Avulsion Model Research Articles

KDM4A facilitates neuropathic pain and microglial M1 polarization by regulating BDNF in a rat model of brachial plexus avulsion

BackgroundMany patients with brachial plexus avulsion (BPA) suffer from neuropathic pain, but the mechanism remains elusive. Modifications of histones, the proteins responsible for organizing DNA, may play an important role...

C286, an orally available retinoic acid receptor β agonist drug, regulates multiple pathways to achieve spinal cord injury repair.

Retinoic acid receptor β2 (RARβ2) is an emerging therapeutic target for spinal cord injuries (SCIs) with a unique multimodal regenerative effect. We have developed a first-in-class RARβ agonist drug, C286, that modulates neuron-glial pathways to induce functional recovery in a rodent model of sensory root avulsion. Here, using genome-wide and pathway enrichment analysis of avulsed rats' spinal cords, we show that C286 also influences the extracellular milieu (ECM). Protein expression studies showed that C286 upregulates tenascin-C, integrin-α9, and osteopontin in the injured cord. Similarly, C286 remodulates these ECM molecules, hampers inflammation and prevents tissue loss in a rodent model of spinal cord contusion C286. We further demonstrate C286's efficacy in human iPSC-derived neurons, with treatment resulting in a significant increase in neurite outgrowth. Additionally, we identify a putative efficacy biomarker, S100B, which plasma levels correlated with axonal regeneration in nerve-injured rats. We also found that other clinically available retinoids, that are not RARβ specific agonists, did not lead to functional recovery in avulsed rats, demonstrating the requirement for RARβ specific pathways in regeneration. In a Phase 1 trial, the single ascending dose (SAD) cohorts showed increases in expression of RARβ2 in white blood cells correlative to increased doses and at the highest dose administered, the pharmacokinetics were similar to the rat proof of concept (POC) studies. Collectively, our data suggests that C286 signalling in neurite/axonal outgrowth is conserved between species and across nerve injuries. This warrants further clinical testing of C286 to ascertain POC in a broad spectrum of neurodegenerative conditions.

5-HT3a receptor contributes to neuropathic pain by regulating central sensitization in a rat with brachial plexus avulsion

PurposeAs a frequently occurring complication resulting from brachial plexus avulsion (BPA), neuropathic pain significantly impacts the quality of life of patients and places a substantial burden on their families. Recent reports have suggested that the 5-HT3a receptor may play a role in the development and regulation of neuropathic pain. The current study aimed to explore the involvement of the 5-HT3a receptor in neuropathic pain resulting from BPA in rats. MethodsA rat model of neuropathic pain was induced through brachial plexus avulsion (BPA). The pain thresholds of the rats were measured after BPA. The spinal dorsal horn (SDH) of rats was collected at day 14 after surgery, and the expression and distribution of the 5-HT3a receptor were analyzed using immunohistochemistry and western blotting. The expression levels of various factors related to central sensitization were measured by western blot, including c-Fos, GFAP, IBA-1, IL-1β and TNF-α. The effects of 5-HT3a receptor antagonists on hyperalgesia were assessed through behavioral tests after intrathecal administration of ondansetron. Additionally, at 120 min postinjection, the SDH of rats was acquired, and the change of expression levels of protiens related to central sensitization were measured by western blot. ResultsBPA induced mechanical and cold hypersensitivity in rats. The 5-HT3a receptor was increased and mainly distributed on neurons and microglia in the SDH after BPA, and the level of central sensitization and expression of inflammatory factors, such as c-Fos, GFAP, IBA-1, IL-1β and TNF-α, were also increased markedly. Ondansetron, which is a selective 5-HT3a receptor antagonist, reversed the behavioral changes caused by BPA. The antagonist also decreased the expression of central sensitization markers and inflammatory factors. ConclusionThe results suggested that the 5-HT3a receptor is involved in neuropathic pain by regulating central nervous system sensitization in a rat brachial plexus avulsion model. Targeting the 5-HT3a receptor may be a promising approach for treating neuropathic pain after brachial plexus avulsion.

Ex Vivo Biomechanical Assessment of Various Repair Techniques in a Rabbit Calcaneal Tendon Avulsion Model: Application of Polycaprolactone Plate

This study was aimed at evaluating the biomechanical properties and gapping characteristics of tendon repair methods using a combination of a three-loop pulley (3LP) pattern, a titanium plate, and a polycaprolactone (PCL) plate in a rabbit gastrocnemius tendon (GT) model (n = 50). GTs were randomly assigned to five groups (n = 10/group). Transected GTs were repaired with a 3LP pattern alone or in conjunction with an epitendinous suture (ES), a 5-hole 1 mm PCL plate, a 5-hole 2 mm PCL plate, or a 5-hole 1.5 mm titanium plate. The yield, peak, and failure force, as well as the occurrence and force of 1-mm and 3-mm gapping were examined. The mean yield, peak, and failure force of the 3LP + titanium plate group were higher than that of other groups. The biomechanical properties of a 3LP + a 2 mm PCL plate were similar to 3LP + ES constructs in this model. In all specimens in all groups, 1 mm gap formation was observed. The frequency of 3 mm gap formation was 70% and 90% in the 3LP + 2 mm PCL plate group and the 3LP + 1.5 mm titanium plate group, respectively. Additional studies evaluating PCL plates to determine the effect on the healing and blood supply of tendon are needed.

Celsr2 Knockout Alleviates Inhibitory Synaptic Stripping and Benefits Motoneuron Survival and Axon Regeneration After Branchial Plexus Avulsion

Axotomy-induced synaptic stripping modulates survival and axon regeneration of injured motoneurons. Celsr2 is supposed to mediate homophilic interactions of neighboring cells during development, and its role in synaptic stripping remains unknow. In a model of brachial plexus avulsion, Celsr2 knockout improved functional recovery, motoneuron survival, and axon regeneration. Celsr2 was indicated to express in spinal motoneurons, excitatory and inhibitory interneurons, astrocytes, and a subset of oligodendrocytes using Celsr2LacZ mice. Double immunostaining showed that the coverage of inhibitory and excitatory vesicles on injured motoneurons were remarkably reduced after injury, whereas more inhibitory vesicles were maintained in Celsr2−/− mutants than control mice. In the ultrastructure, the density of inhibitory F-boutons on injured motoneurons was higher in Celsr2−/− mutants than controls. Conditional knockout of Celsr2 in astrocytes or oligodendrocytes showed the similar axotomy-induced synaptic withdrawal to the control. RNAseq of injured spinal samples identified 12 MHC I molecules with significant changes between Celsr2−/− and control mice. After injury, expression of MHC I surrounding injured motoneurons was increased, particularly high in Celsr2−/− mutants. In conclusion, Celsr2 knockout enhances MHC I signaling, alleviates inhibitory synaptic stripping cell-autonomously, and contributes to motoneuron survival and regeneration, and Celsr2 is a potential target for neural repair.

MiR-506-3p Relieves Neuropathic Pain following Brachial Plexus Avulsion via Mitigating Microglial Activation through Targeting the CCL2-CCR2 Axis

Neuroinflammation results in neuropathic pain (NP) following brachial plexus avulsion (BPA). This research was designed for investigating the function of miR-506-3p in BPA-induced NP. A total brachial plexus root avulsion model was produced in adult rats as well as IL-1β-treated motoneuron-like NSC-34 cells and the LPS-treated microglia cell line BV2 for in vivo and in vitro experiments, respectively. RT-PCR and Western blot were performed to detect the profiles of miR-506-3p, CCL2 and CCR2, NF-κB, FOXO3a, TNF-α, IL-1β, and IL-6 in cells or the spinal cord close to the tBPI lesion. Neuronal apoptosis was evaluated by immunohistochemistry in vivo. CCK8, TUNEL staining, and the lactic dehydrogenase kit were adopted for the evaluation of neuronal viability or damage in vitro. RNA immunoprecipitation and dual luciferase reporter gene assays analyzed the targeted association between miR-506-3p and CCL2. As shown by the data, miR-506-3p was vigorously less expressed, while CCL2-CCR2, NF-κB TNF-α, IL-1β, and IL-6 were upregulated in the spinal cord with tBPI. Overexpression of miR-506-3p attenuated neuronal apoptosis and microglial inflammation. Mechanistically, CCL2 was a downstream target of miR-506-3p. Upregulating miR-506-3p dampened CCL2-CCR2 and NF-κB activation in the spinal cord and microglia. miR-506-3p had neuroprotective and inflammation-fighting functions in the tBPI rat model via CCL2/CCR2/NF-κB axis.

Quercetin enhances survival and axonal regeneration of motoneurons after spinal root avulsion and reimplantation: experiments in a rat model of brachial plexus avulsion

BackgroundBrachial plexus avulsion (BPA) physically involves the detachment of spinal nerve roots themselves and the associated spinal cord segment, leading to permanent paralysis of motor function of the upper limb. Root avulsion induces severe pathological changes, including inflammatory reaction, oxidative damage, and finally massive motoneuron apoptosis. Quercetin (QCN), a polyphenolic flavonoid found in abundance in fruit and vegetables, has been reported to possess anti-oxidative, anti-inflammatory, and neuroprotective effects in many experimental models of both central nervous system (CNS) and peripheral nervous system (PNS) disorders. The purpose of this study was to investigate whether QCN could improve motor function recovery after C5–7 ventral root avulsion and C6 reimplantation in a rat model of BPA.MethodsThe right fifth cervical (C5) to C7 ventral roots were avulsed followed by re-implantation of only C6 to establish the spinal root avulsion plus re-implantation model in rats. After surgery, rats were treated with QCN (25, 50, and 100 mg/kg) by gavage for 2 or 8 consecutive weeks. The effects of QCN were assessed using behavior test (Terzis grooming test, TGT) and histological evaluation. The molecular mechanisms were determined by immunohistochemistry analysis and western blotting.ResultsOur results demonstrated that QCN significantly expedited motor function recovery in the forelimb as shown by the increased Terzis grooming test score, and accelerated motor axon regeneration as evidenced by the ascending number of Fluoro-Ruby-labeled and P75-positive regenerative motoneurons. The raised ChAT-immunopositive and cresyl violet-stained neurons indicated the enhanced survival of motoneurons by QCN administration. Furthermore, QCN treatment markedly alleviated muscle atrophy, restored functional motor endplates in biceps and inhibited the microglial and astroglia activation via modulating Nrf2/HO-1 and neurotrophin/Akt/MAPK signaling pathway.ConclusionsTaken together, these findings have for the first time unequivocally indicated that QCN has promising potential for further development into a novel therapeutic in conjunction with reimplantation surgery for the treatment of BPA.

Decellularized liver extracellular matrix and thrombin loaded biodegradable TOCN/Chitosan nanocomposite for hemostasis and wound healing in rat liver hemorrhage model

During deep noncompressible wound management, surgery, transplantation or post-surgical hemorrhage, rapid blood absorption and hemostasis are the key factors to be taken into consideration to reduce unexpected deaths from severe trauma. In this study, a novel hemostatic biodegradable nanocomposite was fabricated where decellularized liver extracellular matrix (L-ECM) was loaded with two natural polymers (oxidized cellulose and chitosan) in association with thrombin. Plant-derived oxidized cellulose nanofiber (TOCN) and Chitosan (CS) from deacylated chitin were self-assembled with each other by electrostatic interactions. ECM was prepared by the whole tissue decellularization process and incorporated into the composite as a source of collagen and other integrated growth factors to promote wound healing. Thrombin was also anchored with the polymers by freeze drying for enhanced hemostatic efficiency of the composite. This study is the first of its kind to report non-solubilized L-ECM and thrombin loaded TOCN and CS composite, CN/CS/EM-Th for faster hemostasis effect in a rat tail amputation (~71 s) and liver avulsion model (~41 s). Furthermore, excellent liver wound regeneration efficacy was observed in-vivo in comparison to the commercially available oxidized regenerated cellulose product SURGICEL gauge.

Bioinformatic analysis of circular RNA expression profiles in a rat lumbosacral spinal root avulsion model

Lumbosacral spinal root avulsion (LSRA) is a severe nerve injury that results in devastating dysfunction in the lower limb. Circular ribonucleic acids (circRNAs) have been reported to be implicated in a variety of diseases. However, the role of circRNAs in LSRA remains unclear. Here, we performed RNA sequencing (RNA-seq) to determine circRNA expression profiles in a rat LSRA model and further investigated their potential functions and the underlying mechanisms by bioinformatic analyses and in vitro experiments. In all, 1708 circRNAs were found to be differentially expressed in spinal cord tissues after LSRA (|fold change| ≥ 2 and p < 0.05), with 591 up-regulated 1117 down-regulated. Meanwhile, 2263 mRNAs were also indentified to be differentially expressed, of which 1471 were upregulated and 792 were downregulated. Eight randomly selected circRNAs and mRNA were successfully verified to be consistent the RNA-seq results by quantitative real-time polymerase chain reaction. Functional analyses based on gene ontology and Kyoto Encyclopedia of Genes and Genomes predicted the potential roles of differentially expressed circRNAs and mRNAs in LSRA, and circRNA/miRNA/mRNA interaction networks revealed that circRNA_7025, a down-regulated circRNA in LSRA, was targeted by two neuronal apoptosis-related miRNAs, rno-miR-1224 and rno-miR-326-5p. Further in vitro experiments revealed that circRNA_7025 protected against oxygen-glucose deprivation induced neuronal apoptosis via the circRNA_7025/miR-1224/miR-326-5p axis. In summary, our results revealed circRNA expression profiles and their potential functions in LSRA. These findings improve our understanding of the pathogenic mechanisms involved in LSRA and might enable us to identify new molecular targets for LSRA.

Loop modification of the traditional three-loop pulley pattern improves the biomechanical properties and resistance to 3-mm gap formation in a canine common calcanean teno-osseous avulsion model.

To compare the biomechanical properties and gapping characteristics following loop modification of a 3-loop-pulley (3LP) pattern in an ex vivo canine common calcaneal tendon (CCT) avulsion repair model. 56 skeletally mature hindlimbs from 28 canine cadavers. The CCTs were randomized to 1 of 4 experimental groups (n = 14/group) then sharply transected at the teno-osseous junction. Groups consisted of a 3LP, 4-loop-pulley (4LP), 5-loop-pulley (5LP), or 6-loop-pulley (6LP) pattern with loops placed 60° apart using size-0 polypropylene. Yield, peak, and failure loads, construct stiffness, loads to produce a 3-mm teno-osseous gap, and failure mode were evaluated and compared between groups. Yield (P = 0.001), peak (P < 0.001), and failure loads (P < 0.001), construct stiffness (P < 0.001), and loads to 3-mm gap formation (P = 0.005) were all significantly greater for 6LP compared to all other groups. Mode of failure did not differ among groups (P = 0.733) with 75% (42/56) of repairs failing by mechanism of core sutures pulling through the tendinous tissue. Pattern modification by increasing the number of loops increased the repair site strength by 1.4, 1.6, and 1.8 times for 4LP, 5LP, and 6LP compared to 3LP, respectively. Increasing the number of suture loops compared to a traditional 3LP repair is a relatively simple technique modification that significantly increases teno-osseous repair site strength and loads required to cause 3-mm gap formation. The results of this study justify further focused investigation of increasing the number of suture loops in vivo for teno-osseous CCT repair in dogs.

Effect of calcanean bone‐tunnel orientation for teno‐osseous repair in a canine common calcanean tendon avulsion model

ObjectiveTo determine the influence of bone‐tunnel anchoring technique on teno‐osseous repair of the common calcanean tendon (CCT) in dogs.Study designRandomized, ex vivo, biomechanical.PopulationForty‐two skeletally mature canine hindlimbs.MethodsCanine hindlimbs were dissected to produce a model simulating avulsion of the CCT and accessory tendons from the calcaneus. Hindlimbs were randomized to 1 of 3 anchoring techniques (n = 14/group): a single transverse tunnel (TT), vertical tunnels (VT), or modified bone tunnels (MT) for teno‐osseous repair in a 3‐loop‐pulley (3LP) pattern using 0 USP polypropylene. Yield, peak and failure loads, construct stiffness, loads to produce a 3 mm teno‐osseous gap, and failure modes were compared between groups.ResultsThe only difference detected consisted of TT constructs yielding at loads 25% higher than MT constructs (P = .027).ConclusionAlthough yield loads were lower in MT constructs than other groups, the bone‐tunnel anchoring techniques tested here did not appear to influence the biomechanical properties or gapping characteristics of teno‐osseous repairs in this canine CCT avulsion model.Clinical significanceAll drilling techniques and bone‐tunnel orientations tested in the study reported here offer viable options to reattach the CCT to the calcaneus. Surgeons should evaluate how bone‐tunnel orientation may affect placement of adjunctive fixation methods to stabilize the talocrural joint after primary CCT repair in dogs.

GDNF to the rescue: GDNF delivery effects on motor neurons and nerves, and muscle re-innervation after peripheral nerve injuries.

Peripheral nerve injuries commonly occur due to trauma, like a traffic accident. Peripheral nerves get severed, causing motor neuron death and potential muscle atrophy. The current golden standard to treat peripheral nerve lesions, especially lesions with large (≥ 3 cm) nerve gaps, is the use of a nerve autograft or reimplantation in cases where nerve root avulsions occur. If not tended early, degeneration of motor neurons and loss of axon regeneration can occur, leading to loss of function. Although surgical procedures exist, patients often do not fully recover, and quality of life deteriorates. Peripheral nerves have limited regeneration, and it is usually mediated by Schwann cells and neurotrophic factors, like glial cell line-derived neurotrophic factor, as seen in Wallerian degeneration. Glial cell line-derived neurotrophic factor is a neurotrophic factor known to promote motor neuron survival and neurite outgrowth. Glial cell line-derived neurotrophic factor is upregulated in different forms of nerve injuries like axotomy, sciatic nerve crush, and compression, thus creating great interest to explore this protein as a potential treatment for peripheral nerve injuries. Exogenous glial cell line-derived neurotrophic factor has shown positive effects in regeneration and functional recovery when applied in experimental models of peripheral nerve injuries. In this review, we discuss the mechanism of repair provided by Schwann cells and upregulation of glial cell line-derived neurotrophic factor, the latest findings on the effects of glial cell line-derived neurotrophic factor in different types of peripheral nerve injuries, delivery systems, and complementary treatments (electrical muscle stimulation and exercise). Understanding and overcoming the challenges of proper timing and glial cell line-derived neurotrophic factor delivery is paramount to creating novel treatments to tend to peripheral nerve injuries to improve patients’ quality of life.

GRP78 Overexpression Triggers PINK1-IP3R-Mediated Neuroprotective Mitophagy.

An experimental model of spinal root avulsion (RA) is useful to study causal molecular programs that drive retrograde neurodegeneration after neuron-target disconnection. This neurodegenerative process shares common characteristics with neuronal disease-related processes such as the presence of endoplasmic reticulum (ER) stress and autophagy flux blockage. We previously found that the overexpression of GRP78 promoted motoneuronal neuroprotection after RA. After that, we aimed to unravel the underlying mechanism by carrying out a comparative unbiased proteomic analysis and pharmacological and genetic interventions. Unexpectedly, mitochondrial factors turned out to be most altered when GRP78 was overexpressed, and the abundance of engulfed mitochondria, a hallmark of mitophagy, was also observed by electronic microscopy in RA-injured motoneurons after GRP78 overexpression. In addition, GRP78 overexpression increased LC3-mitochondria tagging, promoted PINK1 translocation, mitophagy induction, and recovered mitochondrial function in ER-stressed cells. Lastly, we found that GRP78-promoted pro-survival mitophagy was mediated by PINK1 and IP3R in our in vitro model of motoneuronal death. This data indicates a novel relationship between the GRP78 chaperone and mitophagy, opening novel therapeutical options for drug design to achieve neuroprotection.

GDNF Gene Therapy to Repair the Injured Peripheral Nerve.

A spinal root avulsion is the most severe proximal peripheral nerve lesion possible. Avulsion of ventral root filaments disconnects spinal motoneurons from their target muscles, resulting in complete paralysis. In patients that undergo brachial plexus nerve repair, axonal regeneration is a slow process. It takes months or even years to bridge the distance from the lesion site to the distal targets located in the forearm. Following ventral root avulsion, without additional pharmacological or surgical treatments, progressive death of motoneurons occurs within 2 weeks (Koliatsos et al., 1994). Reimplantation of the avulsed ventral root or peripheral nerve graft can act as a conduit for regenerating axons and increases motoneuron survival (Chai et al., 2000). However, this beneficial effect is transient. Combined with protracted and poor long-distance axonal regeneration, this results in permanent function loss. To overcome motoneuron death and improve functional recovery, several promising intervention strategies are being developed. Here, we focus on GDNF gene-therapy. We first introduce the experimental ventral root avulsion model and discuss its value as a proxy to study clinical neurotmetic nerve lesions. Second, we discuss our recent studies showing that GDNF gene-therapy is a powerful strategy to promote long-term motoneuron survival and improve function when target muscle reinnervation occurs within a critical post-lesion period. Based upon these observations, we discuss the influence of timing of the intervention, and of the duration, concentration and location of GDNF delivery on functional outcome. Finally, we provide a perspective on future research directions to realize functional recovery using gene therapy.

Upregulation of JHDM1D-AS1 alleviates neuroinflammation and neuronal injury via targeting miR-101-3p-DUSP1 in spinal cord after brachial plexus injury

BackgroundNeuroinflammation in the spinal cord following acute brachial plexus injury (BPI) remains a vital cause that leads to motor dysfunction and neuropathic pain. In this study, we aim to explore the role of long non-coding RNA JHDM1D antisense 1 (JHDM1D-AS1) in mediating BPI-induced neuroinflammation and neuronal injury. MethodsA total brachial plexus root avulsion (tBPRA) model in adult rats and IL-1β-treated motor neuron-like NSC-34 cells and LPS-treated microglia cell line BV2 were conducted for in vivo and in vitro experiments, respectively. The expressions of JHDM1D-AS1, miR-101-3p and DUSP1, p38, NF-κB, TNF-α, IL-1β, and IL-6 were detected by RT-PCR and western blot seven days after tBPI. Immunohistochemistry (IHC) was used to detect neuronal apoptosis. CCK8 assay, Tunel assay and LDH kit were used for the detection of neuronal injury. The targeted relationships between JHDM1D-AS1 and miR-101-3p, miR-101-3p and DUSP1 were verified by RNA immunoprecipitation (RIP) and dual-luciferase reporter gene assay. ResultsWe found significant downregulated expression of JHDM1D-AS1 and DUSP1 but upregulated expression of miR-101-3p in the spinal cord after tBPI. Overexpression of JHDM1D-AS1 had a prominent neuroprotective effect by suppressing neuronal apoptosis and microglial inflammation through reactivation of DUSP1. Further exploration revealed that JHDM1D-AS1 may act as a competitive endogenous RNA targeting miR-101-3p, which bound on the 3′UTR of DUSP1 mRNA. In addition, overexpression of miR-101-3p could reverse the neuroprotective effects of JHDM1D-AS1 upregulation by blocking DUSP1. ConclusionsJHDM1D-AS1 exerted neuroprotective and anti-inflammatory effects in a rat model of tBPI by regulating miR-101-3p/DUSP1 axis.

Anterior cruciate ligament repair versus reconstruction: A kinematic analysis

BackgroundThe purpose of this study was to compare the biomechanical properties of an anterior cruciate ligament (ACL) anatomic repair of a true femoral avulsion to an anatomic ACL reconstruction. It was hypothesized that the ACL repair and ACL reconstruction would have comparable biomechanical behavior when compared to the native knee. MethodsTen paired fresh-frozen cadaveric knees (n = 20) were used to investigate knee kinematics when an anterior drawer force, varus, valgus, internal, and external rotational moment were applied at 0, 15, 30, 45, 60, and 90 degrees of flexion. Displacement and rotation were recorded in the following conditions: ACL-intact, ACL-deficient, and ACL-repaired vs reconstructed. ResultsSectioning of the ACL significantly increased anterior tibial translation (0°, 15°, 30° and 45°) compared to the intact state. The mean anterior displacement difference from intact was lower in the ACL-repaired knees compared to reconstructed knees at 30° and 90°. There were no significant differences between conditions in varus, valgus, internal, or external rotations. ConclusionACL repair and ACL reconstruction procedures restored knee anterior tibial translation in matched paired specimens. There were no differences in valgus, varus, internal, or external rotation. Although, ACL-repaired knees (avulsion model) demonstrated less anterior tibial translation when compared to ACL-reconstructed knees, this difference was less than one millimeter. Based on the findings of this study, repair and reconstruction procedures both restored anterior tibial translation in matched-pair specimens. This suggests that the initial functionality of both techniques is similar and that further clinical studies are needed to compare the long-term stability.

Hyperoxygenated solution improves tissue viability in an avulsion injury flap model

Management of avulsion injuries remains a challenge due to necrosis. The aim of the present study is to create an experimental model reproducing an avulsion injury and investigate the effects of hyperoxygenated solution (HOS), a method of oxygen delivery that has been widely used in the therapy of ischaemia-hypoxia diseases, on avulsion injury flap survival in rats. Forty male rats were divided into four groups (n = 10 each). Dorsal random pattern flaps measuring 3 × 9 cm, including the panniculus carnosus, were elevated and run over by the skin avulsion model machine, and the flaps were then sutured into their original places. The sham+HOS and avulsion+HOS groups received intravenous HOS (20 ml/kg) each day for 7 days after the operation. The sham+saline and avulsion+saline groups received intravenous saline solution (20 ml/kg) each day for 7 days after the operation. Percutaneous O2 pressure (TcpO2) measurement, serial examinations of skin flap blood perfusion, skin flap survival evaluation and histopathology were performed to assess the efficacy of HOS on avulsion injury. Compared to the avulsion+saline groups, TcpO2 was significantly higher in the avulsion+HOS groups at 15, 30 and 60 min after infusion (P<0.05). The blood perfusion of flaps in the avulsion+HOS group was higher than in the avulsion+saline group (P<0.05). The survival rate was higher in the avulsion+HOS group than in the avulsion+saline group (P<0.05), and the histopathology assays supported the data. We succeeded in developing an avulsion injury model and demonstrated that HOS could improve the survival of the avulsion injury flaps in rats by effectively increasing the local oxygen content and blood perfusion and ameliorating inflammatory damage.

NIR-II Fluorescence Imaging of Skin Avulsion and Necrosis.

Skin avulsion is commonly seen in individuals exposed to heavy shearing forces. Subcutaneous tissue detachment and bone fractures usually accompany skin avulsion. Thus, the estimation of the extent of damaged tissue is very important. Currently, the viability of skin and subcutaneous tissue is determined by clinical observations, and these observations always underestimate the true extent of the avulsed skin. Herein, we synthesized an innovative probe, CH1055-GRRRDEVDK (CH1055-GK), which can specifically bind to caspase-3 so as to image skin avulsion and define necrotic regions. Our uptake and binding affinity tests in apoptotic cells and evaluation of the probe ex vivo and in vivo showed that the probe has a strong ability to bind caspase-3 in skin avulsion models and that it vividly detected the necrotic area in avulsed skins. Furthermore, the increased fluorescence intensity of the probe in the avulsed skin showed a larger affected area than that determined by clinical observations in live mice. Consequently, our results indicated that observation of the caspase-3-targeted probe CH1055-GK via NIR-II imaging allowed the clear detection of skin avulsion in subjects, indicating its potential as an imaging tool for the early diagnosis of skin avulsion and the determination of necrotic margins.

TrkB agonistic antibodies superior to BDNF: Utility in treating motoneuron degeneration

While Brain-derived Neurotrophic Factor (BDNF) has long been implicated in treating neurological diseases, recombinant BDNF protein has failed in multiple clinical trials. In addition to its unstable and adhesive nature, BDNF can activate p75NTR, a receptor mediating cellular functions opposite to those of TrkB. We have now identified TrkB agonistic antibodies (TrkB-agoAbs) with several properties superior to BDNF: They exhibit blood half-life of days instead of hours, diffuse centimeters in neural tissues instead millimeters, and bind and activate TrkB, but not p75NTR. In addition, TrkB-agoAbs elicit much longer TrkB activation, reduced TrkB internalization and less intracellular degradation, compared with BDNF. More importantly, some of these TrkB-agoAbs bind TrkB epitopes distinct from that by BDNF, and work cooperatively with endogenous BDNF. Unlike BDNF, the TrkB-agoAbs exhibit a half-life of days/weeks and diffused readily in nerve tissues. We tested one of TrkB-agoAbs further and showed that it enhanced motoneuron survival in the spinal-root avulsion model for motoneuron degeneration in vivo. Thus, TrkB-agoAbs are promising drug candidates for the treatment of neural injury.

Development of anatomically based customizable three-dimensional finite-element model of pelvic floor support system: POP-SIM1.0.

To develop an anatomically based customizable finite-element (FE) model of the pelvic floor support system to simulate pelvic organ prolapse (POP): POP-SIM1.0. This new simulation platform allows for the construction of an array of models that objectively represent the key anatomical and functional variation in women with and without prolapse to test pathomechanism hypotheses of the prolapse formation. POP-SIM1.0 consists of anatomically based FE models and a suite of Python-based tools developed to rapidly construct FE models by customizing the base model with desired structural parameters. Each model consists of anatomical structures from three support subsystems which can be customized based on magnetic resonance image measurements in women with and without prolapse. The customizable structural parameters include presence of levator ani (LA) avulsion, hiatus size, anterior vaginal wall dimension, attachment fascia length and apical location in addition to the tissue material properties and intra-abdominal pressure loading. After customization, the FE model was loaded with increasing intra-abdominal pressure (0-100 cmH2O) and solved using ABAQUS explicit solver. We were able to rapidly construct anatomically based FE models with specific structural geometry which reflects the morphology changes often observed in women with prolapse. At maximum loading, simulated structural deformations have similar anatomical characteristics to those observed during clinical exams and stress magnetic resonance images. Simulation results showed the presence of LA muscle avulsion negatively impacts the pelvic floor support. The normal model with intact muscle had the smallest exposed vaginal length of 11 mm, while the bilateral avulsion produced the largest exposed vaginal length at 24 mm. The unilateral avulsion model had an exposed vaginal length of 18 mm and also demonstrated a tipped perineal body similar to that seen in clinical observation. Increasing the hiatus size, vaginal wall length and fascia length also resulted in worse pelvic floor support, increasing the exposed vaginal length from 18 mm in the base model to 33 mm, 54 mm and 23.5 mm, respectively. The developed POP-SIM1.0 can simulate the anatomical structure changes often observed in women with prolapse. Preliminary results showed that the presence of LA avulsion, enlarged hiatus, longer vaginal wall and fascia length can result in larger prolapse at simulated maximum Valsalva.