fully restored following nerve crush injury (Tukey’s HSD post hoc p=0.730), but remained at a significant deficit following VML (~41% lower than injury naïve; Tukey’s HSD post hoc p<0.001). Compared to nerve crush and injury naïve, we found VML results in aberrantly high trophic signaling and numbers of supporting cells. Protein expression of neuregulin-1 and connective tissue growth factor were both highest in the VML group (~1.2-fold and 12-fold increase compared to injury naïve, respectively; Main effect of group, p<0.001). While nerve crush injury elicited a transient increase in terminal Schwann cell numbers (~38% higher than injury naïve at 48dpi; Tukey’s HSD post hoc p<0.001), VML resulted in a chronic increase (~53% and 27% higher than injury naïve at 48 and 112dpi, respectively; Tukey’s HSD post hoc p<0.001). Moreover, nerve crush injury did not alter the number of sub-synaptic nuclei per NMJ, while VML resulted in significant increases at both 48- and 112-days post injury (~27% and 24% higher than injury naïve, respectively; Tukey’s HSD post hoc p=0.002). In some cases, sex differences were detected, including higher rates of innervation in females than males (Main effect of sex p=0.003), particularly following VML-injury (Tukey’s HSD post hoc p=0.004). The findings herein bear a striking resemblance to NMJs found in dystrophic muscle, which are thought to be the result of continuous cycles of muscle fiber degeneration and regeneration. Thus, the NMJ changes observed following VML may be indicative of a larger issue, such as chronic muscle fiber remodeling and pervasive fibrotic accumulation. R01-AR078903 (JAC & SMG); K02-AG081488 (SMG). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Neutrophil peptide 1 accelerates the clearance of degenerative axons during Wallerian degeneration by activating macrophages after peripheral nerve crush injury.

JOURNAL/nrgr/04.03/01300535-202408000-00036/figure1/v/2023-12-16T180322Z/r/image-tiff Macrophages play an important role in peripheral nerve regeneration, but the specific mechanism of regeneration is still unclear. Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide closely involved in peripheral nerve regeneration. However, the mechanism by which neutrophil peptide 1 enhances nerve regeneration remains unclear. This study was designed to investigate the relationship between neutrophil peptide 1 and macrophages in vivo and in vitro in peripheral nerve crush injury. The functions of RAW 264.7 cells were elucidated by Cell Counting Kit-8 assay, flow cytometry, migration assays, phagocytosis assays, immunohistochemistry and enzyme-linked immunosorbent assay. Axonal debris phagocytosis was observed using the CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis) optical clearing technique during Wallerian degeneration. Macrophage inflammatory factor expression in different polarization states was detected using a protein chip. The results showed that neutrophil peptide 1 promoted the proliferation, migration and phagocytosis of macrophages, and CD206 expression on the surface of macrophages, indicating M2 polarization. The axonal debris clearance rate during Wallerian degeneration was enhanced after neutrophil peptide 1 intervention. Neutrophil peptide 1 also downregulated inflammatory factors interleukin-1α, -6, -12, and tumor necrosis factor-α in vivo and in vitro. Thus, the results suggest that neutrophil peptide 1 activates macrophages and accelerates Wallerian degeneration, which may be one mechanism by which neutrophil peptide 1 enhances peripheral nerve regeneration.

Study on the Crush Injury Model of the Sciatic Nerve in Rabbits by Conventional Ultrasound and Elastography.

The crush injury model of the sciatic nerve in rabbits is a common nerve injury model, but there is little literature on the evaluation of nerve stiffness by ultrasound elastography. To explore the value of ultrasound elastography in diagnosing crush injury of the sciatic nerve in rabbits. Forty cases of crush injury model of the sciatic nerve in rabbits were examined by conventional ultrasound and elastography. The sonographic manifestations and stiffness of the model were analyzed, and the regularity of nerve injury with time was summarized. Ultrasound could clearly show the location, range, and thickness of the injured nerve in this model. The thickness of the injured area reached a peak in the 2nd week and returned to normal thickness in the 8th week. The nerve stiffness of the injured area increased gradually with time. Conventional ultrasound combined with elastography can comprehensively and quantitatively evaluate the morphological changes and mechanical properties of the injured sciatic nerve in rabbits, which may be of great significance to the repair and rehabilitation of peripheral nerve crush injuries.

Injectable hydrogel encapsulated with VEGF-mimetic peptide-loaded nanoliposomes promotes peripheral nerve repair in vivo.

Repair of peripheral nerve crush injury remains a major clinical challenge. Currently, oral or intravenous neurotrophic drugs are the main treatment for peripheral nerve crush injury; however, this repair process is slow, and the final effect may be uncertain. The current study aimed at developing an injectable hydrogel with vascular endothelial growth factor (VEGF)-mimetic peptide (QK)-encapsulated nanoliposomes (QK-NLs@Gel) for sustainable drug release that creates an appropriate microenvironment for nerve regeneration. The QK-encapsulated nanoliposomes (QK-NLs) could facilitate the proliferation, migration, and tube formation capacities of human umbilical vein endothelial cells through the VEGF signaling pathway. The QK-NLs@Gel hydrogel encapsulated with QK-NLs showed enhanced physical properties and appropriate biocompatibility in vitro. Thereafter, the QK-NLs@Gel hydrogel was directly injected into the site of peripheral nerve crush injury in a rat model, where it enhanced revascularization and promoted the M2-polarization of the macrophages, thus providing an optimized microenvironment for nerve regeneration. At four weeks post-surgery, the QK-NLs@Gel injected rats exhibited enhanced axon regeneration, remyelination, and better functional recovery in comparison with other groups in vivo. Overall, these findings demonstrate that the composite hydrogel could promote a multicellular pro-regenerative microenvironment at the peripheral nerve injury site, thus revealing great potential for peripheral nerve restoration. STATEMENT OF SIGNIFICANCE: Peripheral nerve injury (PNI) is a leading public health issue, and how to delivery beneficial drugs to injured sites efficiently is still a big challenge. In the current study, an injectable hydrogel with VEGF-mimetic peptide (QK)-encapsulated nanoliposomes (QK-NLs@Gel) was first developed and used to repair a rat crush injury model. Our results showed that QK-NLs promoted the proliferation, migration, and angiogenesis of HUVEC via VEGF signaling pathway in vitro. Furthermore, when injected to the crushed sites in vivo, the QK-NLs@Gel hydrogel could accelerate nerve repair through enhanced revascularization and M2-polarization of macrophages. These results collectively demonstrate that injection of QK-NLs@Gel hydrogel could create an appropriate microenvironment for peripheral nerve regeneration. This strategy is effective, economical, and convenient for clinical applications.

The effects of curcumin and blueberry on axonal regeneration after peripheral nerve injury

The purpose of this study was to analyze the axonal regeneration and therapeutic effects of curcumin and blueberry administration following peripheral nerve injury using stereological, electron microscopic and electrophysiological methods. Animals in were assigned into one of four groups - control (Cont), injury (Inj), injury+curcumin (Cur) and injury+blueberry (Blue). Following the induction of sciatic nerve crush injury (75 Newtons for 5 s) in the Inj, Cur, and Blue groups, the rats in the Cur group received intraperitoneal injection of 30 mg/kg curcumin (Sigma C1386) and the rats in the Blue group received 4 g/kg blueberry by gavage over a four-week period. The rats in the Cont and Inj groups were not exposed to any substance. All animals were given standard chow. Sciatic functional index analyses were performed on the 14th and 28th days after injury, and electromyography (EMG) results were recorded. Stereological analysis of the nerve was performed under light microscopy. Light and electron microscopies were used for the histopathological evaluation of the sciatic nerve. Analysis of myelinated axon numbers revealed no significant differences between the Inj group and the Cur and Blue groups. However, a significant difference was observed between the Blue and Inj groups in terms of axonal areas. EMG test results differed between the Blue and the Inj groups (p < 0.05), but no significant difference was observed between the Inj and Cur groups. Electron microscopic analysis revealed protective effects of curcumin and blueberry treatment after injury. The use of the curcumin and blueberry may represent a supportive approach to the protection of nerve fibers after peripheral nerve crush injury.

The effect of dietary sesame oil and ginger oil as antioxidants in the adult rat dorsal root ganglia after peripheral nerve crush injury

Aim The purpose of this study was to investigate the effect of dietary sesame oil and ginger oil supplements on the dorsal root ganglia following a sciatic nerve crush model in male Wistar albino rats. Materials and methods Crush injury models have been done by means of graded forceps (50 Newton). The animals were given a daily sesame oil (4 ml/kg/day) and ginger oil (400 mg/kg/day) via oral gavage for a period of 28 days. Dorsal root ganglia from the L5 levels were harvested. Processing of tissues was done for electron microscopy and light microscopy. Immunohistochemical staining with active caspase-3 antibody and qualitative ultrastructural analyses of tissues were made by a light and a transmission electron microscope, respectively. Results The results showed that crush injury leads to remarkable ultrastructural changes in sensory neurons, such as swollen mitochondria, disruption of cristae structure, glial cell proliferation and, consequently, phagocytosis of the damaged neuron. These ultrastructural changes were less evident in the treated groups, and both natural compounds reduced the expression of activated caspase-3, which may also affect ultrastructural changes. Conclusion The application of the natural products sesame oil and ginger oil may represent a supportive approach to the protection of sensory neurons against the destructive effects of peripheral nerve crush injury.

Thymoquinone Modulates Local MMP-9, IL-10, and IgG in Sciatic Nerve Crush Injury Animal Model

BACKGROUND: Interleukin (IL)-10 is involved in Wallerian degeneration after peripheral nerve crush injury. Oral thymoquinone was previously observed to decrease local immunoglobulin-G (IgG) in a crush-injured rat model. No study has evaluated the pathway of various thymoquinone dosages on local IgG and IL-10 in this injury.METHODS: This experimental study used 126 Rattus norvegicus Wistar rats that were divided into 18 groups: six groups received a placebo, the other six groups received thymoquinone at 100 mg/kg/day and the last six groups received thymoquinone at 250 mg/kg/day, respectively. Rats were sacrificed at 12, 18, 24, 5x24, 6x24, and 7x24 hours. Matrix metalloproteinase-9 (MMP-9), IL-10, and local IgG levels were assessed by Enzyme-Linked Immunosorbent Assay (ELISA). The nuclear factor KappaB (NF-κB) expressions on Schwann cells were examined by flow cytometry. Path analysis was performed using SmartPLS.RESULTS: The path analysis showed that 100mg/kg/day of thymoquinone significantly decreased NF-κB expression. However, NF-κB did not affect local MMP-9, and MMP-9 had no significant relationship with local IL-10 and IgG. Thymoquinone 250 mg/kg/day also significantly inhibited NF-kB expression, decreased local MMP-9, and, in turn, decreased local IL-10 and IgG.CONCLUSION: Administration of oral thymoquinone 250 mg/kg/day decreases local IgG and IL-10 levels via suppressing NF-κB expression and MMP-9 leveKEYWORDS: thymoquinone, crush injury, IgG, IL-10, MMP-9, NF-κB

Evaluation of the therapeutic effects of calcium dobesilate in sciatic nerve crush injury in rats

IntroductionProinflammatory cytokines released from nerve endings and surrounding injured tissue after nerve damage can prolong the inflammation process, delay nerve healing or result in poor quality nerve healing. In this case, due to the loss of function in the muscles innervated by the damaged nerve, the patient may have neurological and functional difficulties which may reduce the patient's quality of life and create an economic burden. Although the attempts of many pharmacological agents to heal crush injury of peripheral nerves have been recorded in literature, a drug that can provide adequate recovery of the crushed nerve and can be applied in daily life has not been defined as yet. This study aimed to assess the effects of calcium dobesilate on sciatic nerve crush injury in a rat model. MethodsA total of 26 male Wistar albino rats were separated into four groups as follows: CONTROL group (healthy subjects, n=6); SHAM group (crush injury was created, n=6); MP group (after created crush injury, methylprednisolone was administered, n=7); and CAD group (after created crush injury, calcium dobesilate was administered, n=7). A crush injury was created, then the electrophysiological findings and sciatic nerve functional index (SFI) were recorded before euthanasia. After the euthanasia of all the rats, samples of the crushed nerve and gastrocnemius muscle were evaluated histopathologically, immunohistochemically, and biochemically. ResultsBoth pharmacological agents were histopathologically effective in axon regeneration and repair. Calcium dobesilate did not preserve total muscle mass but was seen to prevent atrophy microscopically. Immunohistochemistry and biochemistry results showed that calcium dobesilate and methylprednisolone had anti-inflammatory, anti-oxidant, anti-apoptotic, and anti-autophagic activity in the crushed sciatic nerve. Neither calcium dobesilate nor methylprednisolone improved the nerve conductance level. SFI values obtained on day 30 from the CAD group were numerically closer to the values of the healthy animals but not at a statistically significant level. ConclusionThe study results demonstrated that calcium dobesilate could suppress inflammatory processes and provide histopathological and functional improvements in the injured nerve in rats. Therefore, further clinical studies are recommended to investigate in detail the therapeutic effects of calcium dobesilate on peripheral nerve crush injury.

A Novel Standardized Peripheral Nerve Transection Method and a Novel Digital Pressure Sensor Device Construction for Peripheral Nerve Crush Injury.

Peripheral nerve injury (PNI) is common in all walks of life, and the most common PNIs are nerve crush and nerve transection. While optimal functional recovery after crush injury occurs over weeks, functional recovery after nerve transection with microsurgical repair and grafting is poor, and associated with permanent disability. The gold-standard treatment for nerve transection injury is microsurgical tensionless end-to-end suture repair. Since it is unethical to do experimental PNI studies in humans, it is therefore indispensable to have a simple, reliable, and reproducible pre-clinical animal model for successful evaluation of the efficacy of a novel treatment strategy. The objective of this article is two-fold: (A) To present a novel standardized peripheral nerve transection method in mice, using fibrin glue for modeling peripheral nerve transection injury, with reproducible gap distance between the severed nerve ends, and (B) to document the step-wise description of constructing a pressure sensor device for crush injury pressure measurements. We have successfully established a novel nerve transection model in mice using fibrin glue, and demonstrated that this transection method decreases surgical difficulties and variability by avoiding microsurgical manipulations on the nerve, ensuring the reproducibility and reliability of this animal model. Although it is quite impossible to exactly mimic the pathophysiological changes seen in nerve transection with sutures, we hope that the close resemblance of our novel pre-clinical model with gold-standard suturing can be easily reproduced by any lab, and that the data generated by this method significantly contributes to better understanding of nerve pathophysiology, molecular mechanisms of nerve regeneration, and the development of novel strategies for optimal functional recovery. In case of peripheral nerve crush injury, current methods rely on inter-device and operator precision to limit the variation with applied pressure. While the inability to accurately quantify the crush pressure may result in reduced reproducibility between animals and studies, there is no documentation of a pressure monitoring device that can be readily used for real-time pressure measurements. To address this deficit, we constructed a novel portable device comprised of an Arduino UNO microcontroller board and force sensitive resistor (FSR) capable of reporting the real-time pressure applied to a nerve. This novel digital pressure sensor device is cheap, easy to construct and assemble, and we believe that this device will be useful for any lab performing nerve crush injury in rodents.

Sustained delivery of vascular endothelial growth factor mediated by bioactive methacrylic anhydride hydrogel accelerates peripheral nerve regeneration after crush injury.

Neurotrophic factors, currently administered orally or by intravenous drip or intramuscular injection, are the main method for the treatment of peripheral nerve crush injury. However, the low effective drug concentration arriving at the injury site results in unsatisfactory outcomes. Therefore, there is an urgent need for a treatment method that can increase the effective drug concentration in the injured area. In this study, we first fabricated a gelatin modified by methacrylic anhydride hydrogel and loaded it with vascular endothelial growth factor that allowed the controlled release of the neurotrophic factor. This modified gelatin exhibited good physical and chemical properties, biocompatibility and supported the adhesion and proliferation of RSC96 cells and human umbilical vein endothelial cells. When injected into the epineurium of crushed nerves, the composite hydrogel in the rat sciatic nerve crush injury model promoted nerve regeneration, functional recovery and vascularization. The results showed that the modified gelatin gave sustained delivery of vascular endothelial growth factors and accelerated the repair of crushed peripheral nerves.

Delivery of nitric oxide-releasing silica nanoparticles for in vivo revascularization and functional recovery after acute peripheral nerve crush injury

Nitric oxide (NO) has been shown to promote revascularization and nerve regeneration after peripheral nerve injury. However, in vivo application of NO remains challenging due to the lack of stable carrier materials capable of storing large amounts of NO molecules and releasing them on a clinically meaningful time scale. Recently, a silica nanoparticle system capable of reversible NO storage and release at a controlled and sustained rate was introduced. In this study, NO-releasing silica nanoparticles (NO-SNs) were delivered to the peripheral nerves in rats after acute crush injury, mixed with natural hydrogel, to ensure the effective application of NO to the lesion. Microangiography using a polymer dye and immunohistochemical staining for the detection of CD34 (a marker for revascularization) results showed that NO-releasing silica nanoparticles increased revascularization at the crush site of the sciatic nerve. The sciatic functional index revealed that there was a significant improvement in sciatic nerve function in NO-treated animals. Histological and anatomical analyses showed that the number of myelinated axons in the crushed sciatic nerve and wet muscle weight excised from NO-treated rats were increased. Moreover, muscle function recovery was improved in rats treated with NO-SNs. Taken together, our results suggest that NO delivered to the injured sciatic nerve triggers enhanced revascularization at the lesion in the early phase after crushing injury, thereby promoting axonal regeneration and improving functional recovery.

Evaluation of peripheral nerve acute crush injury in rabbits: comparison among diffusion kurtosis imaging, diffusion tensor imaging and electromyography.

Diffusion kurtosis imaging (DKI) has been proven to provide additional value for assessing many central nervous system diseases compared with conventional diffusion tensor imaging (DTI); however, whether it has the same value in peripheral nerve injury is unclear. This study aimed to investigate the performance of DKI, DTI, and electromyography (EMG) in evaluating peripheral nerve crush injury (PNCI) in rabbits. A total of 27 New Zealand white rabbits were selected to establish a PNCI model. Longitudinal DTI, DKI, and EMG were evaluated before surgery and 1day, 3days, 1week, 2weeks, 4weeks, 6weeks, and 8weeks after surgery. At each time point, two rabbits were randomly selected for pathological examination. The results showed that fractional anisotropy (FA) derived from both DKI and DTI demonstrated a significant difference between injured and control nerves at all time points (all P < 0.005) mean kurtosis of the injured nerve was lower than that on the control side after 2-8weeks (all P < 0.05). No statistically significant difference was found in radial kurtosis, axial kurtosis, and apparent diffusion coefficient at almost every time point. The difference in compound muscle action potential (CMAP) of the bilateral gastrocnemius at each time point was statistically significant (all P < 0.001). CMAP was a sensitive and reliable method to assess acute PNCI without being affected by perineural edema. DKI may not be superior to DTI in evaluating peripheral nerves, DTI with a shorter scanning time was preferred as an effective choice for evaluating acute peripheral nerve traumatic injury.

Antibiotics impair and probiotics improve functional recovery after traumatic peripheral nerve crush injury in mice

Antibiotics (ABX) are widely used for the treatment of life-threatening infections and also for routine surgical prophylaxis. Compelling evidence suggests that ABX-induced alterations of gut microbiota composition and metabolism, termed dysbiosis, are linked with various disease states including gastrointestinal, metabolic, autoimmune, inflammatory, neurological, and neurodegenerative conditions. To combat the consequences of dysbiosis, nutritional interventions consisting of prebiotics and probiotics (PBX) are widely used. ABX-induced dysbiosis has recently been reported to impair neurological function after traumatic spinal cord injury. Traumatic peripheral nerve injury (TPNI) is prevalent and can result in profound neurologic impairment, prolonged recovery time, delayed return to work, and permanent disability. It is unknown whether ABX treatment-induced dysbiosis has any impact on TPNI-induced functional recovery, and if so, what role medical-grade PBX could have on TPNI recovery. In this study, ABX-induced dysbiosis (loss-of-function) and PBX-induced microbiota enrichment (gain-of-function) models were used to explore the potential role of the gut microbiome in the functional recovery of TPNI. An established mouse model of severe sciatic nerve crush injury was used for post-injury functional recovery with or without ABX, PBX, and ABX plus PBX (ABX-PBX) treatments, and stool samples were analyzed for 16S ribosomal RNA (rRNA) gene sequencing and gut microbiota compositions. Our findings demonstrate that pre-injury ABX significantly impaired post-injury functional recovery compared with the vehicle group. Conversely, pre-injury PBX significantly improved TPNI-induced functional recovery. Importantly, post-injury PBX protected against pre-injury ABX-induced functional impairment. 16S rRNA gene sequencing confirmed an ABX-induced dysbiosis and revealed that ABX-induced changes could be partially restored by PBX administration. These interesting findings demonstrate that ABX-induced gut dysbiosis exacerbates post-TPNI functional impairment and that this impairment can be prevented by early PBX administration. Since the gut microbiota are modifiable and no therapeutic treatment exists to improve patient outcomes and prevent long term disability after TPNI, clinically approved commercially available probiotics could play a significant role in the management of TPNIs, especially for those patients receiving ABX.

Novel Real-time Digital Pressure Sensor Reveals Wide Variations in Current Nerve Crush Injury Models.

Peripheral nerve crush injury (PNCI) models are commonly used to study nerve damage and the potential beneficial effects of novel therapeutic strategies. Current models of PNCI rely on inter-device and operator precision to limit the variation with applied pressure. Although the inability to accurately quantify the PNCI pressure may result in reduced reproducibility between animals and studies, there is very limited information on the standardization and quantification of applied pressure with PNCI. To address this deficit, we constructed a novel device comprised of an Arduino UNO microcontroller board and Force Sensitive Resistor capable of reporting the real-time pressure applied to a nerve. Two forceps and two needle drivers were used to perform 30-second PNCIs to the sciatic nerves of mice (n = 5/group). Needle drivers were set to the first notch, and a jig was used to hold the forceps pinch at a reproducible pressure. The Force Sensitive Resistor was interposed in-series between the nerve and instrument during PNCI. Data collected from these procedures displayed average needle driver pressures an order of multitude greater than forceps pressures. Additionally, needle driver inter- and intra-procedure pressure remained more consistent than forceps pressure, with needle driver coefficient of variation equal to 14.5% vs. a forceps coefficient of variation equal to 45.4%. This is the first demonstration of real-time pressure measurements in PNCI models and it reveals that the applied pressures are dependent on the types of device used. The large disparity in pressure represents an inability to apply graded accurate and consistent intermediate pressure gradients in PNCI. These findings indicate a need for documentation of pressure severity as a screening for PNCI in animals, and the real-time pressure sensor could be a useful tool in monitoring and applying consistent pressure, reducing the outcome variability within the same experimental model of PNCI.

Ultrasound-guided platelet-rich plasma injection and multimodality ultrasound examination of peripheral nerve crush injury

Ultrasound-guided platelet-rich plasma (PRP) injection is able to make up for the limitations of applying a single growth factor. The goal of this study was to investigate the effects of serial ultrasound-guided PRP injections of the appropriate concentration on the treatment of sciatic nerve crush injury, and explore the value of multimodality ultrasound techniques in evaluating the prognosis of crushed peripheral nerve. In vitro, optimal concentration of PRP (from 150%, 250%, 450%, and 650%) was screened due for its maximal effect on proliferation and neurotrophic function of Schwann cells (SCs). In vivo, ninety rabbits were equally and randomly divided into normal control, model, PRP-2.5×, PRP-4.5×, and PRP-6.5× groups. The neurological function and electrophysiological recovery evaluation, and the comparison of the multimodality ultrasound evaluation with the histological results of sciatic nerve crush injury were performed to investigate the regenerative effects of PRP at different concentrations on the sciatic nerve crush injury. Our results showed that the PRP with a 4.5-fold concentration of whole blood platelets could significantly stimulate the proliferation and secretion of SCs and nerve repair. The changes in stiffness and blood perfusion were positively correlated with the collagen area percentage and VEGF expression in the injured nerve, respectively. Thus, serial ultrasound-guided PRP injections at an appropriate concentration accelerates the recovery of axonal function. Multimodality ultrasound techniques provide a clinical reference for prognosis by allowing the stiffness and microcirculation perfusion of crush-injured peripheral nerves to be quantitatively evaluated.

Value of Contrast-enhanced Ultrasound in Detection of Peripheral Nerve Crush Injury

Objective To explore the value of contrast-enhanced ultrasound(CEUS)in the detection of peripheral nerve crush injury.Methods Thirty New Zealand white rabbits were randomly divided into normal control group and sciatic nerve crush injury group(which included 3-day,2-week,4-week,and 8-week groups after operation).The morphological structure and blood perfusion of the injured sciatic nerves were detected by high-frequency ultrasound,power Doppler ultrasound(PDUS),CEUS,and histopathology.Results Conventional ultrasound revealed that the internal diameter of nerves showed no significant difference between the 8-week group and the control group [(1.14±0.15)mm vs.(0.92±0.11)mm;t=4.72,P=0.86].Analysis of nerve blood perfusion showed that PDUS had a high sensitivity in displaying fine blood flow signal inside the injured nerve in the acute stage of inflammation(3-day group)but not good enough in the 4-and 8-week groups.CEUS could clearly show the microcirculation perfusion in the 3-day,2-week,4-week,and 8-week groups,and analyses of the area under the curve,the peak time,and the peak intensity showed that the nerve blood perfusion increased significantly 3 days after operation and then decreased gradually.Histopathological examination showed that the median cumulative OD value was 12 035.6(10 566.3,14 805.8)8 weeks after operation,which was still significantly lower than that 18 784.8(15 904.5,21 103.5)in the normal control group(H=6.10, P=0.0003).Conclusions Conventional high-frequency ultrasound and PDUS can not adequately evaluate the microcirculation perfusion in different periods after nerve injury.CEUS can quantitatively evaluate the microvascular perfusion of injured nerve at any period and provide more information for indirect evaluation of nerve regeneration.

Ketogenic Diet Potentiates Electrical Stimulation-Induced Peripheral Nerve Regeneration after Sciatic Nerve Crush Injury in Rats.

Recent findings indicate that the ketogenic diet (KD) is neuroprotective and electrical stimulation (ES) can improve functional recovery from peripheral nerve injury. However, it is not clear whether KD and ES play a synergistical role in the peripheral nerve recovery following injury. A KD consisting of a 3:1 ratio of fat to carbohydrate+protein is used and is coupled with ES treatment in a rat model of peripheral nerve crush injury. Neuromuscular recovery is evaluated by electromyography, and axonal regeneration and myelination by histological methods. The effects on insulin-like growth factor 1 (IGF-1) and IGF-1 receptor expression in peripheral nerve tissue, pre- and post-nerve injury, are also investigated. The combination of KD and ES synergistically increases muscle force in biceps femoris and gluteus maximus and prevents development of hypersensitivity in biceps femoris. It promotes peripheral nerve regeneration by increasing total axons, axon density, and axonal diameter, as well as myelin thickness and axon/fiber ratio. These effects are due to modulation of the IGF system as the treatment expression of IGF-1 and IGF-1 receptor in regenerated nerve tissue. The results establish that KD and ES promote peripheral nerve regeneration. Patients recovering from peripheral nerve injury may benefit from this combinational approach.

Non-thermal plasma accelerates the healing process of peripheral nerve crush injury in rats.

The objective of this study was to evaluate the effect of non-thermal plasma (NTP) on the healing process of peripheral nerve crush injuries, which can occur during dental implant procedures. For this, a rat model of sciatic nerve crush injury (SNCI) was adopted. The rats were divided into three groups: non-nerve damage (non-ND), nerve damage (ND), and ND+NTP group. To evaluate the sciatic nerve (SN) function, the static sciatic index was calculated, and the muscle and SN tissues were subjected to a histologic analysis. The results showed that NTP effectively accelerated the healing process of SNCI in rats. In contrast to the ND group, which showed approximately 60% recovery in the SN function, the NTP-treated rats showed complete recovery. Histologically, the NTP treatments not only accelerated the muscle healing, but also reduced the edema-like phenotype of the damaged SN tissues. In the ND group, the SN tissues had an accumulation of CD68-positive macrophages, partially destroyed axonal fibers and myelinated Schwann cells. Conversely, in the ND+NTP group, the macrophage accumulation was reduced and an overall regeneration of the damaged axon fibers and the myelin sheath was accomplished. The results of this study indicate that NTP can be used for healing of injured peripheral nerves.

Miconazole alleviates peripheral nerve crush injury by mediating a macrophage phenotype change through the NF-κB pathway.

BackgroundPeripheral nerve injury (PNI) causes motor and sensory defects, has strong impact on life quality and still has no effective therapy. Miconazole is one of the most widely used antifungal drugs; the aims of the study were to investigate the effects of miconazole during sciatic nerve regeneration in a mouse model of sciatic nerve crush injury.MethodsWe established peripheral nerve crush model and investigated the effects of miconazole by multiple aspects. We further studied the potential mechanism of action of miconazole by Western blotting, fluorescence immunohistochemistry, and PCR analysis.ResultsMiconazole improves the symptoms of crushed nerve by improving inflammatory cell infiltration and demyelinating myelin of sciatic nerve. Affected by miconazole, the proportion of inflammatory M1 macrophages in the distal part of the sciatic nerve was reduced, and the proportion of anti‐inflammatory M2 macrophages was increased. Finally, the neuroprotective properties of miconazole may be regulated by the nuclear factor (NF)‐κB pathway.ConclusionsOur data suggest that miconazole can effectively alleviate PNI, and the mechanism involves mediating a phenotype change of M1/ M2 macrophages. Thus, miconazole may represent a potential therapeutic intervention for nerve crush injury.

Laminin polymer treatment accelerates repair of the crushed peripheral nerve in adult rats

Promoting axon growth after peripheral nerve injury may support recovery. Soluble laminin polymers formed at pH 4 (aLam) accelerate axon growth from adult dorsal root ganglion neurons in vitro. We used an adult rat model of a peripheral (peroneal) nerve crush to investigate whether an injection of aLam enhances axon growth and functional recovery in vivo. Rats that received an injection of aLam into the crush at 2 days post-injury show significant improvements in hind limb motor function at 2 and 5 weeks after injury compared with control rats that received phosphate-buffered saline. Functional improvement was not associated with changes in sensitivity to thermal or mechanical stimuli. Treatment with aLam decreased the occurrence of autophagia and abolished non-compliance with treadmill walking. Rats treated with aLam showed increased axon presence in the crush site at 2 weeks post-injury and larger axon diameter at 10 weeks post-injury compared with controls. Treatment with aLam did not affect Schwann cell presence or axon myelination. Our results demonstrated that aLam accelerates axon growth and maturity in a crushed peroneal nerve associated with expedited hind limb motor function recovery. Our data support the therapeutic potential of injectable aLam polymers for treatment of peripheral nerve crush injuries. Statement of SignificanceIncidence of peripheral nerve injury has been estimated to be as high as 5% of all cases entering a Level 1 trauma center and the majority of cases are young males. Peripheral nerves have some endogenous repair capabilities, but overall recovery of function remains limited, which typically has devastating effects on the individual, family, and society, as wages are lost and rehabilitation is extended until the nerves can repair. We report here that laminin polymers injected into a crush accelerated repair and recovery, had no adverse effects on sensory function, obliterated non-compliance for walking tests, and decreased the occurrence of autophagia. These data support the use of laminin polymers for safe and effective recovery after peripheral nerve injury.